WorldWideScience

Sample records for 15n resonance assignments

  1. (1)H, (13)C, and (15)N resonance assignments for the pro-inflammatory cytokine interleukin-36α.

    Science.gov (United States)

    Goradia, Nishit; Wißbrock, Amelie; Wiedemann, Christoph; Bordusa, Frank; Ramachandran, Ramadurai; Imhof, Diana; Ohlenschläger, Oliver

    2016-10-01

    Interleukin-36α (IL-36α) is a recently characterised member of the interleukin-1 superfamily. It is involved in the pathogenesis of inflammatory arthritis in one third of psoriasis patients. By binding of IL-36α to its receptor IL-36R via the NF-κB pathway other cytokines involved in inflammatory and apoptotic cascade are activated. The efficacy of complex formation is controlled by N-terminal processing. To obtain a more detailed view on the structure function relationship we performed a heteronuclear multidimensional NMR investigation and here report the (1)H, (13)C, and (15)N resonance assignments for the backbone and side chain nuclei of the pro-inflammatory cytokine interleukin-36α.

  2. Towards fully automated structure-based NMR resonance assignment of 15N-labeled proteins from automatically picked peaks

    KAUST Repository

    Jang, Richard

    2011-03-01

    In NMR resonance assignment, an indispensable step in NMR protein studies, manually processed peaks from both N-labeled and C-labeled spectra are typically used as inputs. However, the use of homologous structures can allow one to use only N-labeled NMR data and avoid the added expense of using C-labeled data. We propose a novel integer programming framework for structure-based backbone resonance assignment using N-labeled data. The core consists of a pair of integer programming models: one for spin system forming and amino acid typing, and the other for backbone resonance assignment. The goal is to perform the assignment directly from spectra without any manual intervention via automatically picked peaks, which are much noisier than manually picked peaks, so methods must be error-tolerant. In the case of semi-automated/manually processed peak data, we compare our system with the Xiong-Pandurangan-Bailey- Kellogg\\'s contact replacement (CR) method, which is the most error-tolerant method for structure-based resonance assignment. Our system, on average, reduces the error rate of the CR method by five folds on their data set. In addition, by using an iterative algorithm, our system has the added capability of using the NOESY data to correct assignment errors due to errors in predicting the amino acid and secondary structure type of each spin system. On a publicly available data set for human ubiquitin, where the typing accuracy is 83%, we achieve 91% accuracy, compared to the 59% accuracy obtained without correcting for such errors. In the case of automatically picked peaks, using assignment information from yeast ubiquitin, we achieve a fully automatic assignment with 97% accuracy. To our knowledge, this is the first system that can achieve fully automatic structure-based assignment directly from spectra. This has implications in NMR protein mutant studies, where the assignment step is repeated for each mutant. © Copyright 2011, Mary Ann Liebert, Inc.

  3. hNCOcanH pulse sequence and a robust protocol for rapid and unambiguous assignment of backbone ((1)H(N), (15)N and (13)C') resonances in (15)N/(13)C-labeled proteins.

    Science.gov (United States)

    Kumar, Dinesh; Hosur, Ramakrishna V

    2011-09-01

    A three-dimensional nuclear magnetic resonance (NMR) pulse sequence named as hNCOcanH has been described to aid rapid sequential assignment of backbone resonances in (15)N/(13)C-labeled proteins. The experiment has been derived by a simple modification of the previously described HN(C)N pulse sequence [Panchal et al., J. Biomol. NMR 20 (2001) 135-147]; t2 evolution is used to frequency label (13)C' rather than (15)N (similar trick has also been used in the design of hNCAnH pulse sequence from hNcaNH [Frueh et al., JACS, 131 (2009) 12880-12881]). The modification results in a spectrum equivalent to HNCO, but in addition to inter-residue correlation peaks (i.e. Hi , Ci-1), the spectrum also contains additional intra-residue correlation peaks (i.e. Hi-1 , Ci-1) in the direct proton dimension which has maximum resolution. This is the main strength of the experiment and thus, even a small difference in amide (1) H chemical shifts (5-6 Hz) can be used for establishing a sequential connectivity. This experiment in combination with the HNN experiment described previously [Panchal et al., J. Biomol. NMR 20 (2001) 135-147] leads to a more robust assignment protocol for backbone resonances ((1) H(N) , (15)N) than could be derived from the combination of HNN and HN(C)N experiments [Bhavesh et al., Biochemistry, 40 (2001) 14727-14735]. Further, this new protocol enables assignment of (13)C' resonances as well. We believe that the experiment and the protocol presented here will be of immense value for structural-and functional-proteomics research by NMR. Performance of this experiment has been demonstrated using (13)C/(15)N labeled ubiquitin.

  4. 1H, 15N and 13C backbone resonance assignments of the archetypal serpin α1-antitrypsin.

    Science.gov (United States)

    Nyon, Mun Peak; Kirkpatrick, John; Cabrita, Lisa D; Christodoulou, John; Gooptu, Bibek

    2012-10-01

    Alpha(1)-antitrypsin is a 45-kDa (394-residue) serine protease inhibitor synthesized by hepatocytes, which is released into the circulatory system and protects the lung from the actions of neutrophil elastase via a conformational transition within a dynamic inhibitory mechanism. Relatively common point mutations subvert this transition, causing polymerisation of α(1)-antitrypsin and deficiency of the circulating protein, predisposing carriers to severe lung and liver disease. We have assigned the backbone resonances of α(1)-antitrypsin using multidimensional heteronuclear NMR spectroscopy. These assignments provide the starting point for a detailed solution state characterization of the structural properties of this highly dynamic protein via NMR methods.

  5. 1H, 13C, and 15N backbone and side chain resonance assignments of thermophilic Geobacillus kaustophilus cyclophilin-A

    Energy Technology Data Exchange (ETDEWEB)

    Holliday, Michael; Zhang, Fengli; Isern, Nancy G.; Armstrong, Geoffrey S.; Eisenmesser, Elan Z.

    2014-04-01

    Cyclophilins catalyze the reversible peptidyl-prolyl isomerization of their substrates and are present across all kingdoms of life from humans to bacteria. Although numerous biological roles have now been discovered for cyclophilins, their function was initially ascribed to their chaperone-like activity in protein folding where they catalyze the often rate-limiting step of proline isomerization. This chaperone-like activity may be especially important under extreme conditions where cyclophilins are often over expressed, such as in tumors for human cyclophilins {Lee, 2010 #1167}, but also in organisms that thrive under extreme conditions, such as theromophilic bacteria. Moreover, the reversible nature of the peptidyl-prolyl isomerization reaction catalyzed by cyclophilins has allowed these enzymes to serve as model systems for probing the role of conformational changes during catalytic turnover {Eisenmesser, 2002 #20;Eisenmesser, 2005 #203}. Thus, we present here the resonance assignments of a thermophilic cyclophilin from Geobacillus kaustophilus derived from deep-sea sediment {Takami, 2004 #1384}. This thermophilic cyclophilin may now be studied at a variety of temperatures to provide insight into the comparative structure, dynamics, and catalytic mechanism of cyclophilins.

  6. 1H, 13C, and 15N backbone and side chain resonance assignments of thermophilic Geobacillus kaustophilus cyclophilin-A.

    Science.gov (United States)

    Holliday, Michael J; Zhang, Fengli; Isern, Nancy G; Armstrong, Geoffrey S; Eisenmesser, Elan Z

    2014-04-01

    Cyclophilins catalyze the reversible peptidyl-prolyl isomerization of their substrates and are present across all kingdoms of life from humans to bacteria. Although numerous biological roles have now been discovered for cyclophilins, their function was initially ascribed to their chaperone-like activity in protein folding where they catalyze the often rate-limiting step of proline isomerization. This chaperone-like activity may be especially important under extreme conditions where cyclophilins are often over expressed, such as in tumors for human cyclophilins (Lee Archiv Pharm Res 33(2): 181-187, 2010), but also in organisms that thrive under extreme conditions, such as theromophilic bacteria. Moreover, the reversible nature of the peptidyl-prolyl isomerization reaction catalyzed by cyclophilins has allowed these enzymes to serve as model systems for probing the role of conformational changes during catalytic turnover (Eisenmesser et al. Science 295(5559): 1520-1523, 2002; Eisenmesser et al. Nature 438(7064): 117-121, 2005). Thus, we present here the resonance assignments of a thermophilic cyclophilin from Geobacillus kaustophilus derived from deep-sea sediment (Takami et al. Extremophiles 8(5): 351-356, 2004). This thermophilic cyclophilin may now be studied at a variety of temperatures to provide insight into the comparative structure, dynamics, and catalytic mechanism of cyclophilins.

  7. Resonance assignments, secondary structure and 15N relaxation data of the human transcriptional coactivator hMBF1 (57-148)

    Energy Technology Data Exchange (ETDEWEB)

    Mishima, Masaki; Ozaki, Jun; Ikegami, Takahisa [Nara Institute of Science and Technology, Graduate School of Biological Sciences (Japan); Kabe, Yasuaki; Goto, Masahide [National Institute of Genetics, Department of Developmental Genetics (Japan); Ueda, Hitoshi; Hirose, Susumu [Tokyo Institute of Technology, Faculty of Bioscience and Biotechnology (Japan); Handa, Hiroshi [National Institute of Genetics, Department of Developmental Genetics (Japan); Shirakawa, Masahiro [Nara Institute of Science and Technology, Graduate School of Biological Sciences (Japan)

    1999-08-15

    Multiprotein bridging factor 1 (MBF1) is a transcriptional coactivator that is thought to bridge between the TATA box-binding protein (TBP) and DNA binding regulatory factors, and is conserved from yeast to human. Human MBF1 (hMBF1) can bind to TBP and to the nuclear receptor Ad4BP, and is suggested to mediate Ad4BP-dependent transcriptional activation. Here we report the resonance assignments and secondary structure of hMBF1 (57-148) that contains both TBP binding and activator binding residues. {sup 15}N relaxation data were also obtained. As a result, hMBF1 (57-148) was shown to consist of flexible N-terminal residues and a C-terminal domain. The C-terminal domain contains four helices and a conserved C-terminal region.

  8. Natural-abundance 15N NMR studies of Turkey ovomucoid third domain. Assignment of peptide 15N resonances to the residues at the reactive site region via proton-detected multiple-quantum coherence

    Science.gov (United States)

    Ortiz-Polo, Gilberto; Krishnamoorthi, R.; Markley, John L.; Live, David H.; Davis, Donald G.; Cowburn, David

    Heteronuclear two-dimensional 1H{ 15N} multiple-quantum (MQ) spectroscopy has been applied to a protein sample at natural abundance: ovomucoid third domain from turkey ( Meleagris gallopavo), a serine proteinase inhibitor of 56 amino acid residues. Peptide amide 1H NMR assignments obtained by two-dimensional 1H{ 1H} NMR methods (R. Krishnamoorthi and J. L. Markley, unpublished data) led to identification of the corresponding 1H{ 15N} MQ coherence cross peaks. From these, 15N NMR chemical shifts were determined for several specific backbone amide groups of amino acid residues located around the reactive site region of the inhibitor. The results suggest that amide 15N chemical shifts, which are readily obtained in this way, may serve as sensitive probes for conformational studies of proteins.

  9. Solid-state NMR [13C,15N] resonance assignments of the nucleotide-Binding Domain of a bacterial Cyclic Nucleotide-Gated Channel

    NARCIS (Netherlands)

    Cukkemane, A.A.; Nand, D.; Gradmann, S.H.E.; Weingarth, M.H.; Kaupp, U.B.; Baldus, M.

    2012-01-01

    Channels regulated by cyclic nucleotides are key signalling proteins in several biological pathways. The regulatory aspect is conferred by a C-terminal cyclic nucleotide-binding domain (CNBD). We report resonance assignments of the CNBD of a bacterial mlCNG channel obtained using 2D and 3D solid-sta

  10. Sequence-specific {sup 1}H, {sup 13}C, and {sup 15}N resonance assignments for intestinal fatty-acid-binding protein complexed with palmitate (15.4 kDA)

    Energy Technology Data Exchange (ETDEWEB)

    Hodsdon, M.E.; Toner, J.J.; Cistola, D.P. [Washington Univ. School of Medicine, St. Louis, MO (United States)

    1994-12-01

    Intestinal fatty-acid-binding protein (I-FABP) belongs to a family of soluble, cytoplasmic proteins that are thought to function in the intracellular transport and trafficking of polar lipids. Individual members of this protein family have distinct specificities and affinities for fatty acids, cholesterol, bile salts, and retinoids. We are comparing several retinol- and fatty-acid-binding proteins from intestine in order to define the factors that control molecular recognition in this family of proteins. We have established sequential resonance assignments for uniformly {sup 13}C/{sup 15}N-enriched I-FABP complexed with perdeuterated palmitate at pH7.2 and 37{degrees}C. The assignment strategy was similar to that introduced for calmodulin. We employed seven three-dimensional NMR experiments to establish scalar couplings between backbone and sidechain atoms. Backbone atoms were correlated using triple-resonance HNCO, HNCA, TOCSY-HMQC, HCACO, and HCA(CO)N experiments. Sidechain atoms were correlated using CC-TOCSY, HCCH-TOCSY, and TOCSY-HMQC. The correlations of peaks between three-dimensional spectra were established in a computer-assisted manner using NMR COMPASS (Molecular Simulations, Inc.) Using this approach, {sup 1}H, {sup 13}C, and {sup 15}N resonance assignments have been established for 120 of the 131 residues of I-FABP. For 18 residues, amide {sup 1}H and {sup 15}N resonances were unobservable, apparently because of the rapid exchange of amide protons with bulk water at pH 7.2. The missing amide protons correspond to distinct amino acid patterns in the protein sequence, which will be discussed. During the assignment process, several sources of ambiguity in spin correlations were observed. To overcome this ambiguity, the additional inter-residue correlations often observed in the HNCA experiment were used as cross-checks for the sequential backbone assignments.

  11. 1H, 13C and 15N resonance assignments and second structure information of Fag s 1: Fagales allergen from Fagus sylvatica.

    Science.gov (United States)

    Moraes, A H; Asam, C; Batista, A; Almeida, F C L; Wallner, M; Ferreira, F; Valente, A P

    2016-04-01

    Fagales allergens belonging to the Bet v 1 family account responsible for the majority of spring pollinosis in the temperate climate zones in the Northern hemisphere. Among them, Fag s 1 from beech pollen is an important trigger of Fagales pollen associated allergic reactions. The protein shares high similarity with birch pollen Bet v 1, the best-characterized member of this allergen family. Of note, recent work on Bet v 1 and its homologues found in Fagales pollen demonstrated that not all allergenic members of this family have the capacity to induce allergic sensitization. Fag s 1 was shown to bind pre-existing IgE antibodies most likely primarily directed against other members of this multi-allergen family. Therefore, it is especially interesting to compare the structures of Bet v 1-like pollen allergens, which have the potential to induce allergic sensitization with allergens that are mainly cross-reactive. This in the end will help to identify allergy eliciting molecular pattern on Bet v 1-like allergens. In this work, we report the (1)H, (15)N and (13)C NMR assignment of beech pollen Fag s 1 as well as the secondary structure information based on backbone chemical shifts.

  12. NMR study of non-structural proteins--part I: (1)H, (13)C, (15)N backbone and side-chain resonance assignment of macro domain from Mayaro virus (MAYV).

    Science.gov (United States)

    Melekis, Efstathios; Tsika, Aikaterini C; Lichière, Julie; Chasapis, Christos T; Margiolaki, Irene; Papageorgiou, Nicolas; Coutard, Bruno; Bentrop, Detlef; Spyroulias, Georgios A

    2015-04-01

    Macro domains are ADP-ribose-binding modules present in all eukaryotic organisms, bacteria and archaea. They are also found in non-structural proteins of several positive strand RNA viruses such as alphaviruses. Here, we report the high yield expression and preliminary structural analysis through solution NMR spectroscopy of the macro domain from New World Mayaro Alphavirus. The recombinant protein was well-folded and in a monomeric state. An almost complete sequence-specific assignment of its (1)H, (15)N and (13)C resonances was obtained and its secondary structure determined by TALOS+.

  13. (1)H, (15)N, (13)C backbone resonance assignments of human soluble catechol O-methyltransferase in complex with S-adenosyl-L-methionine and 3,5-dinitrocatechol.

    Science.gov (United States)

    Czarnota, Sylwia; Baxter, Nicola J; Cliff, Matthew J; Waltho, Jonathan P; Scrutton, Nigel S; Hay, Sam

    2016-12-15

    Catechol O-methyltransferase (COMT) is an enzyme that plays a major role in catechol neurotransmitter deactivation. Inhibition of COMT can increase neurotransmitter levels, which provides a means of treatment for Parkinson's disease, schizophrenia and depression. COMT exists as two isozymes: a soluble cytoplasmic form (S-COMT), expressed in the liver and kidneys and a membrane-bound form (MB-COMT), found mostly in the brain. Here we report the backbone (1)H, (15)N and (13)C chemical shift assignments of S-COMT in complex with S-adenosyl-L-methionine, 3,5-dinitrocatechol and Mg(2+). Assignments were obtained by heteronuclear multidimensional NMR spectroscopy. In total, 97 % of all backbone resonances were assigned in the complex, with 205 out of a possible 215 residues assigned in the (1)H-(15)N TROSY spectrum. Prediction of solution secondary structure from a chemical shift analysis using the TALOS+ webserver is in good agreement with published X-ray crystal structures.

  14. (1)H, (13)C, and (15)N backbone resonance assignments of the full-length 40 kDa S. acidocaldarius Y-family DNA polymerase, dinB homolog.

    Science.gov (United States)

    Moro, Sean L; Cocco, Melanie J

    2015-10-01

    The dinB homolog (Dbh) is a member of the Y-family of translesion DNA polymerases, which are specialized to accurately replicate DNA across from a wide variety of lesions in living cells. Lesioned bases block the progression of high-fidelity polymerases and cause detrimental replication fork stalling; Y-family polymerases can bypass these lesions. The active site of the translesion synthesis polymerase is more open than that of a replicative polymerase; consequently Dbh polymerizes with low fidelity. Bypass polymerases also have low processivity. Short extension past the lesion allows the high-fidelity polymerase to switch back onto the site of replication. Dbh and the other Y-family polymerases have been used as structural models to investigate the mechanisms of DNA polymerization and lesion bypass. Many high-resolution crystal structures of Y-family polymerases have been reported. NMR dynamics studies can complement these structures by providing a measure of protein motions. Here we report the (15)N, (1)H, and (13)C backbone resonance assignments at two temperatures (35 and 50 °C) for Sulfolobus acidocaldarius Dbh polymerase. Backbone resonance assignments have been obtained for 86 % of the residues. The polymerase active site is assigned as well as the majority of residues in each of the four domains.

  15. (1)H, (13)C, (15)N backbone and side-chain resonance assignment of Nostoc sp. C139A variant of the heme-nitric oxide/oxygen binding (H-NOX) domain.

    Science.gov (United States)

    Alexandropoulos, Ioannis I; Argyriou, Aikaterini I; Marousis, Kostas D; Topouzis, Stavros; Papapetropoulos, Andreas; Spyroulias, Georgios A

    2016-10-01

    The H-NOX (Heme-nitric oxide/oxygen binding) domain is conserved across eukaryotes and bacteria. In human soluble guanylyl cyclase (sGC) the H-NOX domain functions as a sensor for the gaseous signaling agent nitric oxide (NO). sGC contains the heme-binding H-NOX domain at its N-terminus, which regulates the catalytic site contained within the C-terminal end of the enzyme catalyzing the conversion of GTP (guanosine 5'-triphosphate) to GMP (guanylyl monophosphate). Here, we present the backbone and side-chain assignments of the (1)H, (13)C and (15)N resonances of the 183-residue H-NOX domain from Nostoc sp. through solution NMR.

  16. (1)H, (13)C and (15)N resonance assignment of the first N-terminal RNA recognition motif (RRM) of the human heterogeneous nuclear ribonucleoprotein H (hnRNP H).

    Science.gov (United States)

    Cabal, Stéphanie; van Heijenoort, Carine; Guittet, Eric

    2007-12-01

    Human heterogeneous nuclear ribonucleoprotein H (hnRNP H) regulates alternative splicing of HIV-1 Tat pre-mRNA. The structure of the first N-terminal domain (residues 1-104) of hnRNP H was solved and its binding to an exonic splicing silencer (pESS2) studied. For this, all backbone and 85% of side-chain resonance frequencies were assigned.

  17. Ner protein of phage Mu: Assignments using {sup 13}C/{sup 15}N-labeled protein

    Energy Technology Data Exchange (ETDEWEB)

    Strzelecka, T.; Gronenborn, A.M.; Clore, G.M. [National Institutes of Health, Bethesda, MD (United States)

    1994-12-01

    The Ner protein is a small (74-amino acid) DNA-binding protein that regulates a switch between the lysogenic and lytic stages of phage Mu. It inhibits expression of the C repressor gene and down-regulates its own expression. Two-dimensional NMR experiments on uniformly {sup 15}N-labeled protein provided most of the backbone and some of the sidechain proton assignments. The secondary structure determination using two-dimensional NOESY experiments showed that Ner consists of five {alpha}-helices. However, because most of the sidechain protons could not be assigned, the full structure was not determined. Using uniformly {sup 13}C/{sup 15}N-labeled Ner and a set of three-dimensional experiments, we were able to assign all of the backbone and 98% of the sidechain protons. In particular, the CBCANH and CBCA(CO)NH experiments were used to sequentially assign the C{alpha} and C{beta} resonances; the HCCH-CTOCSY and HCCH-COSY were used to assign sidechain carbon and proton resonances.

  18. Stereospecific assignments of glycine in proteins by stereospecific deuteration and {sup 15}N labeling

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, A.P.; Curley, R.W. Jr.; Panigot, M.J.; Fesik, S.W. [Ohio State Univ., Columbus, OH (United States)

    1994-12-01

    Stereospecific assignments are important for accurately determining the three-dimensional structures of proteins through the use of multidimensional NMR techniques. It is especially important to stereospecifically assign the glycine {alpha}-protons in proteins because of the potential for different backbone conformations of this residue. These stereospecific assignments are critical for interpreting the {sup 3}J{sub NH,{alpha}H} coupling constants and NOEs involving the glycine {alpha}-protons that determine the conformation of this part of the protein. However, it is often difficult to unambiguously obtain the stereospecific assignments for glycine residues by using only NOE data. In this poster, we present a method for unambiguous, stereospecific assignment of the {alpha}-protons of glycine residues. This method involves synthesis of stereo-specifically deuterated and {sup 15}N-labeled Gly using a slightly modified procedure originally described by Woodard and coworkers for the stereoselective deuteration of glycine. The stereospecifically deuterated and {sup 15}N-labeled Gy has been incorporated into recombinant proteins expressed in both bacterial systems (FKBP) and mammalian cells (u-PA). Two- and three-dimensional isotope-filtered and isotope-edited NMR experiments were used to obtain the stereospecific assignments of the glycine {alpha}-protons for these proteins.

  19. Addendum to "Determination of $\\gamma$-ray widths in $^{15}$N using nuclear resonance fluorescence''

    CERN Document Server

    Szücs, Tamás

    2015-01-01

    The determination of absolute widths of two observed levels above the proton threshold in $^{15}$N has been improved by a combined analysis of our recent $^{15}$N($\\gamma$,$\\gamma'$)$^{15}$N$^\\ast$ photon scattering data, resonance strengths $\\omega\\gamma$ of the $^{14}$C($p$,$\\gamma$)$^{15}$N reaction, and $\\gamma$-ray branchings $b_{\\gamma,i}$ in $^{15}$N. The revised data are compared to the adopted values, and some inconsistencies in the adopted values are illustrated.

  20. MUSIC in triple-resonance experiments: amino acid type-selective (1)H-(15)N correlations

    Science.gov (United States)

    Schubert; Smalla; Schmieder; Oschkinat

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective (1)H-(15)N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH(2) or XH(3) (X can be (15)N or (13)C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains. Copyright 1999 Academic Press.

  1. Complete 1H, 15N and 13C assignment of trappin-2 and 1H assignment of its two domains, elafin and cementoin.

    Science.gov (United States)

    Loth, Karine; Alami, Soha Abou Ibrahim; Habès, Chahrazed; Garrido, Solène; Aucagne, Vincent; Delmas, Agnès F; Moreau, Thierry; Zani, Marie-Louise; Landon, Céline

    2016-04-01

    Trappin-2 is a serine protease inhibitor with a very narrow inhibitory spectrum and has significant anti-microbial activities. It is a 10 kDa cationic protein composed of two distinct domains. The N-terminal domain (38 residues) named cementoin is known to be intrinsically disordered when it is not linked to the elafin. The C-terminal domain (57 residues), corresponding to elafin, is a cysteine-rich domain stabilized by four disulfide bridges and is characterized by a flat core and a flexible N-terminal part. To our knowledge, there is no structural data available on trappin-2. We report here the complete (1)H, (15)N and (13)C resonance assignment of the recombinant trappin-2 and the (1)H assignments of cementoin and elafin, under the same experimental conditions. This is the first step towards the 3D structure determination of the trappin-2.

  2. Combining automated peak tracking in SAR by NMR with structure-based backbone assignment from 15N-NOESY

    KAUST Repository

    Jang, Richard

    2012-03-21

    Background: Chemical shift mapping is an important technique in NMR-based drug screening for identifying the atoms of a target protein that potentially bind to a drug molecule upon the molecule\\'s introduction in increasing concentrations. The goal is to obtain a mapping of peaks with known residue assignment from the reference spectrum of the unbound protein to peaks with unknown assignment in the target spectrum of the bound protein. Although a series of perturbed spectra help to trace a path from reference peaks to target peaks, a one-to-one mapping generally is not possible, especially for large proteins, due to errors, such as noise peaks, missing peaks, missing but then reappearing, overlapped, and new peaks not associated with any peaks in the reference. Due to these difficulties, the mapping is typically done manually or semi-automatically, which is not efficient for high-throughput drug screening.Results: We present PeakWalker, a novel peak walking algorithm for fast-exchange systems that models the errors explicitly and performs many-to-one mapping. On the proteins: hBclXL, UbcH5B, and histone H1, it achieves an average accuracy of over 95% with less than 1.5 residues predicted per target peak. Given these mappings as input, we present PeakAssigner, a novel combined structure-based backbone resonance and NOE assignment algorithm that uses just 15N-NOESY, while avoiding TOCSY experiments and 13C-labeling, to resolve the ambiguities for a one-to-one mapping. On the three proteins, it achieves an average accuracy of 94% or better.Conclusions: Our mathematical programming approach for modeling chemical shift mapping as a graph problem, while modeling the errors directly, is potentially a time- and cost-effective first step for high-throughput drug screening based on limited NMR data and homologous 3D structures. 2012 Jang et al.; licensee BioMed Central Ltd.

  3. Combining ambiguous chemical shift mapping with structure-based backbone and NOE assignment from 15N-NOESY

    KAUST Repository

    Jang, Richard

    2011-01-01

    Chemical shift mapping is an important technique in NMRbased drug screening for identifying the atoms of a target protein that potentially bind to a drug molecule upon the molecule\\'s introduction in increasing concentrations. The goal is to obtain a mapping of peaks with known residue assignment from the reference spectrum of the unbound protein to peaks with unknown assignment in the target spectrum of the bound protein. Although a series of perturbed spectra help to trace a path from reference peaks to target peaks, a one-to-one mapping generally is not possible, especially for large proteins, due to errors, such as noise peaks, missing peaks, missing but then reappearing, overlapped, and new peaks not associated with any peaks in the reference. Due to these difficulties, the mapping is typically done manually or semi-automatically. However, automated methods are necessary for high-throughput drug screening. We present PeakWalker, a novel peak walking algorithm for fast-exchange systems that models the errors explicitly and performs many-to-one mapping. On the proteins: hBclXL, UbcH5B, and histone H1, it achieves an average accuracy of over 95% with less than 1.5 residues predicted per target peak. Given these mappings as input, we present PeakAssigner, a novel combined structure-based backbone resonance and NOE assignment algorithm that uses just 15N-NOESY, while avoiding TOCSY experiments and 13C- labeling, to resolve the ambiguities for a one-toone mapping. On the three proteins, it achieves an average accuracy of 94% or better. Copyright © 2011 ACM.

  4. Binding of thiocyanate to lactoperoxidase: 1H and 15N nuclear magnetic resonance studies

    Energy Technology Data Exchange (ETDEWEB)

    Modi, S.; Behere, D.V.; Mitra, S. (Tata Institute of Fundamental Research, Bombay (India))

    1989-05-30

    The binding of thiocyanate to lactoperoxidase (LPO) has been investigated by 1H and 15N NMR spectroscopy. 1H NMR of LPO shows that the major broad heme methyl proton resonance at about 61 ppm is shifted upfield by addition of the thiocyanate, indicating binding of the thiocyanate to the enzyme. The pH dependence of line width of 15N resonance of SC15N- in the presence of the enzyme has revealed that the binding of the thiocyanate to the enzyme is facilitated by protonation of an ionizable group (with pKa of 6.4), which is presumably distal histidine. Dissociation constants (KD) of SC15N-/LPO, SC15N-/LPO/I-, and SC15N-/LPO/CN- equilibria have been determined by 15N T1 measurements and found to be 90 +/- 5, 173 +/- 20, and 83 +/- 6 mM, respectively. On the basis of these values of KD, it is suggested that the iodide ion inhibits the binding of the thiocyanate but cyanide ion does not. The thiocyanate is shown to bind at the same site of LPO as iodide does, but the binding is considerably weaker and is away from the ferric ion. The distance of 15N of the bound thiocyanate ion from the iron is determined to be 7.2 +/- 0.2 A from the 15N T1 measurements.

  5. Through-space (19) F-(15) N couplings for the assignment of stereochemistry in flubenzimine.

    Science.gov (United States)

    Ghiviriga, Ion; Rubinski, Miles A; Dolbier, William R

    2016-07-01

    Through-space (19) F-(15) N couplings revealed the configuration of flubenzimine, with the CF3 group on N4 pointing towards the lone pair of N5. The (19) F-(15) N coupling constants were measured at natural abundance using a spin-state selective indirect-detection pulse sequence. As (15) N-labelled proteins are routinely synthesized for NMR studies, through-space (19) F-(15) N couplings have the potential to probe the stereochemistry of these proteins by (19) F labelling of some amino acids or can reveal the site of docking of fluorine-containing drugs. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Determination of gamma-ray widths in $^{15}$N using nuclear resonance fluorescence

    OpenAIRE

    Szücs, T.; Bemmerer, D.; Caciolli, A.; Fülöp, Zs.; Massarczyk, R.; Michelagnoli, C.; Reinhardt, T. P.; Schwengner, R.; Takács, M. P.; Ur, C. A.; Wagner, A.; Wagner, L.

    2015-01-01

    The stable nucleus $^{15}$N is the mirror of $^{15}$O, the bottleneck in the hydrogen burning CNO cycle. Most of the $^{15}$N level widths below the proton emission threshold are known from just one nuclear resonance fluorescence (NRF) measurement, with limited precision in some cases. A recent experiment with the AGATA demonstrator array determined level lifetimes using the Doppler Shift Attenuation Method (DSAM) in $^{15}$O. As a reference and for testing the method, level lifetimes in $^{1...

  7. Gamma-ray width measurements in {sup 15}N at the ELBE nuclear resonance fluorescence setup

    Energy Technology Data Exchange (ETDEWEB)

    Szuecs, Tamas [Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden (Germany); MTA ATOMKI, Debrecen/Hungary (Hungary); Bemmerer, Daniel; Schwengner, Ronald [Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden (Germany); Massarczyk, Ralph; Takacs, Marcell; Wagner, Louis [Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden (Germany); TU Dresden (Germany)

    2014-07-01

    The stable nucleus {sup 15}N is the mirror of the astrophysically important {sup 15}O, compound nucleus of the leading reaction of the Bethe-Weizsaecker cycle of hydrogen burning. Most of the {sup 15}N level widths below the neutron and proton emission thresholds are known from just one nuclear resonance fluorescence (NRF) measurement published more than 30 years ago, with unsatisfactory precision on some cases. A recent experiment with the AGATA demonstrator array aimed to determine level widths with the Doppler Shift Attenuation Method (DSAM) in {sup 15}O and {sup 15}N populated in {sup 14}N + {sup 2}H reaction. In order to set a benchmark value for the upcoming AGATA demonstrator data, the widths of several {sup 15}N levels are being studied using the bremsstrahlung facility γELBE at the electron accelerator of Helmholtz-Zentrum Dresden-Rossendorf (HZDR). The γELBE experiment and its preliminary results are presented.

  8. Determination of level widths in 15N using nuclear resonance fluorescence

    Directory of Open Access Journals (Sweden)

    Szücs T.

    2015-01-01

    Full Text Available Level widths in 15N have been measured with the nuclear resonance fluorescence (NRF technique. Solid nitrogen compounds, bremsstrahlung, and HPGe detectors have been used as target, beam, and detectors, respectively. The preliminarily level widths are in agreement with the literature values, but more precise.

  9. Determination of gamma-ray widths in $^{15}$N using nuclear resonance fluorescence

    CERN Document Server

    Szücs, T; Caciolli, A; Fülöp, Zs; Massarczyk, R; Michelagnoli, C; Reinhardt, T P; Schwengner, R; Takács, M P; Ur, C A; Wagner, A; Wagner, L

    2015-01-01

    The stable nucleus $^{15}$N is the mirror of $^{15}$O, the bottleneck in the hydrogen burning CNO cycle. Most of the $^{15}$N level widths below the proton emission threshold are known from just one nuclear resonance fluorescence (NRF) measurement, with limited precision in some cases. A recent experiment with the AGATA demonstrator array determined level lifetimes using the Doppler Shift Attenuation Method (DSAM) in $^{15}$O. As a reference and for testing the method, level lifetimes in $^{15}$N have also been determined in the same experiment. The latest compilation of $^{15}$N level properties dates back to 1991. The limited precision in some cases in the compilation calls for a new measurement in order to enable a comparison to the AGATA demonstrator data. The widths of several $^{15}$N levels have been studied with the NRF method. The solid nitrogen compounds enriched in $^{15}$N have been irradiated with bremsstrahlung. The $\\gamma$-rays following the deexcitation of the excited nuclear levels were dete...

  10. Automatic assignment of protein backbone resonances by direct spectrum inspection in targeted acquisition of NMR data.

    Science.gov (United States)

    Wong, Leo E; Masse, James E; Jaravine, Victor; Orekhov, Vladislav; Pervushin, Konstantin

    2008-10-01

    The necessity to acquire large multidimensional datasets, a basis for assignment of NMR resonances, results in long data acquisition times during which substantial degradation of a protein sample might occur. Here we propose a method applicable for such a protein for automatic assignment of backbone resonances by direct inspection of multidimensional NMR spectra. In order to establish an optimal balance between completeness of resonance assignment and losses of cross-peaks due to dynamic processes/degradation of protein, assignment of backbone resonances is set as a stirring criterion for dynamically controlled targeted nonlinear NMR data acquisition. The result is demonstrated with the 12 kDa (13)C,(15) N-labeled apo-form of heme chaperone protein CcmE, where hydrolytic cleavage of 29 C-terminal amino acids is detected. For this protein, 90 and 98% of manually assignable resonances are automatically assigned within 10 and 40 h of nonlinear sampling of five 3D NMR spectra, respectively, instead of 600 h needed to complete the full time domain grid. In addition, resonances stemming from degradation products are identified. This study indicates that automatic resonance assignment might serve as a guiding criterion for optimal run-time allocation of NMR resources in applications to proteins prone to degradation.

  11. Automatic assignment of protein backbone resonances by direct spectrum inspection in targeted acquisition of NMR data

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Leo E. [Nanyang Technological University, School of Biological Sciences (Singapore); Masse, James E. [National Institutes of Health (United States); Jaravine, Victor [J. W. Goethe-University Frankfurt, Institute of Biophysical Chemistry (Germany); Orekhov, Vladislav [Gothenburg University, Swedish NMR Centre (Sweden); Pervushin, Konstantin [Nanyang Technological University, School of Biological Sciences (Singapore)], E-mail: kpervushin@ntu.edu.sg

    2008-10-15

    The necessity to acquire large multidimensional datasets, a basis for assignment of NMR resonances, results in long data acquisition times during which substantial degradation of a protein sample might occur. Here we propose a method applicable for such a protein for automatic assignment of backbone resonances by direct inspection of multidimensional NMR spectra. In order to establish an optimal balance between completeness of resonance assignment and losses of cross-peaks due to dynamic processes/degradation of protein, assignment of backbone resonances is set as a stirring criterion for dynamically controlled targeted nonlinear NMR data acquisition. The result is demonstrated with the 12 kDa {sup 13}C,{sup 15} N-labeled apo-form of heme chaperone protein CcmE, where hydrolytic cleavage of 29 C-terminal amino acids is detected. For this protein, 90 and 98% of manually assignable resonances are automatically assigned within 10 and 40 h of nonlinear sampling of five 3D NMR spectra, respectively, instead of 600 h needed to complete the full time domain grid. In addition, resonances stemming from degradation products are identified. This study indicates that automatic resonance assignment might serve as a guiding criterion for optimal run-time allocation of NMR resources in applications to proteins prone to degradation.

  12. Assignments of the Pfr-Pr FTIR difference spectrum of cyanobacterial phytochrome Cph1 using 15N and 13C isotopically labeled phycocyanobilin chromophore.

    Science.gov (United States)

    van Thor, Jasper J; Fisher, Nicholas; Rich, Peter R

    2005-11-03

    The reversible red and far-red light-induced transitions of cyanobacterial phytochrome Cph1 from Synechocystis PCC 6803 were investigated by Fourier transform infrared (FTIR) difference spectroscopy. High-quality light-induced Pfr-Pr difference FTIR spectra were recorded for the 58 kDa N-terminal domain of Cph1 by repetitive photochemical cycling and signal averaging. The Pfr-Pr difference spectra in H(2)O and D(2)O were very similar to those previously reported for full-length 85 kDa Cph1.(1) Published assignments were extended by analysis of the effects of (13)C and (15)N isotope substitutions at selected sites in the phycocyanobilin chromophore and by (15)N global labeling of the protein. The Pfr-Pr difference spectra were dominated by an amide I peak/trough at 1653 cm(-1)(+)/1631 cm(-1)(-) and a smaller amide II band at 1554 cm(-1). Labeling effects allowed specific chromophore assignments for the C(1)=O (1736 cm(-1)(-)/1724 cm(-1)(+)) and C(19)=O (1704 cm(-1)(-)) carbonyl vibrations, C=C vibrations at 1589 cm(-1)(+), and bands at 1537(-), 1512(+), 1491(-), 1163(+), 1151(-), 1134(+), 1109(-), and 1072(-) cm(-1) that must involve chromophore C-N bonds. A variety of additional changes were insensitive to isotope labeling of the chromophore. Effects of (15)N labeling of the protein were used to tentatively assign some of these to specific amino acid changes. Those insensitive to (15)N labeling included a protonated aspartic or glutamic acid at 1734 cm(-1)(-)/1722 cm(-1)(+) and a cysteine at 2575 cm(-1)(+)/2557 cm(-1)(-). Bands sensitive to (15)N protein labeling at 1487 cm(-1)(+)/1502 cm(-1)(-) might arise from trytophan and bands at 1261 cm(-1)(+)/1244 cm(-1)(-) and 1107 cm(-1)(-)/1095 cm(-1)(+) might arise from a histidine environment or protonation change. These assignments are discussed in light of the 15Z-E photoisomerization model of phototransformation and the associated protein conformational changes.

  13. Nitrogen-detected CAN and CON experiments as alternative experiments for main chain NMR resonance assignments.

    Science.gov (United States)

    Takeuchi, Koh; Heffron, Gregory; Sun, Zhen-Yu J; Frueh, Dominique P; Wagner, Gerhard

    2010-08-01

    Heteronuclear direct-detection experiments, which utilize the slower relaxation properties of low gamma nuclei, such as (13)C have recently been proposed for sequence-specific assignment and structural analyses of large, unstructured, and/or paramagnetic proteins. Here we present two novel (15)N direct-detection experiments. The CAN experiment sequentially connects amide (15)N resonances using (13)C(alpha) chemical shift matching, and the CON experiment connects the preceding (13)C' nuclei. When starting from the same carbon polarization, the intensities of nitrogen signals detected in the CAN or CON experiments would be expected four times lower than those of carbon resonances observed in the corresponding (13)C-detecting experiment, NCA-DIPAP or NCO-IPAP (Bermel et al. 2006b; Takeuchi et al. 2008). However, the disadvantage due to the lower gamma is counteracted by the slower (15)N transverse relaxation during detection, the possibility for more efficient decoupling in both dimensions, and relaxation optimized properties of the pulse sequences. As a result, the median S/N in the (15)N observe CAN experiment is 16% higher than in the (13)C observe NCA-DIPAP experiment. In addition, significantly higher sensitivity was observed for those residues that are hard to detect in the NCA-DIPAP experiment, such as Gly, Ser and residues with high-field C(alpha) resonances. Both CAN and CON experiments are able to detect Pro resonances that would not be observed in conventional proton-detected experiments. In addition, those experiments are free from problems of incomplete deuterium-to-proton back exchange in amide positions of perdeuterated proteins expressed in D(2)O. Thus, these features and the superior resolution of (15)N-detected experiments provide an attractive alternative for main chain assignments. The experiments are demonstrated with the small model protein GB1 at conditions simulating a 150 kDa protein, and the 52 kDa glutathione S-transferase dimer, GST.

  14. Absolute hydrogen depth profiling using the resonant $^{1}$H($^{15}$N,$\\alpha\\gamma$)$^{12}$C nuclear reaction

    CERN Document Server

    Reinhardt, Tobias P; Bemmerer, Daniel; Stöckel, Klaus; Wagner, Louis

    2016-01-01

    Resonant nuclear reactions are a powerful tool for the determination of the amount and profile of hydrogen in thin layers of material. Usually, this tool requires the use of a standard of well-known composition. The present work, by contrast, deals with standard-less hydrogen depth profiling. This approach requires precise nuclear data, e.g. on the widely used $^{1}$H($^{15}$N,$\\alpha\\gamma$)$^{12}$C reaction, resonant at 6.4\\,MeV $^{15}$N beam energy. Here, the strongly anisotropic angular distribution of the emitted $\\gamma$-rays from this resonance has been re-measured, resolving a previous discrepancy. Coefficients of (0.38$\\pm$0.04) and (0.80$\\pm$0.04) have been deduced for the second and fourth order Legendre polynomials, respectively. In addition, the resonance strength has been re-evaluated to (25.0$\\pm$1.5)\\,eV, 10\\% higher than previously reported. A simple working formula for the hydrogen concentration is given for cases with known $\\gamma$-ray detection efficiency. Finally, the absolute approach i...

  15. Ratios of 15N/12C and 4He/12C inclusive electroproduction cross sections in the nucleon resonance region

    CERN Document Server

    Bosted, P E; Amarian, M; Anefalos, S; Anghinolfi, M; Asryan, G; Avakian, H; Bagdasaryan, H; Baillie, N; Ball, J P; Baltzell, N A; Barrow, S; Batourine, V; Battaglieri, M; Beard, K; Bedlinskiy, I; Bektasoglu, M; Bellis, M; Benmouna, N; Biselli, A S; Bonner, B E; Bouchigny, S; Boiarinov, S; Bradford, R; Branford, D; Brooks, W K; Bültmann, S; Burkert, V D; Butuceanu, C; Calarco, J R; Careccia, S L; Carman, D S; Carnahan, B; Cazes, A; Chen, S; Cole, P L; Collins, P; Coltharp, P; Cords, D; Corvisiero, P; Crabb, D; Crannell, H; Credé, V; Cummings, J P; De Masi, R; De Vita, R; De Sanctis, E; Degtyarenko, P V; Denizli, H; Dennis, L; Deur, A; Djalali, C; Dodge, G E; Donnelly, J; Doughty, D; Dragovitsch, P; Dugger, M; Dharmawardane, K V; Dytman, S; Dzyubak, O P; Egiyan, H; Egiyan, K S; Elouadrhiri, L; Eugenio, P; Fatemi, R; Fedotov, G; Feuerbach, R J; Forest, T A; Fradi, A; Funsten, H; Garçon, M; Gavalian, G; Gilfoyle, G P; Giovanetti, K L; Girod, F X; Goetz, J T; Golovatch, E; Gothe, R W; Griffioen, K A; Guidal, M; Guillo, M; Guler, N; Guo, L; Gyurjyan, V; Hadjidakis, C; Hafidi, K; Hakobyan, R S; Hardie, J; Heddle, D; Hersman, F W; Hicks, K; Hleiqawi, I; Holtrop, M; Huertas, M; Hyde-Wright, C E; Ilieva, Y; Ireland, D G; Ishkhanov, B S; Isupov, E L; Ito, M M; Jenkins, D; Jo, H S; Joo, K; Jüngst, H G; Kalantarians, N; Keith, C; Kellie, J D; Khandaker, M; Kim, K Y; Kim, K; Kim, W; Klein, A; Klein, F J; Klusman, M; Kossov, M; Kramer, L H; Kubarovski, V; Kühn, J; Kuhn, S E; Kuleshov, S V; Lachniet, J; Laget, J M; Langheinrich, J; Lawrence, D; Ji Li; Lima, A C S; Livingston, K; Lü, H; Lukashin, K; MacCormick, M; Markov, N; McAleer, S; McKinnon, B; McNabb, J W C; Mecking, B A; Mestayer, M D; Meyer, C A; Mibe, T; Mikhailov, K; Minehart, R; Mirazita, M; Miskimen, R; Mokeev, V; Morand, L; Morrow, S A; Moteabbed, M; Müller, J; Mutchler, G S; Nadel-Turonski, P; Nasseripour, R; Niccolai, S; Niculescu, G; Niculescu, I; Niczyporuk, B B; Niroula, M R; Niyazov, R A; Nozar, M; O'Rielly, G V; Osipenko, M; Ostrovidov, A I; Park, K; Pasyuk, E; Paterson, C; Philips, S A; Pierce, J; Pivnyuk, N; Pocanic, D; Pogorelko, O; Polli, E; Pozdniakov, S; Preedom, B M; Price, J W; Prok, Y; Protopopescu, D; Qin, L M; Raue, B A; Riccardi, G; Ricco, G; Ripani, M; Rosner, G; Rossi, P; Rowntree, D; Rubin, P D; Sabati, F; Salgado, C; Santoro, J P; Sapunenko, V; Schumacher, R A; Serov, V S; Sharabyan, Yu G; Shaw, J; Shvedunov, N V; Skabelin, A V; Smith, E S; Smith, L C; Sober, D I; Stavinsky, A; Stepanyan, S S; Stepanyan, S; Stokes, B E; Stoler, P; Strauch, S; Suleiman, R; Taiuti, M; Taylor, S; Tedeschi, D J; Thoma, U; Tkabladze, A; Tkachenko, S; Todor, L; Ungaro, M; Vineyard, M F; Vlassov, A V; Weinstein, L B; Weygand, D P; Williams, M; Wolin, E; Wood, M H; Yegneswaran, A; Yun, J; Zana, L; Zhang, J; Zhao, B; Zhao, Z

    2007-01-01

    The ratio of inclusive electron scattering cross sections for 15N/12C was determined in the kinematic range 0.8resonance structure, as predicted by a phenomenological model, and also by quark-hadron duality. Within the super-scaling quasi-elastic model, slight evidence is found for a 1 MeV lower effective nucleon binding energy in 15N than in 12C. Ratios of 4He/12C using 1.6 to 2.5 GeV electrons are in good agreement with the phenomenological model.

  16. 1H, 13C and 15N backbone and side-chain chemical shift assignment of the Fyn SH2 domain and its complex with a phosphotyrosine peptide.

    Science.gov (United States)

    Huculeci, Radu; Buts, Lieven; Lenaerts, Tom; van Nuland, Nico A J

    2011-10-01

    SH2 domains are interaction modules uniquely dedicated to recognize phosphotyrosine sites, playing a central role in for instance the activation of tyrosine kinases or phosphatases. Here we report the (1)H, (15)N and (13)C backbone and side-chain chemical shift assignments of the SH2 domain of the human protein tyrosine kinase Fyn, both in its free state and bound to a high-affinity phosphotyrosine peptide corresponding to a specific sequence in the hamster middle-T antigen. The BMRB accession numbers are 17,368 and 17,369, respectively.

  17. Natural abundance 15N NMR assignments delineate structural differences between intact and reactive-site hydrolyzed Cucurbita maxima trypsin inhibitor III.

    Science.gov (United States)

    Krishnamoorthi, R; Nemmers, S; Tobias, B

    1992-06-15

    15N NMR assignments were made to the backbone amide nitrogen atoms at natural isotopic abundance of intact and reactive-site (Arg5-Ile6) hydrolyzed Cucurbita maxima trypsin inhibitor III (CMTI-III and CMTI-III*, respectively) by means of 2D proton-detected heteronuclear single bond chemical shift correlation (HSBC) spectroscopy, utilizing the previously made sequence-specific 1H NMR assignments (Krishnamoorthi et al. (1992) Biochemistry 31, 898-904). Comparison of the 15N chemical shifts of the two forms of the inhibitor molecule revealed significant changes not only for residues located near the reactive-site region, but also for those distantly located. Residues Cys3, Arg5, Leu7, Met8, Cys10, Cys16, Glu19, His25, Tyr27, Cys28 and Gly29 showed significant chemical shift changes ranging from 0.3 to 6.1 ppm, thus indicating structural perturbations that were transmitted throughout the molecule. These findings confirm the earlier conclusions based on 1H NMR investigations.

  18. Combination of {sup 15}N reverse labeling and afterglow spectroscopy for assigning membrane protein spectra by magic-angle-spinning solid-state NMR: application to the multidrug resistance protein EmrE

    Energy Technology Data Exchange (ETDEWEB)

    Banigan, James R.; Gayen, Anindita; Traaseth, Nathaniel J., E-mail: traaseth@nyu.edu [New York University, Department of Chemistry (United States)

    2013-04-15

    Magic-angle-spinning (MAS) solid-state NMR spectroscopy has emerged as a viable method to characterize membrane protein structure and dynamics. Nevertheless, the spectral resolution for uniformly labeled samples is often compromised by redundancy of the primary sequence and the presence of helical secondary structure that results in substantial resonance overlap. The ability to simplify the spectrum in order to obtain unambiguous site-specific assignments is a major bottleneck for structure determination. To address this problem, we used a combination of {sup 15}N reverse labeling, afterglow spectroscopic techniques, and frequency-selective dephasing experiments that dramatically improved the ability to resolve peaks in crowded spectra. This was demonstrated using the polytopic membrane protein EmrE, an efflux pump involved in multidrug resistance. Residues preceding the {sup 15}N reverse labeled amino acid were imaged using a 3D NCOCX afterglow experiment and those following were recorded using a frequency-selective dephasing experiment. Our approach reduced the spectral congestion and provided a sensitive way to obtain chemical shift assignments for a membrane protein where no high-resolution structure is available. This MAS methodology is widely applicable to the study of other polytopic membrane proteins in functional lipid bilayer environments.

  19. 1H, 15N and 13C backbone assignments of GDP-bound human H-Ras mutant G12V.

    Science.gov (United States)

    Amin, Nader; Chiarparin, Elisabetta; Coyle, Joe; Nietlispach, Daniel; Williams, Glyn

    2016-04-01

    Harvey Ras (H-Ras) is a membrane-associated GTPase with critical functions in cell proliferation and differentiation. The G12V mutant of H-Ras is one of the most commonly encountered oncoproteins in human cancer. This mutation disrupts the GTPase activity of H-Ras, leading to constitutive activation and aberrant downstream signalling. Here we report the backbone resonance assignments of human H-Ras mutant G12V lacking the C-terminal membrane attachment domain.

  20. Identifying the African wintering grounds of hybrid flycatchers using a multi-isotope (δ2H, δ13C, δ15N) assignment approach.

    Science.gov (United States)

    Veen, Thor; Hjernquist, Mårten B; Van Wilgenburg, Steven L; Hobson, Keith A; Folmer, Eelke; Font, Laura; Klaassen, Marcel

    2014-01-01

    Migratory routes and wintering grounds can have important fitness consequences, which can lead to divergent selection on populations or taxa differing in their migratory itinerary. Collared (Ficedula albicollis) and pied (F. hypoleuca) flycatchers breeding in Europe and wintering in different sub-Saharan regions have distinct migratory routes on the eastern and western sides of the Sahara desert, respectively. In an earlier paper, we showed that hybrids of the two species did not incur reduced winter survival, which would be expected if their migration strategy had been a mix of the parent species' strategies potentially resulting in an intermediate route crossing the Sahara desert to different wintering grounds. Previously, we compared isotope ratios and found no significant difference in stable-nitrogen isotope ratios (δ15N) in winter-grown feathers between the parental species and hybrids, but stable-carbon isotope ratios (δ13C) in hybrids significantly clustered only with those of pied flycatchers. We followed up on these findings and additionally analyzed the same feathers for stable-hydrogen isotope ratios (δ2H) and conducted spatially explicit multi-isotope assignment analyses. The assignment results overlapped with presumed wintering ranges of the two species, highlighting the efficacy of the method. In contrast to earlier findings, hybrids clustered with both parental species, though most strongly with pied flycatcher.

  1. Identifying the African wintering grounds of hybrid flycatchers using a multi-isotope (δ2H, δ13C, δ15N assignment approach.

    Directory of Open Access Journals (Sweden)

    Thor Veen

    Full Text Available Migratory routes and wintering grounds can have important fitness consequences, which can lead to divergent selection on populations or taxa differing in their migratory itinerary. Collared (Ficedula albicollis and pied (F. hypoleuca flycatchers breeding in Europe and wintering in different sub-Saharan regions have distinct migratory routes on the eastern and western sides of the Sahara desert, respectively. In an earlier paper, we showed that hybrids of the two species did not incur reduced winter survival, which would be expected if their migration strategy had been a mix of the parent species' strategies potentially resulting in an intermediate route crossing the Sahara desert to different wintering grounds. Previously, we compared isotope ratios and found no significant difference in stable-nitrogen isotope ratios (δ15N in winter-grown feathers between the parental species and hybrids, but stable-carbon isotope ratios (δ13C in hybrids significantly clustered only with those of pied flycatchers. We followed up on these findings and additionally analyzed the same feathers for stable-hydrogen isotope ratios (δ2H and conducted spatially explicit multi-isotope assignment analyses. The assignment results overlapped with presumed wintering ranges of the two species, highlighting the efficacy of the method. In contrast to earlier findings, hybrids clustered with both parental species, though most strongly with pied flycatcher.

  2. Pseudo 5D HN(C)N Experiment to Facilitate the Assignment of Backbone Resonances in Proteins Exhibiting High Backbone Shift Degeneracy

    CERN Document Server

    Kumar, Dinesh; Shukla, Vaibhav Kumar; Pandey, Himanshu; Arora, Ashish; Guleria, Anupam

    2014-01-01

    Assignment of protein backbone resonances is most routinely carried out using triple resonance three dimensional NMR experiments involving amide 1H and 15N resonances. However for intrinsically unstructured proteins, alpha-helical proteins or proteins containing several disordered fragments, the assignment becomes problematic because of high degree of backbone shift degeneracy. In this backdrop, a novel reduced dimensionality (RD) experiment -(5,3)D-hNCO-CANH- is presented to facilitate (and/or to validate) the sequential backbone resonance assignment in such proteins. The proposed 3D NMR experiment makes use of the modulated amide 15N chemical shifts (resulting from the joint sampling along both its indirect dimensions) to resolve the ambiguity involved in connecting the neighboring amide resonances (i.e. HiNi and Hi-1Ni-1) for overlapping amide NH peaks. The experiment -encoding 5D spectral information- leads to a conventional 3D spectrum with significantly reduced spectral crowding and complexity. The impr...

  3. Approaches to the assignment of {sup 19}F resonances from 3-fluorophenylalanine labeled calmodulin using solution state NMR

    Energy Technology Data Exchange (ETDEWEB)

    Kitevski-LeBlanc, Julianne L.; Evanics, Ferenc; Scott Prosser, R., E-mail: scott.prosser@utoronto.c [University of Toronto, Department of Chemistry (Canada)

    2010-06-15

    Traditional single site replacement mutations (in this case, phenylalanine to tyrosine) were compared with methods which exclusively employ {sup 15}N and {sup 19}F-edited two- and three-dimensional NMR experiments for purposes of assigning {sup 19}F NMR resonances from calmodulin (CaM), biosynthetically labeled with 3-fluorophenylalanine (3-FPhe). The global substitution of 3-FPhe for native phenylalanine was tolerated in CaM as evidenced by a comparison of {sup 1}H-{sup 15}N HSQC spectra and calcium binding assays in the presence and absence of 3-FPhe. The {sup 19}F NMR spectrum reveals six resolved resonances, one of which integrates to three 3-FPhe species, making for a total of eight fluorophenylalanines. Single phenylalanine to tyrosine mutants of five phenylalanine positions resulted in {sup 19}F NMR spectra with significant chemical shift perturbations of the remaining resonances, and provided only a single definitive assignment. Although {sup 1}H-{sup 19}F heteronucleclear NOEs proved weak, {sup 19}F-edited {sup 1}H-{sup 1}H NOESY connectivities were relatively easy to establish by making use of the {sup 3}J{sub FH} coupling between the fluorine nucleus and the adjacent fluorophenylalanine {delta} proton. {sup 19}F-edited NOESY connectivities between the {delta} protons and {alpha} and {beta} nuclei in addition to {sup 15}N-edited {sup 1}H, {sup 1}H NOESY crosspeaks proved sufficient to assign 4 of 8 {sup 19}F resonances. Controlled cleavage of the protein into two fragments using trypsin, and a repetition of the above 2D and 3D techniques resulted in unambiguous assignments of all 8 {sup 19}F NMR resonances. Our studies suggest that {sup 19}F-edited NOESY NMR spectra are generally adequate for complete assignment without the need to resort to mutational analysis.

  4. A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins.

    Science.gov (United States)

    Wiedemann, Christoph; Bellstedt, Peter; Häfner, Sabine; Herbst, Christian; Bordusa, Frank; Görlach, Matthias; Ohlenschläger, Oliver; Ramachandran, Ramadurai

    2016-07-04

    The RF pulse scheme RN[N-CA HEHAHA]NH, which provides a convenient approach to the acquisition of different multidimensional chemical shift correlation NMR spectra leading to backbone resonance assignments, including those of the proline residues of intrinsically disordered proteins (IDPs), is experimentally demonstrated. Depending on the type of correlation data required, the method involves the generation of in-phase ((15) N)(x) magnetisation via different magnetisation transfer pathways such as H→N→CO→N, HA→CA→CO→N, H→N→CA→N and H→CA→N, the subsequent application of (15) N-(13) C(α) heteronuclear Hartmann-Hahn mixing over a period of ≈100 ms, chemical-shift labelling of relevant nuclei before and after the heteronuclear mixing step and amide proton detection in the acquisition dimension. It makes use of the favourable relaxation properties of IDPs and the presence of (1) JCαN and (2) JCαN couplings to achieve efficient correlation of the backbone resonances of each amino acid residue "i" with the backbone amide resonances of residues "i-1" and "i+1". It can be implemented in a straightforward way through simple modifications of the RF pulse schemes commonly employed in protein NMR studies. The efficacy of the approach is demonstrated using a uniformly ((15) N,(13) C) labelled sample of α-synuclein. The different possibilities for obtaining the amino-acid-type information, simultaneously with the connectivity data between the backbone resonances of sequentially neighbouring residues, have also been outlined.

  5. Root-mean-square-deviation-based rapid backbone resonance assignments in proteins.

    Science.gov (United States)

    Rout, Ashok K; Barnwal, Ravi P; Agarwal, Geetika; Chary, Kandala V R

    2010-10-01

    We have shown that the methodology based on the estimation of root-mean-square deviation (RMSD) between two sets of chemical shifts is very useful to rapidly assign the spectral signatures of (1)H(N), (13)C(α), (13)C(β), (13)C', (1)H(α) and (15)N spins of a given protein in one state from the knowledge of its resonance assignments in a different state, without resorting to routine established procedures (manual and automated). We demonstrate the utility of this methodology to rapidly assign the 3D spectra of a metal-binding protein in its holo-state from the knowledge of its assignments in apo-state, the spectra of a protein in its paramagnetic state from the knowledge of its assignments in diamagnetic state and, finally, the spectra of a mutant protein from the knowledge of the chemical shifts of the corresponding wild-type protein. The underlying assumption of this methodology is that, it is impossible for any two amino acid residues in a given protein to have all the six chemical shifts degenerate and that the protein under consideration does not undergo large conformational changes in going from one conformational state to another. The methodology has been tested using experimental data on three proteins, M-crystallin (8.5 kDa, predominantly β-sheet, for apo- to holo-state), Calbindin (7.5 kDa, predominantly α-helical, for diamagnetic to paramagnetic state and apo to holo) and EhCaBP1 (14.3 kDa, α-helical, the wild-type protein with one of its mutant). In all the cases, the extent of assignment is found to be greater than 85%.

  6. Resonance strengths in the 14N(p,gamma)15O and 15N(p,alpha gamma)12C reactions

    CERN Document Server

    Marta, Michele; Bemmerer, Daniel; Beyer, Roland; Broggini, Carlo; Caciolli, Antonio; Erhard, Martin; Fülöp, Zsolt; Grosse, Eckart; Gyürky, György; Hannaske, Roland; Junghans, Arnd R; Menegazzo, Roberto; Nair, Chithra; Schwengner, Ronald; Szücs, Tamás; Vezzú, Simone; Wagner, Andreas; Yakorev, Dmitry

    2010-01-01

    The 14N(p,gamma)15O reaction is the slowest reaction of the carbon-nitrogen-oxygen cycle of hydrogen burning in stars. As a consequence, it determines the rate of the cycle. The 15N(p,alpha gamma)12C reaction is frequently used in inverse kinematics for hydrogen depth profiling in materials. The 14N(p,gamma)15O and 15N(p,alpha gamma)12C reactions have been studied simultaneously, using titanium nitride targets of natural isotopic composition and a proton beam. The strengths of the resonances at Ep = 1058 keV in 14N(p,gamma)15O and at Ep = 897 and 430 keV in 15N(p,alpha gamma)12C have been determined with improved precision, relative to the well-known resonance at Ep = 278 keV in 14N(p,gamma)15O. The new recommended values are \\omega\\gamma = 0.352$\\pm$0.018, 362$\\pm$20, and 22.0$\\pm$0.9\\,eV for their respective strengths. In addition, the branching ratios for the decay of the Ep = 1058 keV resonance in 14N(p,gamma)15O have been redetermined. The data reported here should facilitate future studies of off-resona...

  7. Resonance assignments and secondary structure of a phytocystatin from Ananas comosus.

    Science.gov (United States)

    Irene, Deli; Chen, Bo-Jiun; Lo, Si-Hung; Liu, Ting-Hang; Tzen, Jason T-C; Chyan, Chia-Lin

    2012-04-01

    A cDNA encoding a cysteine protease inhibitor, cystatin was cloned from pineapple (Ananas comosus L.) stem. This clone was constructed into an expression vector and expressed in E. coli and purified to homogeneous. The recombinant pineapple cystatins (AcCYS) showed effectively inhibitory activity toward cysteine proteases including papain, bromelain, and cathepsin B. In order to unravel its inhibitory action from structural point of view, multidimensional heteronuclear NMR techniques were used to characterize the structure of AcCYS. The full (1)H, (15)N, and (13)C resonance assignments of AcCYS were determined. The secondary structure of AcCYS was identified by using the assigned chemical shift of (1)Hα, (13)Cα, (13)Cβ, and (13)CO through the consensus chemical shift index (CSI). The results of CSI analysis suggest 5 β-strands (residues 45-47, 84-91, 94-104, 106-117, and 123-130) and one α-helix (residues 55-73).

  8. Titanium carbide, nitride and carbonitrides: a 13C, 14N, 15N and 47,49Ti solid-state nuclear magnetic resonance study.

    Science.gov (United States)

    MacKenzie, K J; Meinhold, R H; McGavin, D G; Ripmeester, J A; Moudrakovski, I

    1995-05-01

    The first 47,49Ti, 13C, 14N and 15N solid-state nuclear magnetic resonance (NMR) spectra of titanium carbide, nitride and a series of cubic carbonitrides have been obtained under both static and magic-angle spinning (MAS) conditions. The 15N samples were isotopically enriched by gas-solid exchange at 1000 degrees C in a closed system. The Ti spectra of the carbide and nitride are sharp, reflecting the well defined cubic symmetry of these compounds, but become considerably broadened in the carbonitride series, with the spectra being approximately the sum of TiC and TiN together with some small electric field gradient (EFG) effects. The resonance positions and widths of all the NMR spectra change as carbon is progressively replaced by nitrogen. A relationship is observed between the 13C chemical shift and the nitrogen content of the carbonitrides, suggesting a possible NMR method for estimating the composition of these compounds. Although electron paramagnetic resonance (EPR) spectra of all these compounds show typically metallic behaviour, the NMR spectra show few effects attributable to conduction electrons, probably due to the lack of s-orbital contributions to the conduction band.

  9. Lactoperoxidase-catalyzed oxidation of thiocyanate by hydrogen peroxide: sup 15 N nuclear magnetic resonance and optical spectral studies

    Energy Technology Data Exchange (ETDEWEB)

    Modi, S.; Deodhar, S.S.; Behere, D.V.; Mitra, S. (Tata Institute of Fundamental Research, Bombay (India))

    1991-01-01

    To establish the agent(s) responsible for the activity of the lactoperoxidase (LPO)/SCN{sup {minus}}/H{sub 2}O{sub 2} system, the oxidation of thiocyanate with hydrogen peroxide, catalyzed by lactoperoxidase, has been studied by {sup 15}N NMR and optical spectroscopy at different concentrations of thiocyanate and hydrogen peroxide and at different pHs. The formation of hypothiocyanite ion (OSCN{sup {minus}}) as one of the oxidation products correlated well with activity of the LPO/SCN{sup {minus}}/H{sub 2}O{sub 2} system and was maximum when the concentrations of the H{sub 2}O{sub 2} and SCN{sup {minus}} were nearly the same and the pH was <6.0. At (H{sub 2}O{sub 2})/(SCN{sup {minus}}) = 1, OSCN{sup {minus}} decomposed very slowly back to thiocyanate. When the ratio (H{sub 2}O{sub 2})/(SCN{sup {minus}}) was above 2, formation of CN{sup {minus}} was observed, which was confirmed by {sup 15}N NMR and also by changes in the optical spectrum of LPO. The oxidation of thiocyanate by H{sub 2}O{sub 2} in the presence of LPO does not take place at pH >8.0. Since thiocyanate does not bind to LPO above this pH, the binding of thiocyanate to LPO is considered to be prerequisite for the oxidation of thiocyanate. Maximum inhibition of oxygen uptake by Streptococcus cremoris 972 bacteria was observed when hydrogen peroxide and thiocyanate were present in equimolar amounts and the pH was below 6.0.

  10. A triple resonance hyperfine sublevel correlation experiment for assignment of electron-nuclear double resonance lines

    Science.gov (United States)

    Potapov, Alexey; Epel, Boris; Goldfarb, Daniella

    2008-02-01

    A new, triple resonance, pulse electron paramagnetic resonance (EPR) sequence is described. It provides spin links between forbidden electron spin transitions (ΔMS=±1, ΔMI≠0) and allowed nuclear spin transitions (ΔMI=±1), thus, facilitating the assignment of nuclear frequencies to their respective electron spin manifolds and paramagnetic centers. It also yields the relative signs of the hyperfine couplings of the different nuclei. The technique is based on the combination of electron-nuclear double resonance (ENDOR) and electron-electron double resonance (ELDOR)-detected NMR experiments in a way similar to the TRIPLE experiment. The feasibility and the information content of the method are demonstrated first on a single crystal of Cu-doped L-histidine and then on a frozen solution of a Cu-histidine complex.

  11. A general Monte Carlo/simulated annealing algorithm for resonance assignment in NMR of uniformly labeled biopolymers

    Science.gov (United States)

    Hu, Kan-Nian; Qiang, Wei; Tycko, Robert

    2011-01-01

    We describe a general computational approach to site-specific resonance assignments in multidimensional NMR studies of uniformly 15N,13C-labeled biopolymers, based on a simple Monte Carlo/simulated annealing (MCSA) algorithm contained in the program MCASSIGN2. Input to MCASSIGN2 includes lists of multidimensional signals in the NMR spectra with their possible residue-type assignments (which need not be unique), the biopolymer sequence, and a table that describes the connections that relate one signal list to another. As output, MCASSIGN2 produces a high-scoring sequential assignment of the multidimensional signals, using a score function that rewards good connections (i.e., agreement between relevant sets of chemical shifts in different signal lists) and penalizes bad connections, unassigned signals, and assignment gaps. Examination of a set of high-scoring assignments from a large number of independent runs allows one to determine whether a unique assignment exists for the entire sequence or parts thereof. We demonstrate the MCSA algorithm using two-dimensional (2D) and three-dimensional (3D) solid state NMR spectra of several model protein samples (α-spectrin SH3 domain and protein G/B1 microcrystals, HET-s218–289 fibrils), obtained with magic-angle spinning and standard polarization transfer techniques. The MCSA algorithm and MCASSIGN2 program can accommodate arbitrary combinations of NMR spectra with arbitrary dimensionality, and can therefore be applied in many areas of solid state and solution NMR. PMID:21710190

  12. 1H, 15N and 13C assignments of domain 5 of Dictyostelium discoideum gelation factor (ABP-120) in its native and 8M urea-denatured states.

    Science.gov (United States)

    Hsu, Shang-Te Danny; Cabrita, Lisa D; Christodoulou, John; Dobson, Christopher M

    2009-06-01

    The gelation factor from Dictyostelium discoideum (ABP-120) is an actin binding protein consisting of six immunoglobulin (Ig) domains in the C-terminal rod domain. We have recently used the pair of domains 5 and 6 of ABP-120 as a model system for studying multi-domain nascent chain folding on the ribosome. Here we present the NMR assignments of domain 5 in its native and 8M urea-denatured states.

  13. An automated framework for NMR resonance assignment through simultaneous slice picking and spin system forming.

    Science.gov (United States)

    Abbas, Ahmed; Guo, Xianrong; Jing, Bing-Yi; Gao, Xin

    2014-06-01

    Despite significant advances in automated nuclear magnetic resonance-based protein structure determination, the high numbers of false positives and false negatives among the peaks selected by fully automated methods remain a problem. These false positives and negatives impair the performance of resonance assignment methods. One of the main reasons for this problem is that the computational research community often considers peak picking and resonance assignment to be two separate problems, whereas spectroscopists use expert knowledge to pick peaks and assign their resonances at the same time. We propose a novel framework that simultaneously conducts slice picking and spin system forming, an essential step in resonance assignment. Our framework then employs a genetic algorithm, directed by both connectivity information and amino acid typing information from the spin systems, to assign the spin systems to residues. The inputs to our framework can be as few as two commonly used spectra, i.e., CBCA(CO)NH and HNCACB. Different from the existing peak picking and resonance assignment methods that treat peaks as the units, our method is based on 'slices', which are one-dimensional vectors in three-dimensional spectra that correspond to certain ([Formula: see text]) values. Experimental results on both benchmark simulated data sets and four real protein data sets demonstrate that our method significantly outperforms the state-of-the-art methods while using a less number of spectra than those methods. Our method is freely available at http://sfb.kaust.edu.sa/Pages/Software.aspx.

  14. Spin and parity assignments for Mo94,95 neutron resonances

    Science.gov (United States)

    Sheets, S. A.; Agvaanluvsan, U.; Becker, J. A.; Bečvář, F.; Bredeweg, T. A.; Haight, R. C.; Krtička, M.; Jandel, M.; Mitchell, G. E.; O'Donnell, J. M.; Parker, W. E.; Reifarth, R.; Rundberg, R. S.; Sharapov, E. I.; Tomandl, I.; Ullmann, J. L.; Vieira, D. J.; Wouters, J. M.; Wilhelmy, J. B.; Wu, C. Y.

    2007-12-01

    The γ rays following the Mo94,95(n,γ) reactions were measured as a function of incident neutron energy by the time-of-flight method with the DANCE (Detector for Advanced Neutron Capture Experiments) array of 160 BaF2 scintillation detectors at the Los Alamos Neutron Science Center. The targets were enriched samples: 91.59% Mo94 and 96.47% Mo95. The γ-ray multiplicities and energy spectra for different multiplicities were measured in s- and p-wave resonances up to En=10 keV for Mo94 and up to En=2 keV for Mo95. Definite spins and parities were assigned in Mo96 for about 60% of the resonances, and tentative spins and parities were assigned for the remaining resonances. In Mo95 the parities were determined for the observed resonances, confirming previously known assignments.

  15. An automated framework for NMR resonance assignment through simultaneous slice picking and spin system forming

    KAUST Repository

    Abbas, Ahmed

    2014-04-19

    Despite significant advances in automated nuclear magnetic resonance-based protein structure determination, the high numbers of false positives and false negatives among the peaks selected by fully automated methods remain a problem. These false positives and negatives impair the performance of resonance assignment methods. One of the main reasons for this problem is that the computational research community often considers peak picking and resonance assignment to be two separate problems, whereas spectroscopists use expert knowledge to pick peaks and assign their resonances at the same time. We propose a novel framework that simultaneously conducts slice picking and spin system forming, an essential step in resonance assignment. Our framework then employs a genetic algorithm, directed by both connectivity information and amino acid typing information from the spin systems, to assign the spin systems to residues. The inputs to our framework can be as few as two commonly used spectra, i.e., CBCA(CO)NH and HNCACB. Different from the existing peak picking and resonance assignment methods that treat peaks as the units, our method is based on \\'slices\\', which are one-dimensional vectors in three-dimensional spectra that correspond to certain (N, H) values. Experimental results on both benchmark simulated data sets and four real protein data sets demonstrate that our method significantly outperforms the state-of-the-art methods while using a less number of spectra than those methods. Our method is freely available at http://sfb.kaust.edu.sa/Pages/Software.aspx. © 2014 Springer Science+Business Media.

  16. Reduced Dimensionality tailored HN(C)N Pulse Sequences for Efficient Backbone Resonance Assignment of Proteins through Rapid Identification of Sequential HSQC peaks

    CERN Document Server

    Kumar, Dinesh

    2013-01-01

    Two novel reduced dimensionality (RD) experiments -(4,3)D-hNCOcaNH and (4,3)D-hNcoCANH- have been presented here to facilitate the backbone resonance assignment of proteins both in terms of its accuracy and speed. The experiments basically represent an improvisation of previously reported HN(C)N experiment [Panchal et. al., J. Biomol. NMR. (2002), 20 (2), 135-147] and exploit the simple reduced dimensionality NMR concept [Szyperski et. al. (2002), Proc. Natl. Acad. Sci. U.S.A. 99(12), 8009-8014] to achieve (a) higher dispersion and resolution along the co-evolved F1 dimension and (b) rapid identification of sequential HSQC peaks on its F2(15N)- F3(1H) planes. The current implementation is based on the fact that the linear combination of 15N and 13CO/13Ca chemical shifts offers relatively better dispersion and randomness compared to the individual chemical shifts; thus enables the assignment of crowded HSQC spectra by resolving the ambiguities generally encountered in HNCN based assignment protocol because of ...

  17. Automated solid-state NMR resonance assignment of protein microcrystals and amyloids

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, Elena [Goethe University Frankfurt am Main, Center for Biomolecular Magnetic Resonance, Institute of Biophysical Chemistry (Germany); Gath, Julia [ETH Zurich, Physical Chemistry (Switzerland); Habenstein, Birgit [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France); Ravotti, Francesco; Szekely, Kathrin; Huber, Matthias [ETH Zurich, Physical Chemistry (Switzerland); Buchner, Lena [Goethe University Frankfurt am Main, Center for Biomolecular Magnetic Resonance, Institute of Biophysical Chemistry (Germany); Boeckmann, Anja, E-mail: a.bockmann@ibcp.fr [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France); Meier, Beat H., E-mail: beme@ethz.ch [ETH Zurich, Physical Chemistry (Switzerland); Guentert, Peter, E-mail: guentert@em.uni-frankfurt.de [Goethe University Frankfurt am Main, Center for Biomolecular Magnetic Resonance, Institute of Biophysical Chemistry (Germany)

    2013-07-15

    Solid-state NMR is an emerging structure determination technique for crystalline and non-crystalline protein assemblies, e.g., amyloids. Resonance assignment constitutes the first and often very time-consuming step to a structure. We present ssFLYA, a generally applicable algorithm for automatic assignment of protein solid-state NMR spectra. Application to microcrystals of ubiquitin and the Ure2 prion C-terminal domain, as well as amyloids of HET-s(218-289) and {alpha}-synuclein yielded 88-97 % correctness for the backbone and side-chain assignments that are classified as self-consistent by the algorithm, and 77-90 % correctness if also assignments classified as tentative by the algorithm are included.

  18. Proton nuclear magnetic resonance studies on the variant-3 neurotoxin from Centruroides sculpturatus Ewing: Sequential assignment of resonances

    Energy Technology Data Exchange (ETDEWEB)

    Nettesheim, D.G.; Klevit, R.E.; Drobny, G.; Watt, D.D.; Krishna, N.R. (Univ. of Alabama, Birmingham (USA))

    1989-02-21

    The authors report the sequential assignment of resonances to specific residues in the proton nuclear magnetic resonance spectrum of the variant-3 neurotoxin from the scorpion Centruroides sculpturatus Ewing (range southwestern U.S.A.). A combination of two-dimensional NMR experiments such as 2D-COSY, 2D-NOESY, and single- and double-RELAY coherence transfer spectroscopy has been employed on samples of the protein dissolved in D{sub 2}O and in H{sub 2}O for assignment purposes. These studies provide a basis for the determination of the solution-phase conformation of this protein and for undertaking detailed structure-function studies of these neurotoxins that modulate the flow of sodium current by binding to the sodium channels of excitable membranes.

  19. 15N NMR Spectroscopic Study on Nitrogen Formsin1mmHumic Substances of Soils

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Nitrogen forms of humic substances from a subalpine meadow soil, alateritic red soil and a weathered coal and the effect of acidhydrolysis on N structures of soil humic substances were studied byusing {15N cross-polarization magic angle spinning nuclearmagnetic resonance (CPMAS NMR) spectroscopy. Of the detectable15N-signal intensity in the spectra of soil humic substances71%79% may be attributed to amide groups, 10%18%to aromatic/aliphatic amines and 6%11% to indole- andpyrrole-like N. Whereas in the spectrum of the fulvic acid fromweathered coal 46%, at least, of the total 15N-signalintensity might be assigned to pyrrole-like N, 14% toaromatic/aliphatic amines, and the remaining intensities could not beassigned with certainty. Data on nonhydrolyzable residue ofprotein-sugar mixture and a 15N-labelled soil fulvic acidconfirm the formation of nonhydrolyzable heterocyclic N during acidhydrolysis.

  20. A novel strategy for NMR resonance assignment and protein structure determination

    Energy Technology Data Exchange (ETDEWEB)

    Lemak, Alexander [University of Toronto, Ontario Cancer Institute and Campbell Family Cancer Research Institute, Department of Medical Biophysics (Canada); Gutmanas, Aleksandras [European Bioinformatics Institute, Protein Data Bank Europe (United Kingdom); Chitayat, Seth; Karra, Murthy [University of Toronto, Ontario Cancer Institute and Campbell Family Cancer Research Institute, Department of Medical Biophysics (Canada); Fares, Christophe [Max-Planck-Institut f. Kohlenforschung (Germany); Sunnerhagen, Maria [Linkoeping University, Division of Molecular Biotechnology, Department of Physics, Chemistry and Biology (Sweden); Arrowsmith, Cheryl H., E-mail: carrow@uhnres.utoronto.ca [University of Toronto, Ontario Cancer Institute and Campbell Family Cancer Research Institute, Department of Medical Biophysics (Canada)

    2011-01-15

    The quality of protein structures determined by nuclear magnetic resonance (NMR) spectroscopy is contingent on the number and quality of experimentally-derived resonance assignments, distance and angular restraints. Two key features of protein NMR data have posed challenges for the routine and automated structure determination of small to medium sized proteins; (1) spectral resolution - especially of crowded nuclear Overhauser effect spectroscopy (NOESY) spectra, and (2) the reliance on a continuous network of weak scalar couplings as part of most common assignment protocols. In order to facilitate NMR structure determination, we developed a semi-automated strategy that utilizes non-uniform sampling (NUS) and multidimensional decomposition (MDD) for optimal data collection and processing of selected, high resolution multidimensional NMR experiments, combined it with an ABACUS protocol for sequential and side chain resonance assignments, and streamlined this procedure to execute structure and refinement calculations in CYANA and CNS, respectively. Two graphical user interfaces (GUIs) were developed to facilitate efficient analysis and compilation of the data and to guide automated structure determination. This integrated method was implemented and refined on over 30 high quality structures of proteins ranging from 5.5 to 16.5 kDa in size.

  1. Backbone, side chain and heme resonance assignments of cytochrome OmcF from Geobacter sulfurreducens.

    Science.gov (United States)

    Dantas, Joana M; Silva E Sousa, Marta; Salgueiro, Carlos A; Bruix, Marta

    2015-10-01

    Gene knockout studies on Geobacter sulfurreducens (Gs) cells showed that the outer membrane cytochrome OmcF is involved in respiratory pathways leading to the extracellular reduction of Fe(III) citrate and U(VI) oxide. In addition, microarray analysis of OmcF-deficient mutant versus the wild-type strain revealed that many of the genes with decreased transcript level were those whose expression is upregulated in cells grown with a graphite electrode as electron acceptor. This suggests that OmcF also regulates the electron transfer to electrode surfaces and the concomitant electrical current production by Gs in microbial fuel cells. Extracellular electron transfer processes (EET) constitute nowadays the foundations to develop biotechnological applications in biofuel production, bioremediation and bioenergy. Therefore, the structural characterization of OmcF is a fundamental step to understand the mechanisms underlying EET. Here, we report the complete assignment of the heme proton signals together with (1)H, (13)C and (15)N backbone and side chain assignments of the OmcF, excluding the hydrophobic residues of the N-terminal predicted lipid anchor.

  2. Sequence specific resonance assignment via Multicanonical Monte Carlo search using an ABACUS approach.

    Science.gov (United States)

    Lemak, Alexander; Steren, Carlos A; Arrowsmith, Cheryl H; Llinás, Miguel

    2008-05-01

    ABACUS [Grishaev et al. (2005) Proteins 61:36-43] is a novel protocol for automated protein structure determination via NMR. ABACUS starts from molecular fragments defined by unassigned J-coupled spin-systems and involves a Monte Carlo stochastic search in assignment space, probabilistic sequence selection, and assembly of fragments into structures that are used to guide the stochastic search. Here, we report further development of the two main algorithms that increase the flexibility and robustness of the method. Performance of the BACUS [Grishaev and Llinás (2004) J Biomol NMR 28:1-101] algorithm was significantly improved through use of sequential connectivities available from through-bond correlated 3D-NMR experiments, and a new set of likelihood probabilities derived from a database of 56 ultra high resolution X-ray structures. A Multicanonical Monte Carlo procedure, Fragment Monte Carlo (FMC), was developed for sequence-specific assignment of spin-systems. It relies on an enhanced assignment sampling and provides the uncertainty of assignments in a quantitative manner. The efficiency of the protocol was validated on data from four proteins of between 68-116 residues, yielding 100% accuracy in sequence specific assignment of backbone and side chain resonances.

  3. Backbone resonance assignments of the micro-RNA precursor binding region of human TRBP.

    Science.gov (United States)

    Benoit, Matthieu P M H; Plevin, Michael J

    2013-10-01

    TAR-RNA binding protein (TRBP) is a multidomain human protein involved in micro-RNA (miRNA) biogenesis. TRBP is a component of both the Dicer complex, which processes precursor miRNAs, and the RNA-induced silencing complex-loading complex. In addition, TRBP is implicated in the human immunodeficiency virus replication cycle and interferon-protein kinase R activity. TRBP contains 3 double-stranded RNA binding domains the first two of which have been shown to interact with miRNA precursors. Here we present the backbone resonance assignments and secondary structure of residues 19-228 of human TRBP2.

  4. Exploring symbiotic nitrogen fixation and assimilation in pea root nodules by in vivo 15N nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry

    DEFF Research Database (Denmark)

    Scharff, A.M.; Egsgaard, H.; Hansen, P.E.

    2003-01-01

    Nitrogen (N) fixation and assimilation in pea (Pisum sativum) root nodules were studied by in vivo N-15 nuclear magnetic resonance (NMR) by exposing detached nodules to N-15, via a perfusion medium, while recording a time course of spectra. In vivo P-31 NMR spectroscopy was used to monitor...... the physiological state of the metabolically active nodules. The nodules were extracted after the NMR studies and analyzed for total soluble amino acid pools and N-15 labeling of individual amino acids by liquid chromatography-mass spectrometry. A substantial pool of free ammonium was observed by N-15 NMR...... labeling of Asn was observed by liquid chromatography-mass spectrometry, which is consistent with the generally accepted role of Asn as the end product of primary N assimilation in pea nodules. However, the Asn N-15 amino signal was absent in in vivo N-15 NMR spectra, which could be because...

  5. Structure and backbone dynamics of vanadate-bound PRL-3: comparison of 15N nuclear magnetic resonance relaxation profiles of free and vanadate-bound PRL-3.

    Science.gov (United States)

    Jeong, Ki-Woong; Kang, Dong-Il; Lee, Eunjung; Shin, Areum; Jin, Bonghwan; Park, Young-Guen; Lee, Chung-Kyoung; Kim, Eun-Hee; Jeon, Young Ho; Kim, Eunice Eunkyeong; Kim, Yangmee

    2014-07-29

    Phosphatases of regenerating liver (PRLs) constitute a novel class of small, prenylated phosphatases with oncogenic activity. PRL-3 is particularly important in cancer metastasis and represents a potential therapeutic target. The flexibility of the WPD loop as well as the P-loop of protein tyrosine phosphatases is closely related to their catalytic activity. Using nuclear magnetic resonance spectroscopy, we studied the structure of vanadate-bound PRL-3, which was generated by addition of sodium orthovanadate to PRL-3. The WPD loop of free PRL-3 extended outside of the active site, forming an open conformation, whereas that of vanadate-bound PRL-3 was directed into the active site by a large movement, resulting in a closed conformation. We suggest that vanadate binding induced structural changes in the WPD loop, P-loop, helices α4-α6, and the polybasic region. Compared to free PRL-3, vanadate-bound PRL-3 has a longer α4 helix, where the catalytic R110 residue coordinates with vanadate in the active site. In addition, the hydrophobic cavity formed by helices α4-α6 with a depth of 14-15 Å can accommodate a farnesyl chain at the truncated prenylation motif of PRL-3, i.e., from R169 to M173. Conformational exchange data suggested that the WPD loop moves between open and closed conformations with a closing rate constant k(close) of 7 s(-1). This intrinsic loop flexibility of PRL-3 may be related to their catalytic rate and may play a role in substrate recognition.

  6. Resonance strengths in the {sup 17,18}O(p, α){sup 14,15}N reactions and background suppression underground. Commissioning of a new setup for charged-particle detection at LUNA

    Energy Technology Data Exchange (ETDEWEB)

    Bruno, C.G.; Scott, D.A.; Aliotta, M.; Davinson, T.; Griffin, C.J. [University of Edinburgh, SUPA, School of Physics and Astronomy, Edinburgh (United Kingdom); Formicola, A.; Best, A.; Junker, M. [Laboratori Nazionali del Gran Sasso, INFN, Assergi (Italy); Anders, M.; Bemmerer, D.; Szuecs, T. [Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden (Germany); Broggini, C.; Menegazzo, R. [INFN, Padova (Italy); Caciolli, A.; Depalo, R. [INFN, Padova (Italy); Universita di Padova, Dipartimento di Fisica e Astronomia, Padova (Italy); Cavanna, F.; Corvisiero, P.; Prati, P. [INFN, Genova (Italy); Dipartimento di Fisica, Universita di Genova, Genova (Italy); Di Leva, A.; Imbriani, G. [Universita di Napoli ' ' Federico II' ' , Dipartimento di Scienze Fisiche, Napoli (Italy); Elekes, Z.; Fueloep, Zs.; Gyuerky, Gy.; Somorjai, E. [MTA Atomki, Institute of Nuclear Research, Debrecen (Hungary); Gervino, G. [Universita degli Studi di Torino, Dipartimento di Fisica Sperimentale, Torino (Italy); Guglielmetti, A.; Trezzi, D. [Universita degli Studi di Milano (Italy); INFN, Milano (Italy); Gustavino, C. [INFN, Roma (Italy); Napolitani, E. [Universita di Padova, Dipartimento di Fisica e Astronomia, Padova (Italy); Straniero, O. [Laboratori Nazionali del Gran Sasso, INFN, Assergi (Italy); Osservatorio Astronomico di Collurania, Teramo (Italy); Strieder, F. [South Dakota School of Mines and Technology, Rapid City, SD (United States); Collaboration: LUNA Collaboration

    2015-08-15

    We report on measurements of resonance strengths and energies for the E{sub p} = 151 and 193 keV resonances in the {sup 18}O(p, α){sup 15}N and {sup 17}O(p, α){sup 14}N reactions, respectively, obtained during commissioning of a new setup for alpha-particle detection studies at the LUNA underground laboratory. Our values, ωγ(151) = 164.2 ± 0.9{sub stat-11.7}{sup +12.1}{sub syst} meV and ωγ (193) = 1.68 ± 0.03{sub stat} ± 0.12{sub syst} meV, are in excellent agreement with those reported in the literature. New values of resonance energies are E{sub p} = 151.2 ± 0.3 keV and E{sub p} = 194.8 ± 0.3 keV, respectively, this latter with the highest precision to date. Comparative background measurements in silicon detectors overground and underground were also carried out, yielding up to a factor of 15 in background suppression at LUNA at energies around 200 keV. This clearly demonstrates the usefulness of underground measurements in charged-particles experiments, especially at low detection energies. (orig.)

  7. Simultaneous acquisition of three NMR spectra in a single experiment for rapid resonance assignments in metabolomics

    Indian Academy of Sciences (India)

    Shivanand M Pudakalakatti; Abhinav Dubey; Hanudatta S Atreya

    2015-06-01

    NMR-based approach to metabolomics typically involves the collection of two-dimensional (2D) heteronuclear correlation spectra for identification and assignment of metabolites. In case of spectral overlap, a 3D spectrum becomes necessary, which is hampered by slow data acquisition for achieving sufficient resolution. We describe here a method to simultaneously acquire three spectra (one 3D and two 2D) in a single data set, which is based on a combination of different fast data acquisition techniques such as G-matrix Fourier transform (GFT) NMR spectroscopy, parallel data acquisition and non-uniform sampling. The following spectra are acquired simultaneously: (1) 13C multiplicity edited GFT (3,2)D HSQC-TOCSY, (2) 2D [1H-1H] TOCSY and (3) 2D [13C-1H] HETCOR. The spectra are obtained at high resolution and provide high-dimensional spectral information for resolving ambiguities. While the GFT spectrum has been shown previously to provide good resolution, the editing of spin systems based on their CH multiplicities further resolves the ambiguities for resonance assignments. The experiment is demonstrated on a mixture of 21 metabolites commonly observed in metabolomics. The spectra were acquired at natural abundance of 13C. This is the first application of a combination of three fast NMR methods for small molecules and opens up new avenues for high-throughput approaches for NMR-based metabolomics.

  8. 15N NMR Spectroscopic Study on Nitrogen Forms in Humic Substances of Soils

    Institute of Scientific and Technical Information of China (English)

    WENQIXAIO; ZHUOSUNENG; 等

    2001-01-01

    Nitrogen forms of humic substances from a subalpine meadow soil,a lateritic red soil and a weathered cola and the effect of acid hydrolysis on N structures of soil humic substances were studied by using 15N cross-polarization magic angle spinning nuclear magnetic resonance(CPMAS NMR) spectroscopy,Of the detectable 15N-signal intensity in the spectra of soil humic substances 71%-79% may be attributed to amide groups ,10%-18% to aromatic/aliphatic amines and 6%-11% to indole-and pyrrole-like N.Whereas in the spectrum of the fulvic acid from weathered coal 46%,at least,of the total 15N-signal intensity might be assigned to pyrrole-like N,14% to aromatic/aliphatic amines,and the reamining intensities could not be assigned with certainty,Data on nonhydrolyzable reside of protein-sugar mixture and a 15N-labelled soil fulvic acid confirm the formation of nonhydrolyzable heterocyclic N during acid hydrolysis.

  9. FLAMEnGO 2.0: an enhanced fuzzy logic algorithm for structure-based assignment of methyl group resonances.

    Science.gov (United States)

    Chao, Fa-An; Kim, Jonggul; Xia, Youlin; Milligan, Michael; Rowe, Nancy; Veglia, Gianluigi

    2014-08-01

    We present an enhanced version of the FLAMEnGO (Fuzzy Logic Assignment of Methyl Group) software, a structure-based method to assign methyl group resonances in large proteins. FLAMEnGO utilizes a fuzzy logic algorithm coupled with Monte Carlo sampling to obtain a probability-based assignment of the methyl group resonances. As an input, FLAMEnGO requires either the protein X-ray structure or an NMR structural ensemble including data such as methyl-methyl NOESY, paramagnetic relaxation enhancement (PRE), methine-methyl TOCSY data. Version 2.0 of this software (FLAMEnGO 2.0) has a user-friendly graphic interface and presents improved modules that enable the input of partial assignments and additional NMR restraints. We tested the performance of FLAMEnGO 2.0 on maltose binding protein (MBP) as well as the C-subunit of the cAMP-dependent protein kinase A (PKA-C). FLAMEnGO 2.0 can be used as a standalone method or to assist in the completion of partial resonance assignments and can be downloaded at www.chem.umn.edu/groups/veglia/forms/flamengo2-form.html.

  10. BEST-TROSY experiments for time-efficient sequential resonance assignment of large disordered proteins

    Energy Technology Data Exchange (ETDEWEB)

    Solyom, Zsofia; Schwarten, Melanie [Universite Grenoble 1, Institut de Biologie Structurale (France); Geist, Leonhard; Konrat, Robert [Max F. Perutz Laboratories, Department of Computational and Structural Biology (Austria); Willbold, Dieter [Commissariat a l' Energie Atomique et aux Energies Alternatives (CEA) (France); Brutscher, Bernhard, E-mail: bernhard.brutscher@ibs.fr [Universite Grenoble 1, Institut de Biologie Structurale (France)

    2013-04-15

    The characterization of the conformational properties of intrinsically disordered proteins (IDPs), and their interaction modes with physiological partners has recently become a major research topic for understanding biological function on the molecular level. Although multidimensional NMR spectroscopy is the technique of choice for the study of IDPs at atomic resolution, the intrinsically low resolution, and the large peak intensity variations often observed in NMR spectra of IDPs call for resolution- and sensitivity-optimized pulse schemes. We present here a set of amide proton-detected 3D BEST-TROSY correlation experiments that yield the required sensitivity and spectral resolution for time-efficient sequential resonance assignment of large IDPs. In addition, we introduce two proline-edited 2D experiments that allow unambiguous identification of residues adjacent to proline that is one of the most abundant amino acids in IDPs. The performance of these experiments, and the advantages of BEST-TROSY pulse schemes are discussed and illustrated for two IDPs of similar length ({approx}270 residues) but with different conformational sampling properties.

  11. Resonance assignments and secondary structure of apolipoprotein E C-terminal domain in DHPC micelles.

    Science.gov (United States)

    Lo, Chi-Jen; Chyan, Chia-Lin; Chen, Yi-Chen; Chang, Chi-Fon; Huang, Hsien-Bin; Lin, Ta-Hsien

    2015-04-01

    Human apolipoprotein E (apoE) has been known to play a key role in the transport of plasma cholesterol and lipoprotein metabolism. It is an apolipoprotein of 299 amino acids with a molecular mass, ~34 kDa. ApoE has three major isoforms, apoE2, apoE3, and apoE4 which differ only at residue 112 or 158. ApoE consists of two independently folded domains (N-terminal and C-terminal domain) separated by a hinge region. The N-terminal domain and C-terminal domain of apoE are responsible for the binding to receptor and to lipid, respectively. Since the high resolution structures of apoE in lipids are still unavailable to date, we therefore aim to resolve the structures in lipids by NMR. Here, we reported the resonance assignments and secondary structure distribution of the C-terminal domain of wild-type human apoE (residue 195-299) in the micelles formed by dihexanoylphosphatidylcholine. Our results may provide a novel structural model of apoE in micelles and may shed new light on the molecular mechanisms underlying the apoE related biological processes.

  12. Resonance assignment of the NMR spectra of disordered proteins using a multi-objective non-dominated sorting genetic algorithm

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Yu; Fritzsching, Keith J.; Hong, Mei, E-mail: mhong@iastate.edu [Iowa State University, Department of Chemistry (United States)

    2013-10-17

    A multi-objective genetic algorithm is introduced to predict the assignment of protein solid-state NMR (SSNMR) spectra with partial resonance overlap and missing peaks due to broad linewidths, molecular motion, and low sensitivity. This non-dominated sorting genetic algorithm II (NSGA-II) aims to identify all possible assignments that are consistent with the spectra and to compare the relative merit of these assignments. Our approach is modeled after the recently introduced Monte-Carlo simulated-annealing (MC/SA) protocol, with the key difference that NSGA-II simultaneously optimizes multiple assignment objectives instead of searching for possible assignments based on a single composite score. The multiple objectives include maximizing the number of consistently assigned peaks between multiple spectra (“good connections”), maximizing the number of used peaks, minimizing the number of inconsistently assigned peaks between spectra (“bad connections”), and minimizing the number of assigned peaks that have no matching peaks in the other spectra (“edges”). Using six SSNMR protein chemical shift datasets with varying levels of imperfection that was introduced by peak deletion, random chemical shift changes, and manual peak picking of spectra with moderately broad linewidths, we show that the NSGA-II algorithm produces a large number of valid and good assignments rapidly. For high-quality chemical shift peak lists, NSGA-II and MC/SA perform similarly well. However, when the peak lists contain many missing peaks that are uncorrelated between different spectra and have chemical shift deviations between spectra, the modified NSGA-II produces a larger number of valid solutions than MC/SA, and is more effective at distinguishing good from mediocre assignments by avoiding the hazard of suboptimal weighting factors for the various objectives. These two advantages, namely diversity and better evaluation, lead to a higher probability of predicting the correct

  13. Resonance assignment of the NMR spectra of disordered proteins using a multi-objective non-dominated sorting genetic algorithm.

    Science.gov (United States)

    Yang, Yu; Fritzsching, Keith J; Hong, Mei

    2013-11-01

    A multi-objective genetic algorithm is introduced to predict the assignment of protein solid-state NMR (SSNMR) spectra with partial resonance overlap and missing peaks due to broad linewidths, molecular motion, and low sensitivity. This non-dominated sorting genetic algorithm II (NSGA-II) aims to identify all possible assignments that are consistent with the spectra and to compare the relative merit of these assignments. Our approach is modeled after the recently introduced Monte-Carlo simulated-annealing (MC/SA) protocol, with the key difference that NSGA-II simultaneously optimizes multiple assignment objectives instead of searching for possible assignments based on a single composite score. The multiple objectives include maximizing the number of consistently assigned peaks between multiple spectra ("good connections"), maximizing the number of used peaks, minimizing the number of inconsistently assigned peaks between spectra ("bad connections"), and minimizing the number of assigned peaks that have no matching peaks in the other spectra ("edges"). Using six SSNMR protein chemical shift datasets with varying levels of imperfection that was introduced by peak deletion, random chemical shift changes, and manual peak picking of spectra with moderately broad linewidths, we show that the NSGA-II algorithm produces a large number of valid and good assignments rapidly. For high-quality chemical shift peak lists, NSGA-II and MC/SA perform similarly well. However, when the peak lists contain many missing peaks that are uncorrelated between different spectra and have chemical shift deviations between spectra, the modified NSGA-II produces a larger number of valid solutions than MC/SA, and is more effective at distinguishing good from mediocre assignments by avoiding the hazard of suboptimal weighting factors for the various objectives. These two advantages, namely diversity and better evaluation, lead to a higher probability of predicting the correct assignment for a

  14. Empirical Equation Based Chirality (n, m) Assignment of Semiconducting Single Wall Carbon Nanotubes from Resonant Raman Scattering Data

    Science.gov (United States)

    Arefin, Md Shamsul

    2012-01-01

    This work presents a technique for the chirality (n, m) assignment of semiconducting single wall carbon nanotubes by solving a set of empirical equations of the tight binding model parameters. The empirical equations of the nearest neighbor hopping parameters, relating the term (2n− m) with the first and second optical transition energies of the semiconducting single wall carbon nanotubes, are also proposed. They provide almost the same level of accuracy for lower and higher diameter nanotubes. An algorithm is presented to determine the chiral index (n, m) of any unknown semiconducting tube by solving these empirical equations using values of radial breathing mode frequency and the first or second optical transition energy from resonant Raman spectroscopy. In this paper, the chirality of 55 semiconducting nanotubes is assigned using the first and second optical transition energies. Unlike the existing methods of chirality assignment, this technique does not require graphical comparison or pattern recognition between existing experimental and theoretical Kataura plot.

  15. Empirical Equation Based Chirality (n, m Assignment of Semiconducting Single Wall Carbon Nanotubes from Resonant Raman Scattering Data

    Directory of Open Access Journals (Sweden)

    Md Shamsul Arefin

    2012-12-01

    Full Text Available This work presents a technique for the chirality (n, m assignment of semiconducting single wall carbon nanotubes by solving a set of empirical equations of the tight binding model parameters. The empirical equations of the nearest neighbor hopping parameters, relating the term (2n, m with the first and second optical transition energies of the semiconducting single wall carbon nanotubes, are also proposed. They provide almost the same level of accuracy for lower and higher diameter nanotubes. An algorithm is presented to determine the chiral index (n, m of any unknown semiconducting tube by solving these empirical equations using values of radial breathing mode frequency and the first or second optical transition energy from resonant Raman spectroscopy. In this paper, the chirality of 55 semiconducting nanotubes is assigned using the first and second optical transition energies. Unlike the existing methods of chirality assignment, this technique does not require graphical comparison or pattern recognition between existing experimental and theoretical Kataura plot.

  16. Assignment of methyl NMR resonances of a 52 kDa protein with residue-specific 4D correlation maps

    Energy Technology Data Exchange (ETDEWEB)

    Mishra, Subrata H.; Frueh, Dominique P., E-mail: dfrueh@jhmi.edu [Johns Hopkins University School of Medicine, Department of Biophysics and Biophysical Chemistry (United States)

    2015-07-15

    Methyl groups have become key probes for structural and functional studies by nuclear magnetic resonance. However, their NMR signals cluster in a small spectral region and assigning their resonances can be a tedious process. Here, we present a method that facilitates assignment of methyl resonances from assigned amide groups. Calculating the covariance between sensitive methyl and amide 3D spectra, each providing correlations to C{sup α} and C{sup β} separately, produces 4D correlation maps directly correlating methyl groups to amide groups. Optimal correlation maps are obtained by extracting residue-specific regions, applying derivative to the dimensions subject to covariance, and multiplying 4D maps stemming from different 3D spectra. The latter procedure rescues weak signals that may be missed in traditional assignment procedures. Using these covariance correlation maps, nearly all assigned isoleucine, leucine, and valine amide resonances of a 52 kDa nonribosomal peptide synthetase cyclization domain were paired with their corresponding methyl groups.

  17. ¹H, ¹³C and ¹⁵N resonance assignments and second structure information of Gad m 1: a β-parvalbumin allergen from Atlantic cod (Gadus morhua).

    Science.gov (United States)

    Moraes, A H; Ackerbauer, D; Kostadinova, M; Bublin, M; Ferreira, F; Almeida, F C L; Breiteneder, H; Valente, A P

    2013-10-01

    Gad m 1 is the major allergen from Atlantic cod. It belongs to β-parvalbumin protein family and is characterized by the presence of two calcium-binding sites so called EF-hand motifs. β-Parvalbumins such as Gad m 1 are the most important fish allergens and their high cross-reactivity is the cause of the observed polysensitization to various fish species in allergic patients. Despite extensive efforts, the complete elucidation of β-parvalbumin-IgE complexes has not been achieved yet. Allergen structural studies are essential for the development of novel immunotherapy strategies, including vaccination with hypoallergenic derivatives and chimeric molecules. Here, we report for the first time the NMR study of a β-parvalbumin: Gad m 1. This report includes: (1)H, (13)C and (15)N resonance assignments of Gad m 1 as well as the second structure information based on the (13)C chemical shifts.

  18. Complete Proton and Carbon Assignment of Triclosan via One- and Two- Dimensional Nuclear Magnetic Resonance Analysis

    Science.gov (United States)

    Students from an upper-division undergraduate spectroscopy class analyzed one- and two-dimensional 400 MHz NMR spectroscopic data from triclosan in CDCl3. Guided assignment of all proton and carbon signals was completed via 1D proton and carbon, nuclear Overhauser effect (nOe), distortionless enhanc...

  19. Resonance assignment for a particularly challenging protein based on systematic unlabeling of amino acids to complement incomplete NMR data sets

    Energy Technology Data Exchange (ETDEWEB)

    Bellstedt, Peter, E-mail: pbell@fli-leibniz.de; Seiboth, Thomas; Haefner, Sabine; Kutscha, Henriette; Ramachandran, Ramadurai; Goerlach, Matthias [Leibniz Institute for Age Research, Fritz Lipmann Institute, Biomolecular NMR Spectroscopy (Germany)

    2013-09-15

    NMR-based structure determination of a protein requires the assignment of resonances as indispensable first step. Even though heteronuclear through-bond correlation methods are available for that purpose, challenging situations arise in cases where the protein in question only yields samples of limited concentration and/or stability. Here we present a strategy based upon specific individual unlabeling of all 20 standard amino acids to complement standard NMR experiments and to achieve unambiguous backbone assignments for the fast precipitating 23 kDa catalytic domain of human aprataxin of which only incomplete standard NMR data sets could be obtained. Together with the validation of this approach utilizing the protein GB1 as a model, a comprehensive insight into metabolic interconversion ('scrambling') of NH and CO groups in a standard Escherichia coli expression host is provided.

  20. Spectral editing of two-dimensional magic-angle-spinning solid-state NMR spectra for protein resonance assignment and structure determination

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt-Rohr, K.; Fritzsching, K. J.; Liao, S. Y.; Hong Mei, E-mail: mhong@iastate.edu [Iowa State University, Department of Chemistry and Ames Laboratory (United States)

    2012-12-15

    Several techniques for spectral editing of 2D {sup 13}C-{sup 13}C correlation NMR of proteins are introduced. They greatly reduce the spectral overlap for five common amino acid types, thus simplifying spectral assignment and conformational analysis. The carboxyl (COO) signals of glutamate and aspartate are selected by suppressing the overlapping amide N-CO peaks through {sup 13}C-{sup 15}N dipolar dephasing. The sidechain methine (CH) signals of valine, lecuine, and isoleucine are separated from the overlapping methylene (CH{sub 2}) signals of long-chain amino acids using a multiple-quantum dipolar transfer technique. Both the COO and CH selection methods take advantage of improved dipolar dephasing by asymmetric rotational-echo double resonance (REDOR), where every other {pi}-pulse is shifted from the center of a rotor period t{sub r} by about 0.15 t{sub r}. This asymmetry produces a deeper minimum in the REDOR dephasing curve and enables complete suppression of the undesired signals of immobile segments. Residual signals of mobile sidechains are positively identified by dynamics editing using recoupled {sup 13}C-{sup 1}H dipolar dephasing. In all three experiments, the signals of carbons within a three-bond distance from the selected carbons are detected in the second spectral dimension via {sup 13}C spin exchange. The efficiencies of these spectral editing techniques range from 60 % for the COO and dynamic selection experiments to 25 % for the CH selection experiment, and are demonstrated on well-characterized model proteins GB1 and ubiquitin.

  1. Assignment of resonances in dissociative recombination of HD+ ions: high-resolution measurements compared with accurate computations

    CERN Document Server

    Tamo, F O Waffeu; Motapon, O; Altevogt, S; Andrianarijaona, V M; Grieser, M; Lammich, L; Lestinsky, M; Motsch, M; Nevo, I; Novotny, S; Orlov, D A; Pedersen, H B; Schwalm, D; Sprenger, F; Urbain, X; Weigel, U; Wolf, A; Schneider, I F

    2011-01-01

    The collision-energy resolved rate coefficient for dissociative recombination of HD+ ions in the vibrational ground state is measured using the photocathode electron target at the heavy-ion storage ring TSR. Rydberg resonances associated with ro-vibrational excitation of the HD+ core are scanned as a function of the electron collision energy with an instrumental broadening below 1 meV in the low-energy limit. The measurement is compared to calculations using multichannel quantum defect theory, accounting for rotational structure and interactions and considering the six lowest rotational energy levels as initial ionic states. Using thermal equilibrium level populations at 300 K to approximate the experimental conditions, close correspondence between calculated and measured structures is found up to the first vibrational excitation threshold of the cations near 0.24 eV. Detailed assignments, including naturally broadened and overlapping Rydberg resonances, are performed for all structures up to 0.024 eV. Resona...

  2. A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins

    Energy Technology Data Exchange (ETDEWEB)

    Plevin, Michael J., E-mail: michael.plevin@ibs.fr [CEA, Institut de Biologie Structurale Jean-Pierre Ebel (France); Hamelin, Olivier [CNRS, Laboratoire de Chimie et Biologie des Metaux (France); Boisbouvier, Jerome; Gans, Pierre [CEA, Institut de Biologie Structurale Jean-Pierre Ebel (France)

    2011-02-15

    A new method for stereospecific assignment of prochiral methyl groups in proteins is presented in which protein samples are produced using U-[{sup 13}C]glucose and subsaturating amounts of 2-[{sup 13}C]methyl-acetolactate. The resulting non-uniform labeling pattern allows proR and proS methyl groups to be easily distinguished by their different phases in a constant-time two-dimensional {sup 1}H-{sup 13}C correlation spectra. Protein samples are conveniently prepared using the same media composition as the main uniformly-labeled sample and contain higher levels of isotope-enrichment than fractional labeling approaches. This new strategy thus represents an economically-attractive, robust alternative for obtaining isotopically-encoded stereospecific NMR assignments of prochiral methyl groups.

  3. Rapid analysis of protein backbone resonance assignments using cryogenic probes, a distributed Linux-based computing architecture, and an integrated set of spectral analysis tools.

    Science.gov (United States)

    Monleón, Daniel; Colson, Kimberly; Moseley, Hunter N B; Anklin, Clemens; Oswald, Robert; Szyperski, Thomas; Montelione, Gaetano T

    2002-01-01

    Rapid data collection, spectral referencing, processing by time domain deconvolution, peak picking and editing, and assignment of NMR spectra are necessary components of any efficient integrated system for protein NMR structure analysis. We have developed a set of software tools designated AutoProc, AutoPeak, and AutoAssign, which function together with the data processing and peak-picking programs NMRPipe and Sparky, to provide an integrated software system for rapid analysis of protein backbone resonance assignments. In this paper we demonstrate that these tools, together with high-sensitivity triple resonance NMR cryoprobes for data collection and a Linux-based computer cluster architecture, can be combined to provide nearly complete backbone resonance assignments and secondary structures (based on chemical shift data) for a 59-residue protein in less than 30 hours of data collection and processing time. In this optimum case of a small protein providing excellent spectra, extensive backbone resonance assignments could also be obtained using less than 6 hours of data collection and processing time. These results demonstrate the feasibility of high throughput triple resonance NMR for determining resonance assignments and secondary structures of small proteins, and the potential for applying NMR in large scale structural proteomics projects.

  4. Resonance assignment of an engineered amino-terminal domain of a major ampullate spider silk with neutralized charge cluster.

    Science.gov (United States)

    Schaal, Daniel; Bauer, Joschka; Schweimer, Kristian; Scheibel, Thomas; Rösch, Paul; Schwarzinger, Stephan

    2016-04-01

    Spider dragline fibers are predominantly made out of the major ampullate spidroins (MaSp) 1 and 2. The assembly of dissolved spidroin into a stable fiber is highly controlled for example by dimerization of its amino-terminal domain (NRN) upon acidification, as well as removal of sodium chloride along the spinning duct. Clustered residues D39, E76 and E81 are the most highly conserved residues of the five-helix bundle, and they are hypothesized to be key residues for switching between a monomeric and a dimeric conformation. Simultaneous replacement of these residues by their non-titratable analogues results in variant D39N/E76Q/E81Q, which is supposed to fold into an intermediate conformation between that of the monomeric and the dimeric state at neutral pH. Here we report the resonance assignment of Latrodectus hesperus NRN variant D39N/E76Q/E81Q at pH 7.2 obtained by high-resolution triple resonance NMR spectroscopy.

  5. Backbone resonance assignments for G protein α(i3) subunit in the GDP-bound state.

    Science.gov (United States)

    Mase, Yoko; Yokogawa, Mariko; Osawa, Masanori; Shimada, Ichio

    2014-10-01

    Guanine-nucleotide binding proteins (G proteins) serve as molecular switches in signaling pathways, by coupling the activation of G protein-coupled receptors (GPCRs) at the cell surface to intracellular responses. In the resting state, G protein forms a heterotrimer, consisting of the G protein α subunit with GDP (Gα·GDP) and the G protein βγ subunit (Gβγ). Ligand binding to GPCRs promotes the GDP-GTP exchange on Gα, leading to the dissociation of the GTP-bound form of Gα (Gα·GTP) and Gβγ. Then, Gα·GTP and Gβγ bind to their downstream effector enzymes or ion channels and regulate their activities, leading to a variety of cellular responses. Finally, Gα hydrolyzes the bound GTP to GDP and returns to the resting state by re-associating with Gβγ. The G proteins are classified with four major families based on the amino acid sequences of Gα: i/o, s, q/11, and 12/13. Here, we established the backbone resonance assignments of human Gαi3, a member of the i/o family with a molecular weight of 41 K, in complex with GDP. The chemical shifts were compared with those of Gα(i3) in complex with a GTP-analogue, GTPγS, which we recently reported, indicating that the residues with significant chemical shift differences are mostly consistent with the regions with the structural differences between the GDP- and GTPγS-bound states, as indicated in the crystal structures. The assignments of Gα(i3)·GDP would be useful for the analyses of the dynamics of Gα(i3) and its interactions with various target molecules.

  6. Sequence-specific Assignment of 1H-NMR Resonance and Determination of the Secondary Structure of Jingzhaotoxin-Ⅰ

    Institute of Scientific and Technical Information of China (English)

    Xiong-Zhi ZENG; Qi ZHU; Song-Ping LIANG

    2005-01-01

    Jingzhaotoxin-Ⅰ (JZTX-Ⅰ) purified from the venom of the spider Chilobrachys jingzhao is a novel neurotoxin preferentially inhibiting cardiac sodium channel inactivation by binding to receptor site 3.The structure of this toxin in aqueous solution was investigated using 2-D 1H-NMR techniques. The complete sequence-specific assignments of proton resonance in the 1H-NMR spectra of JZTX-Ⅰ were obtained by analyzing a series of 2-D spectra, including DQF-COSY, TOCSY and NOESY spectra, in H2O and D2O. All the backbone protons except for Gln4 and more than 95% of the side-chain protons were identified by dαN,dαδ, dβN and dNN connectivities in the NOESY spectrum. These studies provide a basis for the further determination of the solution conformation of JZTX-Ⅰ. Furthermore, the secondary structure of JZTX-Ⅰ was identified from NMR data. It consists mainly of a short triple-stranded antiparallel β-sheet with Trp7-Cys9, Phe20-Lys23 and Leu28-Trp31. The characteristics of the secondary structure of JZTX-Ⅰ are similar to those of huwentoxin-Ⅰ (HWTX-Ⅰ) and hainantoxin-Ⅳ (HNTX-Ⅳ), whose structures in solution have previously been reported.

  7. Kinetic 15N-isotope effects on algal growth

    Science.gov (United States)

    Andriukonis, Eivydas; Gorokhova, Elena

    2017-03-01

    Stable isotope labeling is a standard technique for tracing material transfer in molecular, ecological and biogeochemical studies. The main assumption in this approach is that the enrichment with a heavy isotope has no effect on the organism metabolism and growth, which is not consistent with current theoretical and empirical knowledge on kinetic isotope effects. Here, we demonstrate profound changes in growth dynamics of the green alga Raphidocelis subcapitata grown in 15N-enriched media. With increasing 15N concentration (0.37 to 50 at%), the lag phase increased, whereas maximal growth rate and total yield decreased; moreover, there was a negative relationship between the growth and the lag phase across the treatments. The latter suggests that a trade-off between growth rate and the ability to adapt to the high 15N environment may exist. Remarkably, the lag-phase response at 3.5 at% 15N was the shortest and deviated from the overall trend, thus providing partial support to the recently proposed Isotopic Resonance hypothesis, which predicts that certain isotopic composition is particularly favorable for living organisms. These findings confirm the occurrence of KIE in isotopically enriched algae and underline the importance of considering these effects when using stable isotope labeling in field and experimental studies.

  8. Synthesis of Gemcitabine-13C, 15N2 and Gemcitabine-13C, 15N2 Metabolites

    Directory of Open Access Journals (Sweden)

    ZHU Cheng-gu;YANG Shao-zu;YAN Sheng-wang;FANG Ning-jing;CAI Ding-long;LI Gang

    2014-02-01

    Full Text Available Homemade urea-13C, 15N2 was used to react with 3-methyl acrylonitrile closure to form cytosine-13C, 15N2 (2,which was protected by trimethylsilylation with BSA and condensed with 2-deoxy-2,2-difluoro-D-erythro-pentofuranose-3,5-dibenzoate-1-methanesulfonate at 120 ℃ to afford blocked gemcitabine-13C, 15N2. Hydrolytic removal of the blocking groups of gemcitabine-13C, 15N2 with NaOH gave gemcitabine-13C, 15N2, and its metabolite was obtained by further hydrolytic deamination of gemcitabine-13C, 15N2. The final products were characterized and detected by HPLC, LC-MS and NMR, and confirmed that the chemical purities were higher than 98%, isotopic abundances were 99% 13C, 98% 15N, and they were suitable for drug metabolism studies.

  9. Protein resonance assignment at MAS frequencies approaching 100 kHz: a quantitative comparison of J-coupling and dipolar-coupling-based transfer methods

    Energy Technology Data Exchange (ETDEWEB)

    Penzel, Susanne; Smith, Albert A.; Agarwal, Vipin; Hunkeler, Andreas [ETH Zürich, Physical Chemistry (Switzerland); Org, Mai-Liis; Samoson, Ago, E-mail: ago.samoson@ttu.ee [Tallinn University of Technology, NMR Instituut, Tartu Teadus, Tehnomeedikum (Estonia); Böckmann, Anja, E-mail: a.bockmann@ibcp.fr [UMR 5086 CNRS/Université de Lyon 1, Institut de Biologie et Chimie des Protéines (France); Ernst, Matthias, E-mail: maer@ethz.ch; Meier, Beat H., E-mail: beme@ethz.ch [ETH Zürich, Physical Chemistry (Switzerland)

    2015-10-15

    We discuss the optimum experimental conditions to obtain assignment spectra for solid proteins at magic-angle spinning (MAS) frequencies around 100 kHz. We present a systematic examination of the MAS dependence of the amide proton T{sub 2}′ times and a site-specific comparison of T{sub 2}′ at 93 kHz versus 60 kHz MAS frequency. A quantitative analysis of transfer efficiencies of building blocks, as they are used for typical 3D experiments, was performed. To do this, we compared dipolar-coupling and J-coupling based transfer steps. The building blocks were then combined into 3D experiments for sequential resonance assignment, where we evaluated signal-to-noise ratio and information content of the different 3D spectra in order to identify the best assignment strategy. Based on this comparison, six experiments were selected to optimally assign the model protein ubiquitin, solely using spectra acquired at 93 kHz MAS. Within 3 days of instrument time, the required spectra were recorded from which the backbone resonances have been assigned to over 96 %.

  10. 1H-NMR study of diamagnetic cytochrome P450cam: assignment of heme resonances and substrate dependance of one cysteinate beta proton.

    Science.gov (United States)

    Mouro, C; Bondon, A; Simonneaux, G; Jung, C

    1997-09-08

    The 1H-NMR study of diamagnetic cytochrome P450cam FeII-CO has been performed for the first time. Chemical shifts of the cysteinate fifth ligand protons and of several heme protons have been assigned through 1- and 2-dimensional spectra at 500 MHz. A substrate dependance has been observed for the resonance of the cysteinate proton detected in the high-field region.

  11. HN-NCA heteronuclear TOCSY-NH experiment for {sup 1}H{sup N} and {sup 15}N sequential correlations in ({sup 13}C, {sup 15}N) labelled intrinsically disordered proteins

    Energy Technology Data Exchange (ETDEWEB)

    Wiedemann, Christoph; Goradia, Nishit; Häfner, Sabine [Leibniz Institute for Age Research, Fritz Lipmann Institute, Research Group Biomolecular NMR Spectroscopy (Germany); Herbst, Christian [Ubon Ratchathani University, Department of Physics, Faculty of Science (Thailand); Görlach, Matthias; Ohlenschläger, Oliver; Ramachandran, Ramadurai, E-mail: raman@fli-leibniz.de [Leibniz Institute for Age Research, Fritz Lipmann Institute, Research Group Biomolecular NMR Spectroscopy (Germany)

    2015-10-15

    A simple triple resonance NMR experiment that leads to the correlation of the backbone amide resonances of each amino acid residue ‘i’ with that of residues ‘i−1’ and ‘i+1’ in ({sup 13}C, {sup 15}N) labelled intrinsically disordered proteins (IDPs) is presented. The experimental scheme, {HN-NCA heteronuclear TOCSY-NH}, exploits the favourable relaxation properties of IDPs and the presence of {sup 1}J{sub CαN} and {sup 2}J{sub CαN} couplings to transfer the {sup 15}N{sub x} magnetisation from amino acid residue ‘i’ to adjacent residues via the application of a band-selective {sup 15}N–{sup 13}C{sup α} heteronuclear cross-polarisation sequence of ∼100 ms duration. Employing non-uniform sampling in the indirect dimensions, the efficacy of the approach has been demonstrated by the acquisition of 3D HNN chemical shift correlation spectra of α-synuclein. The experimental performance of the RF pulse sequence has been compared with that of the conventional INEPT-based HN(CA)NH pulse scheme. As the availability of data from both the HCCNH and HNN experiments will make it possible to use the information extracted from one experiment to simplify the analysis of the data of the other and lead to a robust approach for unambiguous backbone and side-chain resonance assignments, a time-saving strategy for the simultaneous collection of HCCNH and HNN data is also described.

  12. H-1 and N-15 resonance assignment of the second fibronectin type III module of the neural cell adhesion molecule

    DEFF Research Database (Denmark)

    Kiselyov, Vladislav V; Berezin, Vladimir; Bock, Elisabeth;

    2008-01-01

    We report here the NMR assignment of the second fibronectin type III module of the neural cell adhesion molecule (NCAM). This module has previously been shown to interact with the fibroblast growth factor receptor (FGFR), and the FGFR-binding site was mapped by NMR to the FG-loop region...... of the module. The FG-loop region also contains a putative nucleotide-binding motif, which was shown by NMR to interact with ATP. Furthermore, ATP was demonstrated to inhibit binding of the second F3 module of NCAM to FGFR....

  13. Synthesis of {sup 15}N labeled glyphosate; Sintese do glifosato enriquecido com {sup 15}N

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Tavares, Glauco Arnold; Rossete, Alexssandra L.R.M.; Tagliassachi, Romulo Barbieri; Prestes, Cleuber Vieira [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Dept. de Isotopos Estaveis]. E-mail: crolivei@cena.usp.br

    2005-07-01

    Amongst the actually commercialized herbicides the Glyphosate is the most used in Brazil. Its efficiency as well as the others herbicides against undesirable weeds is harmed by its final composts left at the environment. Although studies has being carried out to improve the knowledge about the herbicides behavior at the environment its complexity has led them towards innumerous to new significant research work where the use of radiolabeled composts (radiative tracers) are recommended to evaluate their bio-availability in the soil. However is the use, the manipulation and the storage of radiolabeled composts is requires an extra care under chemical safety point of view. The use of non radiolabeled composts is a world tendency especially for field researches. Under this context the presented work describes a method for the synthesis of {sup 15}N labeled glyphosate. The {sup 15}N-herbicide was undertaken by phosphometilation with the phosphit dialquil and {sup 15}N-glycine. The tests where carried out through a micro scale production plant and of equimolars amounts. At these conditions it's was possible to reach approximately a 20% of yield. At the conclusion of a best operational condition its expected to offer another important toll that shall be used in glyphosate behavior at the environment and undesirably weeds. (author)

  14. Robust, integrated computational control of NMR experiments to achieve optimal assignment by ADAPT-NMR.

    Science.gov (United States)

    Bahrami, Arash; Tonelli, Marco; Sahu, Sarata C; Singarapu, Kiran K; Eghbalnia, Hamid R; Markley, John L

    2012-01-01

    ADAPT-NMR (Assignment-directed Data collection Algorithm utilizing a Probabilistic Toolkit in NMR) represents a groundbreaking prototype for automated protein structure determination by nuclear magnetic resonance (NMR) spectroscopy. With a [(13)C,(15)N]-labeled protein sample loaded into the NMR spectrometer, ADAPT-NMR delivers complete backbone resonance assignments and secondary structure in an optimal fashion without human intervention. ADAPT-NMR achieves this by implementing a strategy in which the goal of optimal assignment in each step determines the subsequent step by analyzing the current sum of available data. ADAPT-NMR is the first iterative and fully automated approach designed specifically for the optimal assignment of proteins with fast data collection as a byproduct of this goal. ADAPT-NMR evaluates the current spectral information, and uses a goal-directed objective function to select the optimal next data collection step(s) and then directs the NMR spectrometer to collect the selected data set. ADAPT-NMR extracts peak positions from the newly collected data and uses this information in updating the analysis resonance assignments and secondary structure. The goal-directed objective function then defines the next data collection step. The procedure continues until the collected data support comprehensive peak identification, resonance assignments at the desired level of completeness, and protein secondary structure. We present test cases in which ADAPT-NMR achieved results in two days or less that would have taken two months or more by manual approaches.

  15. Robust, integrated computational control of NMR experiments to achieve optimal assignment by ADAPT-NMR.

    Directory of Open Access Journals (Sweden)

    Arash Bahrami

    Full Text Available ADAPT-NMR (Assignment-directed Data collection Algorithm utilizing a Probabilistic Toolkit in NMR represents a groundbreaking prototype for automated protein structure determination by nuclear magnetic resonance (NMR spectroscopy. With a [(13C,(15N]-labeled protein sample loaded into the NMR spectrometer, ADAPT-NMR delivers complete backbone resonance assignments and secondary structure in an optimal fashion without human intervention. ADAPT-NMR achieves this by implementing a strategy in which the goal of optimal assignment in each step determines the subsequent step by analyzing the current sum of available data. ADAPT-NMR is the first iterative and fully automated approach designed specifically for the optimal assignment of proteins with fast data collection as a byproduct of this goal. ADAPT-NMR evaluates the current spectral information, and uses a goal-directed objective function to select the optimal next data collection step(s and then directs the NMR spectrometer to collect the selected data set. ADAPT-NMR extracts peak positions from the newly collected data and uses this information in updating the analysis resonance assignments and secondary structure. The goal-directed objective function then defines the next data collection step. The procedure continues until the collected data support comprehensive peak identification, resonance assignments at the desired level of completeness, and protein secondary structure. We present test cases in which ADAPT-NMR achieved results in two days or less that would have taken two months or more by manual approaches.

  16. Rapid mass spectrometric analysis of 15N-Leu incorporation fidelity during preparation of specifically labeled NMR samples

    DEFF Research Database (Denmark)

    Truhlar, Stephanie M E; Cervantes, Carla F; Torpey, Justin W

    2008-01-01

    Advances in NMR spectroscopy have enabled the study of larger proteins that typically have significant overlap in their spectra. Specific (15)N-amino acid incorporation is a powerful tool for reducing spectral overlap and attaining reliable sequential assignments. However, scrambling of the label...... during protein expression is a common problem. We describe a rapid method to evaluate the fidelity of specific (15)N-amino acid incorporation. The selectively labeled protein is proteolyzed, and the resulting peptides are analyzed using MALDI mass spectrometry. The (15)N incorporation is determined...... by analyzing the isotopic abundance of the peptides in the mass spectra using the program DEX. This analysis determined that expression with a 10-fold excess of unlabeled amino acids relative to the (15)N-amino acid prevents the scrambling of the (15)N label that is observed when equimolar amounts are used...

  17. Rapid proton-detected NMR assignment for proteins with fast magic angle spinning.

    Science.gov (United States)

    Barbet-Massin, Emeline; Pell, Andrew J; Retel, Joren S; Andreas, Loren B; Jaudzems, Kristaps; Franks, W Trent; Nieuwkoop, Andrew J; Hiller, Matthias; Higman, Victoria; Guerry, Paul; Bertarello, Andrea; Knight, Michael J; Felletti, Michele; Le Marchand, Tanguy; Kotelovica, Svetlana; Akopjana, Inara; Tars, Kaspars; Stoppini, Monica; Bellotti, Vittorio; Bolognesi, Martino; Ricagno, Stefano; Chou, James J; Griffin, Robert G; Oschkinat, Hartmut; Lesage, Anne; Emsley, Lyndon; Herrmann, Torsten; Pintacuda, Guido

    2014-09-03

    Using a set of six (1)H-detected triple-resonance NMR experiments, we establish a method for sequence-specific backbone resonance assignment of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra of 5-30 kDa proteins. The approach relies on perdeuteration, amide (2)H/(1)H exchange, high magnetic fields, and high-spinning frequencies (ωr/2π ≥ 60 kHz) and yields high-quality NMR data, enabling the use of automated analysis. The method is validated with five examples of proteins in different condensed states, including two microcrystalline proteins, a sedimented virus capsid, and two membrane-embedded systems. In comparison to contemporary (13)C/(15)N-based methods, this approach facilitates and accelerates the MAS NMR assignment process, shortening the spectral acquisition times and enabling the use of unsupervised state-of-the-art computational data analysis protocols originally developed for solution NMR.

  18. Sequential NMR resonance assignment and structure determination of the Kunitz-type inhibitor domain of the Alzheimer's beta-amyloid precursor protein.

    Science.gov (United States)

    Heald, S L; Tilton, R F; Hammond, L J; Lee, A; Bayney, R M; Kamarck, M E; Ramabhadran, T V; Dreyer, R N; Davis, G; Unterbeck, A

    1991-10-29

    Certain precursor proteins (APP751 and APP770) of the amyloid beta-protein (AP) present in Alzheimer's disease contain a Kunitz-type serine protease inhibitor domain (APPI). In this study, the domain is obtained as a functional inhibitor through both recombinant (APPIr) and synthetic (APPIs) methodologies, and the solution structure of APPI is determined by 1H 2D NMR techniques. Complete sequence-specific resonance assignments (except for P13 and G37 NH) for both APPIr and APPIs are achieved using standard procedures. Ambiguities arising from degeneracies in the NMR resonances are resolved by varying sample conditions. Qualitative interpretation of short- and long-range NOEs reveals secondary structural features similar to those extensively documented by NMR for bovine pancreatic trypsin inhibitor (BPTI). A more rigorous interpretation of the NOESY spectra yields NOE-derived interresidue distance restraints which are used in conjunction with dynamic simulated annealing to generate a family of APPI structures. Within this family, the beta-sheet and helical regions are in good agreement with the crystal structure of BPTI, whereas portions of the protease-binding loops deviate from those in BPTI. These deviations are consistent with those recently described in the crystal structure of APPI (Hynes et al., 1990). Also supported in the NMR study is the hydrophobic patch in the protease-binding domain created by side chain-side chain NOE contacts between M17 and F34. In addition, the NMR spectra indicate that the rotation of the W21 ring in APPI is hindered, unlike Y21 in BPTI, showing a greater than 90% preference for one orientation in the hydrophobic groove.

  19. Purification, Characterization and 1H NMR Resonance Assignment of an α-Like Neurotoxin BmK 16 from the Venom of Chinese Scorpion Buthus martensii Karsch

    Institute of Scientific and Technical Information of China (English)

    ZHANG, Nai-Xia; WU, Gong; WANG, Zhong-Hua; WU, Hou-Ming

    2003-01-01

    A natural scorpion toxin BmK 16 was purified for the first time from the venom of the Chinese scorpion Buthus martensii Karsch (BmK) by using combined gel-filtration, ion exchange and reversed phase chromatography. The sequence of the Nterminal 8 amino acid residues was determined by Edman degradation. Using the N-terminal sequence as a tag, the database searching revealed a hit in the scorpion cDNA Bank.The sequence for N-terminal 8 amino acid residues, molecular weight and amino acid compositions of BmK 16 were identical with the calculated values according to the first 64 residues' sequence of the precursor peptide alpha-neurotoxin TX16 derived from the sequence of the cDNA AF156597 (EMBL). The sequence-specific resonance assignment of BmK 16 was achieved and the intact sequence of BmK 16 was determined as followings: VRDAY IAKPH NCVYE CARNE YCNDL CTKNGAKSGY CQWVG KYGNG CWCKE LPDNV PIRVP GKCH.Furthermore, the results from the sequence homology analysis and the toxicity assays indicated that BmK 16 was an -like scorpion neurotoxin.

  20. Direct NMR resonance assignments of the active site histidine residue in Escherichia coli thioesterase I/protease I/lysophospholipase L1.

    Science.gov (United States)

    Wu, Wen-Jin; Tyukhtenko, Sergiy I; Huang, Tai-Huang

    2006-11-01

    Owing to the hydrogen-bond interaction and rapid exchange rate with the bulk water, the transverse relaxation time for the N(delta1)-H proton of the catalytic histidine in Escherichia coli thioesterase I/protease I/lysophospholipase L1 (TEP-I) is rather short. Because of its catalytic importance, it is desirable to detect and assign this proton resonance. In this paper, we report the first direct NMR correlation between the short-lived N(delta1)-H proton and its covalently attached N(delta1)-nitrogen of the catalytic His157 residue in E. coli thioesterase/protease I. We have used gradient-enhanced jump-return spin-echo HMQC (GE-JR SE HMQC) to obtain a direct correlation between the short-lived N(delta1)-H proton and its covalently attached N(delta1)-nitrogen. The sensitivity of detection for the short-lived N(delta1)-H proton was enhanced substantially by improved water suppression, in particular, the suppression of radiation damping via pulsed field gradients.

  1. Pentacyclic triterpenoids of Mentha villosa: structural identification and {sup 1}H and {sup 13}C resonance assignments; Triterpenoides pentaciclicos de Mentha villosa: identificacao estrutural e atribuicao dos deslocamentos quimicos dos atomos de hidrogenio e carbono

    Energy Technology Data Exchange (ETDEWEB)

    Monte, Francisco J. Queiroz; Oliveira, Eliete F. de [Ceara Univ., Fortaleza, CE (Brazil). Dept. de Quimica Organica e Inorganica; Braz Filho, Raimundo [Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, RJ (Brazil). Setor de Quimica de Produtos Naturais

    2001-08-01

    The structures of seven oleanene and ursene triterpenoids (1-7) isolated from aerial parts of Mentha villosa were identified. In addition, the complete {sup 1} H and {sup 13} C resonance assignments of these triterpenoids were accomplished using 1D and 2D NMR spectroscopic experiments. (author)

  2. Synthesis of {sup 15}N isotope labeled alanine; Sintese da alanina enriquecida com {sup 15}N

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Sant' Ana, Carlos Roberto; Tagliassachi, Romulo Barbieri; Maximo, Everaldo; Prestes, Clelber Vieira [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Dept. de Isotopos Estaveis]. E-mail: crolivei@cena.usp.br

    2005-07-01

    The application of light chemical elements and their stable isotopes in biological studies have been increased over the last years. The use of {sup 15}N labeled amino acids is an important tool for elucidation of peptides structures. This paper describe a method for the synthesis of {sup 15}N isotope labeled alanine at lower costs than international ones, as well as the details of the recovery system of the nitrogen residues. In the present work an amination of {alpha}-haloacids, with the bromopropionic carboxylic acid and labeled aqua ammonia ({sup 15}NH{sub 3} aq) was carried out. In order to avoid eventually losses of {sup 15}NH{sub 3}, special cares were adopted, since the production cost is high. Although the acquisition cost of the {sup 13}N (radioactive) labeled compounds is lower, the obtained stable tracer will allow the accomplishment of important studies of the nitrogen cycling in living things, less occupational and environment hazards, and the time limitation problems in field studies. The tests took place in triplicates with NH{sub 3} (aq) being employed. With the establishment of the system for {sup 15}NH{sub 3} recovery, an average of 94 % of the ammonia employed in the synthesis process was recovered. The purity of the amino acid was state determined by TLC (Thin Layer Chromatography) and HPLC (High-Performance Liquid Chromatography) with a fluorescence detector. The Rf and the retention time of the synthesized sample were similar the sigma standard. Finally, regarding the established conditions, it was possible to obtain the alanine with a production cost about 40 % lower than the international price. (author)

  3. General method of preparation of uniformly {sup 13}C, {sup 15}N-labeled DNA fragments for NMR analysis of DNA structures

    Energy Technology Data Exchange (ETDEWEB)

    Rene, Brigitte; Masliah, Gregoire; Zargarian, Loussine; Mauffret, Olivier; Fermandjian, Serge [UMR 8113 CNRS - LBPA Ecole Normale Superieure de Cachan, Institut Gustave Roussy, Departement de Biologie et Pharmacologie Structurales (France)], E-mail: sfermand@igr.fr

    2006-11-15

    Summary{sup 13}C, {sup 15}N labeling of biomolecules allows easier assignments of NMR resonances and provides a larger number of NMR parameters, which greatly improves the quality of DNA structures. However, there is no general DNA-labeling procedure, like those employed for proteins and RNAs. Here, we describe a general and widely applicable approach designed for preparation of isotopically labeled DNA fragments that can be used for NMR studies. The procedure is based on the PCR amplification of oligonucleotides in the presence of labeled deoxynucleotides triphosphates. It allows great flexibility thanks to insertion of a short DNA sequence (linker) between two repeats of DNA sequence to study. Size and sequence of the linker are designed as to create restriction sites at the junctions with DNA of interest. DNA duplex with desired sequence and size is released upon enzymatic digestion of the PCR product. The suitability of the procedure is validated through the preparation of two biological relevant DNA fragments.

  4. A tracked approach for automated NMR assignments in proteins (TATAPRO)

    Energy Technology Data Exchange (ETDEWEB)

    Atreya, H.S.; Sahu, S.C.; Chary, K.V.R.; Govil, Girjesh [Tata Institute of Fundamental Research, Department of Chemical Sciences (India)

    2000-06-15

    A novel automated approach for the sequence specific NMR assignments of {sup 1}H{sup N}, {sup 13}C{sup {alpha}}, {sup 13}C{sup {beta}}, {sup 13}C'/{sup 1}H{sup {alpha}} and {sup 15}N spins in proteins, using triple resonance experimental data, is presented. The algorithm, TATAPRO (Tracked AuTomated Assignments in Proteins) utilizes the protein primary sequence and peak lists from a set of triple resonance spectra which correlate {sup 1}H{sup N} and {sup 15}N chemical shifts with those of {sup 13}C{sup {alpha}}, {sup 13}C{sup {beta}} and {sup 13}C'/{sup 1}H{sup {alpha}}. The information derived from such correlations is used to create a 'master{sub l}ist' consisting of all possible sets of {sup 1}H{sup N}{sub i}, {sup 15}N{sub i}, {sup 13}C{sup {alpha}}{sub i}, {sup 13}C{sup {beta}}{sub i}, {sup 13}C'{sub i}/{sup 1}H{sup {alpha}}{sub i}, {sup 13}C{sup {alpha}}{sub i-1}, {sup 13}C{sup {beta}}{sub i-1} and {sup 13}C'{sub i-1}/ {sup 1}H{sup {alpha}}{sub i-1} chemical shifts. On the basis of an extensive statistical analysis of {sup 13}C{sup {alpha}} and {sup 13}C{sup {beta}} chemical shift data of proteins derived from the BioMagResBank (BMRB), it is shown that the 20 amino acid residues can be grouped into eight distinct categories, each of which is assigned a unique two-digit code. Such a code is used to tag individual sets of chemical shifts in the master{sub l}ist and also to translate the protein primary sequence into an array called pps{sub a}rray. The program then uses the master{sub l}ist to search for neighbouring partners of a given amino acid residue along the polypeptide chain and sequentially assigns a maximum possible stretch of residues on either side. While doing so, each assigned residue is tracked in an array called assig{sub a}rray, with the two-digit code assigned earlier. The assig{sub a}rray is then mapped onto the pps{sub a}rray for sequence specific resonance assignment. The program has been tested using

  5. Isolation and measurement of 15N2 from respiratory gases of animals administered 15N-labeled substances.

    Science.gov (United States)

    Springer, D L; Reed, D J; Dost, F N

    1981-01-01

    A method is described for collection of metabolic 15N2 from in vitro preparations or intact rats administered 15N-containing compounds. The methods enables routine collection and mass spectrometric measurement of as little as 10 mumol 15N2 respired by a rat over a 24-h period. A device is described that includes either an animal chamber or a tissue reaction vessel in a closed recycling atmosphere, with automatic O2 replenishment and removal of CO2 and water. It is capable of sustaining moderate vacuum and is coupled to a high-vacuum manifold designed to process the contained atmosphere and respiratory gases. The starting atmosphere is an 80:20 mix of sulfur hexafluoride and O2. Recovery of 15N2 gas from the system without an animal present was 101.3 +/- 5.75%. When 15N2 gas was very slowly infused iv into an animal, recovery was 89.1 +/- 5.38%. Use of the method in studies of the fate of [15N]hydrazine in rats indicated that about 15% of the administered hydrazine is rapidly converted to 15N2, followed by slower conversion of an additional 7-10% over the next several hours.

  6. 15N Hyperpolarization by Reversible Exchange Using SABRE-SHEATH

    OpenAIRE

    2015-01-01

    NMR signal amplification by reversible exchange (SABRE) is a NMR hyperpolarization technique that enables nuclear spin polarization enhancement of molecules via concurrent chemical exchange of a target substrate and parahydrogen (the source of spin order) on an iridium catalyst. Recently, we demonstrated that conducting SABRE in microtesla fields provided by a magnetic shield enables up to 10% 15N-polarization (Theis, T.; et al. J. Am. Chem. Soc. 2015, 137, 1404). Hyperpolarization on 15N (an...

  7. δ15N value does not reflect fasting in mysticetes.

    Directory of Open Access Journals (Sweden)

    Alex Aguilar

    Full Text Available The finding that tissue δ(15N values increase with protein catabolism has led researchers to apply this value to gauge nutritive condition in vertebrates. However, its application to marine mammals has in most occasions failed. We investigated the relationship between δ(15N values and the fattening/fasting cycle in a model species, the fin whale, a migratory capital breeder that experiences severe seasonal variation in body condition. We analyzed two tissues providing complementary insights: one with isotopic turnover (muscle and one that keeps a permanent record of variations in isotopic values (baleen plates. In both tissues δ(15N values increased with intensive feeding but decreased with fasting, thus contradicting the pattern previously anticipated. The apparent inconsistency during fasting is explained by the fact that a individuals migrate between different isotopic isoscapes, b starvation may not trigger significant negative nitrogen balance, and c excretion drops and elimination of 15N-depleted urine is minimized. Conversely, when intensive feeding is resumed in the northern grounds, protein anabolism and excretion start again, triggering 15N enrichment. It can be concluded that in whales and other mammals that accrue massive depots of lipids as energetic reserves and which have limited access to drinking water, the δ15N value is not affected by fasting and therefore cannot be used as an indication of nutritive condition.

  8. Sequential protein NMR assignments in the liquid state via sequential data acquisition

    Science.gov (United States)

    Wiedemann, Christoph; Bellstedt, Peter; Kirschstein, Anika; Häfner, Sabine; Herbst, Christian; Görlach, Matthias; Ramachandran, Ramadurai

    2014-02-01

    Two different NMR pulse schemes involving sequential 1H data acquisition are presented for achieving protein backbone sequential resonance assignments: (i) acquisition of 3D {HCCNH and HNCACONH} and (ii) collection of 3D {HNCOCANH and HNCACONH} chemical shift correlation spectra using uniformly 13C,15N labelled proteins. The sequential acquisition of these spectra reduces the overall experimental time by a factor of ≈2 as compared to individual acquisitions. The suitability of this approach is experimentally demonstrated for the C-terminal winged helix (WH) domain of the minichromosome maintenance (MCM) complex of Sulfolobus solfataricus.

  9. Phenylalanine δ15N in Paleo Archives as a New Proxy for δ15N of Exported Primary Production

    Science.gov (United States)

    McCarthy, M.; Batista, F. C.; Vokhshoori, N. L.; Brown, J. T.; Guilderson, T. P.; Ravelo, A. C.; Sherwood, O.

    2012-12-01

    Compound-specific isotope analysis of individual amino acids (CSI-AA) is emerging as a powerful new tool for studying the paleo nitrogen cycle. Because most detrital organic nitrogen is composed of amino acids, CSI-AA can reveal the mechanistic basis for organic nitrogen diagenesis, preserve a record of past food web structure, and potentially reconstruct the δ15N values of past nitrate and primary production. Within the commonly measured amino acids, the δ15N value of phenylalanine (Phe) appears uniquely promising as a new proxy that reflects the nitrogen isotopic value of the original source. Phe δ15N values remain almost unchanged with trophic transfer through food webs, and also during at least the initial stages of organic matter degradation. Here we synthesize results from both bio-archives and recent sediments, which together suggest that at least in Holocene archives the Phe δ15N value does in fact record the average inorganic nitrogen δ15N value at the base of planktonic food webs. However, several important unknowns also remain. These include the extent of variation in amino acid isotopic fractionation patterns in phylogenetically distinct algal groups. The stability of Phe δ15N values in older sediments where organic matter has undergone extensive diagenesis is also an important research area, which may ultimately establish the temporal limit for application of this approach to study past geological epochs. Together, however, results to date suggest that of Phe δ15N values in paleo archives represent a novel molecular-level proxy which is not tied to any specific organism or group, but rather can provide an integrated estimate of δ15N value of exported primary production.

  10. Solvent effects on 15N NMR coordination shifts.

    Science.gov (United States)

    Kleinmaier, Roland; Arenz, Sven; Karim, Alavi; Carlsson, Anna-Carin C; Erdélyi, Máté

    2013-01-01

    (15)N NMR chemical shift became a broadly utilized tool for characterization of complex structures and comparison of their properties. Despite the lack of systematic studies, the influence of solvent on the nitrogen coordination shift, Δ(15)N(coord), was hitherto claimed to be negligible. Herein, we report the dramatic impact of the local environment and in particular that of the interplay between solvent and substituents on Δ(15)N(coord). The comparative study of CDCl(3) and CD(3)CN solutions of silver(I)-bis(pyridine) and silver(I)-bis(pyridylethynyl)benzene complexes revealed the strong solvent dependence of their (15)N NMR chemical shift, with a solvent dependent variation of up to 40 ppm for one and the same complex. The primary influence of the effect of substituent and counter ion on the (15)N NMR chemical shifts is rationalized by corroborating Density-Functional Theory (nor discrete Fourier transform) calculations on the B3LYP/6-311 + G(2d,p)//B3LYP/6-31G(d) level. Cooperative effects have to be taken into account for a comprehensive description of the coordination shift and thus the structure of silver complexes in solution. Our results demonstrate that interpretation of Δ(15)N(coord) in terms of coordination strength must always consider the solvent and counter ion. The comparable magnitude of Δ(15)N(coord) for reported transition metal complexes makes the principal findings most likely general for a broad scale of complexes of nitrogen donor ligands, which are in frequent use in modern organometallic chemistry.

  11. Side chain NMR assignments in the membrane protein OmpX reconstituted in DHPC micelles.

    Science.gov (United States)

    Hilty, Christian; Fernández, César; Wider, Gerhard; Wüthrich, Kurt

    2002-08-01

    Sequence-specific assignments have been obtained for side chain methyl resonances of Val, Leu and Ile in the outer membrane protein X (OmpX) from Escherichia coli reconstituted in 60 kDa micelles in aqueous solution. Using previously established techniques, OmpX was uniformly 2H,13C,15N-labeled with selectively protonated Val-gamma(1,2), Leu-delta(1,2) and Ile-delta1 methyl groups. The thus labeled protein was studied with the novel experiments 3D (H)C(CC)-TOCSY-(CO)-[15N,1H]-TROSY and 3D H(C)(CC)-TOCSY-(CO)-[15N,1H]-TROSY. Compared to the corresponding conventional experimental schemes, the TROSY-type experiments yielded a sensitivity gain of about 2 at 500 MHz. The overall sensitivity of the experiments was further enhanced more than two-fold by the use of a cryoprobe. Complete assignments of the proton and carbon chemical shifts were obtained for all isopropyl methyl groups of Val and Leu, as well as for the delta1-methyls of Ile. The present approach is applicable for soluble proteins or micelle-reconstituted membrane proteins in structures with overall molecular weights up to about 100 kDa, and adds to the potentialities of solution NMR for de novo structure determination as well as for functional studies, such as ligand screening with proteins in large structures.

  12. Resonances

    DEFF Research Database (Denmark)

    an impetus or drive to that account: change, innovation, rupture, or discontinuity. Resonances: Historical Essays on Continuity and Change explores the historiographical question of the modes of interrelation between these motifs in historical narratives. The essays in the collection attempt to realize...... theoretical consciousness through historical narrative ‘in practice’, by discussing selected historical topics from Western cultural history, within the disciplines of history, literature, visual arts, musicology, archaeology, philosophy, and theology. The title Resonances indicates the overall perspective...

  13. Double resonance capacitance spectroscopy (DORCAS): A new experimental technique for assignment of X-ray absorption peaks to surface sites of semiconductor

    CERN Document Server

    Ishii, M

    2003-01-01

    As a new microspectroscopy for semiconductor surface analysis using an X-ray beam, double resonance capacitance spectroscopy (DORCAS) is proposed. For a microscopic X-ray absorption measurement, a local capacitance change owing to X-ray induced emission of localized electrons is detected by a microprobe. The applied bias voltage V sub b dependence of the capacitance also provides information on the surface density of state. The resonance of the Fermi energy with a surface level by V sub b control makes possible the selection of the observable surface site in the X-ray absorption measurements, i.e. site-specific spectroscopy. The double resonance of the surface site selection (V sub b resonance) and the resonant X-ray absorption of the selected site (photon energy h nu resonance) enhances the capacitance signal. The DORCAS measurement of the GaAs surface shows correlation peaks at h nu=10.402 keV and V sub b =-0.4 V and h nu=10.429 keV and V sub b =+0.1 V, indicating that these resonant X-ray absorption peaks ...

  14. (13)C and (15)N solid-state NMR studies on albendazole and cyclodextrin albendazole complexes.

    Science.gov (United States)

    Ferreira, M João G; García, A; Leonardi, D; Salomon, Claudio J; Lamas, M Celina; Nunes, Teresa G

    2015-06-05

    (13)C and (15)N solid-state nuclear magnetic resonance (NMR) spectra were recorded from albendazole (ABZ) and from ABZ:β-cyclodextrin, ABZ:methyl-β-cyclodextrin, ABZ:hydroxypropyl-β-cyclodextrin and ABZ:citrate-β-cyclodextrin, which were prepared by the spray-drying technique. ABZ signals were typical of a crystalline solid for the pure drug and of an amorphous compound obtained from ABZ:cyclodextrin samples. Relevant spectral differences were correlated with chemical interaction between ABZ and cyclodextrins. The number and type of complexes revealed a strong dependence on the cyclodextrin group substituent. Solid-state NMR data were consistent with the presence of stable inclusion complexes.

  15. Spectral density function mapping using 15N relaxation data exclusively.

    Science.gov (United States)

    Farrow, N A; Zhang, O; Szabo, A; Torchia, D A; Kay, L E

    1995-09-01

    A method is presented for the determination of values of the spectral density function, J(omega), describing the dynamics of amide bond vectors from 15N relaxation parameters alone. Assuming that the spectral density is given by the sum of Lorentzian functions, the approach allows values of J(omega) to be obtained at omega = 0, omega N and 0.870 omega H, where omega N and omega H are Larmor frequencies of nitrogen and proton nuclei, respectively, from measurements of 15N T1, T2 and 1H-15N steady-state NOE values at a single spectrometer frequency. Alternatively, when measurements are performed at two different spectrometer frequencies of i and j MHz, J(omega) can be mapped at omega = 0, omega iN, omega jN, 0.870 omega iH and 0.870 omega iH, where omega iN, for example, is the 15N Larmor frequency for a spectrometer operating at 1 MHz. Additionally, measurements made at two different spectrometer frequencies enable contributions to transverse relaxation from motions on millisecond-microsecond time scales to be evaluated and permit assessment of whether a description of the internal dynamics is consistent with a correlation function consisting of a sum of exponentials. No assumptions about the specific form of the spectral density function describing the dynamics of the 15N-NH bond vector are necessary, provided that dJ(omega)/d omega is relatively constant between omega = omega H + omega N to omega = omega H - omega N. Simulations demonstrate that the method is accurate for a wide range of protein motions and correlation times, and experimental data establish the validity of the methodology. Results are presented for a folded and an unfolded form of the N-terminal SH3 domain of the protein drk.

  16. Millimeter-wave optical double resonance schemes for rapid assignment of perturbed spectra, with applications to the C{sup ~} {sup 1}B{sub 2} state of SO{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Park, G. Barratt, E-mail: barratt@mit.edu, E-mail: barratt.park@gmail.com; Womack, Caroline C.; Jiang, Jun; Field, Robert W., E-mail: rwfield@mit.edu [Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States); Whitehill, Andrew R.; Ono, Shuhei [Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States)

    2015-04-14

    Millimeter-wave detected, millimeter-wave optical double resonance (mmODR) spectroscopy is a powerful tool for the analysis of dense, complicated regions in the optical spectra of small molecules. The availability of cavity-free microwave and millimeter wave spectrometers with frequency-agile generation and detection of radiation (required for chirped-pulse Fourier-transform spectroscopy) opens up new schemes for double resonance experiments. We demonstrate a multiplexed population labeling scheme for rapid acquisition of double resonance spectra, probing multiple rotational transitions simultaneously. We also demonstrate a millimeter-wave implementation of the coherence-converted population transfer scheme for background-free mmODR, which provides a ∼10-fold sensitivity improvement over the population labeling scheme. We analyze perturbations in the C{sup ~} state of SO{sub 2}, and we rotationally assign a b{sub 2} vibrational level at 45 328 cm{sup −1} that borrows intensity via a c-axis Coriolis interaction. We also demonstrate the effectiveness of our multiplexed mmODR scheme for rapid acquisition and assignment of three predissociated vibrational levels of the C{sup ~} state of SO{sub 2} between 46 800 and 47 650 cm{sup −1}.

  17. Reduced Dimensionality (4,3)D-hnCOCANH Experiment: An Efficient Backbone Assignment tool for NMR studies of Proteins

    CERN Document Server

    Kumar, Dinesh

    2013-01-01

    Sequence specific resonance assignment and secondary structure determination of proteins form the basis for variety of structural and functional proteomics studies by NMR. In this context, an efficient standalone method for rapid assignment of backbone (1H, 15N, 13Ca and 13C') resonances and secondary structure determination of proteins has been presented here. Compared to currently available strategies used for the purpose, the method employs only a single reduced dimensionality (RD) experiment -(4,3)D-hnCOCANH and exploits the linear combinations of backbone (13Ca and 13C') chemical shifts to achieve a dispersion relatively better compared to those of individual chemical shifts (see the text) for efficient and rapid data analysis. Further, the experiment leads to the spectrum with direct distinction of self (intra-residue) and sequential (inter-residue) carbon correlation peaks; these appear opposite in signs and therefore can easily be discriminated without using an additional complementary experiment. On ...

  18. Out-and-back {sup 13}C-{sup 13}C scalar transfers in protein resonance assignment by proton-detected solid-state NMR under ultra-fast MAS

    Energy Technology Data Exchange (ETDEWEB)

    Barbet-Massin, Emeline; Pell, Andrew J. [University of Lyon, CNRS/ENS Lyon/UCB Lyon 1, Centre de RMN a Tres Hauts Champs (France); Jaudzems, Kristaps [Latvian Institute of Organic Synthesis (Latvia); Franks, W. Trent; Retel, Joren S. [Leibniz-Institut fuer Molekulare Pharmakologie (Germany); Kotelovica, Svetlana; Akopjana, Inara; Tars, Kaspars [Biomedical Research and Study Center (Latvia); Emsley, Lyndon [University of Lyon, CNRS/ENS Lyon/UCB Lyon 1, Centre de RMN a Tres Hauts Champs (France); Oschkinat, Hartmut [Leibniz-Institut fuer Molekulare Pharmakologie (Germany); Lesage, Anne; Pintacuda, Guido, E-mail: guido.pintacuda@ens-lyon.fr [University of Lyon, CNRS/ENS Lyon/UCB Lyon 1, Centre de RMN a Tres Hauts Champs (France)

    2013-08-15

    We present here {sup 1}H-detected triple-resonance H/N/C experiments that incorporate CO-CA and CA-CB out-and-back scalar-transfer blocks optimized for robust resonance assignment in biosolids under ultra-fast magic-angle spinning (MAS). The first experiment, (H)(CO)CA(CO)NH, yields {sup 1}H-detected inter-residue correlations, in which we record the chemical shifts of the CA spins in the first indirect dimension while during the scalar-transfer delays the coherences are present only on the longer-lived CO spins. The second experiment, (H)(CA)CB(CA)NH, correlates the side-chain CB chemical shifts with the NH of the same residue. These high sensitivity experiments are demonstrated on both fully-protonated and 100 %-H{sup N} back-protonated perdeuterated microcrystalline samples of Acinetobacter phage 205 (AP205) capsids at 60 kHz MAS.

  19. Geomorphic control on the δ15N of mountain forests

    Directory of Open Access Journals (Sweden)

    R. G. Hilton

    2013-03-01

    Full Text Available Mountain forests are subject to high rates of physical erosion which can export particulate nitrogen from ecosystems. However, the impact of geomorphic processes on nitrogen budgets remains poorly constrained. We have used the elemental and isotopic composition of soil and plant organic matter to investigate nitrogen cycling in the mountain forest of Taiwan, from 24 sites with distinct geomorphic (topographic slope and climatic (precipitation, temperature characteristics. The organic carbon to nitrogen ratio of soil organic matter decreased with soil 14C age, providing constraint on average rates of nitrogen loss using a mass balance model. Model predictions suggest that present day estimates of nitrogen deposition exceed contemporary and historic nitrogen losses. We found ∼6‰ variability in the stable isotopic composition (δ15N of soil and plants which was not related to soil 14C age or climatic conditions. Instead, δ15N was significantly, negatively correlated with topographic slope. Using the mass balance model, we demonstrate that the correlation can be explained by an increase in nitrogen loss by non-fractioning pathways on steeper slopes, where physical erosion most effectively removes particulate nitrogen. Published data from forests on steep slopes are consistent with the correlation. Based on our dataset and these observations, we hypothesise that variable physical erosion rates can significantly influence soil δ15N, and suggest particulate nitrogen export is a major, yet underappreciated, loss term in the nitrogen budget of mountain forests.

  20. Probing site-specific 13C/15N-isotope enrichment of spider silk with liquid-state NMR spectroscopy.

    Science.gov (United States)

    Shi, Xiangyan; Yarger, Jeffery L; Holland, Gregory P

    2013-05-01

    Solid-state nuclear magnetic resonance (NMR) has been extensively used to elucidate spider silk protein structure and dynamics. In many of these studies, site-specific isotope enrichment is critical for designing particular NMR methods for silk structure determination. The commonly used isotope analysis techniques, isotope-ratio mass spectroscopy and liquid/gas chromatography-mass spectroscopy, are typically not capable of providing the site-specific isotope information for many systems because an appropriate sample derivatization method is not available. In contrast, NMR does not require any sample derivatization or separation prior to analysis. In this article, conventional liquid-state (1)H NMR was implemented to evaluate incorporation of (13)C/(15)N-labeled amino acids in hydrolyzed spider dragline silk. To determine site-specific (13)C and (15)N isotope enrichments, an analysis method was developed to fit the (1)H-(13)C and (1)H-(15)N J-splitting (J CH and J NH) (1)H NMR peak patterns of hydrolyzed silk fiber. This is demonstrated for Nephila clavipes spiders, where [U-(13)C3,(15)N]-Ala and [1-(13)C,(15)N]-Gly were dissolved in their water supplies. Overall, contents for Ala and Gly isotopomers are extracted for these silk samples. The current methodology can be applied to many fields where site-specific tracking of isotopes is of interest.

  1. HNCA-TOCSY-CANH experiments with alternate {sup 13}C-{sup 12}C labeling: a set of 3D experiment with unique supra-sequential information for mainchain resonance assignment

    Energy Technology Data Exchange (ETDEWEB)

    Takeuchi, Koh; Gal, Maayan [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States); Takahashi, Hideo; Shimada, Ichio [National Institute of Advanced Industrial Science and Technology, Biomedicinal Information Research Center (Japan); Wagner, Gerhard, E-mail: gerhard_wagner@hms.harvard.edu [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States)

    2011-01-15

    Described here is a set of three-dimensional (3D) NMR experiments that rely on CACA-TOCSY magnetization transfer via the weak {sup 3}J(C{sub {alpha}}C{sub {alpha}}) coupling. These pulse sequences, which resemble recently described {sup 13}C detected CACA-TOCSY (Takeuchi et al. 2010) experiments, are recorded in {sup 1}H{sub 2}O, and use {sup 1}H excitation and detection. These experiments require alternate {sup 13}C-{sup 12}C labeling together with perdeuteration, which allows utilizing the small {sup 3}J(C{sub {alpha}}C{sub {alpha}}) scalar coupling that is otherwise masked by the stronger {sup 1}J{sub CC} couplings in uniformly {sup 13}C labeled samples. These new experiments provide a unique assignment ladder-mark that yields bidirectional supra-sequential information and can readily straddle proline residues. Unlike the conventional HNCA experiment, which contains only sequential information to the {sup 13}(C{sub {alpha}}) of the preceding residue, the 3D hnCA-TOCSY-caNH experiment can yield sequential correlations to alpha carbons in positions i-1, i + 1 and i-2. Furthermore, the 3D hNca-TOCSY-caNH and Hnca-TOCSY-caNH experiments, which share the same magnetization pathway but use a different chemical shift encoding, directly couple the {sup 15}N-{sup 1}H spin pair of residue i to adjacent amide protons and nitrogens at positions i-2, i-1, i + 1 and i + 2, respectively. These new experimental features make protein backbone assignments more robust by reducing the degeneracy problem associated with the conventional 3D NMR experiments.

  2. Nitrogen input 15N-signatures are reflected in plant 15N natural abundances of N-rich tropical forest in China

    Science.gov (United States)

    Abdisa Gurmesa, Geshere; Lu, Xiankai; Gundersen, Per; Yunting, Fang; Mo, Jiangming

    2016-04-01

    In this study, we tested the measurement of natural abundance of 15N15N) for its ability to assess changes in N cycling due to increased N deposition in two forest types; namely, an old-growth broadleaved forest and a pine forest, in southern China. We measured δ15N values of inorganic N in input and output fluxes under ambient N deposition, and N concentration and δ15N of major ecosystem compartments under ambient and increased N deposition. Our results showed that N deposition to the forests was 15N-depleted, and was dominated by NH4-N. Plants were 15N-depleted due to imprint from the 15N-depleted atmospheric N deposition. The old-growth forest had larger N concentration and was more 15N-enriched than the pine forest. Nitrogen addition did not significantly affect N concentration, but it significantly increased δ15N values of plants, and slightly more so in the pine forest, toward the 15N signature of the added N in both forests. The result indicates that the pine forest may rely more on the 15N-depleted deposition N. Soil δ15N values were slightly decreased by the N addition. Our result suggests that ecosystem δ15N is more sensitive to the changes in ecosystem N status and N cycling than N concentration in N-saturated sub-tropical forests.

  3. Automated analysis of protein NMR assignments using methods from artificial intelligence.

    Science.gov (United States)

    Zimmerman, D E; Kulikowski, C A; Huang, Y; Feng, W; Tashiro, M; Shimotakahara, S; Chien, C; Powers, R; Montelione, G T

    1997-06-20

    An expert system for determining resonance assignments from NMR spectra of proteins is described. Given the amino acid sequence, a two-dimensional 15N-1H heteronuclear correlation spectrum and seven to eight three-dimensional triple-resonance NMR spectra for seven proteins, AUTOASSIGN obtained an average of 98% of sequence-specific spin-system assignments with an error rate of less than 0.5%. Execution times on a Sparc 10 workstation varied from 16 seconds for smaller proteins with simple spectra to one to nine minutes for medium size proteins exhibiting numerous extra spin systems attributed to conformational isomerization. AUTOASSIGN combines symbolic constraint satisfaction methods with a domain-specific knowledge base to exploit the logical structure of the sequential assignment problem, the specific features of the various NMR experiments, and the expected chemical shift frequencies of different amino acids. The current implementation specializes in the analysis of data derived from the most sensitive of the currently available triple-resonance experiments. Potential extensions of the system for analysis of additional types of protein NMR data are also discussed.

  4. Assignment of congested NMR spectra: Carbonyl backbone enrichment via the Entner Doudoroff pathway

    Science.gov (United States)

    Goldbourt, Amir; Day, Loren A.; McDermott, Ann E.

    2007-12-01

    In NMR spectra of complex proteins, sparse isotope enrichment can be important, in that the removal of many 13C- 13C homonuclear J-couplings can narrow the lines and thereby facilitate the process of spectral assignment and structure elucidation. We present a simple scheme for selective yet extensive isotopic enrichment applicable for production of proteins in organisms utilizing the Entner-Doudoroff (ED) metabolic pathway. An enrichment scheme so derived is demonstrated in the context of a magic-angle spinning solid-state NMR (MAS SSNMR) study of Pf1 bacteriophage, the host of which is Pseudomonas aeruginosa, strain K (PAK), an organism that uses the ED pathway for glucose catabolism. The intact and infectious Pf1 phage in this study was produced by infected PAK cells grown on a minimal medium containing 1- 13C D-glucose ( 13C in position 1) as the sole carbon source, as well as 15NH 4Cl as the only nitrogen source. The 37 MDa Pf1 phage consists of about 93% major coat protein, 1% minor coat proteins, and 6% single-stranded, circular DNA. As a consequence of this composition and the enrichment scheme, the resonances in the MAS SSNMR spectra of the Pf1 sample were almost exclusively due to carbonyl carbons in the major coat protein. Moreover, 3D heteronuclear NCOCX correlation experiments also show that the amino acids leucine, serine, glycine, and tyrosine were not isotopically enriched in their carbonyl positions (although most other amino acids were), which is as expected based upon considerations of the ED metabolic pathway. 3D NCOCX NMR data and 2D 15N- 15N data provided strong verification of many previous assignments of 15N amide and 13C carbonyl shifts in this highly congested spectrum; both the semi-selective enrichment patterns and the narrowed linewidths allowed for greater certainty in the assignments as compared with use of uniformly enriched samples alone.

  5. Nonpolar nitrous oxide dimer: fundamentals of the mixed 14N2O-15N2O dimer and new combination bands of (14N2O)2 and (15N2O)2 involving the Bu, intermolecular bend.

    Science.gov (United States)

    Dehghany, M; Afshari, Mahin; Abusara, Z; Moazzen-Ahmadi, N

    2009-09-21

    Spectra of the nonpolar nitrous oxide dimer in the region of the N2O v1 fundamental band are observed in a pulsed supersonic slit jet expansion probed with a tunable diode laser. Four bands are analysed: two fundamentals of the mixed 14N2O-15N2O dimer and combination bands involving the intermolecular disrotation of the monomers (Bu intermolecular bend) for both (14N2O)2 and (15N2O)2. Because the determination of this intermolecular frequency relies on the experimentally unknown frequency of the (forbidden) symmetric fundamental, we used previously published ab initio results and their proximity to our experimental values to assign the upper state of the combination bands. The resulting intermolecular disrotation frequencies are 42.3(1.0) and 41.6(1.0) cm(-1) for the (14N2O)2 and (15N2O)2, respectively. This represents the first observation of the mixed 14N2O-15N2O dimer, and the direct determination of a second intermolecular frequency for the nonpolar (N2O)2.

  6. 15N NMR chemical shifts in papaverine decomposition products

    Science.gov (United States)

    Czyrski, Andrzej; Girreser, Ulrich; Hermann, Tadeusz

    2013-03-01

    Papaverine can be easily oxidized to papaverinol, papaveraldine and 2,3,9,10-tetramethoxy-12-oxo-12H-indolo[2,1-a]isoquinolinium chloride. On addition of alkali solution the latter compound forms 2-(2-carboxy-4,5-dimethoxyphenyl)-6,7-dimethoxyisoquinolinium inner salt. Together with these structures the interesting 13-(3,4-dimethoxyphenyl)-2,3,8,9-tetramethoxy-6a-12a-diazadibenzo[a,g]fluorenylium chloride is discussed, which is formed in the Gadamer-Schulemann reaction of papaverine as a side product. This letter reports the 15N NMR spectra of the above mentioned compounds.

  7. Characterization of Humic Fractions in a 15N-labelled Soil by Solid by State-State 13C and 15N NMR

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Five humic fractions were obtained from a uniformly 15N-labelled soil by extraction with 0.1 mol L-1 Na4P2O7,0.1 mol L-1 NaOH,and HF/HC1-0.1 mol L-1 NaOH,consecutively,and analyzed by 13C and 15N CPMAS NMR (croas polarization and magic angle spinning nuclear magnetic resonance).Compared with those of native soils humic fractions studied as a whole contained more alkyls,methoxyls and O-alkyls,being 27%~36%,17%~21% and 36%~40%,respectively,but fewer aromatics and carboxyls (being 14%~20% and 13%~90%,respectively).Among those humic fractions,the humic acid (HA) and fulvic acid (FA) extracted by 0.1 mol L-1 Na4P2O7 contained slightly more carboxyls than corresponding humic fractions extracted by 0.1 mol L-1 NaOH,and the HA extracted by 0.1 mol L-1 NaOH after treatment with HF/HC1 contained the least aromatics and carboxyls. The distribution of nitrogen functional groups of soil humic fractions studied was quite similar to each other and also quite similar to that of humic fraction from native soils.More than 75% of total N in each fraction was in amide form,with 9%~13% present as aromatic and/or aliphatic amines and the remainder as heterocyclic N.

  8. 3-DIMENSIONAL N-15-H-1-H-1 AND N-15-C-13-H-1 NUCLEAR-MAGNETIC-RESONANCE STUDIES OF HPR, A CENTRAL COMPONENT OF THE PHOSPHOENOLPYRUVATE-DEPENDENT PHOSPHOTRANSFERASE SYSTEM FROM ESCHERICHIA-COLI - ASSIGNMENT OF BACKBONE RESONANCES

    NARCIS (Netherlands)

    VANNULAND, NAJ; VANDIJK, AA; DIJKSTRA, K; VANHOESEL, FHJ; SCHEEK, RM; ROBILLARD, GT

    1992-01-01

    We have performed three-dimensional NMR studies on a central component of the phosphoenolpyruvate-dependent phosphotransferase system of Escherichia coli, denoted as HPr. The protein was uniformly enriched with N-15 and C-13 to overcome spectral overlap. Complete assignments were obtained for the ba

  9. Multiple-quantum HCN-CCH-TOCSY experiment for 13C/15N labeled RNA oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Hu Weidong; Jiang Licong [Memorial Sloan-Kettering Cancer Center (United States)

    1999-12-15

    A multiple-quantum 3D HCN-CCH-TOCSY experiment is presented for the assignment of RNA ribose resonances. The experiment makes use of the chemical shift dispersion of N1 of pyrimidine and N9 of purine to distinguish the ribose spin systems. It provides an alternative approach for the assignment of ribose resonances to the currently used COSY- and TOCSY-type experiments in which either {sup 13}C or {sup 1}H is utilized to distinguish the different spin systems. Compared to the single-quantum version, the sensitivity of the multiple-quantum HCN-CCH-TOCSY experiment is enhanced on average by a factor of 2 for a 23-mer RNA aptamer complexed with neomycin.

  10. On the Assignment of the Resonance D(2550)%关于共振态D(2550)的安排

    Institute of Scientific and Technical Information of China (English)

    李祯

    2013-01-01

    利用雷吉轨迹唯象模型及其不同夸克味道参数间满足的截距相加性和斜率倒数相加性关系,计算出21S0介子九重态中Dcn的质量为2520MeV.该结果与Babar工作组测出的含有粲夸克的共振态D(2550)的质量符合得很好,也进一步说明李德民等基于衰变宽度的计算结果,将共振态D(2550)安排为10D(2 S )有其合理性.%Based on the Regge trajectory phenomenology and the relations, the parameters for different flavors can be related by the additivity of intercepts and inverse slopes, the mass of the cnD ( 21S0 ) meson is given to be 2520MeV. The result is in excellent agreement with the experimental data of mass of D(2550) from Babar Collaboration, and further shows the rationality that the D(2550 ) is assigned to be 1 0D(2 S ) based on the calculated decay width by Li et al.

  11. Synthesis and biosynthesis of {sup 13}C-, {sup 15}N-labeled deoxynucleosides useful for biomolecular structural determinations

    Energy Technology Data Exchange (ETDEWEB)

    Ashburn, D.A.; Garcia, K.; Hanners, J.L.; Silks, L.A. III; Unkefer, C.J. [Los Alamos National Laboratory, NM (United States)

    1994-12-01

    Currently, there is a great emphasis on elucidating the structure, function, and dynamics of DNA. Much of the research involved in this study uses nuclear magnetic resonance (NMR) spectroscopy. Effective use of NMR spectroscopy for DNA molecules with mw > 10,000 requires stable isotope enrichment. We present strategies for site-specific isotopic labeling of the purine bases adenosine and guanosine and the biosynthesis of (U-{sup 13}C, {sup 15}N) DNA from methylotropic bacteria. With commercially available 6-chloropurine, an effective two-step route leads to 2{prime}-deoxy-(amino-{sup 15}N)adenosine (dA). The resulting d(amino-{sup 15}N)A is used in a series of reactions to synthesize 2{prime}-deoxy-(2-{sup 13}C,1,amino-{sup 15}N{sub 2})guanosine or any combination thereof. An improved biosynthesis of labeled DNA has been accomplished using Methylobacterium extorquens AS1. Each liter of growth medium contains 4 g of methanol to yield 1 g of lyophilized cells. As much as 200 mg of RNA per liter of culture has been obtained. We are currently developing large-scale isolation protocols. General synthetic pathways to oligomeric DNA will be presented.

  12. Stable isotope tracking of endangered sea turtles: validation with satellite telemetry and δ15N analysis of amino acids.

    Directory of Open Access Journals (Sweden)

    Jeffrey A Seminoff

    Full Text Available Effective conservation strategies for highly migratory species must incorporate information about long-distance movements and locations of high-use foraging areas. However, the inherent challenges of directly monitoring these factors call for creative research approaches and innovative application of existing tools. Highly migratory marine species, such as marine turtles, regularly travel hundreds or thousands of kilometers between breeding and feeding areas, but identification of migratory routes and habitat use patterns remains elusive. Here we use satellite telemetry in combination with compound-specific isotope analysis of amino acids to confirm that insights from bulk tissue stable isotope analysis can reveal divergent migratory strategies and within-population segregation of foraging groups of critically endangered leatherback sea turtles (Dermochelys coriacea across the Pacific Ocean. Among the 78 turtles studied, we found a distinct dichotomy in δ(15N values of bulk skin, with distinct "low δ(15N" and "high δ(15N" groups. δ(15N analysis of amino acids confirmed that this disparity resulted from isotopic differences at the base of the food chain and not from differences in trophic position between the two groups. Satellite tracking of 13 individuals indicated that their bulk skin δ(15N value was linked to the particular foraging region of each turtle. These findings confirm that prevailing marine isoscapes of foraging areas can be reflected in the isotopic compositions of marine turtle body tissues sampled at nesting beaches. We use a Bayesian mixture model to show that between 82 and 100% of the 78 skin-sampled turtles could be assigned with confidence to either the eastern Pacific or western Pacific, with 33 to 66% of all turtles foraging in the eastern Pacific. Our forensic approach validates the use of stable isotopes to depict leatherback turtle movements over broad spatial ranges and is timely for establishing wise conservation

  13. Fate of orally administered {sup 15}N-labeled polyamines in rats bearing solid tumors

    Energy Technology Data Exchange (ETDEWEB)

    Kobayashi, Masaki; Samejima, Keijiro; Goda, Hitomi; Niitsu, Masaru [Josai Univ., Sakado, Saitama (Japan). Faculty of Pharmaceutical Sciences; Xu Yongji [Qingdao Univ. of Science and Technology (China). Inst. of Chemical and Molecular Technology; Takahashi, Masakazu [Sasaki Inst., Tokyo (Japan); Hashimoto, Yoshiyuki [Kyoritsu Coll. of Pharmacy, Tokyo (Japan)

    2003-03-01

    We studied absorption, distribution, metabolism, and excretion of polyamines (putrescine, spermidine, and spermine) in the gastrointestinal tract using {sup 15}N-labeled polyamines as tracers and ionspray ionization mass spectrometry (IS-MS). The relatively simple protocol using rats bearing solid tumors provided useful information. Three {sup 15}N-labeled polyamines that were simultaneously administered were absorbed equally from gastrointestinal tract, and distributed within tissues at various concentrations. The uptake of {sup 15}N-spermidine seemed preferential to that of {sup 15}N-spermine since the concentrations of {sup 15}N-spermidine in the liver and tumors were higher, whereas those of {sup 15}N-spermine were higher in the kidney, probably due to the excretion of excess extracellular spermine. Most of the absorbed {sup 15}N-putrescine seemed to be lost, suggesting blood and tissue diamine oxidase degradation. Concentrations of {sup 15}N-spermidine and {sup 15}N-spermine in the tumor were low. We also describe the findings from two rats that were administered with {sup 15}N-spermine. The tissue concentrations of {sup 15}N-spermine were unusually high, and significant levels of {sup 15}N-spermidine were derived from {sup 15}N-spermine in these animals. (author)

  14. Application of rate equations to ELDOR and saturation recovery experiments on 14N: 15N spin-label pairs

    Science.gov (United States)

    Yin, Jun-Jie; Hyde, James S.

    Rate equations describing the time dependence of population differences of the five allowed transitions in an 14N 15N spin-label pair problem are set up. Included in the formulation are the three Heisenberg exchange rate constants and different nitrogen nuclear spin-lattice relaxation rates, electron spin-lattice relaxation rates, and populations for the 14N and 15N moieties. Using matrix algebra, stationary and time-dependent solutions are obtained in a unified theoretical framework. The calculations apply to stationary and pulse electron-electron double resonance and to saturation-recovery ESR. Particular emphasis is placed on short pulse initial excitation, where the transverse relaxation processes are sufficiently slow that only the population difference of the irradiated transition departs significantly from Boltzmann equilibrium during the excitation.

  15. 1H, 13C and 15N assignment of the GNA1946 outer membrane lipoprotein from Neisseria meningitidis

    NARCIS (Netherlands)

    Neumoin, A.; Leonchiks, A.; Petit, P.; Vuillard, L.; Pizza, M.; Soriani, M.; Boelens, R.; Bonvin, A.M.J.J.

    2011-01-01

    GNA1946 (Genome-derived Neisseria Antigen 1946) is a highly conserved exposed outer membrane lipoprotein from Neisseria meningitidis bacteria of 287 amino acid length (31 kDa). Although the structure of NMB1946 has been solved recently by X-Ray crystallography, understanding the behaviour of GNA1946

  16. Side chain NMR assignments in the membrane protein OmpX reconstituted in DHPC micelles

    Energy Technology Data Exchange (ETDEWEB)

    Hilty, Christian; Fernandez, Cesar; Wider, Gerhard; Wuethrich, Kurt [Institut fuer Molekularbiologie und Biophysik, Eidgenoessische Technische Hochschule Zuerich (Switzerland)], E-mail: wuthrich@mol.biol.ethz.ch

    2002-08-15

    Sequence-specific assignments have been obtained for side chain methyl resonances of Val, Leu and Ile in the outer membrane protein X (OmpX) from Escherichia colireconstituted in 60 kDa micelles in aqueous solution. Using previously established techniques, OmpX was uniformly {sup 2}H,{sup 13}C,{sup 15}N-labeled with selectively protonated Val-{gamma}{sup 1,2}, Leu-{delta}{sup 1,2}and Ile-{delta}{sup 1}methyl groups. The thus labeled protein was studied with the novel experiments 3D (H)C(CC)-TOCSY-(CO)-[{sup 15}N,{sup 1}H]-TROSY and 3D H(C)(CC)-TOCSY-(CO)-[{sup 15}N,{sup 1}H]-TROSY. Compared to the corresponding conventional experimental schemes, the TROSY-type experiments yielded a sensitivity gain of about 2 at 500 MHz. The overall sensitivity of the experiments was further enhanced more than two-fold by the use of a cryoprobe. Complete assignments of the proton and carbon chemical shifts were obtained for all isopropyl methyl groups of Val and Leu, as well as for the {delta}{sup 1}-methyls of Ile. The present approach is applicable for soluble proteins or micelle-reconstituted membrane proteins in structures with overall molecular weights up to about 100 kDa, and adds to the potentialities of solution NMR for de novostructure determination as well as for functional studies, such as ligand screening with proteins in large structures.

  17. (13)C, (15)N CPMAS NMR and GIAO DFT calculations of stereoisomeric oxindole alkaloids from Cat's Claw (Uncaria tomentosa).

    Science.gov (United States)

    Paradowska, Katarzyna; Wolniak, Michał; Pisklak, Maciej; Gliński, Jan A; Davey, Matthew H; Wawer, Iwona

    2008-11-01

    Oxindole alkaloids, isolated from the bark of Uncaria tomentosa [Willd. ex Schult.] Rubiaceae, are considered to be responsible for the biological activity of this herb. Five pentacyclic and two tetracyclic alkaloids were studied by solid-state NMR and theoretical GIAO DFT methods. The (13)C and (15)N CPMAS NMR spectra were recorded for mitraphylline, isomitraphylline, pteropodine (uncarine C), isopteropodine (uncarine E), speciophylline (uncarine D), rhynchophylline and isorhynchophylline. Theoretical GIAO DFT calculations of shielding constants provide arguments for identification of asymmetric centers and proper assignment of NMR spectra. These alkaloids are 7R/7S and 20R/20S stereoisomeric pairs. Based on the (13)C CP MAS chemical shifts the 7S alkaloids (delta C3 70-71ppm) can be easily and conveniently distinguished from 7R (deltaC3 74.5-74.9ppm), also 20R (deltaC20 41.3-41.7ppm) from the 20S (deltaC20 36.3-38.3ppm). The epiallo-type isomer (3R, 20S) of speciophylline is characterized by a larger (15)N MAS chemical shift of N4 (64.6ppm) than the allo-type (3S, 20S) of isopteropodine (deltaN4 53.3ppm). (15)N MAS chemical shifts of N1-H in pentacyclic alkaloids are within 131.9-140.4ppm.

  18. Mimicking floodplain reconnection and disconnection using 15N mesocosm incubations

    Directory of Open Access Journals (Sweden)

    W. Wanek

    2012-04-01

    Full Text Available Floodplain restoration changes the nitrate delivery pattern and dissolved organic matter pool in backwaters but other effects are not yet well known. We performed two mesocosm experiments to quantify the nitrate metabolism in two types of floodplains. Rates of denitrification, dissimilatory nitrate reduction to ammonium (DNRA and anammox were measured using 15N tracer additions in mesocosms containing undisturbed floodplain sediments originating from (1 restored and (2 disconnected sites in the Alluvial Zone National Park on the Danube River downstream of Vienna, Austria. DNRA rates were an order of magnitude lower than denitrification and neither rate was affected by changes in nitrate delivery pattern or organic matter quality. Anammox was not detected at any of the sites. Denitrification was out-competed by assimilation which was estimated to use up to 70% of the available nitrate. Overall, denitrification was higher in the restored sites, with mean rates of 5.7±2.8 mmol N m−2 h−1 compared to the disconnected site (0.6±0.5 mmol N m−1 h−1. In addition, ratios of N2O : N2 were lower in the restored site indicating a more complete denitrification. Nitrate addition did not have any effect on denitrification, nor on the N2O : N2 ratio. However, DOM quality significantly changed the N2O : N2 ratio in both sites. Addition of riverine derived organic matter lowered the N2O : N2 ratio in the disconnected site, whereas addition of floodplain derived organic matter increased the N2O : N2 ratio in the restored site. These results demonstrate that increasing floodplains hydrological connection to the main river channel increases nitrogen retention and decreases nitrous oxide emissions.

  19. NMR assignments of the N-terminal domain of Nephila clavipes spidroin 1.

    Science.gov (United States)

    Parnham, Stuart; Gaines, William A; Duggan, Brendan M; Marcotte, William R; Hennig, Mirko

    2011-10-01

    The building blocks of spider dragline silk are two fibrous proteins secreted from the major ampullate gland named spidroins 1 and 2 (MaSp1, MaSp2). These proteins consist of a large central domain composed of approximately 100 tandem copies of a 35-40 amino acid repeat sequence. Non-repetitive N and C-terminal domains, of which the C-terminal domain has been implicated to transition from soluble and insoluble states during spinning, flank the repetitive core. The N-terminal domain until recently has been largely unknown due to difficulties in cloning and expression. Here, we report nearly complete assignment for all (1)H, (13)C, and (15)N resonances in the 14 kDa N-terminal domain of major ampullate spidroin 1 (MaSp1-N) of the golden orb-web spider Nephila clavipes.

  20. Extensive de novo solid-state NMR assignments of the 33 kDa C-terminal domain of the Ure2 prion

    Energy Technology Data Exchange (ETDEWEB)

    Habenstein, Birgit [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France); Wasmer, Christian [Harvard Medical School (United States); Bousset, Luc; Sourigues, Yannick [UPR 3082 CNRS, Laboratoire d' Enzymologie et Biochimie Structurales (France); Schuetz, Anne [ETH Zurich, Physical Chemistry (Switzerland); Loquet, Antoine [Max Planck Institute for Biophysical Chemistry (Germany); Meier, Beat H., E-mail: beme@ethz.ch [ETH Zurich, Physical Chemistry (Switzerland); Melki, Ronald, E-mail: melki@lebs.cnrs-gif.fr [UPR 3082 CNRS, Laboratoire d' Enzymologie et Biochimie Structurales (France); Boeckmann, Anja, E-mail: a.bockmann@ibcp.fr [UMR 5086 CNRS/Universite de Lyon 1, Institut de Biologie et Chimie des Proteines (France)

    2011-11-15

    We present the de novo resonance assignments for the crystalline 33 kDa C-terminal domain of the Ure2 prion using an optimized set of five 3D solid-state NMR spectra. We obtained, using a single uniformly {sup 13}C, {sup 15}N labeled protein sample, sequential chemical-shift information for 74% of the N, C{alpha}, C{beta} triples, and for 80% of further side-chain resonances for these spin systems. We describe the procedures and protocols devised, and discuss possibilities and limitations of the assignment of this largest protein assigned today by solid-state NMR, and for which no solution-state NMR shifts were available. A comparison of the NMR chemical shifts with crystallographic data reveals that regions with high crystallographic B-factors are particularly difficult to assign. While the secondary structure elements derived from the chemical shift data correspond mainly to those present in the X-ray crystal structure, we detect an additional helical element and structural variability in the protein crystal, most probably originating from the different molecules in the asymmetric unit, with the observation of doubled resonances in several parts, including entire stretches, of the protein. Our results provide the point of departure towards an atomic-resolution structural analysis of the C-terminal Ure2p domain in the context of the full-length prion fibrils.

  1. Analysis of the hyperfine structures and Λ doubling of high-resolution LMR spectra of 15N16O (X2Ⅱ)

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    From the theories of the nuclear hyperfine structure (HFS) and Λ doubling of diatomic molecules, several brief algebraic equations for interpretation of HFS and Λ doubling of transitions of diatomic molecule have been developed. A few important parameters of HFS and Λ doubling of 15N16O have been efficiently and accurately obtained from the analysis of the high resolution spectra of 15N16O (X2Ⅱ) observed in our experiments with these equations. This method can provide an effective approach to obtain important hyperfine parameters of novel radicals from their high resolution laser magnetic resonance spectra.

  2. Regulation of [15N]urea synthesis from [5-15N]glutamine. Role of pH, hormones, and pyruvate.

    Science.gov (United States)

    Nissim, I; Yudkoff, M; Brosnan, J T

    1996-12-01

    We have utilized both [5-15N]glutamine and [3-13C] pyruvate as metabolic tracers in order to: (i) examine the effect of pH, glucagon (GLU), or insulin on the precursor-product relationship between 15NH3, [15N]citrulline, and, thereby, [15N]urea synthesis and (ii) elucidate the mechanism(s) by which pyruvate stimulates [15N] urea synthesis. Hepatocytes isolated from rat were incubated at pH 6.8, 7.4, or 7.6 with 1 mM [5-15N]glutamine and 0.1 mM 14NH4Cl in the presence or the absence of [3-13C] pyruvate (2 mM). A separate series of experiments was performed at pH 7.4 in the presence of insulin or GLU. 15NH3 enrichment exceeded or was equal to that of [15N]citrulline under all conditions except for pH 7.6, when the 15N enrichment in citrulline exceeded that in ammonia. The formation of [15N]citrulline (atom % excess) was increased with higher pH. Flux through phosphate-dependent glutaminase (PDG) and [15N]urea synthesis were stimulated (p < 0.05) at pH 7.6 or with GLU and decreased (p < 0.05) at pH 6.8. Insulin had no significant effect on flux through PDG or on [15N]urea synthesis. Decreased [15N]urea production at pH 6.8 was associated with depleted aspartate and glutamate levels. Pyruvate attenuated this decrease in the aspartate and glutamate pools and stimulated [15N]urea synthesis. Production of Asp from pyruvate was increased with increasing medium pH. Approximately 80% of Asp was derived from [3-13C]pyruvate regardless of incubation pH or addition of hormone. Furthermore, approximately 20, 40, and 50% of the mitochondrial N-acetylglutamate (NAG) pool was derived from [3-13C]pyruvate at pH 6.8, 7.4, and 7.6, respectively. Both the concentration and formation of [13C]NAG from [3-13C]pyruvate were increased (p < 0.05) with glucagon and decreased (p < 0.05) with insulin or at pH 6.8. The data suggest a correlation between changes in [15N]urea synthesis and alterations in the level and synthesis of [13C]NAG from pyruvate. The current observations suggest that the

  3. 1H, 13C, 15N and 195Pt NMR studies of Au(III) and Pt(II) chloride organometallics with 2-phenylpyridine.

    Science.gov (United States)

    Pazderski, Leszek; Pawlak, Tomasz; Sitkowski, Jerzy; Kozerski, Lech; Szłyk, Edward

    2009-11-01

    (1)H, (13)C, (15)N and (195)Pt NMR studies of gold(III) and platinum(II) chloride organometallics with N(1),C(2')-chelated, deprotonated 2-phenylpyridine (2ppy*) of the formulae [Au(2ppy*)Cl(2)], trans(N,N)-[Pt(2ppy*)(2ppy)Cl] and trans(S,N)-[Pt(2ppy*)(DMSO-d(6))Cl] (formed in situ upon dissolving [Pt(2ppy*)(micro-Cl)](2) in DMSO-d(6)) were performed. All signals were unambiguously assigned by HMBC/HSQC methods and the respective (1)H, (13)C and (15)N coordination shifts (i.e. differences between chemical shifts of the same atom in the complex and ligand molecules: Delta(1H)(coord) = delta(1H)(complex) - delta(1H)(ligand), Delta(13C)(coord) = delta(13C)(complex) - delta(13C)(ligand), Delta(15N)(coord) = delta(15N)(complex) - delta(15N)(ligand)), as well as (195)Pt chemical shifts and (1)H-(195)Pt coupling constants discussed in relation to the known molecular structures. Characteristic deshielding of nitrogen-adjacent H(6) protons and metallated C(2') atoms as well as significant shielding of coordinated N(1) nitrogens is discussed in respect to a large set of literature NMR data available for related cyclometallated compounds.

  4. Nutrient Status and δ15N Values in Leaves and Soils: A Cross-Biome Comparison

    Science.gov (United States)

    Mayor, J. R.; Schuur, E. A.; Turner, B. L.; Wright, S. J.

    2011-12-01

    Stable nitrogen (N) isotope ratios (δ15N) are often assumed to provide an integrated measure of multiple nitrogen cycling processes. For instance, shifts in the bioavailability of soil N forms are thought to alter plant δ15N values. Demonstrating this relationship is important as ecosystems undergo anthropogenic disturbances. We evaluated patterns and implied mechanisms of the N cycle using ecosystem δ15N values from 16 plots in boreal black spruce (Picea mariana) forest and lowland wet tropical forest. Fertilizer N and phosphorus (P) was applied annually for five and 11 years prior to measurement of ecosystem δ15N values. Full sun canopy foliage and soil extractable nitrate, ammonium, and dissolved organic N (DON) were sampled in fertilized and control plots and analyzed for δ15N. In boreal forest, N fertilization reduced DON concentrations and caused a depletion of δ15N in foliage and fungal sporocarps. Of four species occurring in all plots in the tropical forest, one (Alseis blackiana) had increased foliar δ15N values following N fertilization, one (Tetragastris panamensis) had increased foliar δ15N values following P fertilization, and one (Oenocarpus mapora) had increased foliar δ15N following N+P fertilization. Surprisingly, soil nitrate in the boreal forest became substantially 15N-enriched under P fertilization, whereas nitrate in the tropical forest soil was enriched only under N or N+P fertilization. Collectively, nitrate enrichment is likely due to enhanced rates of soil denitrification as evidenced by elevated resin extractable soil nitrate concentrations and close correlations between δ15N and δ18O values. On average, foliar δ15N in tropical trees corresponded well with δ15N in soil nitrate in control and P fertilized plots, but was 2-3% more enriched than DON under N and N+P fertilization. In boreal forests, N and N+P fertilization increased foliar N concentration and δ15N values indicating substantial use of applied fertilizer. Taken

  5. Proton-detected 3D (15)N/(1)H/(1)H isotropic/anisotropic/isotropic chemical shift correlation solid-state NMR at 70kHz MAS.

    Science.gov (United States)

    Pandey, Manoj Kumar; Yarava, Jayasubba Reddy; Zhang, Rongchun; Ramamoorthy, Ayyalusamy; Nishiyama, Yusuke

    2016-01-01

    Chemical shift anisotropy (CSA) tensors offer a wealth of information for structural and dynamics studies of a variety of chemical and biological systems. In particular, CSA of amide protons can provide piercing insights into hydrogen-bonding interactions that vary with the backbone conformation of a protein and dynamics. However, the narrow span of amide proton resonances makes it very difficult to measure (1)H CSAs of proteins even by using the recently proposed 2D (1)H/(1)H anisotropic/isotropic chemical shift (CSA/CS) correlation technique. Such difficulties due to overlapping proton resonances can in general be overcome by utilizing the broad span of isotropic chemical shifts of low-gamma nuclei like (15)N. In this context, we demonstrate a proton-detected 3D (15)N/(1)H/(1)H CS/CSA/CS correlation experiment at fast MAS frequency (70kHz) to measure (1)H CSA values of unresolved amide protons of N-acetyl-(15)N-l-valyl-(15)N-l-leucine (NAVL).

  6. Factors Controlling the Stable Nitrogen Isotopic Composition (δ15N of Lipids in Marine Animals.

    Directory of Open Access Journals (Sweden)

    Elisabeth Svensson

    Full Text Available Lipid extraction of biomass prior to stable isotope analysis is known to cause variable changes in the stable nitrogen isotopic composition (δ15N of residual biomass. However, the underlying factors causing these changes are not yet clear. Here we address this issue by comparing the δ15N of bulk and residual biomass of several marine animal tissues (fish, crab, cockle, oyster, and polychaete, as well as the δ15N of the extracted lipids. As observed previously, lipid extraction led to a variable offset in δ15N of biomass (differences ranging from -2.3 to +1.8 ‰. Importantly, the total lipid extract (TLE was highly depleted in 15N compared to bulk biomass, and also highly variable (differences ranging from -14 to +0.7 ‰. The TLE consisted mainly of phosphatidylcholines, a group of lipids with one nitrogen atom in the headgroup. To elucidate the cause for the 15N-depletion in the TLE, the δ15N of amino acids was determined, including serine because it is one of the main sources of nitrogen to N-containing lipids. Serine δ15N values differed by -7 to +2 ‰ from bulk biomass δ15N, and correlated well with the 15N depletion in TLEs. On average, serine was less depleted (-3‰ than the TLE (-7 ‰, possibly due to fractionation during biosynthesis of N-containing headgroups, or that other nitrogen-containing compounds, such as urea and choline, or recycled nitrogen contribute to the nitrogen isotopic composition of the TLE. The depletion in 15N of the TLE relative to biomass increased with the trophic level of the organisms.

  7. An optimized method for (15)N R(1) relaxation rate measurements in non-deuterated proteins.

    Science.gov (United States)

    Gairí, Margarida; Dyachenko, Andrey; González, M Teresa; Feliz, Miguel; Pons, Miquel; Giralt, Ernest

    2015-06-01

    (15)N longitudinal relaxation rates are extensively used for the characterization of protein dynamics; however, their accurate measurement is hindered by systematic errors. (15)N CSA/(1)H-(15)N dipolar cross-correlated relaxation (CC) and amide proton exchange saturation transfer from water protons are the two main sources of systematic errors in the determination of (15)N R1 rates through (1)H-(15)N HSQC-based experiments. CC is usually suppressed through a train of 180° proton pulses applied during the variable (15)N relaxation period (T), which can perturb water magnetization. Thus CC cancellation is required in such a way as to minimize water saturation effects. Here we examined the level of water saturation during the T period caused by various types of inversion proton pulses to suppress CC: (I) amide-selective IBURP-2; (II) cosine-modulated IBURP-2; (III) Watergate-like blocks; and (IV) non-selective hard. We additionally demonstrate the effect of uncontrolled saturation of aliphatic protons on (15)N R1 rates. In this paper we present an optimized pulse sequence that takes into account the crucial effect of controlling also the saturation of the aliphatic protons during (15)N R1 measurements in non-deuterated proteins. We show that using cosine-modulated IBURP-2 pulses spaced 40 ms to cancel CC in this optimized pulse program is the method of choice to minimize systematic errors coming from water and aliphatic protons saturation effects.

  8. A fast method to prepare water samples for 15N analysis

    Institute of Scientific and Technical Information of China (English)

    肖化云; 刘丛强

    2001-01-01

    Automatic element analyser is often used to prepare organic matters tor 15N analysis. It is seldom used to prepare water samples. Water samples are conventionally dealt with by Kjeldahl-Rittenberg technique. But it requires tedious and labor-intensive sample preparation. A fast and reliable method is proposed in this paper to prepare water samples for 15N analysis.

  9. Human dietary δ(15)N intake: representative data for principle food items.

    Science.gov (United States)

    Huelsemann, F; Koehler, K; Braun, H; Schaenzer, W; Flenker, U

    2013-09-01

    Dietary analysis using δ(15)N values of human remains such as bone and hair is usually based on general principles and limited data sets. Even for modern humans, the direct ascertainment of dietary δ(15)N is difficult and laborious, due to the complexity of metabolism and nitrogen fractionation, differing dietary habits and variation of δ(15)N values of food items. The objective of this study was to summarize contemporary regional experimental and global literature data to ascertain mean representative δ(15)N values for distinct food categories. A comprehensive data set of more than 12,000 analyzed food samples was summarized from the literature. Data originated from studies dealing with (1) authenticity tracing or origin control of food items, and (2) effects of fertilization or nutrition on δ(15)N values of plants or animals. Regional German food δ(15)N values revealed no major differences compared with the mean global values derived from the literature. We found that, in contrast to other food categories, historical faunal remains of pig and poultry are significantly enriched in (15)N compared to modern samples. This difference may be due to modern industrialized breeding practices. In some food categories variations in agricultural and feeding regimens cause significant differences in δ(15)N values that may lead to misinterpretations when only limited information is available.

  10. δ(15) N from soil to wine in bulk samples and proline.

    Science.gov (United States)

    Paolini, Mauro; Ziller, Luca; Bertoldi, Daniela; Bontempo, Luana; Larcher, Roberto; Nicolini, Giorgio; Camin, Federica

    2016-09-01

    The feasibility of using δ(15) N as an additional isotopic marker able to link wine to its area of origin was investigated. The whole production chain (soil-leaves-grape-wine) was considered. Moreover, the research included evaluation of the effect of the fermentation process, the use of different types of yeast and white and red vinification, the addition of nitrogen adjuvants and ultrasound lysis simulating wine ageing. The δ(15) N of grapes and wine was measured in bulk samples and compounds, specifically in proline, for the first time. Despite isotopic fractionation from soil to wine, the δ(15) N values of leaves, grapes, wine and particularly must and wine proline conserved the variability of δ(15) N in the growing soil. Fermentation and ultrasound treatment did not affect the δ(15) N values of grape must, which was therefore conserved in wine. The addition of inorganic or organic adjuvants was able to influence the δ(15) N of bulk wine, depending on the amount and the difference between the δ(15) N of must and that of the adjuvant. The δ(15) N of wine proline was not influenced by adjuvant addition and is therefore the best marker for tracing the geographical origin of wine. Copyright © 2016 John Wiley & Sons, Ltd.

  11. Disturbance and topography shape nitrogen availability and δ15N over long-term forest succession

    Science.gov (United States)

    Forest disturbance and long-term succession can promote open N cycling that increases N loss and soil δ15N values. We examined soil and foliar patterns in N and δ15N, and soil N mineralization, across a topographically complex montane forest landscape influenced by human logging ...

  12. Syntheses of 15N-labeled pre-queuosine nucleobase derivatives

    Directory of Open Access Journals (Sweden)

    Jasmin Levic

    2014-08-01

    Full Text Available Pre-queuosine or queuine (preQ1 is a guanine derivative that is involved in the biosynthetic pathway of the hypermodified tRNA nucleoside queuosine (Que. The core structure of preQ1 is represented by 7-(aminomethyl-7-deazaguanine (preQ1 base. Here, we report the synthesis of three preQ1 base derivatives with complementary 15N-labeling patterns, utilizing [15N]-KCN, [15N]-phthalimide, and [15N3]-guanidine as cost-affordable 15N sources. Such derivatives are required to explore the binding process of the preQ1 base to RNA targets using advanced NMR spectroscopic methods. PreQ1 base specifically binds to bacterial mRNA domains and thereby regulates genes that are required for queuosine biosynthesis.

  13. 1H and 15N Dynamic Nuclear Polarization Studies of Carbazole

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Jian Zhi; Solum, Mark S.; Wind, Robert A.; Nilsson, Brad L.; Peterson, Matt A.; Pugmire, Ronald J.; Grant, David M.

    2000-01-01

    15N NMR experiments, combined with dynamic nuclear polarization (DNP), are reported on carbazole doped with the stable free radical 1,3 bisdiphenylene-2 phenylally1 (BDPA). Doping shortens the nuclear relaxation times and provides paramagnetic centers that can be used to enhance the nuclear signal by means of DNP so that 15 N NMR experiments can be done in minutes. The factors were measured in a 1.4 T external field, using both unlabeled and 98% 15N labeled carbazole with doping levels varying between 0.65 and 5.0 wt % BDPA. A doping level of approximately 1 wt % produced optimal results. DNP enhancement factors of 35 and 930 were obtained for 1H and 15N, respectively making it possible to perform 15N DNP NMR experiments at the natural abundance level.

  14. Symbiotic nitrogen fixation in an arid ecosystem measured by sup 15 N natural abundance

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, G.V. (Univ. of New Mexico, Albuquerque (USA))

    1990-05-01

    Plants dependent on nitrogen fixation have an {sup 15}N abundance similar to the atmosphere, while non-nitrogen fixing plants usually are enriched in {sup 15}N and are similar to soil nitrogen values. The natural abundance of {sup 15}N in leaf tissues and soils was determined to evaluate symbiotic nitrogen fixation by several legumes and actinorhizal species in the Sevilleta Long-term Ecological Research area in central New Mexico. Comparison of {delta}{sup 15}N values for the legume Prosopis glandulosa (mesquite) to adjacent Atriplex canascens (fourwing saltbush) indicated that P. glandulosa obtained 66% of its nitrogen by fixation. The legume Hoffmanseggia jamesii was found to be utilizing soil nitrogen. The {delta}{sup 15}N values for the actinorhizal plants, Elaeagnus angustifolia and Cercocarpus montanus, while below values for soil nitrogen, did not differ from associated non-fixing plants.

  15. Applying Thymine Isostere 2,4-Difluoro-5-Methylbenzene as a NMR Assignment Tool and Probe of Homopyrimidine/Homopurine Tract Structural Dynamics.

    Science.gov (United States)

    Brinson, Robert G; Miller, Jennifer T; Kahn, Jason D; Le Grice, Stuart F J; Marino, John P

    2016-01-01

    Proton assignment of nuclear magnetic resonance (NMR) spectra of homopyrimidine/homopurine tract oligonucleotides becomes extremely challenging with increasing helical length due to severe cross-peak overlap. As an alternative to the more standard practice of (15)N and (13)C labeling of oligonucleotides, here, we describe a method for assignment of highly redundant DNA sequences that uses single-site substitution of the thymine isostere 2,4-difluoro-5-methylbenzene (dF). The impact of this approach in facilitating the assignment of intractable spectra and analyzing oligonucleotide structure and dynamics is demonstrated using A-tract and TATA box DNA and two polypurine tract-containing RNA:DNA hybrids derived from HIV-1 and the Saccharomyces cerevisiae long-terminal repeat-containing retrotransposon Ty3. Only resonances proximal to the site of dF substitution exhibit sizable chemical shift changes, providing spectral dispersion while still allowing chemical shift mapping of resonances from unaffected residues distal to the site of modification directly back to the unmodified sequence. It is further illustrated that dF incorporation can subtly alter the conformation and dynamics of homopyrimidine/homopurine tract oligonucleotides, and how these NMR observations can be correlated, in the cases of the TATA box DNA, with modulation in the TATA box-binding protein interaction using an orthogonal gel assay.

  16. Compound-specific 15N analysis of amino acids in 15N tracer experiments provide an estimate of newly synthesised soil protein from inorganic and organic substrates

    Science.gov (United States)

    Charteris, Alice; Michaelides, Katerina; Evershed, Richard

    2015-04-01

    Organic N concentrations far exceed those of inorganic N in most soils and despite much investigation, the composition and cycling of this complex pool of SOM remains poorly understood. A particular problem has been separating more recalcitrant soil organic N from that actively cycling through the soil system; an important consideration in N cycling studies and for the soil's nutrient supplying capacity. The use of 15N-labelled substrates as stable isotope tracers has contributed much to our understanding of the soil system, but the complexity and heterogeneity of soil organic N prevents thorough compound-specific 15N analyses of organic N compounds and makes it difficult to examine any 15N-labelled organic products in any detail. As a result, a significant proportion of previous work has either simply assumed that since the majority of soil N is organic, all of the 15N retained in the soil is organic N (e.g. Sebilo et al., 2013) or subtracted 15N-labelled inorganic compounds from bulk values (e.g. Pilbeam et al., 1997). While the latter approach is more accurate, these methods only provide an estimate of the bulk 15N value of an extremely complex and non-uniformly labelled organic pool. A more detailed approach has been to use microbial biomass extraction (Brookes et al., 1985) and subsequent N isotopic analysis to determine the 15N value of biomass-N, representing the fraction of 15N assimilated by microbes or the 15N cycling through the 'living' or 'active' portion of soil organic N. However, this extraction method can only generate estimates and some lack of confidence in its validity and reliability remains. Here, we present an alternative technique to obtain a measure of the assimilation of an applied 15N substrate by the soil microbial biomass and an estimate of the newly synthesized soil protein, which is representative of the magnitude of the active soil microbial biomass. The technique uses a stable isotope tracer and compound-specific 15N analysis, but

  17. Effect of four plant species on soil 15N-access and herbage yield in temporary agricultural grasslands

    DEFF Research Database (Denmark)

    Pirhofter-Walzl, Karin; Eriksen, Jørgen; Rasmussen, Jim;

    2013-01-01

    access to greater amounts of soil 15N compared with a shallow-rooting binary mixture, and if leguminous plants affect herbage yield and soil 15N-access. Methods 15N-enriched ammonium-sulphate was placed at three different soil depths (0.4, 0.8 and 1.2 m) to determine the depth dependent soil 15N...

  18. Resonance-enhanced multiphoton ionization (REMPI) spectroscopy of bromobenzene and its perdeuterated isotopologue: Assignment of the vibrations of the S{sub 0}, S{sub 1}, and D{sub 0}{sup +} states of bromobenzene and the S{sub 0} and D{sub 0}{sup +} states of iodobenzene

    Energy Technology Data Exchange (ETDEWEB)

    Andrejeva, Anna; Tuttle, William D.; Harris, Joe P.; Wright, Timothy G., E-mail: Tim.Wright@nottingham.ac.uk [School of Chemistry, University of Nottingham, University Park, Nottingham NG7 2RD (United Kingdom)

    2015-12-28

    We report vibrationally resolved spectra of the S{sub 1}←S{sub 0} transition of bromobenzene using resonance-enhanced multiphoton ionization spectroscopy. We study bromobenzene-h{sub 5} as well as its perdeuterated isotopologue, bromobenzene-d{sub 5}. The form of the vibrational modes between the isotopologues and also between the S{sub 0} and S{sub 1} electronic states is discussed for each species, allowing assignment of the bands to be achieved and the activity between states and isotopologues to be established. Vibrational bands are assigned utilizing quantum chemical calculations, previous experimental results, and isotopic shifts. Previous work and assignments of the S{sub 1} spectra are discussed. Additionally, the vibrations in the ground state cation, D{sub 0}{sup +}, are considered, since these have also been used by previous workers in assigning the excited neutral state spectra. We also examine the vibrations of iodobenzene in the S{sub 0} and D{sub 0}{sup +} states and comment on the previous assignments of these. In summary, we have been able to assign the corresponding vibrations across the whole monohalobenzene series of molecules, in the S{sub 0}, S{sub 1}, and D{sub 0}{sup +} states, gaining insight into vibrational activity and vibrational couplings.

  19. 15N Content Reflects Development of Mycorrhizae and Nitrogen Dynamics During Primary Succession

    Science.gov (United States)

    Hobbie, E. A.; Jumpponen, A.

    2004-05-01

    Mycorrhizal fungi are ubiquitous symbionts on terrestrial plants that are particularly important for plant nitrogen nutrition. 15N content appears to be a useful marker of the mycorrhizal role in plant nitrogen supply because of an apparent fractionation against 15N during transfer of nitrogen from mycorrhizal fungi to host plants. Because plants developing during primary succession are gradually colonized by mycorrhizal fungi, such situations provide good opportunities to study interactions between mycorrhizal colonization and plant 15N content. Here, we present results of a study of nitrogen isotope patterns in ecosystem components during the first 100 years of ecosystem development after glacial retreat, and compare those patterns with those on adjacent mature terrain. Soils in primary succession were depleted in 15N relative to nitrogen-fixing plants. Nonmycorrhizal plants and plants generally colonized by ectomycorrhizal, ericoid, or arbuscular fungi showed similar 15N content very early in succession (-4 to -6‰ ), corresponding to low colonization levels of all plant species. Subsequent colonization of evergreen plants by ectomycorrhizal and ericoid fungi led to a 5-6‰ decline in 15N content, indicating transfer of 15N-depleted N from fungi to plants. The values recorded (-10 to -14‰ ) are among the lowest yet observed in vascular plants. Nonmycorrhizal plants and plants colonized by arbuscular mycorrhizal fungi did not decline in 15N content. Most ectomycorrhizal and saprotrophic fungi were similar in 15N content in early succession (-1 to -3‰ ), with the notable exception of ectomycorrhizal fungi suspected of proteolytic capabilities, which were 15N enriched relative to all other fungi. 15N contents in both plants and soil from the mature site were 5‰ greater than in recently exposed sites. We conclude that 1) the primary nitrogen source to this ecosystem must be atmospheric deposition, 2) low plant 15N content generally corresponds with greater

  20. Spectral editing at ultra-fast magic-angle-spinning in solid-state NMR: facilitating protein sequential signal assignment by HIGHLIGHT approach.

    Science.gov (United States)

    Wang, Songlin; Matsuda, Isamu; Long, Fei; Ishii, Yoshitaka

    2016-02-01

    This study demonstrates a novel spectral editing technique for protein solid-state NMR (SSNMR) to simplify the spectrum drastically and to reduce the ambiguity for protein main-chain signal assignments in fast magic-angle-spinning (MAS) conditions at a wide frequency range of 40-80 kHz. The approach termed HIGHLIGHT (Wang et al., in Chem Comm 51:15055-15058, 2015) combines the reverse (13)C, (15)N-isotope labeling strategy and selective signal quenching using the frequency-selective REDOR pulse sequence under fast MAS. The scheme allows one to selectively observe the signals of "highlighted" labeled amino-acid residues that precede or follow unlabeled residues through selectively quenching (13)CO or (15)N signals for a pair of consecutively labeled residues by recoupling (13)CO-(15)N dipolar couplings. Our numerical simulation results showed that the scheme yielded only ~15% loss of signals for the highlighted residues while quenching as much as ~90% of signals for non-highlighted residues. For lysine-reverse-labeled micro-crystalline GB1 protein, the 2D (15)N/(13)Cα correlation and 2D (13)Cα/(13)CO correlation SSNMR spectra by the HIGHLIGHT approach yielded signals only for six residues following and preceding the unlabeled lysine residues, respectively. The experimental dephasing curves agreed reasonably well with the corresponding simulation results for highlighted and quenched residues at spinning speeds of 40 and 60 kHz. The compatibility of the HIGHLIGHT approach with fast MAS allows for sensitivity enhancement by paramagnetic assisted data collection (PACC) and (1)H detection. We also discuss how the HIGHLIGHT approach facilitates signal assignments using (13)C-detected 3D SSNMR by demonstrating full sequential assignments of lysine-reverse-labeled micro-crystalline GB1 protein (~300 nmol), for which data collection required only 11 h. The HIGHLIGHT approach offers valuable means of signal assignments especially for larger proteins through reducing the

  1. Spectral editing at ultra-fast magic-angle-spinning in solid-state NMR: facilitating protein sequential signal assignment by HIGHLIGHT approach

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Songlin; Matsuda, Isamu; Long, Fei; Ishii, Yoshitaka, E-mail: yishii@uic.edu [University of Illinois at Chicago, Department of Chemistry (United States)

    2016-02-15

    This study demonstrates a novel spectral editing technique for protein solid-state NMR (SSNMR) to simplify the spectrum drastically and to reduce the ambiguity for protein main-chain signal assignments in fast magic-angle-spinning (MAS) conditions at a wide frequency range of 40–80 kHz. The approach termed HIGHLIGHT (Wang et al., in Chem Comm 51:15055–15058, 2015) combines the reverse {sup 13}C, {sup 15}N-isotope labeling strategy and selective signal quenching using the frequency-selective REDOR pulse sequence under fast MAS. The scheme allows one to selectively observe the signals of “highlighted” labeled amino-acid residues that precede or follow unlabeled residues through selectively quenching {sup 13}CO or {sup 15}N signals for a pair of consecutively labeled residues by recoupling {sup 13}CO–{sup 15}N dipolar couplings. Our numerical simulation results showed that the scheme yielded only ∼15 % loss of signals for the highlighted residues while quenching as much as ∼90 % of signals for non-highlighted residues. For lysine-reverse-labeled micro-crystalline GB1 protein, the 2D {sup 15}N/{sup 13}C{sub α} correlation and 2D {sup 13}C{sub α}/{sup 13}CO correlation SSNMR spectra by the HIGHLIGHT approach yielded signals only for six residues following and preceding the unlabeled lysine residues, respectively. The experimental dephasing curves agreed reasonably well with the corresponding simulation results for highlighted and quenched residues at spinning speeds of 40 and 60 kHz. The compatibility of the HIGHLIGHT approach with fast MAS allows for sensitivity enhancement by paramagnetic assisted data collection (PACC) and {sup 1}H detection. We also discuss how the HIGHLIGHT approach facilitates signal assignments using {sup 13}C-detected 3D SSNMR by demonstrating full sequential assignments of lysine-reverse-labeled micro-crystalline GB1 protein (∼300 nmol), for which data collection required only 11 h. The HIGHLIGHT approach offers valuable

  2. Alkaline Hydrolysis/Polymerization of 2,4,6-Trinitrotoluene: Characterization of Products by 13C and 15N NMR

    Science.gov (United States)

    Thorn, K.A.; Thorne, P.G.; Cox, L.G.

    2004-01-01

    Alkaline hydrolysis has been investigated as a nonbiological procedure for the destruction of 2,4,6-trinitrotoluene (TNT) in explosives contaminated soils and munitions scrap. Nucleophilic substitutions of the nitro and methyl groups of TNT by hydroxide ion are the initial steps in the alkaline degradation of TNT. Potential applications of the technique include both in situ surface liming and ex situ alkaline treatment of contaminated soils. A number of laboratory studies have reported the formation of an uncharacterized polymeric material upon prolonged treatment of TNT in base. As part of an overall assessment of alkaline hydrolysis as a remediation technique, and to gain a better understanding of the chemical reactions underlying the hydrolysis/polymerization process, the soluble and precipitate fractions of polymeric material produced from the calcium hydroxide hydrolysis of unlabeled and 15N-labeled TNT were analyzed by elemental analysis and 13C and 15N nuclear magnetic resonance spectroscopy. Spectra indicated that reactions leading to polymerization included nucleophilic displacement of nitro groups by hydroxide ion, formation of ketone, carboxyl, alcohol, ether, and other aliphatic carbons, conversion of methyl groups to diphenyl methylene carbons, and recondensation of aromatic amines and reduced forms of nitrite, including ammonia and possibly hydroxylamine, into the polymer. Compared to the distribution of carbons in TNT as 14% sp 3- and 86% sp2-hybridized, the precipitate fraction from hydrolysis of unlabeled TNT contained 33% sp3- and 67% sp 2-hybridized carbons. The concentration of nitrogen in the precipitate was 64% of that in TNT. The 15N NMR spectra showed that, in addition to residual nitro groups, forms of nitrogen present in the filtrate and precipitate fractions include aminohydroquinone, primary amide, indole, imine, and azoxy, among others. Unreacted nitrite was recovered in the filtrate fraction. The toxicities and susceptibilities to

  3. Light-mediated 15N fractionation in Caribbean gorgonian octocorals: implications for pollution monitoring

    Science.gov (United States)

    Baker, D. M.; Kim, K.; Andras, J. P.; Sparks, J. P.

    2011-09-01

    The stable nitrogen isotope ratio ( δ 15N) of coral tissue is a useful recorder of anthropogenic pollution in tropical marine ecosystems. However, little is known of the natural environmentally induced fractionations that affect our interpretation of coral δ 15N values. In symbiotic scleractinians, light affects metabolic fractionation of N during photosynthesis, which may confound the identification of N pollution between sites of varied depth or turbidity. Given the superiority of octocorals for δ 15N studies, our goal was to quantify the effect of light on gorgonian δ 15N in the context of monitoring N pollution sources. Using field collections, we show that δ 15N declined by 1.4‰ over 20 m depth in two species of gorgonians, the common sea fan, Gorgonia ventalina, and the slimy sea plume, Pseudopterogorgia americana. An 8-week laboratory experiment with P. americana showed that light, not temperature causes this variation, whereby the lowest fractionation of the N source was observed in the highest light treatment. Finally, we used a yearlong reciprocal depth transplant experiment to quantify the time frame over which δ 15N changes in G. ventalina as a function of light regime . Over the year, δ 15N was unchanged and increased slightly in the deep control colonies and shallow colonies transplanted to the deep site, respectively. Within 6 months, colonies transplanted from deep to shallow became enriched by 0.8‰, mirroring the enrichment observed in the shallow controls, which was likely due to the combined effect of an increase in the source δ 15N and reduced fractionation. We conclude that light affects gorgonian δ 15N fractionation and should be considered in sampling designs for N pollution monitoring. However, these fractionations are small relative to differences observed between natural and anthropogenic N sources.

  4. Measurement of marine productivity using 15N and 13C tracers: Some methodological aspects

    Indian Academy of Sciences (India)

    Naveen Gandhi; Sanjeev Kumar; S Prakash; R Ramesh; M S Sheshshayee

    2011-02-01

    Various experiments involving the measurement of new, regenerated and total productivity using 15N and 13C tracers were carried out in the Bay of Bengal (BOB) and in the Arabian Sea. Results from 15N tracer experiments indicate that nitrate uptake can be underestimated by experiments with incubation time > 4 hours. Indirect evidence suggests pico- and nano-phytoplankton, on their dominance over microphytoplankton, can also influence the f-ratios. Difference in energy requirement for assimilation of different nitrogen compounds decides the preferred nitrogen source during the early hours of incubation. Variation in light intensity during incubation also plays a significant role in the assimilation of nitrogen. Results from time course experiments with both 15N and 13C tracers suggest that photoinhibition appears significant in BOB and the Arabian Sea during noon. A significant correlation has been found in the productivity values obtained using 15N and 13C tracers.

  5. The First in Vivo Observation of 13C- 15N Coupling in Mammalian Brain

    Science.gov (United States)

    Kanamori, Keiko; Ross, Brian D.

    2001-12-01

    [5-13C,15N]Glutamine, with 1J(13C-15N) of 16 Hz, was observed in vivo in the brain of spontaneously breathing rats by 13C MRS at 4.7 T. The brain [5-13C]glutamine peak consisted of the doublet from [5-13C,15N]glutamine and the center [5-13C,14N]glutamine peak, resulting in an apparent triplet with a separation of 8 Hz. The time course of formation of brain [5-13C,15N]glutamine was monitored in vivo with a time resolution of 20-35 min. This [5-13C,15N]glutamine was formed by glial uptake of released neurotransmitter [5-13C]glutamate and its reaction with 15NH3 catalyzed by the glia-specific glutamine synthetase. The neurotransmitter glutamate C5 was selectively13C-enriched by intravenous [2,5-13C]glucose infusion to 13C-label whole-brain glutamate C5, followed by [12C]glucose infusion to chase 13C from the small and rapidly turning-over glial glutamate pool, leaving 13C mainly in the neurotransmitter [5-13C]glutamate pool, which is sequestered in vesicles until release. Hence, the observed [5-13C,15N]glutamine arises from a coupling between 13C of neuronal origin and 15N of glial origin. Measurement of the rate of brain [5-13C,15N]glutamine formation provides a novel noninvasive method of studying the kinetics of neurotransmitter uptake into glia in vivo, a process that is crucial for protecting the brain from glutamate excitotoxicity.

  6. Variable δ(15)N diet-tissue discrimination factors among sharks: implications for trophic position, diet and food web models.

    Science.gov (United States)

    Olin, Jill A; Hussey, Nigel E; Grgicak-Mannion, Alice; Fritts, Mark W; Wintner, Sabine P; Fisk, Aaron T

    2013-01-01

    The application of stable isotopes to characterize the complexities of a species foraging behavior and trophic relationships is dependent on assumptions of δ(15)N diet-tissue discrimination factors (∆(15)N). As ∆(15)N values have been experimentally shown to vary amongst consumers, tissues and diet composition, resolving appropriate species-specific ∆(15)N values can be complex. Given the logistical and ethical challenges of controlled feeding experiments for determining ∆(15)N values for large and/or endangered species, our objective was to conduct an assessment of a range of reported ∆(15)N values that can hypothetically serve as surrogates for describing the predator-prey relationships of four shark species that feed on prey from different trophic levels (i.e., different mean δ(15)N dietary values). Overall, the most suitable species-specific ∆(15)N values decreased with increasing dietary-δ(15)N values based on stable isotope Bayesian ellipse overlap estimates of shark and the principal prey functional groups contributing to the diet determined from stomach content analyses. Thus, a single ∆(15)N value was not supported for this speciose group of marine predatory fishes. For example, the ∆(15)N value of 3.7‰ provided the highest percent overlap between prey and predator isotope ellipses for the bonnethead shark (mean diet δ(15)N = 9‰) whereas a ∆(15)N value shark (mean diet δ(15)N = 15‰). These data corroborate the previously reported inverse ∆(15)N-dietary δ(15)N relationship when both isotope ellipses of principal prey functional groups and the broader identified diet of each species were considered supporting the adoption of different ∆(15)N values that reflect the predators' δ(15)N-dietary value. These findings are critical for refining the application of stable isotope modeling approaches as inferences regarding a species' ecological role in their community will be influenced with consequences for conservation and

  7. The C$^{14}$N/C$^{15}$N Ratio in Diffuse Molecular Clouds

    CERN Document Server

    Ritchey, Adam M; Lambert, David L

    2015-01-01

    We report the first detection of C$^{15}$N in diffuse molecular gas from a detailed examination of CN absorption lines in archival VLT/UVES spectra of stars probing local diffuse clouds. Absorption from the C$^{15}$N isotopologue is confidently detected (at $\\gtrsim4\\sigma$) in three out of the four directions studied and appears as a very weak feature between the main $^{12}$CN and $^{13}$CN absorption components. Column densities for each CN isotopologue are determined through profile fitting, after accounting for weak additional line-of-sight components of $^{12}$CN, which are seen in the absorption profiles of CH and CH$^+$ as well. The weighted mean value of C$^{14}$N/C$^{15}$N for the three sight lines with detections of C$^{15}$N is $274\\pm18$. Since the diffuse molecular clouds toward our target stars have relatively high gas kinetic temperatures and relatively low visual extinctions, their C$^{14}$N/C$^{15}$N ratios should not be affected by chemical fractionation. The mean C$^{14}$N/C$^{15}$N ratio ...

  8. 1H and 15N NMR Analyses on Heparin, Heparan Sulfates and Related Monosaccharides Concerning the Chemical Exchange Regime of the N-Sulfo-Glucosamine Sulfamate Proton

    Directory of Open Access Journals (Sweden)

    Vitor H. Pomin

    2016-09-01

    Full Text Available Heparin and heparan sulfate are structurally related glycosaminoglycans (GAGs. Both GAGs present, although in different concentrations, N-sulfo-glucosamine (GlcNS as one of their various composing units. The conditional fast exchange property of the GlcNS sulfamate proton in these GAGs has been pointed as the main barrier to its signal detection via NMR experiments, especially 1H-15N HSQC. Here, a series of NMR spectra is collected on heparin, heparan sulfate and related monosaccharides. The N-acetyl glucosamine-linked uronic acid types of these GAGs were properly assigned in the 1H-15N HSQC spectra. Dynamic nuclear polarization (DNP was employed in order to facilitate 1D spectral acquisition of the sulfamate 15N signal of free GlcNS. Analyses on the multiplet pattern of scalar couplings of GlcNS 15N has helped to understand the chemical properties of the sulfamate proton in solution. The singlet peak observed for GlcNS happens due to fast chemical exchange of the GlcNS sulfamate proton in solution. Analyses on kinetics of alpha-beta anomeric mutarotation via 1H NMR spectra have been performed in GlcNS as well as other glucose-based monosaccharides. 1D 1H and 2D 1H-15N HSQC spectra recorded at low temperature for free GlcNS dissolved in a proton-rich solution showed signals from all exchangeable protons, including those belonging to the sulfamate group. This work suits well to the current grand celebration of one-century-anniversary of the discovery of heparin.

  9. ¹H and (15)N NMR Analyses on Heparin, Heparan Sulfates and Related Monosaccharides Concerning the Chemical Exchange Regime of the N-Sulfo-Glucosamine Sulfamate Proton.

    Science.gov (United States)

    Pomin, Vitor H

    2016-09-07

    Heparin and heparan sulfate are structurally related glycosaminoglycans (GAGs). Both GAGs present, although in different concentrations, N-sulfo-glucosamine (GlcNS) as one of their various composing units. The conditional fast exchange property of the GlcNS sulfamate proton in these GAGs has been pointed as the main barrier to its signal detection via NMR experiments, especially ¹H-(15)N HSQC. Here, a series of NMR spectra is collected on heparin, heparan sulfate and related monosaccharides. The N-acetyl glucosamine-linked uronic acid types of these GAGs were properly assigned in the ¹H-(15)N HSQC spectra. Dynamic nuclear polarization (DNP) was employed in order to facilitate 1D spectral acquisition of the sulfamate (15)N signal of free GlcNS. Analyses on the multiplet pattern of scalar couplings of GlcNS (15)N has helped to understand the chemical properties of the sulfamate proton in solution. The singlet peak observed for GlcNS happens due to fast chemical exchange of the GlcNS sulfamate proton in solution. Analyses on kinetics of alpha-beta anomeric mutarotation via ¹H NMR spectra have been performed in GlcNS as well as other glucose-based monosaccharides. 1D ¹H and 2D ¹H-(15)N HSQC spectra recorded at low temperature for free GlcNS dissolved in a proton-rich solution showed signals from all exchangeable protons, including those belonging to the sulfamate group. This work suits well to the current grand celebration of one-century-anniversary of the discovery of heparin.

  10. Historical WBAN ID Assignments

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — 4"x6" index cards represent the first written assignments of Weather Bureau Army Navy (WBAN) station identifier numbers by the National Climatic Data Center....

  11. My Favorite Assignment.

    Science.gov (United States)

    Post, Robert E.; Johnson, Jack E.

    1982-01-01

    Presents two assignments that show (1) how George Orwell's "Politics and the English Language" can be applied to business writing and (2) how structured student-teacher conferences can generate enthusiasm for oral expression in a business communication course. (AEA)

  12. True cooking aroma or artefact. {sup 15}N gives the answer; Veritable arome de cuisson ou artefact. {sup 15}N fournit la reponse

    Energy Technology Data Exchange (ETDEWEB)

    Metro, F.; Boudaud, N.; Dumont, J.P. [INRA, 44 - Nantes (France)

    1994-12-31

    In order to determine the respective contributions of the various nitrous precursor families in aroma preparations, the usually added amino acids were substituted with {sup 15}N isotope labelled homologous components. Results concerning isotope ratios for the volatile fraction nitrous components collected from poultry meat aromatic preparations, are presented. Terminal product labelling appears to allow for a better determination of the substrate and functional additive contributions. 4 figs., 6 refs.

  13. Absorption of ammonium sulphate {sup 15}N by coffee plants; Recuperacao do {sup 15}N do sulfato de amonio por plantas de cafe

    Energy Technology Data Exchange (ETDEWEB)

    Fenilli, Tatiele Anete Bergamo; Reichardt, Klaus; Bacchi, Osny Oliveira Santos [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Fisica do Solo]. E-mail: tatiele@cena.usp.br; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis; Dourado Neto, Durval [Sao Paulo Univ., Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz. Dept. de Producao Vegetal

    2005-07-01

    The objective of this study was to quantify the absorption of ammonium sulphate {sup 15}N by coffee plants. Treatments consisted of five sub-plots of 9 plants, of which the three central ones received 280 kg ha{sup -1} of {sup 15}N, applied at four times: 1/4 on 01 Set 03; 1/4 on 03 Nov 03; 1/4 on 15 Dec 03 and 1/4 on 30 Jan 04. The isotopic enrichment was 2,072 {+-} 0,001 atom % {sup 15}N. The dry matter of the shoot was evaluated every 60 days, using one plant per replicate, collected outside the sub-plot. They were as similar as possible to the labeled plants, which were used only for isotopic and Total N analysis, after being dried at 65 deg C until constant weight. At harvest, plants had absorbed 42,88% of the fertilizer N. Leaves accumulated the largest amount of fertilizer N, and were also the compartments that received most N from other parts of the plant. The following partition of the fertilizer N was found at harvest: 23.01% in young leaves, 6.23% in old leaves, 4,46% in stem, 3.46% in fruits, 3.10% in young branches and 2.63% in old branches. (author)

  14. Compound-specific δ15N amino acid measurements in littoral mussels in the California upwelling ecosystem: a new approach to generating baseline δ15N Isoscapes for coastal ecosystems.

    Science.gov (United States)

    Vokhshoori, Natasha L; McCarthy, Matthew D

    2014-01-01

    We explored δ(15)N compound-specific amino acid isotope data (CSI-AA) in filter-feeding intertidal mussels (Mytilus californianus) as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ(15)N gradients in the California Upwelling Ecosystem (CUE), determining bulk δ(15)N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ(15)N values showed a strong linear trend with latitude, increasing from North to South (from ∼ 7‰ to ∼ 12‰, R(2) = 0.759). In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ(15)N trend is therefore most consistent with a baseline δ(15)N gradient, likely due to the mixing of two source waters: low δ(15)N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC), with (15)N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ(15)N values of phenylalanine (δ(15)NPhe), the best AA proxy for baseline δ(15)N values. We hypothesize δ(15)N(Phe) values in intertidal mussels can approximate annual integrated δ(15)N values of coastal phytoplankton primary production. We therefore used δ(15)N(Phe) values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ(15)N values. We propose that δ(15)N(Phe) isoscapes derived from filter feeders can directly characterize baseline δ(15)N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives.

  15. Compound-specific δ15N amino acid measurements in littoral mussels in the California upwelling ecosystem: a new approach to generating baseline δ15N Isoscapes for coastal ecosystems.

    Directory of Open Access Journals (Sweden)

    Natasha L Vokhshoori

    Full Text Available We explored δ(15N compound-specific amino acid isotope data (CSI-AA in filter-feeding intertidal mussels (Mytilus californianus as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ(15N gradients in the California Upwelling Ecosystem (CUE, determining bulk δ(15N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ(15N values showed a strong linear trend with latitude, increasing from North to South (from ∼ 7‰ to ∼ 12‰, R(2 = 0.759. In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ(15N trend is therefore most consistent with a baseline δ(15N gradient, likely due to the mixing of two source waters: low δ(15N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC, with (15N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ(15N values of phenylalanine (δ(15NPhe, the best AA proxy for baseline δ(15N values. We hypothesize δ(15N(Phe values in intertidal mussels can approximate annual integrated δ(15N values of coastal phytoplankton primary production. We therefore used δ(15N(Phe values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ(15N values. We propose that δ(15N(Phe isoscapes derived from filter feeders can directly characterize baseline δ(15N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives.

  16. 42 CFR 433.146 - Rights assigned; assignment method.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false Rights assigned; assignment method. 433.146 Section 433.146 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN... Assignment of Rights to Benefits § 433.146 Rights assigned; assignment method. (a) Except as specified...

  17. Measurement of {sup 1}H-{sup 15}N and {sup 1}H-{sup 13}C residual dipolar couplings in nucleic acids from TROSY intensities

    Energy Technology Data Exchange (ETDEWEB)

    Ying Jinfa [National Institutes of Health, Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases (United States); Wang Jinbu [National Cancer Institute, National Institutes of Health, Structural Biophysics Laboratory (United States); Grishaev, Alex [National Institutes of Health, Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases (United States); Yu Ping; Wang Yunxing [National Cancer Institute, National Institutes of Health, Structural Biophysics Laboratory (United States); Bax, Ad, E-mail: bax@nih.gov [National Institutes of Health, Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases (United States)

    2011-09-15

    Analogous to the recently introduced ARTSY method for measurement of one-bond {sup 1}H-{sup 15}N residual dipolar couplings (RDCs) in large perdeuterated proteins, we introduce methods for measurement of base {sup 13}C-{sup 1}H and {sup 15}N-{sup 1}H RDCs in protonated nucleic acids. Measurements are based on quantitative analysis of intensities in {sup 1}H-{sup 15}N and {sup 13}C-{sup 1}H TROSY-HSQC spectra, and are illustrated for a 71-nucleotide adenine riboswitch. Results compare favorably with those of conventional frequency-based measurements in terms of completeness and convenience of use. The ARTSY method derives the size of the coupling from the ratio of intensities observed in two TROSY-HSQC spectra recorded with different dephasing delays, thereby minimizing potential resonance overlap problems. Precision of the RDC measurements is limited by the signal-to-noise ratio, S/N, achievable in the 2D TROSY-HSQC reference spectrum, and is approximately given by 30/(S/N) Hz for {sup 15}N-{sup 1}H and 65/(S/N) Hz for {sup 13}C-{sup 1}H. The signal-to-noise ratio of both {sup 1}H-{sup 15}N and {sup 1}H-{sup 13}C spectra greatly benefits when water magnetization during the experiments is not perturbed, such that rapid magnetization transfer from bulk water to the nucleic acid, mediated by rapid amino and hydroxyl hydrogen exchange coupled with {sup 1}H-{sup 1}H NOE transfer, allows for fast repetition of the experiment. RDCs in the mutated helix 1 of the riboswitch are compatible with nucleotide-specifically modeled, idealized A-form geometry and a static orientation relative to the helix 2/3 pair, which differs by ca 6 Degree-Sign relative to the X-ray structure of the native riboswitch.

  18. Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1.

    Science.gov (United States)

    Barb, Adam W

    2015-01-20

    Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. However, widely applicable methods do not yet exist. To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. Thus, conformational restriction increases as Fc N-glycan maturation proceeds. Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of

  19. Intramolecular N-Glycan/Polypeptide Interactions Observed at Multiple N-Glycan Remodeling Steps through [13C,15N]-N-Acetylglucosamine Labeling of Immunoglobulin G1

    Science.gov (United States)

    2014-01-01

    Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. Attempts to characterize the structure–activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. However, widely applicable methods do not yet exist. To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [13C,15N]-N-acetylglucosamine (GlcNAc). UDP-[13C,15N]GlcNAc was synthesized enzymatically in a one-pot reaction from [13C]glucose and [15N-amido]glutamine. Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[13C,15N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [13C,15N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. Thus, conformational restriction increases as Fc N-glycan maturation proceeds. Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein

  20. Heteronuclear 2D-correlations in a uniformly [13C, 15N] labeled membrane-protein complex at ultra-high magnetic fields

    Energy Technology Data Exchange (ETDEWEB)

    Egorova-Zachernyuk, T.A.; Hollander, J. [Gorlaeus Laboratories (Netherlands); Fraser, N. [University of Glasgow, Division of Biochemistry and Molecular Biology (United Kingdom); Gast, P.; Hoff, A.J. [Leiden University, Huygens Laboratories (Netherlands); Cogdell, R. [University of Glasgow, Division of Biochemistry and Molecular Biology (United Kingdom); Groot, H.J.M. de; Baldus, M. [Gorlaeus Laboratories (Netherlands)

    2001-03-15

    One- and two-dimensional solid-state NMR experiments on a uniformly labeled intrinsic membrane-protein complex at ultra-high magnetic fields are presented. Two-dimensional backbone and side-chain correlations for a [U-{sup 13}C,{sup 15}N] labeled version of the LH2 light-harvesting complex indicate significant resolution at low temperatures and under Magic Angle Spinning. Tentative assignments of some of the observed correlations are presented and attributed to the {alpha}-helical segments of the protein, mostly found in the membrane interior.

  1. A new method to track seed dispersal and recruitment using 15N isotope enrichment.

    Science.gov (United States)

    Carlo, Tomás A; Tewksbury, Joshua J; Martínez Del Río, Carlos

    2009-12-01

    Seed dispersal has a powerful influence on population dynamics, genetic structuring, evolutionary rates, and community ecology. Yet, patterns of seed dispersal are difficult to measure due to methodological shortcomings in tracking dispersed seeds from sources of interest. Here we introduce a new method to track seed dispersal: stable isotope enrichment. It consists of leaf-feeding plants with sprays of 15N-urea during the flowering stage such that seeds developed after applications are isotopically enriched. We conducted a greenhouse experiment with Solanum americanum and two field experiments with wild Capsicum annuum in southern Arizona, USA, to field-validate the method. First, we show that plants sprayed with 15N-urea reliably produce isotopically enriched progeny, and that delta 15N (i.e., the isotopic ratio) of seeds and seedlings is a linear function of the 15N-urea concentration sprayed on mothers. We demonstrate that three urea dosages can be used to distinctly enrich plants and unambiguously differentiate their offspring after seeds are dispersed by birds. We found that, with high urea dosages, the resulting delta 15N values in seedlings are 10(3) - 10(4) times higher than the delta 15N values of normal plants. This feature allows tracking not only where seeds arrive, but in locations where seeds germinate and recruit, because delta 15N enrichment is detectable in seedlings that have increased in mass by at least two orders of magnitude before fading to normal delta 15N values. Last, we tested a mixing model to analyze seed samples in bulk. We used the delta 15N values of batches (i.e., combined seedlings or seeds captured in seed traps) to estimate the number of enriched seeds coming from isotopically enriched plants in the field. We confirm that isotope enrichment, combined with batch-sampling, is a cheap, reliable, and user-friendly method for bulk-processing seeds and is thus excellent for the detection of rare dispersal events. This method could

  2. Rivermouth alteration of agricultural impacts on consumer tissue δ(15N.

    Directory of Open Access Journals (Sweden)

    James H Larson

    Full Text Available Terrestrial agricultural activities strongly influence riverine nitrogen (N dynamics, which is reflected in the δ(15N of riverine consumer tissues. However, processes within aquatic ecosystems also influence consumer tissue δ(15N. As aquatic processes become more important terrestrial inputs may become a weaker predictor of consumer tissue δ(15N. In a previous study, this terrestrial-consumer tissue δ(15N connection was very strong at river sites, but was disrupted by processes occurring in rivermouths (the 'rivermouth effect'. This suggested that watershed indicators of N loading might be accurate in riverine settings, but could be inaccurate when considering N loading to the nearshore of large lakes and oceans. In this study, the rivermouth effect was examined on twenty-five sites spread across the Laurentian Great Lakes. Relationships between agriculture and consumer tissue δ(15N occurred in both upstream rivers and at the outlets where rivermouths connect to the nearshore zone, but agriculture explained less variation and had a weaker effect at the outlet. These results suggest that rivermouths may sometimes be significant sources or sinks of N, which would cause N loading estimates to the nearshore zone that are typically made at discharge gages further upstream to be inaccurate. Identifying definitively the controls over the rivermouth effect on N loading (and other nutrients will require integration of biogeochemical and hydrologic models.

  3. The {sup 15}N isotope to evaluate fertilizer nitrogen absorption efficiency by the coffee plant

    Energy Technology Data Exchange (ETDEWEB)

    Fenilli, Tatiele A.B. [Universidade Regional de Blumenau, (FURB), SC (Brazil); Reichart, Klaus; Bacchi, Osny O.S.; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil)]. E-mail: klaus@cena.usp.br; Dourado-Neto, Durval [Universidade de Sao Paulo (USP), Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz (ESALQ)

    2007-12-15

    The use of the {sup 15}N label for agronomic research involving nitrogen (N) cycling and the fate of fertilizer-N is well established, however, in the case of long term experimentation with perennial crops like citrus, coffee and rubber tree, there are still shortcomings mainly due to large plant size, sampling procedures, detection levels and interferences on the system. This report tries to contribute methodologically to the design and development of {sup 15}N labeled fertilizer experiments, using as an example a coffee crop fertilized with {sup 15}N labeled ammonium sulfate, which was followed for two years. The N of the plant derived from the fertilizer was studied in the different parts of the coffee plant in order to evaluate its distribution within the plant and the agronomic efficiency of the fertilizer application practice. An enrichment of the fertilizer-N of the order of 2% {sup 15}N abundance was sufficient to study N absorption rates and to establish fertilizer-N balances after one and two years of coffee cropping. The main source of errors in the estimated values lies in the inherent variability among field replicates and not in the measurements of N contents and {sup 15}N enrichments of plant material by mass-spectrometry. (author)

  4. The 15N isotope to evaluate fertilizer nitrogen absorption efficiency by the coffee plant.

    Science.gov (United States)

    Fenilli, Tatiele A B; Reichart, Klaus; Bacchi, Osny O S; Trivelin, Paulo C O; Dourado-Neto, Durval

    2007-12-01

    The use of the 15N label for agronomic research involving nitrogen (N) cycling and the fate of fertilizer-N is well established, however, in the case of long term experimentation with perennial crops like citrus, coffee and rubber tree, there are still shortcomings mainly due to large plant size, sampling procedures, detection levels and interferences on the system. This report tries to contribute methodologically to the design and development of 15N labeled fertilizer experiments, using as an example a coffee crop fertilized with 15N labeled ammonium sulfate, which was followed for two years. The N of the plant derived from the fertilizer was studied in the different parts of the coffee plant in order to evaluate its distribution within the plant and the agronomic efficiency of the fertilizer application practice. An enrichment of the fertilizer-N of the order of 2% 15N abundance was sufficient to study N absorption rates and to establish fertilizer-N balances after one and two years of coffee cropping. The main source of errors in the estimated values lies in the inherent variability among field replicates and not in the measurements of N contents and 15N enrichments of plant material by mass-spectrometry.

  5. Using a macroalgal δ15N bioassay to detect cruise ship waste water effluent inputs.

    Science.gov (United States)

    Kaldy, James

    2011-08-01

    Green macroalgae bioassays were used to determine if the δ15N signature of cruise ship waste water effluent (CSWWE) could be detected in a small harbor. Opportunistic green macroalgae (Ulva spp.) were collected, cultured under nutrient depleted conditions and characterized with regard to N content and δ15N. Samples of algae were used in controlled incubations to evaluate the direction of isotope shift from exposure to CSWWE. Algae samples exposed to CSWWE exhibited an increase of 1-2.5‰ in δ15N values indicating that the CSWWE had an enriched isotope signature. In contrast, algae samples exposed to field conditions exhibited a significant decrease in the observed δ15N indicating that a light N source was used. Isotopically light, riverine nitrogen derived from N2-fixing trees in the watershed may be a N source utilized by algae. These experiments indicate that the δ15N CSWWE signature was not detectable under the CSWWE loading conditions of this experiment.

  6. Total N difference method and 15N isotope dilution methode - A comparative study on N-fixation

    OpenAIRE

    2002-01-01

    In the study, the 15N fixation of a number of green manure crops were studied using either the 15N dilution technique, or the simple total N difference method. The results of the two methods were not very different, and the total N difference method seemed to give as good results as the more complicated and expencive 15N dillution method.

  7. Random Cell Identifiers Assignment

    Directory of Open Access Journals (Sweden)

    Robert Bestak

    2012-01-01

    Full Text Available Despite integration of advanced functions that enable Femto Access Points (FAPs to be deployed in a plug-and-play manner, the femtocell concept still cause several opened issues to be resolved. One of them represents an assignment of Physical Cell Identifiers (PCIs to FAPs. This paper analyses a random based assignment algorithm in LTE systems operating in diverse femtocell scenarios. The performance of the algorithm is evaluated by comparing the number of confusions for various femtocell densities, PCI ranges and knowledge of vicinity. Simulation results show that better knowledge of vicinity can significantly reduce the number of confusions events.

  8. NR 512 All Assignments

    OpenAIRE

    Laynebaril1

    2017-01-01

     NR 512 All Assignments Click Link Below To Buy: http://hwcampus.com/shop/nr-512-assignments/ Or Visit www.hwcampus.com   NR512 Week 4 Scavenger Hunt Latest 2017 W4_NR512 Second Life (SL) Scavenger Hunt Work Sheet Student Name: Dorinda Ezell Avatar Name: Dvezellnr512 INSTRUCTIONS Instructions: 1. As you complete the Scavenger Hunt, please write your answers to the questions on this worksheet. 2. Save it as a MS Word document in the following format: ...

  9. Synthesis and NMR of {sup 15}N-labeled DNA fragments

    Energy Technology Data Exchange (ETDEWEB)

    Jones, R.A. [Rutgers, The State Univ. of New Jersey, Piscataway, NJ (United States)

    1994-12-01

    DNA fragments labeled with {sup 15}N at the ring nitrogens and at the exocyclic amino groups can be used to obtain novel insight into interactions such as base pairing, hydration, drug binding, and protein binding. A number of synthetic routes to {sup 15}N-labeled pyrimidine nucleosides, purines, and purine nucleosides have been reported. Moreover, many of these labeled bases or monomers have been incorporated into nucleic acids, either by chemical synthesis or by biosynthetic procedures. The focus of this chapter will be on the preparation of {sup 15}N-labeled purine 2{prime}-deoxynucleosides, their incorporation into DNA fragments by chemical synthesis, and the results of NMR studies using these labeled DNA fragments.

  10. Accurate proteome-wide protein quantification from high-resolution 15N mass spectra.

    Science.gov (United States)

    Khan, Zia; Amini, Sasan; Bloom, Joshua S; Ruse, Cristian; Caudy, Amy A; Kruglyak, Leonid; Singh, Mona; Perlman, David H; Tavazoie, Saeed

    2011-12-19

    In quantitative mass spectrometry-based proteomics, the metabolic incorporation of a single source of 15N-labeled nitrogen has many advantages over using stable isotope-labeled amino acids. However, the lack of a robust computational framework for analyzing the resulting spectra has impeded wide use of this approach. We have addressed this challenge by introducing a new computational methodology for analyzing 15N spectra in which quantification is integrated with identification. Application of this method to an Escherichia coli growth transition reveals significant improvement in quantification accuracy over previous methods.

  11. Constraints on oceanic N balance/imbalance from sedimentary 15N records

    Directory of Open Access Journals (Sweden)

    M. A. Altabet

    2007-01-01

    Full Text Available According to current best estimates, the modern ocean's N cycle is in severe deficit. N isotope budgeting provides an independent geochemical constraint in this regard as well as the only means for past reconstruction. Overall, it is the relative proportion of N2 fixation consumed by water column denitrification that sets average oceanic δ15N under steady-state conditions. Several factors (conversion of organic N to N2, Rayleigh closed and open system effects likely reduce the effective fractionation factor (ε for water column denitrification to about half the inherent microbial value for εden. If so, the average oceanic δ15N of ~5‰ is consistent with a canonical contribution from water column denitrification of 50% of the source flux from N2 fixation. If an imbalance in oceanic N sources and sinks changes this proportion then a transient in average oceanic δ15N would occur. Using a simple model, changing water column denitrification by ±30% or N2 fixation by ±15% produces detectable (>1‰ changes in average oceanic δ15N over one residence time period or more with corresponding changes in oceanic N inventory. Changing sedimentary denitrification produces no change in δ15N but does change N inventory. Sediment δ15N records from sites thought to be sensitive to oceanic average δ15N all show no detectible change over the last 3 kyr or so implying a balanced marine N budget over the latest Holocene. A mismatch in time scales is the most likely meaningful interpretation of the apparent conflict with modern flux estimates. Decadal to centennial scale oscillations between net N deficit and net surplus may occur but on the N residence timescale of several thousand years, net balance is achieved in sum. However, sediment δ15N records from the literature covering the period since the last glacial maximum show excursions of up to several ‰ that are consistent with sustained N deficit during the deglaciation followed by readjustment

  12. 15N Fractionation in Star-Forming Regions and Solar System Objects

    Science.gov (United States)

    Wirstrom, Eva; Milam, Stefanie; Adande, GIlles; Charnley, Steven; Cordiner, Martin

    2015-01-01

    A central issue for understanding the formation and evolution of matter in the early Solar System is the relationship between the chemical composition of star-forming interstellar clouds and that of primitive Solar System materials. The pristinemolecular content of comets, interplanetary dust particles and carbonaceous chondrites show significant bulk nitrogen isotopic fractionation relative to the solar value, 14N15N 440. In addition, high spatial resolution measurements in primitive materials locally show even more extreme enhancements of 14N15N 100.

  13. Task assignment and coaching

    NARCIS (Netherlands)

    Dominguez-Martinez, S.

    2009-01-01

    An important task of a manager is to motivate her subordinates. One way in which a manager can give incentives to junior employees is through the assignment of tasks. How a manager allocates tasks in an organization, provides information to the junior employees about his ability. Without coaching fr

  14. Binding of oxytocin and 8-arginine-vasopressin to neurophysin studied by /sup 15/N NMR using magnetization transfer and indirect detection via protons

    Energy Technology Data Exchange (ETDEWEB)

    Live, D.H.; Cowburn, D.

    1987-10-06

    NMR was used to monitor the binding to neurophysin of oxytocin and 8-arginine-vasopressin, /sup 15/N labeling being used to identify specific backbone /sup 15/N and /sup 1/H signals. The most significant effects of binding were large downfield shifts in the amino nitrogen resonance of Phe-3 of vasopressin and in its associated proton, providing evidence that the peptide bond between residues 2 and 3 of the hormones is hydrogen-bonded to the protein within hormone-neurophysin complexes. Suggestive evidence for hydrogen bonding of the amino nitrogen of Tyr-2 was also obtained in the form of decreased proton exchange rates on binding; however, the chemical shift changes of this nitrogen and its associated proton indicated that such hydrogen bonding, if present, is probably weak. Shifts in the amino nitrogen of Asn-5 and in the -NH protons of both Asn-5 and Cys-6 demonstrated that these residues are significantly perturbed by binding, suggesting conformational changes of the ring on binding and/or the presence of binding sites on the hormone outside the 1-3 region. No support was obtained for the thesis that there is a significant second binding site for vasopressin on each neutrophysin chain. The behavior of both oxytocin and vasopressin on binding was consistent with formation of 1:1 complexes in slow exchange with the free state under most pH conditions. At low pH there was evidence of an increased exchange rate. Additionally, broadening of /sup 15/N resonances in the bound state at low pH occurred without a corresponding change in the resonances of equilibrating free hormone. The results suggest significant conformational alteration in neurophysin-hormone complexes at low pH possibly associated with protonation of the carboxyl group of the hormone-protein salt bridge.

  15. The degree of urbanization across the globe is not reflected in the δ(15)N of seagrass leaves.

    Science.gov (United States)

    Christiaen, Bart; Bernard, Rebecca J; Mortazavi, Behzad; Cebrian, Just; Ortmann, Alice C

    2014-06-30

    Many studies show that seagrass δ(15)N ratios increase with the amount of urbanization in coastal watersheds. However, there is little information on the relationship between urbanization and seagrass δ(15)N ratios on a global scale. We performed a meta-analysis on seagrass samples from 79 independent locations to test if seagrass δ(15)N ratios correlate with patterns of population density and fertilizer use within a radius of 10-200 km around the sample locations. Our results show that seagrass δ(15)N ratios are more influenced by intergeneric and latitudinal differences than the degree of urbanization or the amount of fertilizer used in nearby watersheds. The positive correlation between seagrass δ(15)N ratios and latitude hints at an underlying pattern in discrimination or a latitudinal gradient in the (15)N isotopic signature of nitrogen assimilated by the plants. The actual mechanisms responsible for the correlation between δ(15)N and latitude remain unknown.

  16. Carbon-rich presolar grains from massive stars. Subsolar 12C/13C and 14N/15N ratios and the mystery of 15N

    CERN Document Server

    Pignatari, M; Hoppe, P; Jordan, C J; Gibson, B K; Trappitsch, R; Herwig, F; Fryer, C; Hirschi, R; Timmes, F X

    2015-01-01

    Carbon-rich grains with isotopic anomalies compared to the Sun are found in primitive meteorites. They were made by stars, and carry the original stellar nucleosynthesis signature. Silicon carbide grains of Type X and C, and low-density graphites condensed in the ejecta of core-collapse supernovae. We present a new set of models for the explosive He shell and compare them with the grains showing 12C/13C and 14N/15N ratios lower than solar. In the stellar progenitor H was ingested into the He shell and not fully destroyed before the explosion. Different explosion energies and H concentrations are considered. If the SN shock hits the He-shell region with some H still present, the models can reproduce the C and N isotopic signatures in C-rich grains. Hot-CNO cycle isotopic signatures are obtained, including a large production of 13C and 15N. The short-lived radionuclides 22Na and 26Al are increased by orders of magnitude. The production of radiogenic 22Ne from the decay of 22Na in the He shell might solve the pu...

  17. Rotational spectrum of deuterated and 15N ethyl cyanides: CH3CHDCN and CH2DCH2CN and of CH3CH2C15N

    CERN Document Server

    Margulès, Laurent; Demyk, Karine; Tercero, Belen; Cernicharo, Jose; Sheng, M; Weidmann, M; Gripp, J; Mäder, H; Demaison, J

    2008-01-01

    Ethyl cyanide is an abundant molecule in hot molecular clouds. Lines from 13C isotopically substituted ethyl cyanide were identified in Orion. To enable the search and the possible detection of other isotopologues of ethyl cyanide in interstellar objects, we have studied the rotational spectrum of deuterated ethyl cyanide: CH2DCH2CN (in-plane and out-of-plane) and CH3CHDCN and the spectrum of15N substituted ethyl cyanide CH3CH2C15N. The rotational spectrum of each species in the ground state was measured in the microwave and millimeter-submillimeter wavelength range using a waveguide Fourier transform spectrometer (8 - 17 GHz) and a source-modulated spectrometer employing backward-wave oscillators (BWOs) (150 - 260 and 580 - 660 GHz). From the fitting procedure, accurate spectroscopic constants were derived for each of the species. These new sets of spectroscopic constants enable us to predict reliably the rotational spectrum (lines frequencies and intensities) in the 4-1000 GHz frequency range and for J and ...

  18. Monitoring the refinement of crystal structures with (15)N solid-state NMR shift tensor data.

    Science.gov (United States)

    Kalakewich, Keyton; Iuliucci, Robbie; Mueller, Karl T; Eloranta, Harriet; Harper, James K

    2015-11-21

    The (15)N chemical shift tensor is shown to be extremely sensitive to lattice structure and a powerful metric for monitoring density functional theory refinements of crystal structures. These refinements include lattice effects and are applied here to five crystal structures. All structures improve based on a better agreement between experimental and calculated (15)N tensors, with an average improvement of 47.0 ppm. Structural improvement is further indicated by a decrease in forces on the atoms by 2-3 orders of magnitude and a greater similarity in atom positions to neutron diffraction structures. These refinements change bond lengths by more than the diffraction errors including adjustments to X-Y and X-H bonds (X, Y = C, N, and O) of 0.028 ± 0.002 Å and 0.144 ± 0.036 Å, respectively. The acquisition of (15)N tensors at natural abundance is challenging and this limitation is overcome by improved (1)H decoupling in the FIREMAT method. This decoupling dramatically narrows linewidths, improves signal-to-noise by up to 317%, and significantly improves the accuracy of measured tensors. A total of 39 tensors are measured with shifts distributed over a range of more than 400 ppm. Overall, experimental (15)N tensors are at least 5 times more sensitive to crystal structure than (13)C tensors due to nitrogen's greater polarizability and larger range of chemical shifts.

  19. An improved method for delta 15N measurements in ice cores

    Directory of Open Access Journals (Sweden)

    M. Leuenberger

    2008-02-01

    Full Text Available The use of isotopic ratios of nitrogen gas (δ15N trapped in ice cores as a paleothermometer to characterise abrupt climate changes is becoming a widespread technique. The versatility of the technique could be enhanced, for instance in quantifying small temperature changes during the last glacial period in Antarctic ice cores, by using high precision methods. In this paper, we outline a method for measuring δ15N to a precision of 0.006permil (1σ, n=9 from replicate ice core samples. The high precision results from removing oxygen, carbon dioxide and water vapour from the air extracted from ice cores. The advantage of the technique is that it does not involve correction for isobaric interference due to CO+ ions. We also highlight the importance of oxygen removal from the sample, and how it influences δ15N measurements. The results show that a small amount of oxygen in the sample can be detrimental to achieving an optimum precision in δ15N measurements of atmospheric nitrogen trapped ice core samples.

  20. Nitrate Removal in Two Relict Oxbow Urban Wetlands: A 15N Mass-balance Approach

    Science.gov (United States)

    A 15N-tracer method was used to quantify nitrogen (N) removal processes in two relict oxbow wetlands located adjacent to the Minebank Run restored stream reach in Baltimore County (Maryland, USA) during summer 2009 and early spring 2010. A mass-balance approach was used to determ...

  1. Scaffolding students’ assignments

    DEFF Research Database (Denmark)

    Slot, Marie Falkesgaard

    2013-01-01

    This article discusses scaffolding in typical student assignments in mother tongue learning materials in upper secondary education in Denmark and the United Kingdom. It has been determined that assignments do not have sufficient scaffolding end features to help pupils understand concepts and build...... objects. The article presents the results of empirical research on tasks given in Danish and British learning materials. This work is based on a further development of my PhD thesis: “Learning materials in the subject of Danish” (Slot 2010). The main focus is how cognitive models (and subsidiary explicit...... learning goals) can help students structure their argumentative and communica-tive learning processes, and how various multimodal representations can give more open-ended learning possibilities for collaboration. The article presents a short introduction of the skills for 21st century learning and defines...

  2. Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.

    Science.gov (United States)

    Meola, Annalisa; Deville, Célia; Jeffers, Scott A; Guardado-Calvo, Pablo; Vasiliauskaite, Ieva; Sizun, Christina; Girard-Blanc, Christine; Malosse, Christian; van Heijenoort, Carine; Chamot-Rooke, Julia; Krey, Thomas; Guittet, Eric; Pêtres, Stéphane; Rey, Félix A; Bontems, François

    2014-10-01

    Nuclear magnetic resonance spectroscopy is a powerful tool to study structural and functional properties of proteins, provided that they can be enriched in stable isotopes such as (15)N, (13)C and (2)H. This is usually easy and inexpensive when the proteins are expressed in Escherichiacoli, but many eukaryotic (human in particular) proteins cannot be produced this way. An alternative is to express them in insect cells. Labeled insect cell growth media are commercially available but at prohibitive prices, limiting the NMR studies to only a subset of biologically important proteins. Non-commercial solutions from academic institutions have been proposed, but none of them is really satisfying. We have developed a (15)N-labeling procedure based on the use of a commercial medium depleted of all amino acids and supplemented with a (15)N-labeled yeast autolysate for a total cost about five times lower than that of the currently available solutions. We have applied our procedure to the production of a non-polymerizable mutant of actin in Sf9 cells and of fragments of eukaryotic and viral membrane fusion proteins in S2 cells, which typically cannot be produced in E. coli, with production yields comparable to those obtained with standard commercial media. Our results support, in particular, the putative limits of a self-folding domain within a viral glycoprotein of unknown structure.

  3. [Characteristics of urea 15N absorption, allocation, and utilization by sweet-cherry (Prunus avium L.)].

    Science.gov (United States)

    Zhao, Feng-Xia; Jiang, Yuan-Mao; Peng, Fu-Tian; Gao, Xiang-Bin; Liu, Bing-Hua; Wang, Hai-Yun; Zhao, Lin

    2008-03-01

    With five-year old 'Zaodaguo' sweet-cherry (Prunus avium L.) as test material, this paper studied the characteristics of its urea 15N absorption, allocation, and utilization when applied before bud-break. The results showed that the Ndff of different organs increased gradually with time, and was higher in fine roots and storage organs at full-blooming stage. At fruit core-hardening stage, the Ndff of long shoots and leaves increased quickly, reaching to 0.72% and 0.59% , respectively. From fruit core-hardening to harvesting stage, the Ndff of fruit had a rapid increase, with the peak (1.78%) at harvesting stage. After harvest, the Ndff of neonatal organs increased slowly while that of storage organs increased quickly. At full-blooming stage, the absorbed 15N in roots was firstly allocated to storage organs, with the highest allocation rate (54.91%) in large roots. At fruit core-hardening stage, the allocation rate in fine roots and storage organs decreased from 85.43% to 55.11%, while that in neonatal branches and leaves increased to 44.89%. At harvesting stage, the allocation rate in different organs had no significant change, but after harvest, the absorbed 15N had a rapid translocation to storage organs, and the allocation rate in fine roots and storage organs reached the highest (72.26%) at flower bud differentiation stage. The 15N allocation rate in neonatal branches and leaves at flower bud differentiation stage was decreased by 19.31%, compared with that at harvesting stage. From full-blooming to flower bud differentiation stage, the utilization rate of urea 15N was increasing, and reached the peak (16.86%) at flower bud differentiation stage.

  4. Araucaria cunninghamii Seedling Response to Different Forms and Rates of 15N-Labelled Fertiliser

    Institute of Scientific and Technical Information of China (English)

    T.J.BLUMFIELD; XU Zhi-Hong

    2006-01-01

    Nitrogenous fertilisers are under consideration for promoting the growth of nursery-reared hoop pine (Araucaria cunninghamii Aiton ex A. Cunn) seedlings in the establishment phase of second rotation (2R) plantations. Using 15Nlabelled fertilisers, we investigated the effect of different forms (ammonium sulphate, ammonium nitrate, potassium nitrate and urea) and rates of application (0, 150 and 300 mg N kg-1 dried soil) of fertilisers on the growth, 15N recovery and carbon isotope composition (δ13C) of hoop pine seedlings in a 12-month glasshouse trial in southeast Queensland,Australia. The 15N-labelled fertilisers were applied to nursery-reared hoop pine seedlings, which were then grown in pots,containing ca. 1.2 kg dried soil, under well watered conditions for 12 months. Four seedlings from each treatment were harvested at 4-month intervals, divided into roots, stem and foliage, with a further subdivision for new and old foliage,and then analysed for 15N, total N, δ13C and total C. There was no significant response in the seedling growth to the form or rate of application of nitrogen (N) fertiliser within the 12-month period, indicating that the seedlings did not experience N deficiency when grown on second rotation hoop pine soils. While the combined 15N recovery from soil and plant remained at around 70% throughout the experiment, the proportion of 15N recovered from the plants increasing steadily over time. Nitrate containing fertilisers at 150 mg N kg-1 soil gradually increased seedling foliage δ13C over the 12-month period, indicating an increase in seedling water use efficiency.

  5. Variable δ(15N diet-tissue discrimination factors among sharks: implications for trophic position, diet and food web models.

    Directory of Open Access Journals (Sweden)

    Jill A Olin

    Full Text Available The application of stable isotopes to characterize the complexities of a species foraging behavior and trophic relationships is dependent on assumptions of δ(15N diet-tissue discrimination factors (∆(15N. As ∆(15N values have been experimentally shown to vary amongst consumers, tissues and diet composition, resolving appropriate species-specific ∆(15N values can be complex. Given the logistical and ethical challenges of controlled feeding experiments for determining ∆(15N values for large and/or endangered species, our objective was to conduct an assessment of a range of reported ∆(15N values that can hypothetically serve as surrogates for describing the predator-prey relationships of four shark species that feed on prey from different trophic levels (i.e., different mean δ(15N dietary values. Overall, the most suitable species-specific ∆(15N values decreased with increasing dietary-δ(15N values based on stable isotope Bayesian ellipse overlap estimates of shark and the principal prey functional groups contributing to the diet determined from stomach content analyses. Thus, a single ∆(15N value was not supported for this speciose group of marine predatory fishes. For example, the ∆(15N value of 3.7‰ provided the highest percent overlap between prey and predator isotope ellipses for the bonnethead shark (mean diet δ(15N = 9‰ whereas a ∆(15N value < 2.3‰ provided the highest percent overlap between prey and predator isotope ellipses for the white shark (mean diet δ(15N = 15‰. These data corroborate the previously reported inverse ∆(15N-dietary δ(15N relationship when both isotope ellipses of principal prey functional groups and the broader identified diet of each species were considered supporting the adoption of different ∆(15N values that reflect the predators' δ(15N-dietary value. These findings are critical for refining the application of stable isotope modeling approaches as inferences regarding a species

  6. Fertilizer {sup 15}N balance in a coffee cropping system: a case study in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Fenilli, Tatiele Anete Bergamo [Universidade Regional de Blumenau (URB), SC (Brazil). Dept. de Engenharia Florestal]. E-mail: tfenilli@furb.br; Reichardt, Klaus; Bacchi, Osny Oliveira Santos [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Fisica do Solo]. E-mails: klaus@cena.usp.br; osny@cena.usp.br; Favarin, Jose Laercio [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Producao Vegetal; Silva, Adriana Lucia [Centro de Tecnologia Canavieira (CTC), Piracicaba, SP (Brazil). Fazenda Santo Antonio]. E-mail: adriana.silva@ctc.com.br; Timm, Luis Carlos [Universidade Federal de Pelotas (UFPel), RS (Brazil). Dept. de Engenharia Rural]. E-mail: lcartimm@yahoo.com.br

    2008-07-15

    Knowledge about the fate of fertilizer nitrogen in agricultural systems is essential for the improvement of management practices in order to maximize nitrogen (N) recovery by the crop and reduce N losses from the system to a minimum. This study involves fertilizer management practices using the {sup 15}N isotope label applied in a single rate to determine the fertilizer-N balance in a particular soil-coffee-atmosphere system and to deepen the understanding of N plant dynamics. Five replicates consisting of plots of about 120 plants each were randomly defined within a 0.2 ha coffee plantation planted in 2001, in Piracicaba, SP, Brazil. Nine plants of each plot were separated in sub-plots for the {sup 15}N balance studies and treated with N rates of 280 and 350 kg ha-1 during 2003/2004 and 2004/ 2005, respectively, both of them as ammonium sulfate enriched to a {sup 15}N abundance of 2.072 atom %. Plant shoots were considered as separate parts: the orthotropic central branch, productive branches, leaves of productive branches, vegetative branches, leaves of vegetative branches and fruit. Litter, consisting of dead leaves accumulated below the plant canopy, was measured by the difference between leaves at harvest and at the beginning of the following flowering. Roots and soil were sampled down to a depth of 1.0 at intervals of 0.2 m. Samples from the isotopic sub-plots were used to evaluate total N and {sup 15}N, and plants outside sub-plots were used to evaluate dry matter. Volatilization losses of NH{sub 3} were estimated using special collectors. Leaching of fertilizer-N was estimated from deep drainage water fluxes and {sup 15}N concentrations of the soil solution at 1 m soil depth. At the end of the 2-year evaluation, the recovery of {sup 15}N applied as ammonium sulfate was 19.1 % in aerial plant parts, 9.4 % in the roots, 23.8 % in the litter, 26.3 % in the fruit and 12.6 % remaining in the 0-1.0 m soil profile. Annual leaching and volatilization losses were

  7. Nitrogen cycling in an extreme hyperarid environment inferred from δ(15)N analyses of plants, soils and herbivore diet.

    Science.gov (United States)

    Díaz, Francisca P; Frugone, Matías; Gutiérrez, Rodrigo A; Latorre, Claudio

    2016-03-09

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ(15)N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ(15)N and δ(13)C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ(15)N values span the entire gradient, soil δ(15)N values show a positive correlation with aridity as expected. In contrast, foliar δ(15)N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ(15)N values.

  8. Nitrogen cycling in an extreme hyperarid environment inferred from δ15N analyses of plants, soils and herbivore diet

    Science.gov (United States)

    Díaz, Francisca P.; Frugone, Matías; Gutiérrez, Rodrigo A.; Latorre, Claudio

    2016-03-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ15N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ15N and δ13C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ15N values span the entire gradient, soil δ15N values show a positive correlation with aridity as expected. In contrast, foliar δ15N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ15N values.

  9. Interactions of /sup 14/N:/sup 15/N stearic acid spin-label pairs: effects of host lipid alkyl chain length and unsaturation

    Energy Technology Data Exchange (ETDEWEB)

    Feix, J.B.; Yin, J.J.; Hyde, J.S.

    1987-06-30

    Electron-electron double resonance (ELDOR) and saturation recovery electron paramagnetic resonance (EPR) spectroscopy have been employed to examine the interactions of /sup 14/N:/sup 15/N stearic acid spin-label pairs in fluid-phase model membrane bilayers composed of a variety of phospholipids. The (/sup 14/N)-16-doxylstearate:(/sup 15/N)-16-doxylstearate (16:16) pair was utilized to measure lateral diffusion of the spin-labels, while the (/sup 14/N)-16-doxylstearate:(/sup 15/N)-5-doxylstearate (16:5) pair provided information on vertical fluctuations of the 16-doxylstearate nitroxide moiety toward the membrane surface. Three saturated host lipids of varying alkyl chain length (dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), and distearoylphosphatidylcholine (DSPC)), an ..cap alpha..-saturated, ..beta..-unsaturated lipid (1-palmitoyl-2-oleoylphosphatidylcholine (POPC)), and phosphatidylcholine from a natural source (egg yolk phosphatidylcholine (egg PC)) were utilized as host lipids. Lateral diffusion of the stearic acid spin-labels was only slightly affected by alkyl chain length at a given reduced temperature (T/sub r/) in the saturated host lipids but was significantly decreased in POPC at the same T/sub r/. Lateral diffusion in DMPC, POPC, and egg PC was quite similar at 37/sup 0/C. A strong correlation was noted between lateral diffusion constants and rotational mobility of (/sup 14/N)-16-doxylstearate. Vertical fluctuations were likewise only slightly influenced by alklyl chain length but were strongly diminished in POPC and egg PC relative to the saturated systems. This diminution of the 16:5 interaction was observed even under conditions where no differences were discernible by conventional EPR.

  10. Fate of 15N and 14C from labelled plant material

    DEFF Research Database (Denmark)

    Rasmussen, Jim; Gjettermann, Birgitte; Eriksen, Jørgen;

    2008-01-01

    strength of labelled plant residues in dissolved inorganic N (DIN) and dissolved organic N (DON) in pore water from the plough layer, and (ii) the plant uptake of organically bound N. Litterbags containing 14C- and 15N-labelled ryegrass or clover roots or leaves were inserted into the sward of a ryegrass......–clover mixture in early spring. The fate of the released 14C and 15N was monitored in harvested biomass, roots, soil, and pore water percolating from the plough layer. No evidence of plant uptake of dual-labelled organic compounds from the dual-labelled residues could be observed. N in pore water from the plough...... water originating from plant residues only constituted 1.5% of the total dissolved N from the plough layer....

  11. Fertilizer nitrogen recovery of rice: /sup 15/N field studies (a short review)

    Energy Technology Data Exchange (ETDEWEB)

    Singh, B.; Katyal, J.C. (Punjab Agricultural Univ., Ludhiana (India). Dept. of Soils)

    1980-12-01

    Reliable quantitative estimates of fertilizer nitrogen recovery by rice are obtained in field investigations with /sup 15/N-labelled materials. Values obtained by conventional 'difference method' of comparing fertilized and unfertilized plots are typically larger than the actual values. Estimating the recovery of fertilizer nitrogen is not a goal in itself. Although it has been an essential component of limited number of /sup 15/N-field experiments conducted with rice so far; these provide little or no information about crop growth stages when fertilizer N is most efficiently utilized by rice plant. Recently, the path coefficient analysis has been used to analyse the effect of N uptake on the development of yield components and their contribution to grain yield. Nitrogen-15 fertilizers along with path coefficient analysis can prove particularly useful in comparing the efficiency of different N fertilizers and in the development of new and more efficient nitrogen sources and management practices.

  12. Study of organic N transformation in red soils by 15N tracer method

    Institute of Scientific and Technical Information of China (English)

    YeQing-Fu; ZhangQin-Zheng; 等

    1997-01-01

    Uniformly 15N-labelled ryegrass was used to investigate NH4+-production,microbial transformation and humification of organic N in two types of red soils by incubating the soils amended with labelled material.The results showed that there was little significant difference in biomass N transformation in the tested solis between 15N tracer method and conventional method,but the amount of NH4++-N released form the ryegrass in the clayey soil than in the sandy soil at all sampling time .By 120d of incubation,humified N was less than 10% of the amount of the applied N in two types of red soils and the amount of residual N in the clayey red soil was obviously higher than that in the sandy red soil.

  13. Chemienzymatic synthesis of Uridine. Nucleotides labeled with [15N] and [13C

    DEFF Research Database (Denmark)

    Gilles, Anne-Marie; Cristea, Ioan; Palibroda, Nicolae

    1995-01-01

    +necessary for the oxidation of glucose 6-phosphate and 6-phosphogluconate was recycled by glutamate dehydrogenase and excess of ammonia and a-oxoglutarate. Despite the number and complexity of the enzymatic steps, the synthesis of [15N,13C]UTP is straightforward with an overall yield exceeding 60%. This method, extended...... and diversified to the synthesis of all natural ribonucleotides, is a more economical alternative for obtaining nucleic acids for structural analysis by heteronuclear NMR spectroscopy....

  14. ~(15)N Isotope Used for Study of Groundwater Nitrogen Pollution in Shijiazhuang City, China

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    Shijiazhuang City is the capital of Hebei province, China. Groundwater is the major water supply source for living and industry need of the city. Due to a rapid increase of population and development of industry and agriculture, a series of groundwater environmental problems are created. In the paper, the situation of groundwater pollution in Shijiazhuang city is reported. Based on the groundwater chemical data and ~(15)N measurement results both on groundwater and soils, the reason of groundwater nitra...

  15. Determination of the δ15N of total nitrogen in solids; RSIL lab code 2893

    Science.gov (United States)

    Revesz, Kinga; Qi, Haiping; Coplen, Tyler B.

    2006-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2893 is to determine the δ(15N/14N), abbreviated as δ15N , of total nitrogen in solid samples. A Carlo Erba NC 2500 elemental analyzer (EA) is used to convert total nitrogen in a solid sample into N2 gas. The EA is connected to a continuous flow isotope-ratio mass spectrometer (CF-IRMS), which determines relative difference in the isotope-amount ratios of stable nitrogen isotopes (15N/14N)of the product N2 gas. The combustion is quantitative; no isotopic fractionation is involved. Samples are placed in a tin capsule and loaded into the Costech Zero Blank Autosampler of the EA. Under computer control, samples are dropped into a heated reaction tube that contains an oxidant, where the combustion takes place in a helium atmosphere containing an excess of oxygen gas. Combustion products are transported by a helium carrier through a reduction tube to remove excess oxygen and convert all nitrous oxides into N2 and through a drying tube to remove water. The gas-phase products, mainly CO2 and N2, are separated by a gas chromatograph. The gas is then introduced into the isotope-ratio mass spectrometer (IRMS) through a Finnigan MAT (now Thermo Scientific) ConFlo II interface, which also is used to inject N2 reference gas and helium for sample dilution. The IRMS is a Thermo Scientific Delta V Plus CF-IRMS. It has a universal triple collector, two wide cups with a narrow cup in the middle, capable of measuring mass/charge (m/z) 28, 29, 30, simultaneously. The ion beams from N2 are as follows: m/z 28 = N2 = 14N14N; m/z 29 = N2 = 14N15N primarily; m/z 30 = NO = 14N16O primarily, which is a sign of contamination or incomplete reduction.

  16. {sup 15}N uptake from manure and fertilizer sources by three consecutive crops under controlled conditions

    Energy Technology Data Exchange (ETDEWEB)

    Quiroga Garza, Hector Mario, E-mail: quiroga.mario@inifap.gob.m [Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias (INIFAP), Coahuila (Mexico); Delgado, Jorge A.; Wong, Jose Antonio Cueto, E-mail: jorge.delgado@ars.usda.go, E-mail: cueto.jose@inifap.gob.m [United States Dept. of Agriculture (USDA), Fort Collins, CO (United States). Agricultural Research Service. Soil Plant Nutrient Research Unit; Lindemann, William C., E-mail: wlindema@nmsu.ed [New Mexico State University (NMST), Las Cruces, NM (United States). Dept. Agronomy and Horticulture

    2009-09-15

    There are several regions of the world where soil N analysis and/or N budgets are not used to determine how much N to apply, resulting in higher than needed N inputs, especially when manure is used. One such region is the North Central 'La Comarca Lagunera', one of the most important dairy production areas of Mexico. We conducted a unique controlled greenhouse study using {sup 15}N fertilizer and {sup 15}N isotopic-labeled manure that was labeled under local conditions to monitor N cycling and recovery under higher N inputs. The manure-N treatment was applied only once and was incorporated in the soil before planting the first forage crop at an equivalent rate of 30, 60 and 120 Mg ha{sup -1} dry manure. The {sup 15}N treatments were equivalent to 120 and 240 kg ha{sup -1} (NH{sub 4}){sub 2}SO{sub 4}-N for each crop. The total N fertilizer for each N fertilized treatment were 360, and 720 kg ha{sup -1} N. We found very low N recoveries: about 9 % from the manure N inputs, lower than the 22 to 25 % from the fertilizer N inputs. The manure N recovered below ground in soil and roots ranged from 82 to 88 %. The low recoveries of N by the aboveground and low soil inorganic nitrate (NO{sup 3}-N) and ammonium (NH{sub 4}-N) content after the third harvested suggested that most of the {sup 15}N recovered below ground was in the soil organic form. The losses from manure N inputs ranged from 3 to 11 %, lower than the 34 to 39 % lost from fertilizer N sources. Our study shows that excessive applications of manure or fertilizer N that are traditionally used in this region will not increase the rate of N uptake by aboveground compartment but will increase the potential for N losses to the environment. (author)

  17. Capture cross sections of 15N(n, {\\gamma})16N at astrophysical energies

    CERN Document Server

    Fan, Guang-wei; Sheng, Zong-qiang; Tian, Feng; Wang, Jun; Zhang, Chao

    2016-01-01

    We have reanalyzed reaction cross sections of 16N on 12C target. The nucleon density distribution of 16N, especially surface density distribution, was extracted using the modified Glauber model. On the basis of dilute surface densities, the discussion of 15N(n, {\\gamma})16N reaction was performed within the framework of the direct capture reaction mechanism. The calculations agreed quite well with the experimental data.

  18. δ15N in the turtle grass from the Mexican Caribbean

    Science.gov (United States)

    Talavera-Saenz, A.; Sanchez, A.; Ortiz-Hernandez, M.

    2013-05-01

    Nutrient inputs associated with population growth threaten the integrity of coastal ecosystems. To assess the rapid increase in tourism, we compared the δ15N from Thalassia testudinum collected at sites with different levels of tourism development and population to detect the N inputs of wastewater discharge (WD) along the coast of Quintana Roo. The contributions of nitrogen enriched in 15N are directly related to the increase of WD inputs in areas of high tourism development (Nichupte Lagoon in Cancun, >3 million tourists per year from 2007 to 2011 and 0.7 million of resident population) and decreased towards Bahia Akumal and Tulum (>3 million tourists per year from 2007 to 2011 and 0.15 million of resident population). The δ15N from T. testudinum was significantly lower at Mahahual and Puerto Morelos (about 0.4 million tourists per year in 2007 to 2011 and 0.25 million of resident population) than other the sites. In areas of the lowest development and with tourist activity restricted and small population, such as the Yum Balam Reserve and Sian Ka'an Biosphere Reserve, the δ15N values were in much higher enrichment that Mahahual and Puerto Morelos. Therefore is suggested that Mahahual and Puerto Morelos may be used for baseline isotopic monitoring, over environmental pressure on the reef lagoon ecosystem, where tourist activities and population are growing very slow rate. The anthropogenic N input has the potential to impact, both environmentally and economically, the seagrass meadows and the coral reefs along the coast of Quintana Roo and the Caribbean.

  19. Real-time pure shift {sup 15}N HSQC of proteins: a real improvement in resolution and sensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Kiraly, Peter; Adams, Ralph W.; Paudel, Liladhar; Foroozandeh, Mohammadali [University of Manchester, School of Chemistry (United Kingdom); Aguilar, Juan A. [Durham University, Department of Chemistry (United Kingdom); Timári, István [University of Debrecen, Department of Inorganic and Analytical Chemistry (Hungary); Cliff, Matthew J. [University of Manchester, Manchester Institute of Biotechnology (United Kingdom); Nilsson, Mathias [University of Manchester, School of Chemistry (United Kingdom); Sándor, Péter [Agilent Technologies R& D and Marketing GmbH & Co. KG (Germany); Batta, Gyula [University of Debrecen, Department of Organic Chemistry (Hungary); Waltho, Jonathan P. [University of Manchester, Manchester Institute of Biotechnology (United Kingdom); Kövér, Katalin E. [University of Debrecen, Department of Inorganic and Analytical Chemistry (Hungary); Morris, Gareth A., E-mail: g.a.morris@manchester.ac.uk [University of Manchester, School of Chemistry (United Kingdom)

    2015-05-15

    Spectral resolution in proton NMR spectroscopy is reduced by the splitting of resonances into multiplets due to the effect of homonuclear scalar couplings. Although these effects are often hidden in protein NMR spectroscopy by low digital resolution and routine apodization, behind the scenes homonuclear scalar couplings increase spectral overcrowding. The possibilities for biomolecular NMR offered by new pure shift NMR methods are illustrated here. Both resolution and sensitivity are improved, without any increase in experiment time. In these experiments, free induction decays are collected in short bursts of data acquisition, with durations short on the timescale of J-evolution, interspersed with suitable refocusing elements. The net effect is real-time (t{sub 2}) broadband homodecoupling, suppressing the multiplet structure caused by proton–proton interactions. The key feature of the refocusing elements is that they discriminate between the resonances of active (observed) and passive (coupling partner) spins. This can be achieved either by using band-selective refocusing or by the BIRD element, in both cases accompanied by a nonselective 180° proton pulse. The latter method selects the active spins based on their one-bond heteronuclear J-coupling to {sup 15}N, while the former selects a region of the {sup 1}H spectrum. Several novel pure shift experiments are presented, and the improvements in resolution and sensitivity they provide are evaluated for representative samples: the N-terminal domain of PGK; ubiquitin; and two mutants of the small antifungal protein PAF. These new experiments, delivering improved sensitivity and resolution, have the potential to replace the current standard HSQC experiments.

  20. Evaluation of a 15N plot design for estimating plant recovery of fertilizer nitrogen applied to sugar cane

    OpenAIRE

    1994-01-01

    Two experiments were conducted on commercial sugar cane fields cropped with the variety SP70-1143, with the objective of evaluating a single row microplot design to determine plant recovery of 15N fertilizer nitrogen. One of them used 15N-aqua ammonia and 15N-urea applied to two linear meter microplots of a ratoon crop (four replicates). The second used one linear meter microplots (three replicates) which received 15N-aqua ammonia only. The fertilizers were applied on 15cm deep furrows, locat...

  1. A novel method for trapping and analyzing 15N in NO for tracing NO sources

    Science.gov (United States)

    Kang, Ronghua; Mulder, Jan; Dörsch, Peter

    2016-04-01

    15N isotope tracing is an effective and direct approach to investigate the biological and chemical sources of nitric oxide (NO) in soil. However, NO is highly reactive and rapidly converted to nitrogen dioxide (NO2) in the presence of ozone. Various chemical conversions of NO to the more stable solutes nitrite (NO2-) and nitrate (NO3-) have been proposed, which allow analysing the 15N abundance without major fractionation. However, NO emissions from soils are usually small, posing major challenges to conversion efficiency and background contamination. Here we present a novel method in which NO is oxidized to NO2- by chromium trioxide (CrO3) prior to conversion to NO2- and NO3- in an alkaline hydrogen peroxide (H2O2) solution. Immediately following trapping, manganese dioxide (MnO2) and 5M HCl are added to remove excess H2O2, and to adjust the pH to around 6.0-7.0, respectively. The resulting solution can be stored until analysis and is none-toxic, allowing to use a modified denitrifier method (Zhu et al., submitted), where NO2- and NO3- are reduced quantitatively to nitrous oxide (N2O). Optimum NO conversion rates of > 90% even at extremely low initial NO concentration were obtained with 4% H2O2, 0.5 M NaOH, and 0.5 L min-1 gas flow rate. In a laboratory test, using NO gas with different 15N signals produced from unlabelled and labelled NO2-, we found an overall precision of 0.4‰ for unlabelled and 49.7‰ for NO enriched with 1.0 atom% 15N, respectively. This indicates that this method can be used for both natural abundance studies of NO, as well as in labelling studies tracing NO sources. Zhu J, Yu L, Bakken LR, Mørkved PT, Mulder J, Dörsch P. Controlled induction of denitrification in Pseudomonas aureofaciens: a modified denitrifier method for 15N and 18O analysis in NO3- from natural water samples by IRMS. Submitted.

  2. Paleoenvironmental implications of taxonomic variation among δ 15 N values of chloropigments

    Science.gov (United States)

    Higgins, Meytal B.; Wolfe-Simon, Felisa; Robinson, Rebecca S.; Qin, Yelun; Saito, Mak A.; Pearson, Ann

    2011-11-01

    Natural variations in the ratios of nitrogen isotopes in biomass reflect variations in nutrient sources utilized for growth. In order to use δ 15N values of chloropigments of photosynthetic organisms to determine the corresponding δ 15N values of biomass - and by extension, surface waters - the isotopic offset between chlorophyll and biomass must be constrained. Here we examine this offset in various geologically-relevant taxa, grown using nutrient sources that may approximate ocean conditions at different times in Earth's history. Phytoplankton in this study include cyanobacteria (diazotrophic and non-diazotrophic), eukaryotic algae (red and green), and anoxygenic photosynthetic bacteria (Proteobacteria), as well as environmental samples from sulfidic lake water. Cultures were grown using N 2, NO 3-, and NH 4+ as nitrogen sources, and were examined under different light regimes and growth conditions. We find surprisingly high variability in the isotopic difference (δ 15N biomass - δ 15N chloropigment) for prokaryotes, with average values for species ranging from -12.2‰ to +11.7‰. We define this difference as ɛpor, a term that encompasses diagenetic porphyrins and chlorins, as well as chlorophyll. Negative values of ɛpor reflect chloropigments that are 15N-enriched relative to biomass. Notably, this enrichment appears to occur only in cyanobacteria. The average value of ɛpor for freshwater cyanobacterial species is -9.8 ± 1.8‰, while for marine cyanobacteria it is -0.9 ± 1.3‰. These isotopic effects group environmentally but not phylogenetically, e.g., ɛpor values for freshwater Chroococcales resemble those of freshwater Nostocales but differ from those of marine Chroococcales. Our measured values of ɛpor for eukaryotic algae (range = 4.7-8.7‰) are similar to previous reports for pure cultures. For all taxa studied, values of ɛpor do not depend on the type of nitrogen substrate used for growth. The observed environmental control of

  3. Delta15N values of tropical savanna and monsoon forest species reflect root specialisations and soil nitrogen status.

    Science.gov (United States)

    Schmidt, S; Stewart, G R

    2003-03-01

    A large number of herbaceous and woody plants from tropical woodland, savanna, and monsoon forest were analysed to determine the impact of environmental factors (nutrient and water availability, fire) and biological factors (microbial associations, systematics) on plant delta(15)N values. Foliar delta(15)N values of herbaceous and woody species were not related to growth form or phenology, but a strong relationship existed between mycorrhizal status and plant delta(15)N. In woodland and savanna, woody species with ectomycorrhizal (ECM) associations and putative N(2)-fixing species with ECM/arbuscular (AM) associations had lowest foliar delta(15)N values (1.0-0.6 per thousand ), AM species had mostly intermediate delta(15)N values (average +0.6 per thousand ), while non-mycorrhizal Proteaceae had highest delta(15)N values (+2.9 to +4.1 per thousand ). Similar differences in foliar delta(15)N were observed between AM (average 0.1 and 0.2 per thousand ) and non-mycorrhizal (average +0.8 and +0.3 per thousand ) herbaceous species in woodland and savanna. Leguminous savanna species had significantly higher leaf N contents (1.8-2.5% N) than non-fixing species (0.9-1.2% N) indicating substantial N acquisition via N(2) fixation. Monsoon forest species had similar leaf N contents (average 2.4% N) and positive delta(15)N values (+0.9 to +2.4 per thousand ). Soil nitrification and plant NO(3)(-) use was substantially higher in monsoon forest than in woodland or savanna. In the studied communities, higher soil N content and nitrification rates were associated with more positive soil delta(15)N and plant delta(15)N. In support of this notion, Ficus, a high NO(3)(-) using taxa associated with NO(3)(-) rich sites in the savanna, had the highest delta(15)N values of all AM species in the savanna. delta(15)N of xylem sap was examined as a tool for studying plant delta(15)N relations. delta(15)N of xylem sap varied seasonally and between differently aged Acacia and other savanna

  4. The Influence of Seed-borne N in 15N Isotope Dilution Studies with Legumes The Influence of Seed-borne N in 15N Isotope Dilution Studies with Legumes

    DEFF Research Database (Denmark)

    Jensen, Erik Steen; Andersen, A. J.; Thomsen, J. D.

    1985-01-01

    The distriution of seed-borne N in shoot and root of pea and field bean was studied using three methods: 1) determination of the N content in shoot and root of plants grown in sand culture without other N sources. 2) 15N isotope dilution in plants grown in Rhizobium-free medium supplied with 15N-...

  5. Job Assignments under Moral Hazard

    DEFF Research Database (Denmark)

    Koch, Alexander; Nafziger, Julia

    Inefficient job assignments are usually explained with incomplete information about employees' abilities or contractual imperfections. We show that inefficient assignments arise even without uncertainty about the employee's ability and with complete contracts. Building on this result we provide a...

  6. Unusually negative nitrogen isotopic compositions (δ15N of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

    Directory of Open Access Journals (Sweden)

    I. Romero

    2008-12-01

    Full Text Available Extremes in δ15N values in mangrove tissues and lichens (range =+4 to −22‰ were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, and atmospheric ammonia, and the δ15N of lichens, mangrove leaves, roots, stems, and wood were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Dwarfed Rhizophora mangle trees had the most negative δ15N, whereas fringing Rhizophora trees, the most positive δ15N values. Porewater ammonium concentrations had little relationship to N isotopic fractionation in mangrove tissues. In dwarfed mangroves, the δ15N of fine and coarse roots were 6–9‰ more positive than leaf tissue from the same tree, indicating different sources of N for root and leaf tissues. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year from −12‰ to −2‰, approaching the δ15N of porewater ammonium (δ15N=+4‰. Isotopically depleted ammonia in the atmosphere (δ15N=−19‰ and in rainwater (δ15N=−10‰ were found on Twin Cays. We propose that foliar uptake of these atmospheric sources by P-stressed, dwarfed mangrove trees and lichens can explain their very negative δ15N values. In environments where P is limiting for growth, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

  7. Long-term 15N tracking from biological N fixation across different plant and humus components of the boreal forest

    Science.gov (United States)

    Arroniz-Crespo, Maria; Jones, David L.; Zackrisson, Olle; Nilsson, Marie-Charlotte; DeLuca, Thomas H.

    2014-05-01

    Biological N2 fixation by cyanobacteria associated with feather mosses is an important cog in the nitrogen (N) cycle of boreal forests; still, our understanding of the turnover and fate of N fixed by this association remains greatly incomplete. The 15N signature of plants and soil serves as a powerful tool to explore N dynamics in forest ecosystems. In particular, in the present study we aimed to investigate the contribution of N2 fixation to δ15N signatures of plants and humus component of the boreal forest. Here we present results from a long-term (7 years) tacking of labelled 15N2 across the humus layer, seedlings of the tree species Pinus sylvestris, two common dwarf shrub species (Empetrum hermaphroditum and Vaccinium vitis-idaea) and the feather moss Pleurozium schreibery. The enriched experiment was conducted in 2005 in a natural boreal forest in northern Sweden. Two different treatments (10% 15N2 headspace enrichment and control) were setup in nine different plots (0.5 x 0.5 m) within the forest. We observed a significant reduction of δ15N signature of the 15N-enriched moss that could be explained by a growth dilution effect. Nevertheless, after 5 years since 15N2 enrichment some of the label 15N was still detected on the moss and in particular in the dead tissue. We could not detect a clear transfer of the labelled 15N2 from the moss-cyanobacteria system to other components of the ecosystem. However, we found consistence relationship through time between increments of δ15N signature of some of the forest components in plots which exhibited higher N fixation rates in the moss. In particular, changes in natural abundance δ15N that could be associated with N fixation were more apparent in the humus layer, the dwarf shrub Vaccinium vitis-idaea and the pine seedlings when comparing across plots and years.

  8. CACA-TOCSY with alternate {sup 13}C-{sup 12}C labeling: a {sup 13}C{sup {alpha}} direct detection experiment for mainchain resonance assignment, dihedral angle information, and amino acid type identification

    Energy Technology Data Exchange (ETDEWEB)

    Takeuchi, Koh [National Institute of Advanced Industrial Science and Technology (AIST), Biomedicinal Information Research Center (BIRC) (Japan); Frueh, Dominique P.; Sun, Zhen-Yu J.; Hiller, Sebastian; Wagner, Gerhard, E-mail: gerhard_wagner@hms.harvard.ed [Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology (United States)

    2010-05-15

    We present a {sup 13}C direct detection CACA-TOCSY experiment for samples with alternate {sup 13}C-{sup 12}C labeling. It provides inter-residue correlations between {sup 13}C{sup {alpha}} resonances of residue i and adjacent C{sup {alpha}s} at positions i - 1 and i + 1. Furthermore, longer mixing times yield correlations to C{sup {alpha}} nuclei separated by more than one residue. The experiment also provides C{sup {alpha}}-to-side chain correlations, some amino acid type identifications and estimates for {psi} dihedral angles. The power of the experiment derives from the alternate {sup 13}C-{sup 12}C labeling with [1,3-{sup 13}C] glycerol or [2-{sup 13}C] glycerol, which allows utilizing the small scalar {sup 3}J{sub CC} couplings that are masked by strong {sup 1}J{sub CC} couplings in uniformly {sup 13}C labeled samples.

  9. Intestinal renal metabolism of L-citrulline and L-arginine following enteral or parenteral infusion of L-alanyl-L-[2,15N]glutamine or L-[2,15N]glutamine in mice.

    Science.gov (United States)

    Boelens, Petra G; van Leeuwen, Paul A M; Dejong, Cornelis H C; Deutz, Nicolaas E P

    2005-10-01

    Previously, we observed increased plasma arginine (ARG) concentrations after glutamine (GLN)-enriched diets, in combination with clinical benefits. GLN delivers nitrogen for ARG synthesis, and the present study was designed to quantify the interorgan relationship of exogenous L-GLN or GLN dipeptide, by enteral or parenteral route, contributing to intestinal citrulline (CIT) and renal de novo ARG synthesis in mice. To study this, we used a multicatheterized mouse model with Swiss mice (n = 43) in the postabsorptive state. Stable isotopes were infused into the jugular vein or into the duodenum {per group either free L-[2,(15)N]GLN or dipeptide L-ALA-L-[2,(15)N]GLN, all with L-[ureido-(13)C-(2)H(2)]CIT and L-[guanidino-(15)N(2)-(2)H(2)]ARG} to establish renal and intestinal ARG and CIT metabolism. Blood flow was measured using (14)C-para-aminohippuric acid. Net intestinal CIT release, renal uptake of CIT, and net renal ARG efflux was found, as assessed by arteriovenous flux measurements. Quantitatively, more de novo L-[2,(15)N]CIT was produced when free L-[2,(15)N]GLN was given than when L-ALA-L-[2,(15)N]GLN was given, whereas renal de novo L-[2,(15)N]ARG was similar in all groups. In conclusion, the intestinal-renal axis is hereby proven in mice in that L-[2,(15)N]GLN or dipeptide were both converted into de novo renal L-[2,(15)N]ARG; however, not all was derived from intestinal L-[2,(15)N]CIT production. In this model, the feeding route and form of GLN did not influence de novo renal ARG production derived from GLN.

  10. Growth and foliar d15N of a Mojave desert shrub in relation to soil hydrological dynamics

    Science.gov (United States)

    Foliar 15N ratios (del15N), % N, and canopy volumes were measured in the two Mojave Desert dominant shrubs, the evergreen Larrea tridentata and drought deciduous Ambrosia dumosa growing across a geomorphically determined soil mosaic. Across three soils with increasingly strong age-dependent surface...

  11. Carbon (δ13C) and Nitrogen (δ15N) Stable Isotope Signatures in Bat Fur Indicate Swarming Sites Have Catchment Areas for Bats from Different Summering Areas.

    Science.gov (United States)

    Segers, Jordi L; Broders, Hugh G

    2015-01-01

    Migratory patterns of bats are not well understood and traditional methods to study this, like capture-mark-recapture, may not provide enough detail unless there are many records. Stable isotope profiles of many animal species have been used to make inferences about migration. Each year Myotis lucifugus and M. septentrionalis migrate from summering roosts to swarming caves and mines in the fall, but the pattern of movement between them is not well understood. In this study, fur δ13C and δ15N values of 305 M. lucifugus and 200 M. septentrionalis were analyzed to make inferences about migration patterns between summering areas and swarming sites in Nova Scotia, Canada. We expected that there would be greater variability in δ13C and δ15N among individuals at swarming sites because it was believed that these sites are used by individuals originating from many summering areas. There was extensive overlap in the standard ellipse area, corrected for small sample sizes (SEAc), of bats at swarming sites and much less overlap in SEAc among groups sampled at summering areas. Meaningful inference could not be made on M. septentrionalis because their low variation in SEAc may have been the result of sampling only 3 summering areas. However, for M. lucifugus, swarming sites had larger SEAc than summering areas and predictive discriminant analysis assigned swarming bats to multiple summering areas, supporting the contention that swarming bats are mixed aggregations of bats from several summering areas. Together, these data support the contention that swarming sites have catchment areas for bats from multiple summering areas and it is likely that the catchment areas for swarming sites overlap. These data suggest that δ13C and δ15N profiling of bat fur offer some potential to make inferences about regional migration in bats.

  12. Interactive Assignments for Online Students

    Directory of Open Access Journals (Sweden)

    Pam Lowry

    2009-04-01

    Full Text Available Students can experience first hand through interactive assignments what is involved in teaching an online course. Most students develop a whole new appreciation for the student learning process. Faculty are beginning to realize that online instruction is more than a series of readings posted to a course management system. This paper summarizes the faculty member's instructional strategies involved when creating student interaction assignments. The paper also summarizes the assignments, discussion board, and trends in education from the student's perspective. In summary, it concludes with the faculty's overall perspective concerning these assignments and how the assignments could be more effective for the student.

  13. Macroalgae δ15N values in well-mixed estuaries: Indicator of anthropogenic nitrogen input or macroalgae metabolism?

    Science.gov (United States)

    Raimonet, Mélanie; Guillou, Gaël; Mornet, Françoise; Richard, Pierre

    2013-03-01

    Although nitrogen stable isotope ratio (δ15N) in macroalgae is widely used as a bioindicator of anthropogenic nitrogen inputs to the coastal zone, recent studies suggest the possible role of macroalgae metabolism in δ15N variability. Simultaneous determinations of δ15N of dissolved inorganic nitrogen (DIN) along the land-sea continuum, inter-species variability of δ15N and its sensitivity to environmental factors are necessary to confirm the efficiency of macroalgae δ15N in monitoring nitrogen origin in mixed-use watersheds. In this study, δ15N of annual and perennial macroalgae (Ulva sp., Enteromorpha sp., Fucus vesiculosus and Fucus serratus) are compared to δ15N-DIN along the Charente Estuary, after characterizing δ15N of the three main DIN sources (i.e. cultivated area, pasture, sewage treatment plant outlet). During late winter and spring, when human activities produce high DIN inputs, DIN sources exhibit distinct δ15N signals in nitrate (NO) and ammonium (NH): cultivated area (+6.5 ± 0.6‰ and +9.0 ± 11.0‰), pasture (+9.2 ± 1.8‰ and +12.4‰) and sewage treatment plant discharge (+16.9 ± 8.7‰ and +25.4 ± 5.9‰). While sources show distinct δN- in this multiple source catchment, the overall mixture of NO sources - generally >95% DIN - leads to low variations of δN-NO at the mouth of the estuary (+7.7 to +8.4‰). Even if estuarine δN-NO values are not significantly different from pristine continental and oceanic site (+7.3‰ and +7.4‰), macroalgae δ15N values are generally higher at the mouth of the estuary. This highlights high anthropogenic DIN inputs in the estuary, and enhanced contribution of 15N-depleted NH in oceanic waters. Although seasonal variations in δN-NO are low, the same temporal trends in macroalgae δ15N values at estuarine and oceanic sites, and inter-species differences in δ15N values, suggest that macroalgae δ15N values might be modified by the metabolic response of macroalgae to environmental parameters (e

  14. Sinks for nitrogen inputs in terrestrial ecosystems: a meta-analysis of 15N tracer field studies

    Science.gov (United States)

    Templer, P.H.; Mack, M.C.; Chapin, F. S.; Christenson, L.M.; Compton, J.E.; Crook, H.D.; Currie, W.S.; Curtis, C.J.; Dail, D.B.; D'Antonio, C. M.; Emmett, B.A.; Epstein, H.E.; Goodale, C.L.; Gundersen, P.; Hobbie, S.E.; Holland, K.; Hooper, D.U.; Hungate, B.A.; Lamontagne, S.; Nadelhoffer, K.J.; Osenberg, C.W.; Perakis, S.S.; Schleppi, P.; Schimel, J.; Schmidt, I.K.; Sommerkorn, M.; Spoelstra, J.; Tietema, A.; Wessel, W.W.; Zak, D.R.

    2012-01-01

    Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (15N tracer application), total ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3–18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C: N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N·ha-1·yr-1 above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer addition.

  15. A 15N-poor isotopic composition for the solar system as shown by Genesis solar wind samples.

    Science.gov (United States)

    Marty, B; Chaussidon, M; Wiens, R C; Jurewicz, A J G; Burnett, D S

    2011-06-24

    The Genesis mission sampled solar wind ions to document the elemental and isotopic compositions of the Sun and, by inference, of the protosolar nebula. Nitrogen was a key target element because the extent and origin of its isotopic variations in solar system materials remain unknown. Isotopic analysis of a Genesis Solar Wind Concentrator target material shows that implanted solar wind nitrogen has a (15)N/(14)N ratio of 2.18 ± 0.02 × 10(-3) (that is, ≈40% poorer in (15)N relative to terrestrial atmosphere). The (15)N/(14)N ratio of the protosolar nebula was 2.27 ± 0.03 × 10(-3), which is the lowest (15)N/(14)N ratio known for solar system objects. This result demonstrates the extreme nitrogen isotopic heterogeneity of the nascent solar system and accounts for the (15)N-depleted components observed in solar system reservoirs.

  16. Enrichment of natural (15)N abundance during soil N losses under 20years of continuous cereal cropping.

    Science.gov (United States)

    Jones, Andrew R; Dalal, Ram C

    2017-01-01

    It is generally accepted that the enrichment of natural (15)N abundance in soil over time is reflective of historic N cycling and loss, but this process in cropping soils is not yet clear. In this study, we identified an enrichment gradient of natural (15)N abundance during 20-year chronosequence of cereal cropping on Alfisols in southwest Queensland, Australia, that have no history of fertilisation. We demonstrate that the increase in soil (15)N abundance is explained by isotopic fractionation of (15)N during organic N mineralisation and nitrification, which lead to isotopically heavier ammonium retained in the soil and isotopically lighter soil nitrate taken up and removed by seasonal crops during harvest. Here we present a framework for natural (15)N isotopic fractionation co-occurring with N losses during long-term cultivation.

  17. Localization of 15N uptake in a Tibetan alpine Kobresia pasture

    Science.gov (United States)

    Schleuß, Per-Marten; Kuzyakov, Yakov

    2014-05-01

    The Kobresia Pygmea ecotone covers approximately 450.000 km2 and is of large global and regional importance due several socio-ecological aspects. For instance Kobresia pastures store high amounts of carbon, nitrogen and other nutrients, represent large grazing areas for herbivores, provide a fast regrowth after grazing events and protect against mechanical degradation and soil erosion. However, Kobresia pastures are assumed to be a grazing induced and are accompanied with distinct root mats varying in thickness between 5-30 cm. Yet, less is known about the morphology and the functions of this root mats, especially in the background of a progressing degradation due to changes of climate and management. Thus we aimed to identify the importance of single soil layers for plant nutrition. Accordingly, nitrogen uptake from different soil depths and its remain in above-ground biomass (AGB), belowground biomass (BGB) and soil were determined by using a 15N pulse labeling approach during the vegetation period in summer 2012. 15N urea was injected into six different soil depths (0.5 cm, 2.5 cm, 7.5 cm, 12.5 cm, 17.5 cm, 22.5 cm / for each 4 replicates) and plots were sampled 45 days after the labeling. For soil and BGB samples were taken in strict sample intervals of 0-1 cm, 1-5 cm, 5-10 cm, 10-15 cm, 15-20 cm, 20-25 cm. Results indicate that total recovery (including AGB, BGB and soil) was highest, if tracer was injected into the top 5 cm and subsequently decreased with decreasing injection depth. This is especially the case for the 15N recovery of BGB, which is clearly attributed to the root density and strongly decreased with soil depth. In contrast, the root activity derived from the 15N content of roots increased with soil depth, which is primary associated to a proportionate increase of living roots related to dead roots. However, most 15N was captured in plant biomass (67.5-85.3 % of total recovery), indicating high 15N uptake efficiency possibly due to N limitation

  18. Provenancing Archaeological Wool Textiles from Medieval Northern Europe by Light Stable Isotope Analysis (δ13C, δ15N, δ2H)

    Science.gov (United States)

    von Holstein, Isabella C. C.; Walton Rogers, Penelope; Craig, Oliver E.; Penkman, Kirsty E. H.; Newton, Jason; Collins, Matthew J.

    2016-01-01

    We investigate the origin of archaeological wool textiles preserved by anoxic waterlogging from seven medieval archaeological deposits in north-western Europe (c. 700–1600 AD), using geospatial patterning in carbon (δ13C), nitrogen (δ15N) and non-exchangeable hydrogen (δ2H) composition of modern and ancient sheep proteins. δ13C, δ15N and δ2H values from archaeological wool keratin (n = 83) and bone collagen (n = 59) from four sites were interpreted with reference to the composition of modern sheep wool from the same regions. The isotopic composition of wool and bone collagen samples clustered strongly by settlement; inter-regional relationships were largely parallel in modern and ancient samples, though landscape change was also significant. Degradation in archaeological wool samples, examined by elemental and amino acid composition, was greater in samples from Iceland (Reykholt) than in samples from north-east England (York, Newcastle) or northern Germany (Hessens). A nominal assignment approach was used to classify textiles into local/non-local at each site, based on maximal estimates of isotopic variability in modern sheep wool. Light element stable isotope analysis provided new insights into the origins of wool textiles, and demonstrates that isotopic provenancing of keratin preserved in anoxic waterlogged contexts is feasible. We also demonstrate the utility of δ2H analysis to understand the location of origin of archaeological protein samples. PMID:27764106

  19. Capture cross sections of 15N(n, γ)16N at astrophysical energies

    Science.gov (United States)

    Fan, Guang-Wei; Ma, Jun-Bing; Sheng, Zong-Qiang; Shi, Guo-Zhu; Tian, Feng; Wang, Jun; Zhang, Chao

    2016-12-01

    We have reanalyzed reaction cross sections of 16N on a 12C target. The nucleon density distribution of 16N, especially surface density distribution, was extracted using the modified Glauber model. On the basis of dilute surface densities, the 15N(n, γ)16N reaction is discussed within the framework of the direct capture reaction mechanism. The calculations agree quite well with the experimental data. Support given by National Natural Science Foundation of China (11447236, 11505002, 11247001) and Foundation of Anhui University of Science and Technology (11130, 12608)

  20. Comparison of {sup 15}N- and {sup 13}C-determined parameters of mobility in melittin

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Lingyang [University Indianapolis, Department of Physics, Indiana University Purdue (United States); Prendergast, Franklyn G. [Mayo Foundation, Department of Pharmacology (United States); Kemple, Marvin D. [University Indianapolis, Department of Physics, Indiana University Purdue (United States)

    1998-07-15

    Backbone and tryptophan side-chain mobilities in the 26-residue, cytolytic peptide melittin (MLT) were investigated by {sup 15}N and {sup 13}C NMR. Specifically, inverse-detected {sup 15}N T{sub 1} and steady-state NOE measurements were made at 30 and 51 MHz on MLT at 22 deg. C enriched with {sup 15}N at six amide positions and in the Trp{sup 19} side chain. Both the disordered MLT monomer (1.2 mM peptide at pH 3.6 in neat water) and {alpha}-helical MLT tetramer (4.0 mM peptide at pH 5.2 in 150 mM phosphate buffer) were examined. The relaxation data were analyzed in terms of the Lipari and Szabo model-free formalism with three parameters: {tau}{sub m}, the correlation time for the overall rotation; S{sup 2}, a site-specific order parameter which is a measure of the amplitude of the internal motion; and {tau}{sub e}, a local, effective correlation time of the internal motion. A comparison was made of motional parameters from the {sup 15}N measurements and from {sup 13}C measurements on MLT, the latter having been made here and previously [Kemple et al. (1997) Biochemistry, 36, 1678-1688]. {tau}{sub m} and {tau}{sub e} values were consistent from data on the two nuclei. In the MLT monomer, S{sup 2} values for the backbone N-H and C{alpha}-H vectors in the same residue were similar in value but in the tetramer the N-H order parameters were about 0.2 units larger than the C{alpha}-H order parameters. The Trp side-chain N-H and C-H order parameters, and {tau}{sub e} values were generally similar in both the monomer and tetramer. Implications of these results regarding the dynamics of MLT are examined.

  1. Determination of the δ15N of nitrate in solids; RSIL lab code 2894

    Science.gov (United States)

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2894 is to determine the δ15N of nitrate (NO3-) in solids. The nitrate fraction of the nitrogen species is dissolved by water (called leaching) and can be analyzed by the bacterial method covered in RSIL lab code 2899. After leaching, the δ15N of the dissolved NO3- is analyzed by conversion of the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  2. Determination of the δ15N of nitrate in water; RSIL lab code 2899

    Science.gov (United States)

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2899 is to determine the δ15N of nitrate (NO3-) in water. The δ15N of the dissolved NO3- is analyzed by conversion of the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of the NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  3. Uptake of stormwater nitrogen in bioretention systems demonstrated from 15N tracer techniques

    Science.gov (United States)

    Houdeshel, D.; Hultine, K. R.; Pomeroy, C. A.

    2012-12-01

    Bioretention stormwater management systems are engineered ecosystems that capture urban stormwater in order to reduce the harmful effects of stormwater pollution on receiving waters. Bioretention systems have been shown to be effective at reducing the volume of runoff, and thereby reduce the nutrient loading to receiving waters from urban areas. However, little work has been done to evaluate the treatment processes that are responsible for reductions in effluent nitrogen (N). We hypothesize that the pulses of inorganic nitrogen associated with urban runoff events are captured in the plat tissues within these systems and not adsorbed to the soil media, thus creating a long-term, sustainable treatment approach to reducing the total nutrient loading to receiving waters. Nitrogen treatment performance was tested on two bioretention systems in Salt Lake City, UT: 1) an upland native community that does not require irrigation in semi-arid climates, and 2) a wetland community that requires 250 l of daily irrigation to offset the relatively high evaporative demand in the region. Each cell is sized to treat a 2.5 cm storm from a 140 m2 impervious surface: the area of the bioretention system is 10 m2. To test the N removal performance of each system, runoff events were simulated to represent an average precipitation regime using a synthetic stormwater blend starting in January, 2012. Effluent was collected from an underdrain and analyzed for total nitrogen (TN); mass removal was calculated for each month by subtracting the TN mass added to the garden minus the TN mass that flowed out of the garden. To test the hypothesis that plants assimilate stormwater N, 4 g of 100 atom% 15N NH4NO3 tracer was used as the N source in the synthetic stormwater during the first 2,000 l synthetic storm event in May. This isotopic label was calculated to enrich the total N pool of each garden to 100‰ 15N/14Nair. New growth was harvested from each plant in both cells and analyzed for 15N

  4. Optical Microscopy Characterization for Borehole U-15n#12 in Support of NCNS Source Physics Experiment

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, Jennifer E. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Sussman, Aviva Joy [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2015-05-22

    Optical microscopy characterization of thin sections from corehole U-15n#12 is part of a larger material characterization effort for the Source Physics Experiment (SPE). The SPE program was conducted in Nevada with a series of explosive tests designed to study the generation and propagation of seismic waves inside Stock quartz monzonite. Optical microscopy analysis includes the following: 1) imaging of full thin sections (scans and mosaic maps); 2) high magnification imaging of petrographic texture (grain size, foliations, fractures, etc.); and 3) measurement of microfracture density.

  5. Sinks for nitrogen inputs in terrestrial ecosystems: a meta-analysis of 15N tracer field studies.

    Science.gov (United States)

    Templer, P H; Mack, M C; Chapin, F S; Christenson, L M; Compton, J E; Crook, H D; Currie, W S; Curtis, C J; Dail, D B; D'Antonio, C M; Emmett, B A; Epstein, H E; Goodale, C L; Gundersen, P; Hobbie, S E; Holland, K; Hooper, D U; Hungate, B A; Lamontagne, S; Nadelhoffer, K J; Osenberg, C W; Perakis, S S; Schleppi, P; Schimel, J; Schmidt, I K; Sommerkorn, M; Spoelstra, J; Tietema, A; Wessel, W W; Zak, D R

    2012-08-01

    Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3-18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C:N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N x ha(-1) x yr(-1) above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer addition.

  6. The signatures of stable isotopes δ 15N and δ 13C in anadromous and non-anadromous Coilia nasus living in the Yangtze River, and the adjacent sea waters

    Science.gov (United States)

    Wang, Lei; Tang, Wenqiao; Dong, Wenxia

    2015-12-01

    Stable isotopes are increasingly used to investigate seasonal migrations of aquatic organisms. This study employed stable isotopes ( δ 13C and δ 15N) for Coilia nasus from the lower Yangtze River and the adjacent East China Sea to distinguish different ecotypic groups, ascertain trophic nutrition positions, and reflect environmental influences on C. nasus. δ 13C signatures of C. nasus sampled from Zhoushan (ZS), Chongming (CM), and Jingjiang (JJ) waters were significantly higher than those from the Poyang Lake (PYL) ( P nutrition positions in the marine and Poyang Lake food webs, respectively. C. nasus in Poyang Lake were significantly more enriched in δ 15N but depleted in δ 13C, suggesting that anthropogenic nutrient inputs and terrigenous organic carbon are important to the Poyang Lake food web. This study is the first to apply δ 15N and δ 13C to population assignment studies of C. nasus in the Yangtze River and its affiliated waters. Analysis of stable isotopes ( δ 15N and δ 13C) is shown to be a useful tool for discriminating anadromous and non-anadromous C. nasus.

  7. Study on Nitrogen Forms in Phenolic Polymers Incorporating Protien by 15N CP—MAS NMR

    Institute of Scientific and Technical Information of China (English)

    CHENGLILI; WENQIXIAO; 等

    1996-01-01

    Phenolic polymers synthesized by reactions by reactions of p-benzoquinone with 15N-labelled protein or (15NH4)2SO4 were studied by using 15N CP-MAS NMR technique in combination with chemical approaches.Results showed that more than 80% of nitrogen in quinone-protein polymers was in the form of amide with some present as aromatic and /or aliphatic amine and less than 10% of nitrogen occurred as heterocyclic N.The nitrogen distribution in the non-hydrolyzable residue of the quinone-protein polymers was basically similar to that of soil humic acid reported in literature with the exception that a higher proportion of N as heterocyclic N and aromatic amine and a lower proportion of N as amide and aliphatic amine were found in the former than in the latter,More than 70% of total nitrogen in quinone-(NH4)2OS4 polymer was acid resistant ,of which about 53% occurred as pyrrole,nitrile and imion type N.The possible roles of the reactions of phenols or quinones with proteins in the formation of humic acid.especially the non-hydrolyzable nitrogen in humicacid,are discussed.

  8. UV-visible and (1)H-(15)N NMR spectroscopic studies of colorimetric thiosemicarbazide anion sensors.

    Science.gov (United States)

    Farrugia, Kristina N; Makuc, Damjan; Podborska, Agnieszka; Szaciłowski, Konrad; Plavec, Janez; Magri, David C

    2015-02-14

    Four model thiosemicarbazide anion chemosensors containing three N-H bonds, substituted with phenyl and/or 4-nitrophenyl units, were synthesised and studied for their anion binding abilities with hydroxide, fluoride, acetate, dihydrogen phosphate and chloride. The anion binding properties were studied in DMSO and 9 : 1 DMSO-H2O by UV-visible absorption and (1)H/(13)C/(15)N NMR spectroscopic techniques and corroborated with DFT studies. Significant changes were observed in the UV-visible absorption spectra with all anions, except for chloride, accompanied by dramatic colour changes visible to the naked eye. These changes were determined to be due to the deprotonation of the central N-H proton and not due to hydrogen bonding based on (1)H/(15)N NMR titration studies with acetate in DMSO-d6-0.5% water. Direct evidence for deprotonation was confirmed by the disappearance of the central thiourea proton and the formation of acetic acid. DFT and charge distribution calculations suggest that for all four compounds the central N-H proton is the most acidic. Hence, the anion chemosensors operate by a deprotonation mechanism of the central N-H proton rather than by hydrogen bonding as is often reported.

  9. Oxygen determination in materials by 18O(p,αγ)15N nuclear reaction

    Science.gov (United States)

    Kumar, Sanjiv; Sunitha, Y.; Reddy, G. L. N.; Sukumar, A. A.; Ramana, J. V.; Sarkar, A.; Verma, Rakesh

    2016-07-01

    The paper presents a proton induced γ-ray emission method based on 18O(p,αγ)15N nuclear reaction to determine bulk oxygen in materials. The determination involves the measurement of 5.27 MeV γ-rays emitted following the de-excitation of 15N nuclei. A description of the energetics of the reaction is given to provide an insight into the origin of 5.27 MeV γ-rays. In addition, thick target γ-ray yields and the limits of detection are measured to ascertain the analytical potential of the reaction. The thick-target γ-ray yields are measured with a high purity germanium detector and a bismuth germanate detector at 0° as well as 90° angles in 3.0-4.2 MeV proton energy region. The best limit of detection of about 1.3 at.% is achieved at 4.2 MeV proton energy for measurements at 0° as well 90° angles with the bismuth germanate detector while the uncertainty in quantitative analysis is methodology is demonstrated by determining oxygen in several oxide as well as non-oxide materials.

  10. Growth, development, and fertilizer-{sup 15}N recovery by the coffee plant

    Energy Technology Data Exchange (ETDEWEB)

    Fenilli, Tatiele Anete Bergamo [Fundacao Universidade Regional de Blumenau (FURB), Blumenau, SC (Brazil). Dept. de Engenharia Florestal; Reichardt, Klaus; Bacchi, Osny Oliveira Santos [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Fisica do Solo]. E-mail: klaus@cena.usp.br; Dourado-Neto, Durval; Favarin, Jose Laercio [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Producao Vegetal; Trivelim, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis; Costa, Flavio Murilo Pereira da [Ministerio do Desenvolvimento Agrario, Brasilia, DF (Brazil). Secretaria de Assuntos Fundiarios - SEAF

    2007-09-15

    The relationship between growth and fertilizer nitrogen recovery by perennial crops such as coffee is poorly understood and improved understanding of such relations is important for the establishment of rational crop management practices. In order to characterize the growth of a typical coffee crop in Brazil and quantify the recovery of {sup 15}N labeled ammonium sulfate, and improve information for fertilizer management practices this study presents results for two consecutive cropping years, fertilized with 280 and 350 kg ha{sup -1} of N, respectively, applied in four splittings, using five replicates. Shoot dry matter accumulation was evaluated every 60 days, separating plants into branches, leaves and fruits. Labeled sub-plots were used to evaluate N-total and {sup 15}N abundance by mass spectrometry. During the first year the aerial part reached a recovery of 71% of the fertilizer N applied up to February, but this value was reduced to 34% at harvest and 19% at the beginning of the next flowering period due to leaf fall and fruit export. For the second year the aerial part absorbed 36% of the fertilizer N up to March, 47% up to harvest and 19% up to the beginning of the next flowering period. The splitting into four applications of the used fertilizer rates was adequate for the requirements of the crop at these growth stages of the coffee crop. (author)

  11. The Origin of Nitrogen on Jupiter and Saturn from the $^{15}$N/$^{14}$N Ratio

    CERN Document Server

    Fletcher, Leigh N; Orton, Glenn S; Irwin, Patrick G J; Mousis, Olivier; Sinclair, James A; Giles, Rohini S

    2014-01-01

    The Texas Echelon cross Echelle Spectrograph (TEXES), mounted on NASA's Infrared Telescope Facility (IRTF), was used to map mid-infrared ammonia absorption features on both Jupiter and Saturn in February 2013. Ammonia is the principle reservoir of nitrogen on the giant planets, and the ratio of isotopologues ($^{15}$N/$^{14}$N) can reveal insights into the molecular carrier (e.g., as N$_2$ or NH$_3$) of nitrogen to the forming protoplanets, and hence the source reservoirs from which these worlds accreted. We targeted two spectral intervals (900 and 960 cm$^{-1}$) that were relatively clear of terrestrial atmospheric contamination and contained close features of $^{14}$NH$_3$ and $^{15}$NH$_3$, allowing us to derive the ratio from a single spectrum without ambiguity due to radiometric calibration (the primary source of uncertainty in this study). We present the first ground-based determination of Jupiter's $^{15}$N/$^{14}$N ratio (in the range from $1.4\\times10^{-3}$ to $2.5\\times10^{-3}$), which is consistent...

  12. Visualizing the principal component of {sup 1}H,{sup 15}N-HSQC NMR spectral changes that reflect protein structural or functional properties: application to troponin C

    Energy Technology Data Exchange (ETDEWEB)

    Robertson, Ian M.; Boyko, Robert F.; Sykes, Brian D., E-mail: brian.sykes@ualberta.ca [University of Alberta, Department of Biochemistry (Canada)

    2011-09-15

    Laboratories often repeatedly determine the structure of a given protein under a variety of conditions, mutations, modifications, or in a number of states. This approach can be cumbersome and tedious. Given then a database of structures, identifiers, and corresponding {sup 1}H,{sup 15}N-HSQC NMR spectra for homologous proteins, we investigated whether structural information could be ascertained for a new homolog solely from its {sup 1}H,{sup 15}N-HSQC NMR spectrum. We addressed this question with two different approaches. First, we used a semi-automated approach with the program, ORBplus. ORBplus looks for patterns in the chemical shifts and correlates these commonalities to the explicit property of interest. ORBplus ranks resonances based on consistency of the magnitude and direction of the chemical shifts within the database, and the chemical shift correlation of the unknown protein with the database. ORBplus visualizes the results by a histogram and a vector diagram, and provides residue specific predictions on structural similarities with the database. The second method we used was partial least squares (PLS), which is a multivariate statistical technique used to correlate response and predictor variables. We investigated the ability of these methods to predict the tertiary structure of the contractile regulatory protein troponin C. Troponin C undergoes a closed-to-open conformational change, which is coupled to its function in muscle. We found that both ORBplus and PLS were able to identify patterns in the {sup 1}H,{sup 15}N-HSQC NMR data from different states of troponin C that correlated to its conformation.

  13. Alkaline Hydrolysis/Polymerization of 2,4,6-Trinitrotoluene:  Characterization of Products by 13C and 15N NMR

    Science.gov (United States)

    Thorn, Kevin A.; Thorne, Philip G.; Cox, Larry G.

    2004-01-01

    Alkaline hydrolysis has been investigated as a nonbiological procedure for the destruction of 2,4,6-trinitrotoluene (TNT) in explosives contaminated soils and munitions scrap. Nucleophilic substitutions of the nitro and methyl groups of TNT by hydroxide ion are the initial steps in the alkaline degradation of TNT. Potential applications of the technique include both in situ surface liming and ex situ alkaline treatment of contaminated soils. A number of laboratory studies have reported the formation of an uncharacterized polymeric material upon prolonged treatment of TNT in base. As part of an overall assessment of alkaline hydrolysis as a remediation technique, and to gain a better understanding of the chemical reactions underlying the hydrolysis/polymerization process, the soluble and precipitate fractions of polymeric material produced from the calcium hydroxide hydrolysis of unlabeled and 15N-labeled TNT were analyzed by elemental analysis and 13C and 15N nuclear magnetic resonance spectroscopy. Spectra indicated that reactions leading to polymerization included nucleophilic displacement of nitro groups by hydroxide ion, formation of ketone, carboxyl, alcohol, ether, and other aliphatic carbons, conversion of methyl groups to diphenyl methylene carbons, and recondensation of aromatic amines and reduced forms of nitrite, including ammonia and possibly hydroxylamine, into the polymer. Compared to the distribution of carbons in TNT as 14% sp3- and 86% sp2-hybridized, the precipitate fraction from hydrolysis of unlabeled TNT contained 33% sp3- and 67% sp2-hybridized carbons. The concentration of nitrogen in the precipitate was 64% of that in TNT. The 15N NMR spectra showed that, in addition to residual nitro groups, forms of nitrogen present in the filtrate and precipitate fractions include aminohydroquinone, primary amide, indole, imine, and azoxy, among others. Unreacted nitrite was recovered in the filtrate fraction. The toxicities and susceptibilities to

  14. Isotopic analysis of bulk, LMW, and HMW DON d15N indicates recycled nitrogen release from marine DON

    Science.gov (United States)

    Knapp, A. N.; Sigman, D. M.; Lipschultz, F.; Kustka, A.; Capone, D. G.

    2010-12-01

    Nitrogen (N) concentration and stable isotope ratio (d15N) measurements were made on bulk and size fractionated surface ocean dissolved organic nitrogen (DON) samples collected in the oligotrophic North Atlantic and Pacific Oceans. The bulk DON concentration in the upper 100 m is similar between the North Atlantic and North Pacific, between 4.5 and 5.0 uM, but the average d15N of bulk DON is significantly different, 3.9 per mil vs. air in the North Atlantic and 4.7 per mil in the North Pacific. The d15N of both bulk and HMW DON from the western tropical North Atlantic are similar to previous measurements, ~4.0 to 4.5 per mil. We report the first measurements of LMW DON d15N, which is consistently lower than HMW DON d15N. Neither the concentration nor d15N of bulk or size-fractionated DON varied with in situ N2 fixation rate, although significant variation in bulk and LMW DON d15N was observed between January and July of the same year in the western tropical North Atlantic. We propose a conceptual model to explain 1) the elevated d15N of bulk DON relative to other surface ocean N pools and fluxes, 2) the elevation of HMW DON d15N relative to LMW DON d15N, and 3) the inter-basin difference in the d15N of bulk DON. In this model, DON is produced from suspended particulate organic nitrogen (PON) without isotope fractionation because the conversion from PON to DON largely does not involve N-bearing bonds. In contrast, deamination and amide hydrolysis, with N isotope effects of 3 to 10 per mil, are major mechanisms by which DON is converted to ammonia and/or to other simple N compounds (e.g., amino acids). Thus these N-specific DON loss reactions result in an elevated d15N of residual DON relative to the parent DON and therefore also to the PON source. Moreover, the ammonium and simple organic N compounds released by microbial DON degradation are efficiently reassimilated back into the PON pool, as an integral part of the regenerated N cycle that further lowers the d15N

  15. {delta}{sup 15}N of seagrass leaves for monitoring anthropogenic nutrient increases in coral reef ecosystems

    Energy Technology Data Exchange (ETDEWEB)

    Yamamuro, M.; Kayanne, H.; Yamano, H

    2003-04-01

    In a coral reef environment, a slight increase in dissolved inorganic nitrogen (DIN;{>=}1.0 {mu}M) can alter the ecosystem via macroalgal blooms. We collected seagrass leaves from the tropical and subtropical Pacific Ocean in five countries and examined the interactions between nutrient concentrations (C, N, P), molar ratios of nutrients, and {delta}{sup 15}N to find a possible indicator of the DIN conditions. Within most sites, the concentrations of nutrients and their molar ratios showed large variations owing to species-specific values. On the other hand, almost identical {delta}{sup 15}N values were found in seagrass leaves of several species at each site. The correlations between {delta}{sup 15}N and nutrient concentrations and between {delta}{sup 15}N and molar ratios of nutrients suggested that nutrient availability did not affect the {delta}{sup 15}N value of seagrass leaves by altering the physiological condition of the plants. Increases in {delta}{sup 15}N of seagrass leaves mostly matched increases in DIN concentrations in the bottom water. We suggest that {delta}{sup 15}N in seagrass leaves can be a good tool to monitor time-integrated decrease/increase of DIN concentrations at a site, both in the water column and the interstitial water.

  16. {sup 15}N-labeled nitrogen from green manure and ammonium sulfate utilization by the sugarcane ratoon

    Energy Technology Data Exchange (ETDEWEB)

    Ambrosano, Edmilson Jose; Rossi, Fabricio, E-mail: ambrosano@apta.sp.gov.b [Agencia Paulista de Tecnologia dos Agronegocios (APTA), Piracicapa, SP (Brazil). Polo Rigional Centro Sul; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis; Cantarella, Heitor [Agencia Paulista de Tecnologia dos Agronegocios (APTA/IAC), Campinas, SP (Brazil). Instituto Agronomico de Campinas. Centro de Solos e Recursos Agroambientais; Ambrosano, Glaucia Maria Bovi [Universidade de Campinas (UNICAMP/FOP), Piracicaba, SP (Brazil). Fac. de Odontologia de Piracicaba. Dept. de Odontologia Social, Bioestatistica; Schammass, Eliana Aparecida [Agencia Paulista de Tecnologia dos Agronegocios (APTA/IZ), Nova Odessa, SP (Brazil). Instituto de Zootecnia; Muraoka, Takashi [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Fertilidade do solo

    2011-05-15

    Legumes as green manure are alternative sources of nitrogen (N) for crops and can supplement or even replace mineral nitrogen fertilization due to their potential for biological nitrogen fixation (BNF). The utilization of nitrogen by sugarcane (Saccharum spp.) fertilized with sunn hemp (Crotalaria juncea L.) and ammonium sulfate (AS) was evaluated using the {sup 15}N tracer technique. N was added at the rate of 196 and 70 kg ha{sup -1} as {sup 15}N-labeled sunn hemp green manure (SH) and as ammonium sulfate (AS), respectively. Treatments were: (I) Control; (II) AS{sup 15}N; (III) SH{sup 15}N + AS; (IV) SH{sup 15}N; and (V) AS{sup 15}N + SH. Sugarcane was cultivated for five years and was harvested three times. {sup 15}N recovery was evaluated in the two first harvests. In the sum of the three harvests, the highest stalk yields were obtained with a combination of green manure and inorganic N fertilizer; however, in the second cutting the yields were higher where SH was used than in plots with AS. The recovery of N by the first two consecutive harvests accounted for 19 to 21% of the N applied as leguminous green manure and 46 to 49% of the N applied as AS. The amounts of inorganic N, derived from both N sources, present in the 0-0.4 m layer of soil in the first season after N application and were below 1 kg ha{sup -1}. (author)

  17. Homogeneity of delta{sup 15}N in needles of Masson pine (Pinus massoniana L.) was altered by air pollution

    Energy Technology Data Exchange (ETDEWEB)

    Kuang Yuanwen, E-mail: kuangyw@scbg.ac.c [Institute of Ecology, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Pearl River Delta Research Center of Environmental Pollution and Control, Chinese Academy of Sciences, Guangzhou 510650 (China); Wen Dazhi; Li Jiong [Institute of Ecology, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Pearl River Delta Research Center of Environmental Pollution and Control, Chinese Academy of Sciences, Guangzhou 510650 (China); Sun Fangfang; Hou Enqing [Institute of Ecology, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Graduate University of the Chinese Academy of Sciences, Beijing 100049 (China); Zhou Guoyi; Zhang Deqiang [Institute of Ecology, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650 (China); Huang Longbin [Centre for Mined Land Rehabilitation, Sustainable Minerals Institute, the University of Queensland, QLD 4072 (Australia)

    2010-05-15

    The present study investigated the changes of delta{sup 15}N values in the tip, middle and base section (divided by the proportion to needle length) of current- and previous-year needles of Masson pine (Pinus massoniana L.) from two declining forest stands suffering from air pollution, in comparison with one healthy stand. At the healthy stand, delta{sup 15}N in the three sections of both current- and previous-year needles were found evenly distributed, while at the polluted stands, delta{sup 15}N values in the needles were revealed significantly different from the tip to the base sections. The results implied that the distribution of delta{sup 15}N among different parts or sections in foliages was not always homogeneous and could be affected by air pollution. We suggested that the difference of delta{sup 15}N values among pine needle sections should be reconsidered and should not be primarily ignored when the needle delta{sup 15}N values were used to assess plant responses to air pollution. - Values of delta{sup 15}N in needles of Masson pine (Pinus massoniana L.) were uneven and affected by air pollution.

  18. Diffusion technique for 15N and inorganic N analysis of low-N aqueous solutions and Kjeldahl digests.

    Science.gov (United States)

    Chen, Rui Rui; Dittert, Klaus

    2008-06-01

    Diffusion of ammonia is a common sample preparation method for the stable isotope analysis of inorganic nitrogen in aqueous solution. Classical diffusion methods usually require 6-12 days of diffusion and often focus on (15)N/(14)N analysis only. More recent studies have discussed whether complete N recovery was necessary for the precise analysis of stable N isotope ratios. In this paper we present a newly revised diffusion technique that allows correct and simultaneous determination of total N and (15)N at% from aqueous solutions and Kjeldahl digests, with N concentrations down to sub-0.5-mg N L(-1) levels, and it is tested under different conditions of (15)N isotope labelling. With the modification described, the diffusion time was reduced to 72 h, while the ratios of measured and expected (15)N at% were greater than 99% and the simultaneous recovery of total N was >95%. Analysis of soil microbial biomass N and its (15)N/(14)N ratio is one of the most important applications of this diffusion technique. An experiment with soil extracts spiked with (15)N-labelled yeast showed that predigestion was necessary to prevent serious N loss during Kjeldahl digestion of aqueous samples (i.e. soil extracts). The whole method of soil microbial biomass N preparation for (15)N/(14)N analysis included chloroform fumigation, predigestion, Kjeldahl digestion and diffusion. An experiment with soil spiked with (15)N-labelled yeast was carried out to evaluate the method. Results showed a highly significant correlation of recovered and added N, with the same recovery rate (0.21) of both total N and (15)N. A k(N) value of 0.25 was obtained based on the data. In conclusion, the diffusion method works for soil extracts and microbial biomass N determination and hence could be useful in many types of soil/water studies.

  19. Foliar δ15N is affected by foliar nitrogen uptake, soil nitrogen, and mycorrhizae along a nitrogen deposition gradient.

    Science.gov (United States)

    Vallano, Dena M; Sparks, Jed P

    2013-05-01

    Foliar nitrogen isotope (δ(15)N) composition patterns have been linked to soil N, mycorrhizal fractionation, and within-plant fractionations. However, few studies have examined the potential importance of the direct foliar uptake of gaseous reactive N on foliar δ(15)N. Using an experimental set-up in which the rate of mycorrhizal infection was reduced using a fungicide, we examined the influence of mycorrhizae on foliar δ(15)N in potted red maple (Acer rubrum) seedlings along a regional N deposition gradient in New York State. Mycorrhizal associations altered foliar δ(15)N values in red maple seedlings from 0.06 to 0.74 ‰ across sites. At the same sites, we explored the predictive roles of direct foliar N uptake, soil δ(15)N, and mycorrhizae on foliar δ(15)N in adult stands of A. rubrum, American beech (Fagus grandifolia), black birch (Betula lenta), and red oak (Quercus rubra). Multiple regression analysis indicated that ambient atmospheric nitrogen dioxide (NO2) concentration explained 0, 69, 23, and 45 % of the variation in foliar δ(15)N in American beech, red maple, red oak, and black birch, respectively, after accounting for the influence of soil δ(15)N. There was no correlation between foliar δ(13)C and foliar %N with increasing atmospheric NO2 concentration in most species. Our findings suggest that total canopy uptake, and likely direct foliar N uptake, of pollution-derived atmospheric N deposition may significantly impact foliar δ(15)N in several dominant species occurring in temperate forest ecosystems.

  20. Optimizing Marine Security Guard Assignments

    Science.gov (United States)

    2011-06-01

    Bangkok , Thailand East Asia and Pacific 18 4 Fort Lauderdale, Florida Western Hemisphere - South 13 5 Frankfurt, Germany Western Europe and Scandinavia 15...2008). Each 7 stationing plan satisfies a myriad of unit requirements, such as building and land availability. Similarly, each assignment solution...optimize the assignment of enlisted Marines to billets. EAM-GLOBAL seeks to assign the best Marine-billet fit while balancing staffing shortages

  1. Balanced input-output assignment

    Science.gov (United States)

    Gawronski, W.; Hadaegh, F. Y.

    1989-01-01

    Actuator/sensor locations and balanced representations of linear systems are considered for a given set of controllability and observability grammians. The case of equally controlled and observed states is given special attention. The assignability of grammians is examined, and the conditions for their existence are presented, along with several algorithms for their determination. Although an arbitrary positive semidefinite matrix is not always assignable, the identity grammian is shown to be always assignable. The results are extended to the case of flexible structures.

  2. d15N dynamics of ammonium and particulate nitrogen during the growth season of a eutrophic estuary

    DEFF Research Database (Denmark)

    De Brabandere, Loreto; Brion, N.; Elskens, M.;

    2007-01-01

    We monitored the stable nitrogen isotopic composition (d15N) of suspended matter and ammonium in the freshwater stretch of the Scheldt estuary (Belgium) over a full year to investigate for seasonal evolution and possible co-variation between isotopic signatures. The d15N value of ammonium remained...... rather constant during winter (average = +11.4 pro mille) but increased significantly with the spring and summer bloom, reaching values as high as +70 pro mille. This enrichment of the ammonium pool in 15N coincided with significant ammonium depletion during summer period, suggesting a close causal...

  3. Novel labeling technique illustrates transfer of 15N2 from Sphagnum moss to vascular plants via diazotrophic nitrogen fixation

    Science.gov (United States)

    Thorp, N. R.; Vile, M. A.; Wieder, R.

    2013-12-01

    We used 15N2 gas to trace nitrogen (N) from biological N2-fixation to vascular plant uptake in an Alberta bog in order to determine if neighboring bog plants acquire recently fixed N from diazotrophs associating with Sphagnum mosses. Recent evidence indicates high rates of N2-fixation in Sphagnum mosses of Alberta bogs (Vile et al. 2013). Our previous work has shown that mosses can assimilate fixed N from associated diazotrophs as evidenced by the high N content of mosses despite minimal inputs from atmospheric deposition, retranslocation, and N mineralization. Therefore, the potential exists for vascular plants to obtain N from ';leaky' tissues of live mosses, however, this phenomenon has not been tested previously. Here we document the potential for relatively rapid transfer to vascular plants of N fixed by Sphagnum moss-associated diazotrophs. We utilized the novel approach of incubating mosses in 15N2 to allow the process of diazotrophic N2-fixation to mechanistically provide the 15N label, which is subsequently transferred to Sphagnum mosses. The potential for vascular bog natives to tap this N was assessed by planting the vascular plants in the labeled moss. Sphagnum mosses (upper 3 cm of live plants) were incubated in the presence of 98 atom % 15N2 gas for 48 hours. Two vascular plants common to Alberta bogs; Picea mariana and Vaccinium oxycoccus were then placed in the labeled mosses, where the mosses served as the substrate. Tissue samples from these plants were collected at three time points during the incubation; prior to 15N2 exposure (to determine natural abundance 15N), and at one and two months after 15N2 exposure. Roots and leaves were separated and run separately on a mass spectrometer to determine 15N concentrations. Sphagnum moss capitula obtained N from N2-fixation (δ15N of -2.43 × 0.40, 122.76 × 23.78, 224.92 × 68.37, 143.74 × 54.38 prior to, immediately after, and at 1 and 2 months after exposure to 15N2, respectively). Nitrogen was

  4. Creating 13C- and 15N-enriched tree leaf litter for decomposition experiments

    Science.gov (United States)

    Szlavecz, K. A.; Pitz, S.; Chang, C.; Bernard, M.

    2013-12-01

    Labeling plant material with heavy isotopes of carbon and nitrogen can produce a traceable nutrient signal that can be followed into the different trophic levels and decomposer food web. We treated 60 tree saplings with 13C-enriched CO2 gas and 15N-enriched ammonium nitrate over a three-month period to create dually-labeled plant material for future decomposition experiments. The trees included both early (Red maple, Sweetgum, Tulip poplar) and late (American beech, White oak) successional deciduous tree species, and a conifer, White pine. We constructed a 2.4 m × 2.4 m × 2.4 m environmental chamber that was climate-controlled using an air conditioning system. An Arduino microcontroller interfaced with a Vaisala GMP343 CO2 probe maintained a CO2 concentration between 500-520 ppm by controlling a solenoid valve on the CO2 tank regulator. The trees were placed into the chamber in August 2012 and remained until senescence unless they were lost to death or disease. Ammonium nitrate was added twice, in September and October. Leaf samples were collected prior to the start of the experiment and after senescence, whereas root samples were collected only in December. Samples were dried, ground and analyzed using an isotope ratio mass spectrometer. American beech and White oak had 40% mortality, and 34% of tulip poplar trees were removed because of powdery mildew overgrowth or death. Most tulip poplar trees exhibited a second leaf out following senescence in late September. Nearly 1 kg of litter was produced with tulip poplar representing over half of the total mass. Levels of enrichment varied greatly by species. Beech (-14.2‰) and White oak (-4.8‰) had low levels of enrichment in comparison to early successional species such as Sweetgum (41.7‰) and Tulip poplar (30.7‰ [first leaf fall] and 238.0‰ [second leaf fall]). Leaf enrichment with 15N followed a similar pattern, though it was achieved at a higher level with δ15N values varying from 271.6‰ to 1354.2

  5. 15N-Labelled Fertilizer Recovery by Sweet Sorghum in Mediterranean Climate

    Directory of Open Access Journals (Sweden)

    Michele Perniola

    2008-12-01

    Full Text Available A 15N-labelled fertilization trial was carried out on sweet sorghum, grown in semi-arid environments of southern Europe with the aim to monitor the efficiency and effectiveness of the N-fertilisation technique under irrigation and different nitrogen fertilization rates, in factorial combination. A rainfed condition was compared with a full irrigation treatment (100% restoration of total crop water consumption, in a similar way, an unfertilized control was tested with respect to N application rates of 60 and 120 Kg ha-1, respectively. The fertilisation efficiency measured directly through the isotope discrimination technique was on average equal to 15%. The aliquot of nitrogen released by the fertiliser into the soil and not absorbed by the plant becomes part of the different components of the soil nitrogen balance, regardless of its origin.

  6. Nitrogen use efficiency evaluation of aerobic rice under field capacity water potential using {sup 15}N isotopic tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Wahid, Ahmad Nazrul Abd, E-mail: a-nazrul@nuclearmalaysia.gov.my [Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, 43600, Selangor (Malaysia); Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia); Rahim, Sahibin Abd, E-mail: haiyan@ukm.edu.my [Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, 43600, Selangor (Malaysia); Rahim, Khairuddin Abdul; Harun, Abdul Rahim [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia)

    2015-09-25

    This study was carried out to evaluate the efficiency use of the nitrogen fertilizer on aerobic rice varieties MR219-4 and MR219-9 which were grown aerobically under field capacity water potential at the controlled environment area or shield house. Direct {sup 15}N isotope tracer method was used in this study, whereby the {sup 15}N isotope was utilized as a tracer for nitrogen nutrient uptake. {sup 15}N isotope presence in the samples is determined by using emission spectrometer analysis and percentage of total nitrogen is determined by using Kjeldahl method. {sup 15}N atom access value contained in the sample will be used in determining the effectiveness of the use of nitrogen in fertilizers through the specific calculation formulas. In this work, the data several data of nitrogen derived from fertilizer (Ndff), total nitrogen, nitrogen uptake and nitrogen use efficiency was obtained.

  7. Nitrogen use efficiency evaluation of aerobic rice under field capacity water potential using 15N isotopic tracer technique

    Science.gov (United States)

    Wahid, Ahmad Nazrul Abd; Rahim, Sahibin Abd; Rahim, Khairuddin Abdul; Harun, Abdul Rahim

    2015-09-01

    This study was carried out to evaluate the efficiency use of the nitrogen fertilizer on aerobic rice varieties MR219-4 and MR219-9 which were grown aerobically under field capacity water potential at the controlled environment area or shield house. Direct 15N isotope tracer method was used in this study, whereby the 15N isotope was utilized as a tracer for nitrogen nutrient uptake. 15N isotope presence in the samples is determined by using emission spectrometer analysis and percentage of total nitrogen is determined by using Kjeldahl method. 15N atom access value contained in the sample will be used in determining the effectiveness of the use of nitrogen in fertilizers through the specific calculation formulas. In this work, the data several data of nitrogen derived from fertilizer (Ndff), total nitrogen, nitrogen uptake and nitrogen use efficiency was obtained.

  8. δ15N constraints on long-term nitrogen balances in temperate forests

    Science.gov (United States)

    Perakis, S.S.; Sinkhorn, E.R.; Compton, J.E.

    2011-01-01

    Biogeochemical theory emphasizes nitrogen (N) limitation and the many factors that can restrict N accumulation in temperate forests, yet lacks a working model of conditions that can promote naturally high N accumulation. We used a dynamic simulation model of ecosystem N and δ15N to evaluate which combination of N input and loss pathways could produce a range of high ecosystem N contents characteristic of forests in the Oregon Coast Range. Total ecosystem N at nine study sites ranged from 8,788 to 22,667 kg ha−1 and carbon (C) ranged from 188 to 460 Mg ha−1, with highest values near the coast. Ecosystem δ15N displayed a curvilinear relationship with ecosystem N content, and largely reflected mineral soil, which accounted for 96–98% of total ecosystem N. Model simulations of ecosystem N balances parameterized with field rates of N leaching required long-term average N inputs that exceed atmospheric deposition and asymbiotic and epiphytic N2-fixation, and that were consistent with cycles of post-fire N2-fixation by early-successional red alder. Soil water δ15NO3 − patterns suggested a shift in relative N losses from denitrification to nitrate leaching as N accumulated, and simulations identified nitrate leaching as the primary N loss pathway that constrains maximum N accumulation. Whereas current theory emphasizes constraints on biological N2-fixation and disturbance-mediated N losses as factors that limit N accumulation in temperate forests, our results suggest that wildfire can foster substantial long-term N accumulation in ecosystems that are colonized by symbiotic N2-fixing vegetation.

  9. δ15N constraints on long-term nitrogen balances in temperate forests.

    Science.gov (United States)

    Perakis, Steven S; Sinkhorn, Emily R; Compton, Jana E

    2011-11-01

    Biogeochemical theory emphasizes nitrogen (N) limitation and the many factors that can restrict N accumulation in temperate forests, yet lacks a working model of conditions that can promote naturally high N accumulation. We used a dynamic simulation model of ecosystem N and δ(15)N to evaluate which combination of N input and loss pathways could produce a range of high ecosystem N contents characteristic of forests in the Oregon Coast Range. Total ecosystem N at nine study sites ranged from 8,788 to 22,667 kg ha(-1) and carbon (C) ranged from 188 to 460 Mg ha(-1), with highest values near the coast. Ecosystem δ(15)N displayed a curvilinear relationship with ecosystem N content, and largely reflected mineral soil, which accounted for 96-98% of total ecosystem N. Model simulations of ecosystem N balances parameterized with field rates of N leaching required long-term average N inputs that exceed atmospheric deposition and asymbiotic and epiphytic N(2)-fixation, and that were consistent with cycles of post-fire N(2)-fixation by early-successional red alder. Soil water δ(15)NO(3)(-) patterns suggested a shift in relative N losses from denitrification to nitrate leaching as N accumulated, and simulations identified nitrate leaching as the primary N loss pathway that constrains maximum N accumulation. Whereas current theory emphasizes constraints on biological N(2)-fixation and disturbance-mediated N losses as factors that limit N accumulation in temperate forests, our results suggest that wildfire can foster substantial long-term N accumulation in ecosystems that are colonized by symbiotic N(2)-fixing vegetation.

  10. An optimized method for {sup 15}N R{sub 1} relaxation rate measurements in non-deuterated proteins

    Energy Technology Data Exchange (ETDEWEB)

    Gairí, Margarida, E-mail: mgairi@rmn.ub.edu [University of Barcelona (CCiTUB), NMR Facility, Scientific and Technological Centers (Spain); Dyachenko, Andrey [Institute for Research in Biomedicine (IRB) (Spain); González, M. Teresa; Feliz, Miguel [University of Barcelona (CCiTUB), NMR Facility, Scientific and Technological Centers (Spain); Pons, Miquel [University of Barcelona, Biomolecular NMR Laboratory and Organic Chemistry Department (Spain); Giralt, Ernest, E-mail: ernest.giralt@irbbarcelona.org [Institute for Research in Biomedicine (IRB) (Spain)

    2015-06-15

    {sup 15}N longitudinal relaxation rates are extensively used for the characterization of protein dynamics; however, their accurate measurement is hindered by systematic errors. {sup 15}N CSA/{sup 1}H–{sup 15}N dipolar cross-correlated relaxation (CC) and amide proton exchange saturation transfer from water protons are the two main sources of systematic errors in the determination of {sup 15}N R{sub 1} rates through {sup 1}H–{sup 15}N HSQC-based experiments. CC is usually suppressed through a train of 180° proton pulses applied during the variable {sup 15}N relaxation period (T), which can perturb water magnetization. Thus CC cancellation is required in such a way as to minimize water saturation effects. Here we examined the level of water saturation during the T period caused by various types of inversion proton pulses to suppress CC: (I) amide-selective IBURP-2; (II) cosine-modulated IBURP-2; (III) Watergate-like blocks; and (IV) non-selective hard. We additionally demonstrate the effect of uncontrolled saturation of aliphatic protons on {sup 15}N R{sub 1} rates. In this paper we present an optimized pulse sequence that takes into account the crucial effect of controlling also the saturation of the aliphatic protons during {sup 15}N R{sub 1} measurements in non-deuterated proteins. We show that using cosine-modulated IBURP-2 pulses spaced 40 ms to cancel CC in this optimized pulse program is the method of choice to minimize systematic errors coming from water and aliphatic protons saturation effects.

  11. Variability in δ{sup 15}N of intertidal brown algae along a salinity gradient: Differential impact of nitrogen sources

    Energy Technology Data Exchange (ETDEWEB)

    Viana, Inés G., E-mail: inesgviana@gmail.com; Bode, Antonio

    2015-04-15

    While it is generally agreed that δ{sup 15}N of brown macroalgae can discriminate between anthropogenic and natural sources of nitrogen, this study provides new insights on net fractionation processes occurring in some of these species. The contribution of continental and marine sources of nitrogen to benthic macroalgae in the estuary-ria system of A Coruña (NW Spain) was investigated by analyzing the temporal (at a monthly and annual basis) and spatial (up to 10 km) variability of δ{sup 15}N in the macroalgae Ascophyllum nodosum and three species of the genus Fucus (F. serratus, F. spiralis and F. vesiculosus). Total nitrate and ammonium concentrations and δ{sup 15}N-DIN, along with salinity and temperature in seawater were also studied to address the sources of such variability. Macroalgal δ{sup 15}N and nutrient concentrations decreased from estuarine to marine waters, suggesting larger dominance of anthropogenic nitrogen sources in the estuary. However, δ{sup 15}N values of macroalgae were generally higher than those of ambient nitrogen at all temporal and spatial scales considered. This suggests that the isotopic composition of these macroalgae is strongly affected by fractionation during uptake, assimilation or release of nitrogen. The absence of correlation between macroalgal and water samples suggests that the δ{sup 15}N of the species considered cannot be used for monitoring short-term changes. But their long lifespan and slow turnover rates make them suitable to determine the impact of the different nitrogen sources integrated over long-time periods. - Highlights: • Variability of Fucacean δ{sup 15}N indicates N sources along a salinity gradient. • δ{sup 15}N of Fucaceae and seawater are not correlated at short time scales. • Isotopic fractionation in macroalgal tissue varies at seasonal and at local scale. • Fucacean species are suitable for monitoring chronic N loadings.

  12. Species specific and environment induced variation of δ13C and δ15N in alpine plants

    Directory of Open Access Journals (Sweden)

    Yang eYang

    2015-06-01

    Full Text Available Stable carbon and nitrogen isotope signals in plant tissues integrate plant-environment interactions over long periods. In this study, we hypothesized that humid alpine life conditions are narrowing the scope for significant deviations from common carbon, water and nitrogen relations as captured by stable isotope signals. We explored the variation in δ13C and δ15N in 32 plant species from tissue type to ecosystem scale across a suite of locations at c. 2500 m elevation in the Swiss Alps. Foliar δ13C and δ15N varied among species by about 3-4 ‰ and 7-8 ‰ respectively. However, there was no overall difference in means of δ13C and δ15N for species sampled in different plant communities or when bulk plant dry matter harvests of different plant communities were compared. δ13C was found to be highly species specific, so that the ranking among species was mostly maintained across 11 habitats. However, δ15N varied significantly from place to place in all species (a range of 2.7 ‰ except in Fabaceae (Trifolium alpinum and Juncaceae (Luzula lutea. There was also a substantial variation among individuals of the same species collected next to each other. No difference was found in foliar δ15N of non-legumes, which were either collected next to or away from the most common legume, T. alpinum. δ15N data place Cyperaceae and Juncaceae, just like Fabaceae, in a low discrimination category, well separated from other families. Soil δ15N was higher than in plants and increased with soil depth. The results indicate a high functional diversity in alpine plants that is similar to that reported for low elevation plants. We conclude that the surprisingly high variation in δ13C and δ15N signals in the studied high elevation plants is largely species specific (genetic and insensitive to obvious environmental cues.

  13. Leaf δ15N as a temporal integrator of nitrogen-cycling processes at the Mojave Desert FACE experiment

    Science.gov (United States)

    Sonderegger, D.; Koyama, A.; Jin, V.; Billings, S. A.; Ogle, K.; Evans, R. D.

    2011-12-01

    Ecosystem response to elevated carbon dioxide (CO2) in arid environments is regulated primarily by water, which may interact with nitrogen availability. Leaf nitrogen isotope composition (δ15N) can serve as an important indicator of changes in nitrogen dynamics by integrating changes in plant physiology and ecosystem biogeochemical processes. Because of this temporal integration, careful modeling of the antecedent conditions is necessary for understanding the processes driving variation in leaf δ15N. We measured leaf δ15N of Larrea tridentata (creosotebush) over the 10-year lifetime of the Nevada Desert Free-Air CO2 Enrichment (FACE) experiment. Leaf δ15N exhibited two patterns. First, elevated atmospheric CO2 significantly increased Larrea leaf δ15N by approximately 2 to 3 % compared to plants exposed to ambient CO2 concentrations Second, plants in both CO2 treatments exhibited significant seasonal cycles in leaf δ15N, with higher values during the fall and winter seasons. We modeled leaf δ15N using a hierarchical Bayesian framework that incorporated soil moisture, temperature, and the Palmer Drought Severity Index (PDSI) covariates in addition to a CO2 treatment effect and plot random effects. Antecedent moisture effects were modeled by using a combination of the previous season's aggregated conditions and a smoothly varying weighted average of the months or weeks directly preceding the observation. The time lag between the driving antecedent condition and the observed change in leaf δ15N indicates a significant and unobserved process mechanism. Preliminary results suggest a CO2 treatment interaction with the lag effect, indicating a treatment effect on the latent process.

  14. 15N Abundance of Nodules as an Indicator of N Metabolism in N2-Fixing Plants 1

    Science.gov (United States)

    Shearer, Georgia; Feldman, Lori; Bryan, Barbara A.; Skeeters, Jerri L.; Kohl, Daniel H.; Amarger, Nöelle; Mariotti, Françoise; Mariotti, André

    1982-01-01

    This paper expands upon previous reports of 15N elevation in nodules (compared to other tissues) of N2-fixing plants. N2-Fixing nodules of Glycine max (soybeans), Vigna unguiculata (cowpea), Phaseolus vulgaris (common bean), Phaseolus coccineus (scarlet runner bean), Prosopis glandulosa (mesquite), and Olneya tesota (desert ironwood) were enriched in 15N. Nodules of Vicia faba (fava beans), Arachis hypogaea (peanut), Trifolium pratense (red clover), Pisum sativum (pea), Lathyrus sativus (grass pea), Medicago sativa (alfalfa), and Lupinus mutabilis (South American lupine) were not; nor were the nodules of nine species of N2-fixing nonlegumes. The nitrogen of ineffective nodules of soybeans and cowpeas was not enriched in 15N. Thus, 15N elevation in nodules of these plants depends on active N2-fixation. Results obtained so far on the generality of 15N enrichment in N2-fixing nodules suggest that only the nodules of plants which actively fix N2 and which transport allantoin or allantoic acid exhibit 15N enrichment. PMID:16662517

  15. Single Particle Strengths and Mirror States in $^{15}$N$-^{15}$O below 12.0 MeV

    CERN Document Server

    Mertin, C E; Crisp, A M; Keeley, N; Kemper, K W; Momtyuk, O; Roeder, B T; Volya, A

    2014-01-01

    New $^{14}$N(d,p) angular distribution data were taken at a deuteron bombarding energy of 16 MeV to locate all narrow single particle neutron states up to 15 MeV in excitation. A new shell model calculation is able to reproduce all levels in $^{15}$N up to 11.5 MeV and is used to characterize a narrow single particle level at 11.236 MeV and to provide a map of the single particle strengths. The known levels in $^{15}$N are then used to determine their mirrors in the lesser known nucleus $^{15}$O. The 2s$_{1/2}$ and 1d$_{5/2}$ single particle centroid energies are determined for the $^{15}$N$-^{15}$O mirror pair as: $^{15}$N $(\\text{2s}_{1/2}) = 8.08$ MeV, $^{15}$O $(\\text{2s}_{1/2}) = 7.43$ MeV, $^{15}$N $(\\text{1d}_{5/2}) = 7.97$ MeV, and $^{15}$O $(\\text{1d}_{5/2}) = 7.47$ MeV. These results confirm the degeneracy of these orbits and that the $^{15}$N$-^{15}$O nuclei are where the transition between the $\\text{2s}_{1/2}$ lying below the $\\text{1d}_{5/2}$ to lying above it, takes place. The $\\text{1d}_{3/2}$...

  16. Elastic and inelastic scattering of {sup 15}N ions by {sup 9}Be at 84 MeV

    Energy Technology Data Exchange (ETDEWEB)

    Rudchik, A.T., E-mail: rudchik@kinr.kiev.ua [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine); Chercas, K.A. [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine); Kemper, K.W. [Physics Department, Florida State University, Tallahassee, FL 32306-4350 (United States); Rusek, K. [Heavy Ion Laboratory of Warsaw University, ul. L. Pasteura 5A, PL-02-093 Warsaw (Poland); Rudchik, A.A.; Herashchenko, O.V. [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine); Koshchy, E.I. [Kharkiv National University, pl. Svobody 4, 61077 Kharkiv (Ukraine); Pirnak, Val.M. [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine); Piasecki, E.; Trzcińska, A. [Heavy Ion Laboratory of Warsaw University, ul. L. Pasteura 5A, PL-02-093 Warsaw (Poland); Sakuta, S.B. [Russian Research Center “Kurchatov Institute”, Kurchatov Sq. 1, 123182 Moscow (Russian Federation); Siudak, R. [H. Niewodniczański Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 152, PL-31-342 Cracow (Poland); Strojek, I. [National Center for Nuclear Researches, ul. Hoża 69, PL-00-681 Warsaw (Poland); Stolarz, A. [Heavy Ion Laboratory of Warsaw University, ul. L. Pasteura 5A, PL-02-093 Warsaw (Poland); Ilyin, A.P.; Ponkratenko, O.A.; Stepanenko, Yu.M.; Shyrma, Yu.O. [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine); Szczurek, A. [H. Niewodniczański Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 152, PL-31-342 Cracow (Poland); Uleshchenko, V.V. [Institute for Nuclear Research, Ukrainian Academy of Sciences, Prospect Nauki 47, 03680 Kyiv (Ukraine)

    2016-03-15

    Angular distributions of the {sup 9}Be + {sup 15}N elastic and inelastic scattering were measured at E{sub lab}({sup 15}N) = 84 MeV (E{sub c.m.} = 31.5 MeV) for the 0–6.76 MeV states of {sup 9}Be and 0–6.32 MeV states of {sup 15}N. The data were analyzed within the optical model and coupled-reaction-channels method. The elastic and inelastic scattering, spin reorientations of {sup 9}Be in ground and excited states and {sup 15}N in excited states as well as the most important one- and two-step transfer reactions were included in the channels-coupling scheme. The parameters of the {sup 9}Be + {sup 15}N optical potential of Woods–Saxon form as well as deformation parameters of these nuclei were deduced. The analysis showed that the {sup 9}Be + {sup 15}N pure potential elastic scattering dominates at the forward angles whereas the ground state spin reorientation of {sup 9}Be gives a major contribution to the elastic scattering cross sections at the large angles. Contributions from particle transfers are found to be negligible for the present scattering system.

  17. Correlation between the synthetic origin of methamphetamine samples and their 15N and 13C stable isotope ratios.

    Science.gov (United States)

    Billault, Isabelle; Courant, Frédérique; Pasquereau, Léo; Derrien, Solène; Robins, Richard J; Naulet, Norbert

    2007-06-12

    The active ingredient of ecstasy, N-methyl-3,4-methyldioxyphenylisopropylamine (MDMA) can be manufactured by a number of easy routes from simple precursors. We have synthesised 45 samples of MDMA following the five most common routes using N-precursors from 12 different origins and three different precursors for the aromatic moiety. The 13C and 15N contents of both the precursors and the MDMA samples derived therefrom were measured by isotope ratio mass spectrometry coupled to an elemental analyser (EA-IRMS). We show that within-pathway correlation between the 15N content of the precursor and that of the derived MDMA can be strong but that no general pattern of correlation can be defined. Rather, it is evident that the delta15N values of MDMA are strongly influenced by a combination of the delta15N values of the source of nitrogen used, the route by which the MDMA is synthesised, and the experimental conditions employed. Multivariate analysis (PCA) based on the delta15N values of the synthetic MDMA and of the delta15N and delta13C values of the N-precursors leads to good discrimination between the majority of the reaction conditions tested.

  18. δ15N Values in Crassostrea virginica Shells Provides Early Direct Evidence for Nitrogen Loading to Chesapeake Bay

    Science.gov (United States)

    Black, H. D.; Andrus, C. F. T.; Lambert, W. J.; Rick, T. C.; Gillikin, D. P.

    2017-01-01

    Crassostrea virginica is one of the most common estuarine bivalves in the United States’ east coast and is frequently found in archaeological sites and sub-fossil deposits. Although there have been several sclerochronological studies on stable carbon and oxygen isotopes in the shells of this species, less is known about δ15N values within their shells, which could be a useful paleoenvironmental proxy to assess estuarine nitrogen dynamics. Modern C. virginica samples were collected in Chesapeake Bay for comparison with archaeological shells from nearby sites ranging in age from ~100 to 3,200 years old. Left valves were sampled by milling the hinge area and the resulting powder was analyzed for %N and δ15N values. Comparison of δ15N values between C. virginica shells shows relatively constant values from ~1250 BC to ~1800 AD. After ~1800 AD, there are rapid increases in 15N enrichment in the shells, which continue to increase in value up to the modern shell values. The increase in δ15N values is evidence of early anthropogenic impact in Chesapeake Bay. These results corroborate the observation that coastal nitrogen pollution occurred earlier than the 19th century and support the use of oyster shell δ15N values as a useful environmental proxy. PMID:28281649

  19. Correlation between the synthetic origin of methamphetamine samples and their {sup 15}N and {sup 13}C stable isotope ratios

    Energy Technology Data Exchange (ETDEWEB)

    Billault, Isabelle [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France)]. E-mail: Isabelle.Billault@univ-nantes.fr; Courant, Frederique [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France); Pasquereau, Leo [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France); Derrien, Solene [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France); Robins, Richard J. [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France); Naulet, Norbert [Laboratoire d' Analyse Isotopique et Electrochimique de Metabolismes, CNRS UMR6006, University of Nantes, BP 92208, 44322 Nantes (France)

    2007-06-12

    The active ingredient of ecstasy, N-methyl-3,4-methyldioxyphenylisopropylamine (MDMA) can be manufactured by a number of easy routes from simple precursors. We have synthesised 45 samples of MDMA following the five most common routes using N-precursors from 12 different origins and three different precursors for the aromatic moiety. The {sup 13}C and {sup 15}N contents of both the precursors and the MDMA samples derived therefrom were measured by isotope ratio mass spectrometry coupled to an elemental analyser (EA-IRMS). We show that within-pathway correlation between the {sup 15}N content of the precursor and that of the derived MDMA can be strong but that no general pattern of correlation can be defined. Rather, it is evident that the {delta} {sup 15}N values of MDMA are strongly influenced by a combination of the {delta} {sup 15}N values of the source of nitrogen used, the route by which the MDMA is synthesised, and the experimental conditions employed. Multivariate analysis (PCA) based on the {delta} {sup 15}N values of the synthetic MDMA and of the {delta} {sup 15}N and {delta} {sup 13}C values of the N-precursors leads to good discrimination between the majority of the reaction conditions tested.

  20. Appraisal of {sup 15}N enrichment and {sup 15}N natural abundance methods for estimating N{sub 2} fixation by understorey Acacia leiocalyx and A. disparimma in a native forest of subtropical Australia

    Energy Technology Data Exchange (ETDEWEB)

    Bai, Shahla Hosseini; Xu, Zhihong; Blumfield, Timothy J. [Griffith Univ., Nathan, Brisbane, QLD (Australia). School of Biomolecular and Physical Sciences, Environmental Futures Centre; Sun, Fangfang [Guangdong Academy of Agricultural Sciences, Guangzhou (China). Research Centre for Quality, Safety and Standard of Agricultural Products; Chen, Chengrong [Griffith Univ., Nathan, Brisbane, QLD (Australia). School of Environment, Environmental Futures Centre; Wild, Clyde [Griffith Univ., Gold Coast, QLD (Australia). School of Environment, Environmental Futures Centre

    2012-05-15

    Purpose: It is anticipated that global climate change will increase the frequency of wildfires in native forests of eastern Australia. Understorey legumes such as Acacia species play an important role in maintaining ecosystem nitrogen (N) balance through biological N fixation (BNF). This is particularly important in Australian native forests with soils of low nutrient status and frequent disturbance of the nutrient cycles by fires. This study aimed to examine {sup 15}N enrichment and {sup 15}N natural abundance techniques in terms of their utilisation for evaluation of N{sub 2} fixation of understorey acacias and determine the relationship between species ecophysiological traits and N{sub 2} fixation. Materials and methods: A trial was established at sites 1 and 2 located at Toohey Forest, Queensland, Australia, a eucalypt-dominated native forest, to examine the determination of BNF using {sup 15}N enrichment and {sup 15}N natural abundance methods. Toohey Forest is an urban forest and subjected to frequent fuel reduction burns to protect the adjacent properties. Plant physiological status was measured to determine the relationship between physiological and N{sub 2} fixation activities. Results and discussion: Both {sup 15}N enrichment and {sup 15}N natural abundance techniques may be used to estimate N{sub 2} fixation of acacia tree species. The estimation of BNF using {sup 15}N enrichment was higher than those of the {sup 15}N natural abundance method. A grass reference plant, Themeda triandra, as well as tree reference plants provided an appropriate {delta}{sup 15}N signal. Potential B values for Acacia spp. between -0.3 permille and 1.0 permille provided an acceptable BNF estimation. This suburban forest is located nearby a busy highway leading to N deposition over time with consequent negative {delta}{sup 15}N signal. This N deposition may explain the separation between the {delta}{sup 15}N signal of the acacias and that of the reference plants which led to

  1. O potencial da rotulação metabólica de 15N para a pesquisa de esquizofrenia The potential of 15N metabolic labeling for schizophrenia research

    Directory of Open Access Journals (Sweden)

    Michaela D Filiou

    2012-01-01

    Full Text Available Pesquisas em psiquiatria ainda necessitam de estudos não dirigidos por hipóteses para revelar fundamentos neurobiológicos e biomarcadores moleculares para distúrbios psiquiátricos. Metodologias proteômicas disponibilizam uma série de ferramentas para esses fins. Apresentamos o princípio de rotulação metabólica utilizando 15N para proteômica quantitativa e suas aplicações em modelos animais de fenótipos psiquiátricos com um foco particular em esquizofrenia. Exploramos o potencial de rotulação metabólica por 15N em diferentes tipos de experimentos, bem como suas considerações metodológicas.Psychiatric research is in need of non-hypothesis driven approaches to unravel the neurobiological underpinnings and identify molecular biomarkers for psychiatric disorders. Proteomics methodologies constitute a state-of-the-art toolbox for biomarker discovery in psychiatric research. Here we present the principle of in vivo 15N metabolic labeling for quantitative proteomics experiments and applications of this method in animal models of psychiatric phenotypes, with a particular focus on schizophrenia. Additionally we explore the potential of 15N metabolic labeling in different experimental set-ups as well as methodological considerations of 15N metabolic labeling-based quantification studies.

  2. O potencial da rotulação metabólica de 15N para a pesquisa de esquizofrenia The potential of 15N metabolic labeling for schizophrenia research

    Directory of Open Access Journals (Sweden)

    Michaela D. Filiou

    2013-01-01

    Full Text Available Pesquisas em psiquiatria ainda necessitam de estudos não dirigidos por hipóteses para revelar fundamentos neurobiológicos e biomarcadores moleculares para distúrbios psiquiátricos. Metodologias proteômicas disponibilizam uma série de ferramentas para esses fins. Apresentamos o princípio de rotulação metabólica utilizando 15N para proteômica quantitativa e suas aplicações em modelos animais de fenótipos psiquiátricos com um foco particular em esquizofrenia. Exploramos o potencial de rotulação metabólica por 15N em diferentes tipos de experimentos, bem como suas considerações metodológicas.Psychiatric research is in need of non-hypothesis driven approaches to unravel the neurobiological underpinnings and identify molecular biomarkers for psychiatric disorders. Proteomics methodologies constitute a state-of-the-art toolbox for biomarker discovery in psychiatric research. Here we present the principle of in vivo 15N metabolic labeling for quantitative proteomics experiments and applications of this method in animal models of psychiatric phenotypes, with a particular focus on schizophrenia. Additionally we explore the potential of 15N metabolic labeling in different experimental set-ups as well as methodological considerations of 15N metabolic labeling-based quantification studies.

  3. NMR assignments of SPOC domain of the human transcriptional corepressor SHARP in complex with a C-terminal SMRT peptide.

    Science.gov (United States)

    Mikami, Suzuka; Kanaba, Teppei; Ito, Yutaka; Mishima, Masaki

    2013-10-01

    The transcriptional corepressor SMRT/HDAC1-associated repressor protein (SHARP) recruits histone deacetylases. Human SHARP protein is thought to function in processes involving steroid hormone responses and the Notch signaling pathway. SHARP consists of RNA recognition motifs (RRMs) in the N-terminal region and the spen paralog and ortholog C-terminal (SPOC) domain in the C-terminal region. It is known that the SPOC domain binds the LSD motif in the C-terminal tail of corepressors silencing mediator for retinoid and thyroid receptor (SMRT)/nuclear receptor corepressor (NcoR). We are interested in delineating the mechanism by which the SPOC domain recognizes the LSD motif of the C-terminal tail of SMRT/NcoR. To this end, we are investigating the tertiary structure of the SPOC/SMRT peptide using NMR. Herein, we report on the (1)H, (13)C and (15)N resonance assignments of the SPOC domain in complex with a SMRT peptide, which contributes towards a structural understanding of the SPOC/SMRT peptide and its molecular recognition.

  4. Structural Dynamics of the Potassium Channel Blocker ShK: SRLS Analysis of (15)N Relaxation.

    Science.gov (United States)

    Meirovitch, Eva; Tchaicheeyan, Oren; Sher, Inbal; Norton, Raymond S; Chill, Jordan H

    2015-12-10

    The 35-residue ShK peptide binds with high affinity to voltage-gated potassium channels. The dynamics of the binding surface was studied recently with (microsecond to millisecond) (15)N relaxation dispersion and (picosecond to nanosecond) (15)N spin relaxation of the N-H bonds. Relaxation dispersion revealed microsecond conformational-exchange-mediated exposure of the functionally important Y23 side chain to the peptide surface. The spin relaxation parameters acquired at 14.1 and 16.45 T have been subjected to model-free (MF) analysis, which yielded a squared generalized order parameter, S(2), of approximately 0.85 for virtually all of the N-H bonds. Only a "rigid backbone" evaluation could be inferred. We ascribe this limited information to the simplicity of MF in the context of challenging data. To improve the analysis, we apply the slowly relaxing local structure (SRLS) approach, which is a generalization of MF. SRLS describes N-H bond dynamics in ShK in terms of a local potential, u, ranging from 10 to 18.5 kBT, and a local diffusion rate, D2, ranging from 4.2 × 10(8) to 2.4 × 10(10) s(-1). This analysis shows that u is outstandingly strong for Y23 and relatively weak for K22, whereas D2 is slow for Y23 and fast for K22. These observations are relevant functionally because of the key role of the K22-Y23 dyad in ShK binding to potassium channels. The disulfide-bond network exhibits a medium-strength potential and an alternating wave-like D2 pattern. This is indicative of moderate structural restraints and motional plasticity, in support of, although not directly correlated with, the microsecond binding-related conformational exchange process detected previously. Thus, new information on functionally important residues in ShK and its overall conformational stability emerged from the SRLS analysis, as compared with the previous MF-based estimate of backbone dynamics as backbone rigidity.

  5. Job assignment with multivariate skills

    OpenAIRE

    Brilon, Stefanie

    2010-01-01

    This paper analyzes the job assignment problem faced by a firm when workers’ skills are distributed along several dimensions and jobs require different skills to varying extent. I derive optimal assignment rules with and without slot constraints, and show that under certain circumstances workers may get promoted although in their new job they are expected to be less productive than in their old job. This can be interpreted as a version of the Peter Principle which states that workers get prom...

  6. In vivo uniform 15N-isotope labelling of plants: Using the greenhouse for structural proteomics

    NARCIS (Netherlands)

    Ippel, J.H.; Pouvreau, L.A.M.; Kroef, T.; Gruppen, H.; Versteeg, G.; Putten, van der P.E.L.; Struik, P.C.; Mierlo, van C.P.M.

    2004-01-01

    Isotope labelling of proteins is important for progress in the field of structural proteomics. It enables the utilisation of the power of nuclear magnetic resonance spectroscopy (NMR) for the characterisation of the three-dimensional structures and corresponding dynamical features of proteins. The u

  7. Production of 15N-enriched nitric acid (H15NO3

    Directory of Open Access Journals (Sweden)

    C. R. Sant Ana Filho

    2008-12-01

    Full Text Available Techniques that employ 15N have proved to be an important tool in many areas of the agronomic and biomedical sciences. Nevertheless, their use is limited by methodological difficulties and by the price of compounds in the international market. Nitric compounds (15NO3- have attracted the interest of researchers. However, these compounds are not currently produced in Brazil. Thus, in the present work H15NO3 was obtained from the oxidation of anhydrous 15NH3. The method we used differs from the industrial process in that the absorption tower is replaced with a polytetrafluoroethylene-lined, stainless-steel hydration reactor. The process output was evaluated based on the following parameters: reaction temperature; ratio of reagents; pressure and flow of 15NH3(g through the catalyst (Pt/Rh. The results showed that, at the best conditions (500 ºC; 50 % excess O2; 0.4 MPa; and 3.39 g.min-1 of 15NH3, a conversion percentage (N-15NH3 to N-15NO3- of 62.2 %, an overall nitrogen balance (N-15NH3 + N-15NO3- of 86.8 %, and purity higher than 99 % could be obtained.

  8. Tracing sewage water by 15N in a mangrove ecosystem to test its bioremediation ability.

    Science.gov (United States)

    Lambs, Luc; Léopold, Audrey; Zeller, Bernd; Herteman, Mélanie; Fromard, Francois

    2011-10-15

    Mangrove forests could be a simple and effective alternative to conventional sewage treatment, particularly for island communities given its low cost and low maintenance. Due to their high adaptation capacity, these plants are able to tolerate and bioremediate the high levels of nutrients and pollutants found in sewage water. This solution could be applied to small tropical islands with high population density such as Mayotte in the Indian Ocean. This paper reports on a trial by stable isotopic (15)N tracing of such a bioremediation process on pre-treated wastewater near the village of Malamani, in the middle of the large coastal mangrove in the bay near Chirongui. The first results show a boost in the mangrove growth, but a longer period of observation is needed to confirm the beneficial effects, and also to clarify the role of the local crab population, whose engineering activities play an important part in the ecosystem. The exact denitrification process is not yet understood, and the mass balance equation also reveals loss of nitrogen-containing compounds, which needs to be analyzed more closely.

  9. 13C and 15N natural isotope abundance reflects breast cancer cell metabolism

    Science.gov (United States)

    Tea, Illa; Martineau, Estelle; Antheaume, Ingrid; Lalande, Julie; Mauve, Caroline; Gilard, Francoise; Barillé-Nion, Sophie; Blackburn, Anneke C.; Tcherkez, Guillaume

    2016-09-01

    Breast cancer is the most common cancer in women worldwide. Despite the information provided by anatomopathological assessment and molecular markers (such as receptor expression ER, PR, HER2), breast cancer therapies and prognostics depend on the metabolic properties of tumor cells. However, metabolomics have not provided a robust and congruent biomarker yet, likely because individual metabolite contents are insufficient to encapsulate all of the alterations in metabolic fluxes. Here, we took advantage of natural 13C and 15N isotope abundance to show there are isotopic differences between healthy and cancer biopsy tissues or between healthy and malignant cultured cell lines. Isotope mass balance further suggests that these differences are mostly related to lipid metabolism, anaplerosis and urea cycle, three pathways known to be impacted in malignant cells. Our results demonstrate that the isotope signature is a good descriptor of metabolism since it integrates modifications in C partitioning and N excretion altogether. Our present study is thus a starting point to possible clinical applications such as patient screening and biopsy characterization in every cancer that is associated with metabolic changes.

  10. Effects of 15N application frequency on nitrogen uptake efficiency in citrus trees.

    Science.gov (United States)

    Quiñones, Ana; Bañuls, Josefina; Millo, Eduardo Primo; Legaz, Francisco

    2003-12-01

    Two irrigation systems were used to compare nitrogen uptake efficiency in citrus trees and to evaluate the NO3- runoff in "Navelina" orange trees [Citrus sinensis (L.) Osbeck] on Carrizo citrange rootstock (Citrus sinensis x Poncirus trifoliata Raf.). These were fertilized with 125 g N as labelled K15NO3 and grown outdoors in containers filled with a sand-loamy soil. Two groups of 3 trees received this N dose either in five equally split applications by a flooding irrigation system or in 66 applications by drip. Trees were harvested at the end of the vegetative cycle (December) and the isotopic ratios of 15N/14N were measured in the soil-plant system. The N uptake efficiency of the whole tree was higher with drip irrigation (75%) than with flooding system (64%). In the 0-90 cm soil profile, the N immobilized in the organic fraction was similar for both irrigation methods (around 13 %), whereas the N retained as NO3- was 1% of the N applied under drip and 10% under flooding. In the last case, most of NO3- remained under root system and it could be lost to leaching either by heavy rainfalls or excessive water applications. These results showed that a drip irrigation system was more efficient for improving water use and N uptake from fertilizer, in addition to potentially reduced leaching losses.

  11. Simultaneous quantification of depolymerization and mineralization rates by a novel 15N tracing model

    Science.gov (United States)

    Andresen, Louise C.; Björsne, Anna-Karin; Bodé, Samuel; Klemedtsson, Leif; Boeckx, Pascal; Rütting, Tobias

    2016-09-01

    The depolymerization of soil organic matter, such as proteins and (oligo-)peptides, into monomers (e.g. amino acids) is currently considered to be the rate-limiting step for nitrogen (N) availability in terrestrial ecosystems. The mineralization of free amino acids (FAAs), liberated by the depolymerization of peptides, is an important fraction of the total mineralization of organic N. Hence, the accurate assessment of peptide depolymerization and FAA mineralization rates is important in order to gain a better process-based understanding of the soil N cycle. In this paper, we present an extended numerical 15N tracing model Ntrace, which incorporates the FAA pool and related N processes in order to provide a more robust and simultaneous quantification of depolymerization and gross mineralization rates of FAAs and soil organic N. We discuss analytical and numerical approaches for two forest soils, suggest improvements of the experimental work for future studies, and conclude that (i) when about half of all depolymerized peptide N is directly mineralized, FAA mineralization can be as important a rate-limiting step for total gross N mineralization as peptide depolymerization rate; (ii) gross FAA mineralization and FAA immobilization rates can be used to develop FAA use efficiency (NUEFAA), which can reveal microbial N or carbon (C) limitation.

  12. STELLAR ORIGINS OF EXTREMELY {sup 13}C- AND {sup 15}N-ENRICHED PRESOLAR SIC GRAINS: NOVAE OR SUPERNOVAE?

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Nan; Nittler, Larry R.; Alexander, Conel M. O’D.; Wang, Jianhua [Department of Terrestrial Magnetism, Carnegie Institution for Science, Washington, DC 20015 (United States); Pignatari, Marco [E.A. Milne Centre for Astrophysics, Department of Physics and Mathematics, University of Hull, HU6 7RX (United Kingdom); José, Jordi [Department de Fisica, EUETIB, Universitat Politécnica de Catalunya, E-08036 Barcelona (Spain); Nguyen, Ann, E-mail: nliu@carnegiescience.edu [Robert M. Walker Laboratory for Space Science, Astromaterials Research and Exploration Science Directorate, NASA Johnson Space Center, Houston, TX 77058 (United States)

    2016-04-01

    Extreme excesses of {sup 13}C ({sup 12}C/{sup 13}C < 10) and {sup 15}N ({sup 14}N/{sup 15}N < 20) in rare presolar SiC grains have been considered diagnostic of an origin in classical novae, though an origin in core collapse supernovae (CCSNe) has also been proposed. We report C, N, and Si isotope data for 14 submicron- to micron-sized {sup 13}C- and {sup 15}N-enriched presolar SiC grains ({sup 12}C/{sup 13}C < 16 and {sup 14}N/{sup 15}N < ∼100) from Murchison, and their correlated Mg–Al, S, and Ca–Ti isotope data when available. These grains are enriched in {sup 13}C and {sup 15}N, but with quite diverse Si isotopic signatures. Four grains with {sup 29,30}Si excesses similar to those of type C SiC grains likely came from CCSNe, which experienced explosive H burning occurred during explosions. The independent coexistence of proton- and neutron-capture isotopic signatures in these grains strongly supports heterogeneous H ingestion into the He shell in pre-supernovae. Two of the seven putative nova grains with {sup 30}Si excesses and {sup 29}Si depletions show lower-than-solar {sup 34}S/{sup 32}S ratios that cannot be explained by classical nova nucleosynthetic models. We discuss these signatures within the CCSN scenario. For the remaining five putative nova grains, both nova and supernova origins are viable because explosive H burning in the two stellar sites could result in quite similar proton-capture isotopic signatures. Three of the grains are sub-type AB grains that are also {sup 13}C enriched, but have a range of higher {sup 14}N/{sup 15}N. We found that {sup 15}N-enriched AB grains (∼50 < {sup 14}N/{sup 15}N < ∼100) have distinctive isotopic signatures compared to putative nova grains, such as higher {sup 14}N/{sup 15}N, lower {sup 26}Al/{sup 27}Al, and lack of {sup 30}Si excess, indicating weaker proton-capture nucleosynthetic environments.

  13. Glacial–interglacial dynamics of Antarctic firn columns: comparison between simulations and ice core air-δ15N measurements

    Directory of Open Access Journals (Sweden)

    E. Capron

    2013-05-01

    Full Text Available Correct estimation of the firn lock-in depth is essential for correctly linking gas and ice chronologies in ice core studies. Here, two approaches to constrain the firn depth evolution in Antarctica are presented over the last deglaciation: outputs of a firn densification model, and measurements of δ15N of N2 in air trapped in ice core, assuming that δ15N is only affected by gravitational fractionation in the firn column. Since the firn densification process is largely governed by surface temperature and accumulation rate, we have investigated four ice cores drilled in coastal (Berkner Island, BI, and James Ross Island, JRI and semi-coastal (TALDICE and EPICA Dronning Maud Land, EDML Antarctic regions. Combined with available ice core air-δ15N measurements from the EPICA Dome C (EDC site, the studied regions encompass a large range of surface accumulation rates and temperature conditions. Our δ15N profiles reveal a heterogeneous response of the firn structure to glacial–interglacial climatic changes. While firn densification simulations correctly predict TALDICE δ15N variations, they systematically fail to capture the large millennial-scale δ15N variations measured at BI and the δ15N glacial levels measured at JRI and EDML – a mismatch previously reported for central East Antarctic ice cores. New constraints of the EDML gas–ice depth offset during the Laschamp event (~41 ka and the last deglaciation do not favour the hypothesis of a large convective zone within the firn as the explanation of the glacial firn model–δ15N data mismatch for this site. While we could not conduct an in-depth study of the influence of impurities in snow for firnification from the existing datasets, our detailed comparison between the δ15N profiles and firn model simulations under different temperature and accumulation rate scenarios suggests that the role of accumulation rate may have been underestimated in the current description of firnification

  14. Evaluation of N sub 2 fixation by applying sup 15 N labeled plant material and ammonium sulfate

    Energy Technology Data Exchange (ETDEWEB)

    Chiu, C.Y.; Yoshida, T.

    1990-06-01

    Effect of different {sup 15}N labeled sources on the estimation of N{sub 2} fixation was investigated. The combination of {sup 15}N labeled ammonium sulfate, {sup 15}N labeled plant material, and {sup 15}N labeled ammonium sulfate with unlabeled plant material, was examined in pot experiments. Two cultivars of soybean (Glycine max) and one of mungbean (Vigna radiata) were used. No significant difference was observed among the treatments for the estimation of N{sub 2} fixation. This was due to the homogeneity and stability of the {sup 15}N abundance in soil which resulted in a similar N uptake from the soil by the N{sub 2} fixing and reference crops. The plant yield, total N uptake and amount of N{sub 2} fixed were higher in the Yellow Soil than in the Andosol. The amount of N{sub 2} fixed was strongly influenced by the plant growth and consequently it affected the plant yield. The slow decomposition of plant material in the Andosol resulted in a low yield in both the N{sub 2} fixing and reference crops. Thus, the artificial decrease of the available N content in soil, by application of plant material, did not stimulate N{sub 2} fixation but suppressed plant growth and N{sub 2} fixation.

  15. Evaluation of a {sup 15} N plot design for estimating plant recovery of fertilizer nitrogen applied to sugar cane

    Energy Technology Data Exchange (ETDEWEB)

    Trivelin, P.C.O.; Cabezas, W.A.R.L.; Victoria, R.L.; Reichardt, K. [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

    1994-05-01

    Two experiments were conducted on commercial sugar cene fields cropped with the variety SP70-1143, with the objective of evaluating a single row microplot design to determine plant recovery of {sup 15} N fertilizer nitrogen. One of them used {sup 15} N-aqua ammonia and {sup 15} N-urea applied to two linear meter microplots of a ratoon crop (four replicates.) The second used one linear meter microplots (three replicates) which received {sup 15} N-aqua ammonia only. The fertilizers were applied on 15cm deep furrows, located 25 cm from both sides of the cane row. One linear meter of ratoon cane, inside and outside of the microplot, and on the same and adjacent rows were harvested twelve months after fertilization. The results indicate the feasibility of using single row regments of ratoon cane with {sup 15} N-fertilizer. The main advantages of this microplot design, when compared to the classical 3 contiguous row segments, is that only one third of the labeled fertilizer is needed. (author). 25 refs, 3 figs, 5 tabs.

  16. Stellar Origins of Extremely $^{\\text{13}}C$- and $^{15}N$-enriched Presolar SiC Grains: Novae or Supernovae?

    CERN Document Server

    Liu, Nan; Alexander, Conel M O'D; Wang, Jianhuan; Pignatari, Marco; Jose, Jordi; Nguyen, Ann

    2016-01-01

    Extreme excesses of $^{13}C$ ($^{12}C$/$^{13}C$<10) and $^{15}N$ ($^{14}N$/$^{15}N$<20) in rare presolar SiC grains have been considered diagnostic of an origin in classical novae, though an origin in core collapse supernovae (CCSNe) has also been proposed. We report C, N, and Si isotope data for 14 submicron- to micron-sized $^{13}C$- and $^{15}N$-enriched presolar SiC grains ($^{12}C$/$^{13}C$<16 and $^{14}N$/$^{15}N$<~100) from Murchison, and their correlated Mg-Al, S, and Ca-Ti isotope data when available. These grains are enriched in $^{13}C$ and $^{15}N$, but with quite diverse Si isotopic signatures. Four grains with $^{29,30}Si$ excesses similar to those of type C SiC grains likely came from CCSNe, which experienced explosive H burning occurred during explosions. The independent coexistence of proton- and neutron-capture isotopic signatures in these grains strongly supports heterogeneous H ingestion into the He shell in pre-supernovae. Two of the seven putative nova grains with $^{30}Si$ e...

  17. {sup 15}N/{sup 14}N isotopic ratio and statistical analysis: an efficient way of linking seized Ecstasy tablets

    Energy Technology Data Exchange (ETDEWEB)

    Palhol, Fabien; Lamoureux, Catherine; Chabrillat, Martine; Naulet, Norbert

    2004-05-10

    In this study, the {sup 15}N/{sup 14}N isotopic ratios of 106 samples of 3,4-methylenedioxymethamphetamine (MDMA) extracted from Ecstasy tablets are presented. These ratios, measured using gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS), show a large discrimination between samples with a range of {delta}{sup 15}N values between -17 and +19%o, depending on the precursors and the method used in clandestine laboratories. Thus, {delta}{sup 15}N values can be used in a statistical analysis carried out in order to link Ecstasy tablets prepared with the same precursors and synthetic pathway. The similarity index obtained after principal component analysis and hierarchical cluster analysis appears to be an efficient way to group tablets seized in different places.

  18. 15N-glycine absorption in the colon of the short bowel rats%短肠大鼠结肠对15N-甘氨酸的吸收研究

    Institute of Scientific and Technical Information of China (English)

    靳大勇; 许剑民; 吴海福; 吴肇汉; 夏立均; 王锦华; 胡耀明

    2001-01-01

    Objective To investigate 15N-glycine absorption in the colon of the short bowel rats.Methods Rats undergoing resection of 80% small intestine and were treated by enteral nutrition for 21 days.The concentration of 15N-glycine in perfusate samples was measured by the technique of HPLC.The atom percent enrichment of 15N-glycine in blood plasma was measured by the technique of isotopic dilution method.Results After 3 hours,the colonic water absorption was 1±0.5 ml,15N-glycine absorption was zero,and the atom percent enrichment was near zero in normal group,vs.(2.6±0.82) ml (P<0.05),(4.39±1.2)% (P<0.01),and (0.614±0.517)% (P<0.01) in short bowel rats,respectively.The mucosal thickness,villous height and surface area of the colon in enteral nutrition group were greater than that in controls (P<0.01).Conclusion The short bowel rats had significant colonic amino acids and water absorption adaptation,though not enough to fulfil the need of the body.%目的探讨短肠大鼠的结肠对氨基酸代偿吸收能力。方法切除80%小肠的短肠大鼠,经肠喂养21天后,测定结肠对15N-甘氨酸的吸收率和血15N-甘氨酸丰度及形态学改变。结果正常大鼠3h水吸收为(1±0.5)ml,对15N-甘氨酸无吸收,血中15N-甘氨酸丰度为零。短肠大鼠3h水吸收为(2.6±0.82)ml (P<0.05),15N-甘氨酸为(4.39±1.2)% (P<0.01),血中15N-甘氨酸丰度为(0.614±0.517)% (P<0.01)。形态学检测显示: 喂养组与对照组相比结肠直径差异有显著意义(P<0.05),而结肠壁厚度、粘膜厚度、皱襞高度和皱襞表面积差异有极显著的意义(P<0.01)。结论短肠大鼠结肠对水和15N-甘氨酸有一定代偿性吸收,组织形态学也有一定程度的代偿。

  19. A 15N-Labeling Study of the Capture of Deep Soil Nitrate from Different Plant Systems

    Institute of Scientific and Technical Information of China (English)

    YANG Zhi-xin; WANG Jue; DI Hong-jie; ZHANG Li-juan; JU Xiao-tang

    2014-01-01

    The objective of this study was to determine the efifciency of different plant systems in capturing deep soil nitrate (NO3-) to reduce NO3-leaching in a ifeld plot experiment using 15N labelling. The study was conducted on a calcareous alluvial soil on the North China Plains and the plant systems evaluated included alfalfa (Medicago sativa), American black poplar (Populus nigra) and cocksfoot (Dactylis). 15N-labelled N fertilizer was injected to 90 cm depth to determine the recovery of 15N by the plants. With conventional water and nutrient management, the total recovery of 15N-labeled NO3--N was 23.4% by alfalfa after two consecutive growth years. The recovery was signiifcantly higher than those by American black poplar (12.3%) and cocksfoot (11.4%). The highest proportion of soil residual 15N from the labeled fertilizer N (%Ndff) was detected around 90 cm soil depth at the time of the 1st year harvest and at 110-130 cm soil depth at time of the 2nd year harvest. Soil%Ndff in 0-80 cm depth was signiifcantly higher in the alfalfa treatment than those in all the other treatments. The soil%Ndff below 100 cm depth was much lower in the alfalfa than those in all the other treatments. These results indicated that 15N leaching losses in the alfalfa treatment were signiifcantly lower than by those in the black poplar and cocksfoot treatments, due to the higher root density located in nitrate labeling zone of soil proifle. In conclusion, alfalfa may be used as a plant to capture deep soil NO3-left from previous crops to reduce NO3-leaching in high intensity crop cultivation systems of North China Plain.

  20. Organic vs. conventional grassland management: do (15N and (13C isotopic signatures of hay and soil samples differ?

    Directory of Open Access Journals (Sweden)

    Valentin H Klaus

    Full Text Available Distinguishing organic and conventional products is a major issue of food security and authenticity. Previous studies successfully used stable isotopes to separate organic and conventional products, but up to now, this approach was not tested for organic grassland hay and soil. Moreover, isotopic abundances could be a powerful tool to elucidate differences in ecosystem functioning and driving mechanisms of element cycling in organic and conventional management systems. Here, we studied the δ(15N and δ(13C isotopic composition of soil and hay samples of 21 organic and 34 conventional grasslands in two German regions. We also used Δδ(15N (δ(15N plant - δ(15N soil to characterize nitrogen dynamics. In order to detect temporal trends, isotopic abundances in organic grasslands were related to the time since certification. Furthermore, discriminant analysis was used to test whether the respective management type can be deduced from observed isotopic abundances. Isotopic analyses revealed no significant differences in δ(13C in hay and δ(15N in both soil and hay between management types, but showed that δ(13C abundances were significantly lower in soil of organic compared to conventional grasslands. Δδ(15N values implied that management types did not substantially differ in nitrogen cycling. Only δ(13C in soil and hay showed significant negative relationships with the time since certification. Thus, our result suggest that organic grasslands suffered less from drought stress compared to conventional grasslands most likely due to a benefit of higher plant species richness, as previously shown by manipulative biodiversity experiments. Finally, it was possible to correctly classify about two third of the samples according to their management using isotopic abundances in soil and hay. However, as more than half of the organic samples were incorrectly classified, we infer that more research is needed to improve this approach before it can be

  1. Organic vs. conventional grassland management: do (15)N and (13)C isotopic signatures of hay and soil samples differ?

    Science.gov (United States)

    Klaus, Valentin H; Hölzel, Norbert; Prati, Daniel; Schmitt, Barbara; Schöning, Ingo; Schrumpf, Marion; Fischer, Markus; Kleinebecker, Till

    2013-01-01

    Distinguishing organic and conventional products is a major issue of food security and authenticity. Previous studies successfully used stable isotopes to separate organic and conventional products, but up to now, this approach was not tested for organic grassland hay and soil. Moreover, isotopic abundances could be a powerful tool to elucidate differences in ecosystem functioning and driving mechanisms of element cycling in organic and conventional management systems. Here, we studied the δ(15)N and δ(13)C isotopic composition of soil and hay samples of 21 organic and 34 conventional grasslands in two German regions. We also used Δδ(15)N (δ(15)N plant - δ(15)N soil) to characterize nitrogen dynamics. In order to detect temporal trends, isotopic abundances in organic grasslands were related to the time since certification. Furthermore, discriminant analysis was used to test whether the respective management type can be deduced from observed isotopic abundances. Isotopic analyses revealed no significant differences in δ(13)C in hay and δ(15)N in both soil and hay between management types, but showed that δ(13)C abundances were significantly lower in soil of organic compared to conventional grasslands. Δδ(15)N values implied that management types did not substantially differ in nitrogen cycling. Only δ(13)C in soil and hay showed significant negative relationships with the time since certification. Thus, our result suggest that organic grasslands suffered less from drought stress compared to conventional grasslands most likely due to a benefit of higher plant species richness, as previously shown by manipulative biodiversity experiments. Finally, it was possible to correctly classify about two third of the samples according to their management using isotopic abundances in soil and hay. However, as more than half of the organic samples were incorrectly classified, we infer that more research is needed to improve this approach before it can be efficiently

  2. Recovery and Leaching of 15N-Labeled Coated Urea in a Lysimeter System in the North China Plain

    Institute of Scientific and Technical Information of China (English)

    LI Gui-Hua; ZHAO Lin-Ping; ZHANG Shu-Xiang; Y. HOSEN; K. YAGI3

    2011-01-01

    The effectiveness of polyolefin-coated urea (Meister-5 and Meister-10; CU) in a wheat (Triticum aestivum L.)-maize (Zea mays L.) rotation system was studied in lysimeter plots located in the North China Plain for three consecutive maize- wheat-maize cropping seasons.An isotopic method was used to compare the fate of CU to that of non-coated urea (NCU),and N application rates of 0,100,150 and 225 kg N ha-1 were evaluated.The results showed that the nitrogen use efficiency (15NUE) of CU was 13.3% 21.4% greater than that of NCU for the first crop.Alternatively,when the difference method was applied (apparent NUE),no significant variations were observed among treatments in all three seasons.Although inorganic N leached from the 1.3 m layer was less than 1% of the total applied N,unidentified losses of 15N (losses of 15N =15N applied as fertilizer - 15N absorbed by crops - 15N remaining in the 0-0.2 m layer 15N leached from the 1.3 m layer)in CU-treated plots were 24.2%-26.5% lower than those of NCU-treated plots.The nitrate concentration in the 0-1.3 m layer of CU plots at the end of the experiment was 53% lower than that of NCU-treated plots.Thus,CU increased crop N uptake from fertilizer and reduced unidentified losses of applied N,which can reduce the risk of groundwater pollution.

  3. Nitrate removal in stream ecosystems measured by 15N addition experiments: Denitrification

    Science.gov (United States)

    Mulholland, P.J.; Hall, R.O.; Sobota, D.J.; Dodds, W.K.; Findlay, S.E.G.; Grimm, N. B.; Hamilton, S.K.; McDowell, W.H.; O'Brien, J. M.; Tank, J.L.; Ashkenas, L.R.; Cooper, L.W.; Dahm, Clifford N.; Gregory, S.V.; Johnson, S.L.; Meyer, J.L.; Peterson, B.J.; Poole, G.C.; Valett, H.M.; Webster, J.R.; Arango, C.P.; Beaulieu, J.J.; Bernot, M.J.; Burgin, A.J.; Crenshaw, C.L.; Helton, A.M.; Johnson, L.T.; Niederlehner, B.R.; Potter, J.D.; Sheibley, R.W.; Thomasn, S.M.

    2009-01-01

    We measured denitrification rates using a field 15N-NO- 3 tracer-addition approach in a large, cross-site study of nitrate uptake in reference, agricultural, and suburban-urban streams. We measured denitrification rates in 49 of 72 streams studied. Uptake length due to denitrification (SWden) ranged from 89 m to 184 km (median of 9050 m) and there were no significant differences among regions or land-use categories, likely because of the wide range of conditions within each region and land use. N2 production rates far exceeded N2O production rates in all streams. The fraction of total NO-3 removal from water due to denitrification ranged from 0.5% to 100% among streams (median of 16%), and was related to NHz 4 concentration and ecosystem respiration rate (ER). Multivariate approaches showed that the most important factors controlling SWden were specific discharge (discharge / width) and NO-3 concentration (positive effects), and ER and transient storage zones (negative effects). The relationship between areal denitrification rate (Uden) and NO- 3 concentration indicated a partial saturation effect. A power function with an exponent of 0.5 described this relationship better than a Michaelis-Menten equation. Although Uden increased with increasing NO- 3 concentration, the efficiency of NO-3 removal from water via denitrification declined, resulting in a smaller proportion of streamwater NO-3 load removed over a given length of stream. Regional differences in stream denitrification rates were small relative to the proximate factors of NO-3 concentration and ecosystem respiration rate, and land use was an important but indirect control on denitrification in streams, primarily via its effect on NO-3 concentration. ?? 2009.

  4. Nitrate retention and removal in Mediterranean streams with contrasting land uses: a 15N tracer study

    Directory of Open Access Journals (Sweden)

    J. L. Riera

    2008-08-01

    Full Text Available We used 15N-labelled nitrate (NO−3 additions to investigate nitrogen (N cycling at the whole-reach scale in three Mediterranean streams subjected to contrasting land uses (i.e. forested, urban and agricultural. Our aim was to examine: i the magnitude and relative importance of NO−3 retention (i.e. assimilatory uptake, and removal, (i.e. denitrification, ii the relative contribution of the different primary uptake compartments to NO−3 retention, and iii the regeneration, transformation and export pathways of the retained N. The concentration of NO−3 increased and that of dissolved oxygen (DO decreased from the forested to the agricultural stream, with intermediate values in the urban stream. Standing stocks of primary uptake compartments were similar among streams and dominated by detritus compartments (i.e. fine and coarse benthic organic matter. In agreement, metabolism was net heterotrophic in all streams, although the degree of heterotrophy increased from the forested to the agricultural stream. The NO−3 uptake length decreased along this gradient, whereas the NO−3 mass-transfer velocity and the areal NO−3 uptake rate were highest in the urban stream. Denitrification was not detectable in the forested stream, but accounted for 9% and 68% of total NO−3 uptake in the urban and the agricultural stream, respectively. The relative contribution of detritus compartments to NO−3 assimilatory uptake was highest in the forested and lowest in the agricultural stream. In all streams, the retained N was rapidly transferred to higher trophic levels and regenerated back to the water column. Due to a strong coupling between regeneration and nitrification, most retained N was exported from the experimental reaches in the form of NO−3. This study evidences fast N cycling in Mediterranean streams. Moreover, results indicate that permanent NO−3 removal via denitrification may be enhanced over temporary NO−3 retention via assimilatory

  5. Multi-Isotopic (δ2H, δ13C, δ15N) Tracing of Molt Origin for Red-Winged Blackbirds Associated with Agro-Ecosystems

    Science.gov (United States)

    Werner, Scott J.; Hobson, Keith A.; Van Wilgenburg, Steven L.; Fischer, Justin W.

    2016-01-01

    We analyzed stable-hydrogen (δ2H), carbon (δ13C) and nitrogen (δ 15N) isotope ratios in feathers to better understand the molt origin and food habits of Red-winged Blackbirds (Agelaius phoeniceus) near sunflower production in the Upper Midwest and rice production in the Mid-South of the United States. Outer primary feathers were used from 661 after-second-year (ASY) male blackbirds collected in Minnesota, Montana, North Dakota and South Dakota (spring collection), and Arkansas, Louisiana, Mississippi, Missouri and Texas (winter collection). The best-fit model indicated that the combination of feather δ2H, δ13C and δ15N best predicted the state of sample collections and thus supported the use of this approach for tracing molt origins in Red-winged Blackbirds. When considering only birds collected in spring, 56% of birds were classified to their collection state on the basis of δ2H and δ13C alone. We then developed feather isoscapes for δ13C based upon these data and for δ2H based upon continental patterns of δ2H in precipitation. We used 81 birds collected at the ten independent sites for model validation. The spatially-explicit assignment of these 81 birds to the δ2H isoscape resulted in relatively high rates (~77%) of accurate assignment to collection states. We also modeled the spatial extent of C3 (e.g. rice, sunflower) and C4 (corn, millet, sorghum) agricultural crops grown throughout the Upper Midwest and Mid-South United States to predict the relative use of C3- versus C4-based foodwebs among sampled blackbirds. Estimates of C3 inputs to diet ranged from 50% in Arkansas to 27% in Minnesota. As a novel contribution to blackbird conservation and management, we demonstrate how such feather isoscapes can be used to predict the molt origin and interstate movements of migratory blackbirds for subsequent investigations of breeding biology (e.g. sex-specific philopatry), agricultural depredation, feeding ecology, physiology of migration and sensitivity to

  6. Potential for assessing long-term dynamics in soil nitrogen availability from variations in delta15N of tree rings.

    Science.gov (United States)

    Hart, S C; Classen, A T

    2003-03-01

    Numerous researchers have used the isotopic signatures of C, H, and O in tree rings to provide a long-term record of changes in the physiological status, climate, or water-source use of trees. The frequently limiting element N is also found in tree rings, and variation in its isotopic signature may provide insight into long-term changes in soil N availability of a site. However, research has suggested that N is readily translocated among tree ring of different years; such infidelity between the isotopic compositions of the N taken up from the soil and the N contained in the ring of that growth year would obscure the long-term N isotopic record. We used a 15-year 15N-tracer study to assess the degree of N translocation among tree rings in ponderosa pine (Pinus ponderosa) trees growing in a young, mixed-conifer plantation. We also measured delta13C and delta15N values in unlabeled trees to assess the degree of their covariance in wood tissue, and to explore the potential for a biological linkage between them. We found that the maximum delta15N values in rings from the labeled trees occurred in the ring formed one-year after the 15N was applied to the roots. The delta15N value of rings from labeled trees declined exponentially and bidirectionally from this maximum peak, toward younger and older rings. The unlabeled trees showed considerable interannual variation in the delta15N values of their rings (up to 3 and 5 per thousand), but these values correlated poorly between trees over time and differed by as much as 6 per thousand. Removal of extractives from the wood reduced their delta15N value, but the change was fairly small and consistent among unlabeled trees. The delta13C and delta15N values of tree rings were correlated over time in only one of the unlabeled trees. Across all trees, both delta13C values of tree rings and annual stem wood production were well correlated with annual precipitation, suggesting that soil water balance is an important environmental

  7. Compound specific amino acid δ15N in marine sediments: A new approach for studies of the marine nitrogen cycle

    Science.gov (United States)

    Batista, Fabian C.; Ravelo, A. Christina; Crusius, John; Casso, Michael A.; McCarthy, Matthew D.

    2014-10-01

    The nitrogen (N) isotopic composition (δ15N) of bulk sedimentary N (δ15Nbulk) is a common tool for studying past biogeochemical cycling in the paleoceanographic record. Empirical evidence suggests that natural fluctuations in the δ15N of surface nutrient N are reflected in the δ15N of exported planktonic biomass and in sedimentary δ15Nbulk. However, δ15Nbulk is an analysis of total combustible sedimentary N, and therefore also includes mixtures of N sources and/or selective removal or preservation of N-containing compounds. Compound-specific nitrogen isotope analyses of individual amino acids (δ15NAA) are novel measurements with the potential to decouple δ15N changes in nutrient N from trophic effects, two main processes that can influence δ15Nbulk records. As a proof of concept study to examine how δ15NAA can be applied in marine sedimentary systems, we compare the δ15NAA signatures of surface and sinking POM sources with shallow surface sediments from the Santa Barbara Basin, a sub-oxic depositional environmental that exhibits excellent preservation of sedimentary organic matter. Our results demonstrate that δ15NAA signatures of both planktonic biomass and sinking POM are well preserved in such surface sediments. However, we also observed an unexpected inverse correlation between δ15N value of phenylalanine (δ15NPhe; the best AA proxy for N isotopic value at the base of the food web) and calculated trophic position. We used a simple N isotope mass balance model to confirm that over long time scales, δ15NPhe values should in fact be directly dependent on shifts in ecosystem trophic position. While this result may appear incongruent with current applications of δ15NAA in food webs, it is consistent with expectations that paleoarchives will integrate N dynamics over much longer timescales. We therefore propose that for paleoceanographic applications, key δ15NAA parameters are ecosystem trophic position, which determines relative partitioning of 15N

  8. Direct measurement of the 15N(p,gamma)16O total cross section at novae energies

    CERN Document Server

    Bemmerer, D; Bonetti, R; Broggini, C; Confortola, F; Corvisiero, P; Costantini, H; Elekes, Z; Formicola, A; Fülöp, Z; Gervino, G; Guglielmetti, A; Gustavino, C; Gyurky, Gy; Junker, M; Limata, B; Marta, M; Menegazzo, R; Prati, P; Roca, V; Rolfs, C; Alvarez, C Rossi; Somorjai, E; Straniero, O

    2009-01-01

    The 15N(p,gamma)16O reaction controls the passage of nucleosynthetic material from the first to the second carbon-nitrogen-oxygen (CNO) cycle. A direct measurement of the total 15N(p,gamma)16O cross section at energies corresponding to hydrogen burning in novae is presented here. Data have been taken at 90-230 keV center-of-mass energy using a windowless gas target filled with nitrogen of natural isotopic composition and a bismuth germanate summing detector. The cross section is found to be a factor two lower than previously believed.

  9. Glacial-interglacial dynamics of Antarctic firn columns: comparison between simulations and ice core air-δ15N measurements

    Directory of Open Access Journals (Sweden)

    R. Mulvaney

    2012-12-01

    Full Text Available Correct estimate of the firn lock-in depth is essential for correctly linking gas and ice chronologies in ice cores studies. Here, two approaches to constrain the firn depth evolution in Antarctica are presented over the last deglaciation: output of a firn densification model and measurements of δ15N of N2 in air trapped in ice core. Since the firn densification process is largely governed by surface temperature and accumulation rate, we have investigated four ice cores drilled in coastal (Berkner Island, BI, and James Ross Island, JRI and semi coastal (TALDICE and EPICA Dronning Maud Land, EDML Antarctic regions. Combined with available δ15N measurements performed from the EPICA Dome C (EDC site, the studied regions encompass a large range of surface accumulation rate and temperature conditions. While firn densification simulations are able to correctly represent most of the δ15N trends over the last deglaciation measured in the EDC, BI, TALDICE and EDML ice cores, they systematically fail to capture BI and EDML δ15N glacial levels, a mismatch previously seen for Central East Antarctic ice cores. Using empirical constraints of the EDML gas-ice depth offset during the Laschamp event (~ 41 ka, we can rule out the existence of a large convective zone as the explanation of the glacial firn model-δ15N data mismatch for this site. The good match between modelled and measured δ15N at TALDICE as well as the lack of any clear correlation between insoluble dust concentration in snow and δ15N records in the different ice cores suggest that past changes in loads of impurities are not the only main driver of glacial-interglacial changes in firn lock-in depth. We conclude that firn densification dynamics may instead be driven mostly by accumulation rate changes. The mismatch between modelled and measured δ15N may be due to inaccurate reconstruction of past accumulation rate or underestimated influence of accumulation rate in firnification models.

  10. Glacial–interglacial dynamics of Antarctic firn columns: comparison between simulations and ice core air-δ15N measurements

    OpenAIRE

    2013-01-01

    Correct estimation of the firn lock-in depth is essential for correctly linking gas and ice chronologies in ice core studies. Here, two approaches to constrain the firn depth evolution in Antarctica are presented over the last deglaciation: outputs of a firn densification model, and measurements of δ15N of N2 in air trapped in ice core, assuming that δ15N is only affected by gravitational fractionation in the firn column. Since the firn densification process is largely governed by...

  11. Impact of charring on cereal grain characteristics: linking prehistoric manuring practice to 15N signatures in archaeobotanical material

    DEFF Research Database (Denmark)

    Kanstrup, Marie; Thomsen, Ingrid Kaag; Mikkelsen, Peter Hambro;

    2012-01-01

    . However, despite attempts to deliberately tamper and distort the grain δ15N signature, the changes observed in this study were too small to be of any consequences for the archaeobotanical applicability of the method. Thus the isotope method offers unique evidence about prehistoric manuring practice....... in either a systematic or substantial way and conclude that manuring most likely will be detectable in archaeobotanical charred grains. As certain within-grain variability in δ15N existed, especially in the intensively manured grains, the resolution of this kind of information should be carefully considered...

  12. Analysis of carbon and nitrogen co-metabolism in yeast by ultrahigh-resolution mass spectrometry applying {sup 13}C- and {sup 15}N-labeled substrates simultaneously

    Energy Technology Data Exchange (ETDEWEB)

    Blank, Lars M. [TU Dortmund University, Laboratory of Chemical Biotechnology, Department of Biochemical and Chemical Engineering, Dortmund (Germany); RWTH Aachen University, Institute of Applied Microbiology - iAMB, Aachen Biology and Biotechnology - ABBt, Aachen (Germany); Desphande, Rahul R. [TU Dortmund University, Laboratory of Chemical Biotechnology, Department of Biochemical and Chemical Engineering, Dortmund (Germany); Michigan State University, Department of Plant Biology, East Lansing, MI (United States); Schmid, Andreas [TU Dortmund University, Laboratory of Chemical Biotechnology, Department of Biochemical and Chemical Engineering, Dortmund (Germany); Hayen, Heiko [Leibniz-Institut fuer Analytische Wissenschaften-ISAS-e.V, Dortmund (Germany); University of Wuppertal, Department of Food Chemistry, Wuppertal (Germany)

    2012-06-15

    Alternative metabolic pathways inside a cell can be deduced using stable isotopically labeled substrates. One prerequisite is accurate measurement of the labeling pattern of targeted metabolites. Experiments are generally limited to the use of single-element isotopes, mainly {sup 13}C. Here, we demonstrate the application of direct infusion nanospray, ultrahigh-resolution Fourier transform ion cyclotron resonance-mass spectrometry (FTICR-MS) for metabolic studies using differently labeled elemental isotopes simultaneously - i.e., {sup 13}C and {sup 15}N - in amino acids of a total protein hydrolysate. The optimized strategy for the analysis of metabolism by a hybrid linear ion trap-FTICR-MS comprises the collection of multiple adjacent selected ion monitoring scans. By limiting both the width of the mass range and the number of ions entering the ICR cell with automated gain control, sensitive measurements of isotopologue distribution were possible without compromising mass accuracy and isotope intensity mapping. The required mass-resolving power of more than 60,000 is only achievable on a routine basis by FTICR and Orbitrap mass spectrometers. Evaluation of the method was carried out by comparison of the experimental data to the natural isotope abundances of selected amino acids and by comparison to GC/MS results obtained from a labeling experiment with {sup 13}C-labeled glucose. The developed method was used to shed light on the complexity of the yeast Saccharomyces cerevisiae carbon-nitrogen co-metabolism by administering both {sup 13}C-labeled glucose and {sup 15}N-labeled alanine. The results indicate that not only glutamate but also alanine acts as an amino donor during alanine and valine synthesis. Metabolic studies using FTICR-MS can exploit new possibilities by the use of multiple-labeled elemental isotopes. (orig.)

  13. A Statistical Programme Assignment Model

    DEFF Research Database (Denmark)

    Rosholm, Michael; Staghøj, Jonas; Svarer, Michael

    When treatment effects of active labour market programmes are heterogeneous in an observable way  across the population, the allocation of the unemployed into different programmes becomes a particularly  important issue. In this paper, we present a statistical model designed to improve the present...... assignment  mechanism, which is based on the discretionary choice of case workers. This is done in a duration model context, using the timing-of-events framework to identify causal effects. We compare different assignment  mechanisms, and the results suggest that a significant reduction in the average...... duration of unemployment spells may result if a statistical programme assignment model is introduced. We discuss several issues regarding the  plementation of such a system, especially the interplay between the statistical model and  case workers....

  14. Recovery of {sup 15}N-urea in soil-plant system of tanzania grass pasture; Recuperacao de {sup 15}N-ureia no sistema solo-planta de pastagem de capim-Tanzania

    Energy Technology Data Exchange (ETDEWEB)

    Martha Junior, Geraldo Bueno; Vilela, Lourival [EMBRAPA Cerrados, Planaltina, DF (Brazil)], e-mail: gbmartha@cpac.embrapa.br; Corsi, Moacyr [Universidade de Sao Paulo (ESALQ/USP), Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz. Dept. de Zootecnica], e-mail: moa@esalq.usp.br; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis], e-mail: pcotrive@cena.usp.br

    2009-01-15

    The economic attractiveness and negative environmental impact of nitrogen (N) fertilization in pastures depend on the N use efficiency in the soil-plant system. However, the recovery of urea-{sup 15}N by Panicum maximum cv. Tanzania pastures, one of the most widely used forage species in intensified pastoral systems, is still unknown. This experiment was conducted in a randomized complete block design with four treatments (0, 40, 80 and 120 kg ha-1 of N-urea) and three replications, to determine the recovery of {sup 15}N urea by Tanzania grass. Forage production, total N content and N yield were not affected by fertilization (p > 0.05), reflecting the high losses of applied N under the experimental conditions. The recovery of {sup 15}N urea (% of applied N) in forage and roots was not affected by fertilization levels (p > 0.05), but decreased exponentially in the soil and soil-plant system (p < 0.05) with increasing urea doses. The amount of {sup 15}N (kg ha{sup -1}) in forage and roots (15 to 30 cm) increased with increasing urea doses (p < 0.05). (author)

  15. AC410 Unit 9 Assignment

    OpenAIRE

    Laynebaril1

    2017-01-01

     AC410 Unit 9 Assignment Click Link Below To Buy: http://hwcampus.com/shop/ac410-unit-9-assignment/   18–29. The CPA firm of Carson & Boggs LLP is performing an internal control audit in accordance with PCAOB Standard No. 5. The partner in charge of the engagement has asked you to explain the process of determining which controls to test. Describe the process, presenting each of the links in this process and a short summary of how the auditors approach each of them. v ...

  16. CARBON-RICH PRESOLAR GRAINS FROM MASSIVE STARS: SUBSOLAR {sup 12}C/{sup 13}C AND {sup 14}N/{sup 15}N RATIOS AND THE MYSTERY OF {sup 15}N

    Energy Technology Data Exchange (ETDEWEB)

    Pignatari, M. [Konkoly Observatory, Research Centre for Astronomy and Earth Sciences, Hungarian Academy of Sciences, Konkoly Thege Miklos ut 15-17, H-1121 Budapest (Hungary); Zinner, E. [Laboratory for Space Sciences and Physics Department, Washington University, St. Louis, MO 63130 (United States); Hoppe, P. [Max Planck Institute for Chemistry, D-55128 Mainz (Germany); Jordan, C. J.; Gibson, B. K. [E.A. Milne Centre for Astrophysics, Dept of Physics and Mathematics, University of Hull, HU6 7RX (United Kingdom); Trappitsch, R. [Department of the Geophysical Sciences and Chicago Center for Cosmochemistry, Chicago, IL 60637 (United States); Herwig, F. [Department of Physics and Astronomy, University of Victoria, Victoria, BC V8P5C2 (Canada); Fryer, C. [Computational Physics and Methods (CCS-2), LANL, Los Alamos, NM, 87545 (United States); Hirschi, R. [Keele University, Keele, Staffordshire ST5 5BG (United Kingdom); Timmes, F. X. [The Joint Institute for Nuclear Astrophysics, Notre Dame, IN 46556 (United States)

    2015-08-01

    Carbon-rich grains with isotopic anomalies compared to the Sun are found in primitive meteorites. They were made by stars, and carry the original stellar nucleosynthesis signature. Silicon carbide grains of Type X and C and low-density (LD) graphites condensed in the ejecta of core-collapse supernovae. We present a new set of models for the explosive He shell and compare them with the grains showing {sup 12}C/{sup 13}C and {sup 14}N/{sup 15}N ratios lower than solar. In the stellar progenitor H was ingested into the He shell and not fully destroyed before the explosion. Different explosion energies and H concentrations are considered. If the supernova shock hits the He-shell region with some H still present, the models can reproduce the C and N isotopic signatures in C-rich grains. Hot-CNO cycle isotopic signatures are obtained, including a large production of {sup 13}C and {sup 15}N. The short-lived radionuclides {sup 22}Na and {sup 26}Al are increased by orders of magnitude. The production of radiogenic {sup 22}Ne from the decay of {sup 22}Na in the He shell might solve the puzzle of the Ne-E(L) component in LD graphite grains. This scenario is attractive for the SiC grains of type AB with {sup 14}N/{sup 15}N ratios lower than solar, and provides an alternative solution for SiC grains originally classified as nova grains. Finally, this process may contribute to the production of {sup 14}N and {sup 15}N in the Galaxy, helping to produce the {sup 14}N/{sup 15}N ratio in the solar system.

  17. Human baby hair amino acid natural abundance 15N-isotope values are not related to the 15N-isotope values of amino acids in mother's breast milk protein.

    Science.gov (United States)

    Romek, Katarzyna M; Julien, Maxime; Frasquet-Darrieux, Marine; Tea, Illa; Antheaume, Ingrid; Hankard, Régis; Robins, Richard J

    2013-12-01

    Since exclusively breast-suckled infants obtain their nutrient only from their mother's milk, it might be anticipated that a correlation will exist between the (15)N/(14)N isotope ratios of amino acids of protein of young infants and those supplied by their mother. The work presented here aimed to determine whether amino nitrogen transfer from human milk to infant hair protein synthesized within the first month of life conserves the maternal isotopic signature or whether post-ingestion fractionation dominates the nitrogen isotope spectrum. The study was conducted at 1 month post-birth on 100 mother-infant pairs. Isotope ratios (15)N/(14)N and (13)C/(12)C were measured using isotope ratio measurement by Mass Spectrometry (irm-MS) for whole maternal milk, and infant hair and (15)N/(14)N ratios were also measured by GC-irm-MS for the N-pivaloyl-O-isopropyl esters of amino acids obtained from the hydrolysis of milk and hair proteins. The δ(15)N and δ(13)C (‰) were found to be significantly higher in infant hair than in breast milk (δ(15)N, P amino acids in infant hair was also significantly higher than that in maternal milk (P < 0.001). By calculation, the observed shift in isotope ratio was shown not to be accounted for by the amino acid composition of hair and milk proteins, indicating that it is not simply due to differences in the composition in the proteins present. Rather, it would appear that each pool-mother and infant-turns over independently, and that fractionation in infant N-metabolism even in the first month of life dominates over the nutrient N-content.

  18. Assigning agents to a line

    DEFF Research Database (Denmark)

    Hougaard, Jens Leth; Moreno-Ternero, Juan D.; Østerdal, Lars Peter Raahave

    2014-01-01

    minimizing modification of the classic random priority method to solve this class of problems. We also provide some logical relations in our setting among standard axioms in the literature on assignment problems, and explore the robustness of our results to several extensions of our setting....

  19. Who Benefits from Homework Assignments?

    Science.gov (United States)

    Ronning, Marte

    2011-01-01

    Using Dutch data on pupils in elementary school this paper is the first empirical study to analyze whether assigning homework has a heterogeneous impact on pupil achievement. Addressing potential biases by using a difference-in-difference approach, I find that the test score gap is larger in classes where everybody gets homework than in classes…

  20. Assessing Contributions to Group Assignments

    Science.gov (United States)

    Johnston, Lucy; Miles, Lynden

    2004-01-01

    We report the use of a combination of self- and peer-assessment in an undergraduate social psychology laboratory course. Students worked in small groups on a self-directed empirical project that they each wrote up independently as a laboratory report. Marks for the written assignment were moderated by a contribution index measure based on the…

  1. Heteronuclear transverse and longitudinal relaxation in AX4 spin systems: application to (15)N relaxations in (15)NH4(+).

    Science.gov (United States)

    Werbeck, Nicolas D; Hansen, D Flemming

    2014-09-01

    The equations that describe the time-evolution of transverse and longitudinal (15)N magnetisations in tetrahedral ammonium ions, (15)NH4(+), are derived from the Bloch-Wangsness-Redfield density operator relaxation theory. It is assumed that the relaxation of the spin-states is dominated by (1) the intra-molecular (15)N-(1)H and (1)H-(1)H dipole-dipole interactions and (2) interactions of the ammonium protons with remote spins, which also include the contribution to the relaxations that arise from the exchange of the ammonium protons with the bulk solvent. The dipole-dipole cross-correlated relaxation mechanisms between each of the (15)N-(1)H and (1)H-(1)H interactions are explicitly taken into account in the derivations. An application to (15)N-ammonium bound to a 41kDa domain of the protein DnaK is presented, where a comparison between experiments and simulations show that the ammonium ion rotates rapidly within its binding site with a local correlation time shorter than approximately 1ns. The theoretical framework provided here forms the basis for further investigations of dynamics of AX4 spin systems, with ammonium ions in solution and bound to proteins of particular interest.

  2. Range expansion of the jumbo squid in the NE Pacific: δ15N decrypts multiple origins, migration and habitat use.

    Science.gov (United States)

    Ruiz-Cooley, Rocio I; Ballance, Lisa T; McCarthy, Matthew D

    2013-01-01

    Coincident with climate shifts and anthropogenic perturbations, the highly voracious jumbo squid Dosidicus gigas reached unprecedented northern latitudes along the NE Pacific margin post 1997-98. The physical or biological drivers of this expansion, as well as its ecological consequences remain unknown. Here, novel analysis from both bulk tissues and individual amino acids (Phenylalanine; Phe and Glutamic acid; Glu) in both gladii and muscle of D. gigas captured in the Northern California Current System (NCCS) documents for the first time multiple geographic origins and migration. Phe δ(15)N values, a proxy for habitat baseline δ(15)N values, confirm at least three different geographic origins that were initially detected by highly variable bulk δ(15)N values in gladii for squid at small sizes (60 cm) converged, indicating feeding in a common ecosystem. The strong latitudinal gradient in Phe δ(15)N values from composite muscle samples further confirmed residency at a point in time for large squid in the NCCS. These results contrast with previous ideas, and indicate that small squid are highly migratory, move into the NCCS from two or more distinct geographic origins, and use this ecosystem mainly for feeding. These results represent the first direct information on the origins, immigration and habitat use of this key "invasive" predator in the NCCS, with wide implications for understanding both the mechanisms of periodic D. gigas population range expansions, and effects on ecosystem trophic structure.

  3. Range expansion of the jumbo squid in the NE Pacific: δ15N decrypts multiple origins, migration and habitat use.

    Directory of Open Access Journals (Sweden)

    Rocio I Ruiz-Cooley

    Full Text Available Coincident with climate shifts and anthropogenic perturbations, the highly voracious jumbo squid Dosidicus gigas reached unprecedented northern latitudes along the NE Pacific margin post 1997-98. The physical or biological drivers of this expansion, as well as its ecological consequences remain unknown. Here, novel analysis from both bulk tissues and individual amino acids (Phenylalanine; Phe and Glutamic acid; Glu in both gladii and muscle of D. gigas captured in the Northern California Current System (NCCS documents for the first time multiple geographic origins and migration. Phe δ(15N values, a proxy for habitat baseline δ(15N values, confirm at least three different geographic origins that were initially detected by highly variable bulk δ(15N values in gladii for squid at small sizes (60 cm converged, indicating feeding in a common ecosystem. The strong latitudinal gradient in Phe δ(15N values from composite muscle samples further confirmed residency at a point in time for large squid in the NCCS. These results contrast with previous ideas, and indicate that small squid are highly migratory, move into the NCCS from two or more distinct geographic origins, and use this ecosystem mainly for feeding. These results represent the first direct information on the origins, immigration and habitat use of this key "invasive" predator in the NCCS, with wide implications for understanding both the mechanisms of periodic D. gigas population range expansions, and effects on ecosystem trophic structure.

  4. Determination of the natural abundance δ15N of taurine by gas chromatography-isotope ratio measurement mass spectrometry.

    Science.gov (United States)

    Tea, Illa; Antheaume, Ingrid; Besnard, Jorick; Robins, Richard J

    2010-12-15

    The measurement of the nitrogen isotope ratio of taurine (2-aminoethanesulphonic acid) in biological samples has a large number of potential applications. Taurine is a small water-soluble molecule which is notoriously difficult to analyze due to its polarity and functionality. A method is described which allows the determination of the natural abundance δ(15)N values of taurine and structural analogues, such as 3-amino-1-propanesulphonic acid (APSA), by isotope ratio mass spectrometry interfaced to gas chromatography (GC-irm-MS). The one-step protocol exploits the simultaneous derivatization of both functionalities of these aminosulphonic acids by reaction with triethylorthoacetate (TEOA). Conditions have been established which ensure quantitative reaction thus avoiding any nitrogen isotope fractionation during derivatization and workup. The differences in the δ(15)N values of derivatized and non-derivatized taurine and APSA all fall within the working range of 0.4‰ (-0.02 to 0.39‰). When applied to four sources of taurine with various δ(15)N values, the method achieved excellent reproducibility and accuracy. The optimized method enables the determination of the natural abundance δ(15)N values of taurine over the concentration range 1.5-7.84 µmol.mL(-1) in samples of biological origin.

  5. Determination of 15N abundance in nanogram pools of NO3 - and NO2 - by denitrification bioassay and mass spectrometry

    DEFF Research Database (Denmark)

    Højberg, Ole; Johansen, H. S.; Sorensen, J.

    1994-01-01

    Suspensions of two strains of Pseudomonas aeruginosa (ON12 and ON12-1) were used to reduce NO3 - and NO2 -, respectively, to N2O. The evolved N2O was quantified by gas chromatography with electron capture detection, and the 15N abundance was determined by mass spectrometry with a special inlet sy...

  6. The influence of fish cage culture on δ13C and δ15N of filter-feeding Bivalvia (Mollusca).

    Science.gov (United States)

    Benedito, E; Figueroa, L; Takeda, A M; Manetta, G I

    2013-11-01

    The objective of this study was to evaluate the effect of Oreochromis niloticus cage culture promoted variations in the δ13C and δ15N in Corbicula fluminea (Mollusca; Bivalvia) and in the sediment of an aquatic food web. Samples were taken before and after net cage installation in the Rosana Reservoir (Paranapanema River, PR-SP). Samples of specimens of the bivalve filterer C. fluminea and samples of sediment were collected using a modified Petersen grab. All samples were dried in an oven (60 °C) for 72 hours, macerated to obtain homogenous fine powders and sent for carbon (δ13C) and nitrogen (δ15N) isotopic value analysis in a mass spectrometer. There were significant differences in the δ13C and δ15N values of the invertebrate C. fluminea between the beginning and the end of the experiment. There were no differences between the δ13C and δ15N values of sediment. These results indicate that the installation of fish cage culture promoted impacts in the isotopic composition of the aquatic food web organisms, which could exert influence over the native species and the ecosystem.

  7. Variation in hair δ13C and δ15N values in long-tailed macaques (Macaca fascicularis) from Singapore

    Science.gov (United States)

    Schillaci, Michael A.; Castellini, J. Margaret; Stricker, Craig A.; Jones-Engel, Lisa; Lee, Benjamin P.Y.-H.

    2014-01-01

    Much of the primatology literature on stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) has focused on African and New World species, with comparatively little research published on Asian primates. Here we present hair δ13C and δ15N isotope values for a sample of 33 long-tailed macaques from Singapore. We evaluate the suggestion by a previous researcher that forest degradation and biodiversity loss in Singapore have led to a decline in macaque trophic level. The results of our analysis indicated significant spatial variability in δ13C but not δ15N. The range of variation in δ13C was consistent with a diet based on C3 resources, with one group exhibiting low values consistent with a closed canopy environment. Relative to other macaque species from Europe and Asia, the macaques from Singapore exhibited a low mean δ13C value but mid-range mean δ15N value. Previous research suggesting a decline in macaque trophic level is not supported by the results of our study.

  8. Analysis of 15N incorporation into D-alanine: a new method for tracing nitrogen uptake by bacteria

    NARCIS (Netherlands)

    Veuger, B.; Middelburg, J.J.; Boschker, H.T.S.; Houtekamer, M.J.

    2005-01-01

    The quantitative contribution of bacteria to total microbial uptake of nitrogenous substrates is an aspect of the aquatic nitrogen cycle that is still largely unclear, mainly because existing methods are generally inadequate. We investigated the feasibility of measuring 15N incorporation into bacter

  9. Contrasting food web linkages for the grazing pathway in 3 temperate forested streams using {sup 15}N as a tracer

    Energy Technology Data Exchange (ETDEWEB)

    Tank, J.L.; Mulholland, P.J.; Meyer, J.L.; Bowden, W.B.; Webster, J.R.; Peterson, B.J.

    1998-11-01

    Nitrogen is a critical element controlling the productivity and dynamics of stream ecosystems and many streams are limited by the supply of biologically available nitrogen. The authors are learning more about the fate of inorganic nitrogen entering streams through {sup 15}N tracer additions. The Lotic Intersite Nitrogen Experiment (LINX) is studying the uptake, cycling, and fate of {sup 15}N-NH{sub 4} in the stream food web of 10 streams draining different biomes. Using the {sup 15}N tracer method and data from 3 sites in the study, the authors can differentiate patterns in the cycling of nitrogen through the grazing pathway (N from the epilithon to grazing macroinvertebrates) for 3 temperate forested streams. Here, they quantify the relationship between the dominant grazer and its proposed food resource, the epilithon, by comparing {sup 15}N levels of grazers with those of the epilithon, as well as the biomass, nitrogen content, and chlorophyll a standing stocks of the epilithon in 3 streams.

  10. Hyperpolarized (13) C,(15) N2 -Urea MRI for assessment of the urea gradient in the porcine kidney

    DEFF Research Database (Denmark)

    Hansen, Esben S S; Stewart, Neil J; Wild, Jim M;

    2016-01-01

    function in healthy porcine kidneys resembling the human physiology. METHODS: Five healthy female Danish domestic pigs (weight 30 kg) were scanned at 3 Tesla (T) using a (13) C 3D balanced steady-state MR pulse sequence following injection of hyperpolarized (13) C,(15) N2 -urea via a femoral vein catheter...

  11. Radiocarbon, 13C and 15N analysis of fossil bone: Removal of humates with XAD-2 resin

    Science.gov (United States)

    Stafford, Thomas W., Jr.; Brendel, Klaus; Duhamel, Raymond C.

    1988-09-01

    Humic acids are the predominant source of error in the 14C and stable isotope analysis of fossil bone organic matter. XAD-2 resin will quantitatively remove humates and give the highest yields of protein from bones with variable types of preservation. Decalcified bone, gelatin and base-leached residues can vary up to 5%. for δ 13C and by 1%. on δ 15N relative to XAD-treated fractions. Simultaneous analysis of 14C age, δ 13C and δ 15N is recommended because each isotope value can be independently affected by the bone's diagenetic history. Radiocarbon analysis is the most sensitive and δ 15N is least sensitive for detecting exogenous organic matter. The uncertainty of analyses on the best pretreated protein is ±0.5%. for both δ 13C and δ 15N and is larger than previous estimates. The accuracy for all isotope analyses will be better assessed by using individual amino acids instead of total collagenous residues. Inaccurate 14C dates on severely degraded bone are an indication that this class of fossils may be unsuitable for any isotopic analysis.

  12. Proton-decoupled CPMG: a better experiment for measuring (15)N R2 relaxation in disordered proteins.

    Science.gov (United States)

    Yuwen, Tairan; Skrynnikov, Nikolai R

    2014-04-01

    (15)N R2 relaxation is one of the most informative experiments for characterization of intrinsically disordered proteins (IDPs). Small changes in nitrogen R2 rates are often used to determine how IDPs respond to various biologically relevant perturbations such as point mutations, posttranslational modifications and weak ligand interactions. However collecting high-quality (15)N relaxation data can be difficult. Of necessity, the samples of IDPs are often prepared with low protein concentration and the measurement time can be limited because of rapid sample degradation. Furthermore, due to hardware limitations standard experiments such as (15)N spin-lock and CPMG can sample the relaxation decay only to ca. 150ms. This is much shorter than (15)N T2 times in disordered proteins at or near physiological temperature. As a result, the sampling of relaxation decay profiles in these experiments is suboptimal, which further lowers the precision of the measurements. Here we report a new implementation of the proton-decoupled (PD) CPMG experiment which allows one to sample (15)N R2 relaxation decay up to ca. 0.5-1s. The new experiment has been validated through comparison with the well-established spin-lock measurement. Using dilute samples of denatured ubiquitin, we have demonstrated that PD-CPMG produces up to 3-fold improvement in the precision of the data. It is expected that for intrinsically disordered proteins the gains may be even more substantial. We have also shown that this sequence has a number of favorable properties: (i) the spectra are recorded with narrow linewidth in nitrogen dimension; (ii) (15)N offset correction is small and easy to calculate; (iii) the experiment is immune to various spurious effects arising from solvent exchange; (iv) the results are stable with respect to pulse miscalibration and rf field inhomogeneity; (v) with minimal change, the pulse sequence can also be used to measure R2 relaxation of (15)N(ε) spins in arginine side chains. We

  13. Retrospective characterization of ontogenetic shifts in killer whale diets via δ13C and δ15N analysis of teeth

    Science.gov (United States)

    Newsome, Seth D.; Etnier, Michael A.; Monson, Daniel H.; Fogel, Marilyn L.

    2009-01-01

    Metabolically inert, accretionary structures such as the dentin growth layers in teeth provide a life history record of individual diet with near-annual resolution. We constructed ontogenetic δ13C and δ15N profiles by analyzing tooth dentin growth layers from 13 individual killer whales Orcinus orca collected in the eastern northeast Pacific Ocean between 1961 and 2003. The individuals sampled were 6 to 52 yr old, representing 2 ecotypes—resident and transient—collected across ~25° of latitude. The average isotopic values of transient individuals (n = 10) are consistent with a reliance on mammalian prey, while the average isotopic values of residents (n = 3) are consistent with piscivory. Regardless of ecotype, most individuals show a decrease in δ15N values of ~2.5‰ through the first 3 yr of life, roughly equivalent to a decrease of one trophic level. We interpret this as evidence of gradual weaning, after which, ontogenetic shifts in isotopic values are highly variable. A few individuals (n = 2) maintained relatively stable δ15N and δ13C values throughout the remainder of their lives, whereas δ15N values of most (n = 11) increased by ~1.5‰, suggestive of an ontogenetic increase in trophic level. Significant differences in mean δ13C and δ15N values among transients collected off California suggest that individuality in prey preferences may be prevalent within this ecotype. Our approach provides retrospective individual life history and dietary information that cannot be obtained through traditional field observations of free-ranging and elusive species such as killer whales, including unique historic ecological information that pre-dates modern studies. By providing insights into individual diet composition, stable isotope analysis of teeth and/or bones may be the only means of evaluating a number of hypothesized historical dietary shifts in killer whales of the northeast Pacific Ocean

  14. A 15N stable isotope semen label to detect mating in the malaria mosquito Anopheles arabiensis Patton

    Directory of Open Access Journals (Sweden)

    Gludovacz Doris

    2008-07-01

    Full Text Available Abstract In previous studies it was determined that the stable isotope 13-carbon can be used as a semen label to detect mating events in the malaria mosquito Anopheles arabiensis. In this paper we describe the use of an additional stable isotope, 15-nitrogen (15N, for that same purpose. Both stable isotopes can be analysed simultaneously in a mass spectrometer, offering the possibility to detect both labels in one sample in order to study complex and difficult-to-detect mating events, such as multiple mating. 15N-glycine was added to larval rearing water and the target enrichment was 5 atom% 15N. Males from these trays were mated with unlabelled virgin females, and spiked spermathecae were analysed for isotopic composition after mating using mass spectrometry. Results showed that spermathecae positive for semen could be distinguished from uninseminated or control samples using the raw δ15N‰ values. The label persisted in spermathecae for up to 5 days after insemination, and males aged 10 days transferred similar amounts of label as males aged 4 days. There were no negative effects of the label on larval survival and male longevity. Enrichment of teneral mosquitoes after emergence was 4.85 ± 0.10 atom% 15N. A threshold value defined as 3 standard deviations above the mean of virgin (i.e. uninseminated spermathecae samples was successful in classifying a large proportion of samples correctly (i.e. on average 95%. We conclude that alongside 13C, 15N can be used to detect mating in Anopheles and the suitability of both labels is briefly discussed.

  15. Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies.

    Science.gov (United States)

    Martínez-Alcántara, Belén; Martínez-Cuenca, Mary-Rus; Fernández, Carlos; Legaz, Francisco; Quiñones, Ana

    2016-01-01

    Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale) two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively) fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L.) grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form) to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95%) ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency.

  16. Bradyrhizobium strain and the {sup 15}N natural abundance quantification of biological N{sub 2} fixation in soybean

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes, Ana Paula [Universidade Estadual do Norte Fluminense (UENF), Campos dos Goytacases, RJ (Brazil). Centro de Ciencias e Tecnologias Agropecuarias (CCTA). Dept. de Producao Vegetal; Morais, Rafael Fiusa de; Urquiaga, Segundo; Boddey, Robert Michael; Alves, Bruno Jose Rodrigues [EMBRAPA Agrobiologia, Seropedica, RJ (Brazil)]. E-mail: bruno@cnpab.embrapa.br

    2008-09-15

    In commercial plantations of soybean in both the Southern and the Cerrado regions, contributions from biological nitrogen fixation (BNF) are generally proportionately high. When using the {sup 15}N natural abundance technique to quantify BNF inputs, it is essential to determine, with accuracy, the {sup 15}N abundance of the N derived from BNF (the 'B' value). This study aimed to determine the effect of four recommended strains of Bradyrhizobium spp. (two B. japonicum and two B. elkanii) on the 'B' value of soybean grown in pots in an open field using an equation based on the determination of {delta}{sup 15}N natural abundance in a non-labelled soil, and estimate of the contribution of BNF derived from the use of {sup 15}N-isotope dilution in soils enriched with {sup 15}N. To evaluate N{sub 2} fixation by soybean, three non-N{sub 2}-fixing reference crops were grown under the same conditions. Regardless of Bradyrhizobium strain, no differences were observed in dry matter, nodule weight and total N between labelled and non-labelled soil. The N{sub 2} fixation of the soybeans grown in the two soil conditions were similar. The mean 'B' values of the soybeans inoculated with the B. japonicum strains were -1.84 per mille and -0.50 per mille, while those inoculated with B. elkanii were -3.67 per mille and -1.0 per mille, for the shoot tissue and the whole plant, respectively. Finally, the 'B' value for the soybean crop varied considerably in function of the inoculated Bradyrhizobium strain, being most important when only the shoot tissue was utilised to estimate the proportion of N in the plant derived from N{sub 2} fixation. (author)

  17. Transporte do 15N e produtividade do tomateiro enxertado irrigado com água carbonatada Transport of 15N and yield of the grafted tomato irrigated with carbonated water

    Directory of Open Access Journals (Sweden)

    Roberto Botelho Ferraz Branco

    2007-03-01

    Full Text Available Verificou-se a ação do dióxido de carbono dissolvido na água de irrigação e da enxertia do tomateiro no transporte de 15N nos tecidos da planta e na produção da cultura. Os tratamentos foram constituídos pela aplicação de CO2, 5 e 10 L min-1, mais a testemunha (dose zero e plantas enxertadas e pés-francos de tomateiro. O experimento foi conduzido no delineamento inteiramente casualizado, em esquema fatorial 3x2, com três repetições. As variáveis analisadas nas folhas e nos frutos da planta foram a produção de massa seca, o nitrogênio total, o excesso de 15N, a quantidade de nitrogênio proveniente do fertilizante e a porcentagem de recuperação do fertilizante e a produção de frutos comerciais. De acordo com os resultados estatísticos não houve diferença significativa entre os tratamentos do experimento em todas as variáveis analisadas. O CO2 dissolvido na água de irrigação e a enxertia do tomateiro não interferiram no transporte de 15N para os tecidos da parte aérea do tomateiro e tampouco em sua produção.We evaluated the action of the carbon dioxide dissolved in the irrigation water and the grafting of the tomato in the transport of 15N in the tissue of the plant and in the production of the culture. The treatments were the CO2 doses (0; 5 and 10 L min-1 and grafted and ungrafted tomato plant. These treatments were arranged in a 3x2 factorial scheme, in completely randomized design, with three replications. The variables analysed in the leaves and fruits were dry mass production, total nitrogen, excess of 15N, amount of nitrogen originated from the fertilizer, percentage of recovery of the fertilizer and commercial fruits production. There was no significant statistical difference among the treatments for any variable. The carbon dioxide dissolved in the irrigation water and the grafting of the tomato did not interfere in the transport of 15N to the shoots tissue neither in the yield.

  18. Graphical interpretation of Boolean operators for protein NMR assignments

    Energy Technology Data Exchange (ETDEWEB)

    Verdegem, Dries [Universite des Sciences et Technologies de Lille, Unite de Glycobiologie Structurale et Fonctionelle, UMR 8576 CNRS, IFR 147 (France); Dijkstra, Klaas [University of Groningen, Department of Biophysical Chemistry (Netherlands); Hanoulle, Xavier; Lippens, Guy [Universite des Sciences et Technologies de Lille, Unite de Glycobiologie Structurale et Fonctionelle, UMR 8576 CNRS, IFR 147 (France)], E-mail: guy.lippens@univ-lille1.fr

    2008-09-15

    We have developed a graphics based algorithm for semi-automated protein NMR assignments. Using the basic sequential triple resonance assignment strategy, the method is inspired by the Boolean operators as it applies 'AND'-, 'OR'- and 'NOT'-like operations on planes pulled out of the classical three-dimensional spectra to obtain its functionality. The method's strength lies in the continuous graphical presentation of the spectra, allowing both a semi-automatic peaklist construction and sequential assignment. We demonstrate here its general use for the case of a folded protein with a well-dispersed spectrum, but equally for a natively unfolded protein where spectral resolution is minimal.

  19. Selective excitation for spectral editing and assignment in separated local field experiments of oriented membrane proteins

    Science.gov (United States)

    Koroloff, Sophie N.; Nevzorov, Alexander A.

    2017-01-01

    Spectroscopic assignment of NMR spectra for oriented uniformly labeled membrane proteins embedded in their native-like bilayer environment is essential for their structure determination. However, sequence-specific assignment in oriented-sample (OS) NMR is often complicated by insufficient resolution and spectral crowding. Therefore, the assignment process is usually done by a laborious and expensive "shotgun" method involving multiple selective labeling of amino acid residues. Presented here is a strategy to overcome poor spectral resolution in crowded regions of 2D spectra by selecting resolved "seed" residues via soft Gaussian pulses inserted into spin-exchange separated local-field experiments. The Gaussian pulse places the selected polarization along the z-axis while dephasing the other signals before the evolution of the 1H-15N dipolar couplings. The transfer of magnetization is accomplished via mismatched Hartmann-Hahn conditions to the nearest-neighbor peaks via the proton bath. By optimizing the length and amplitude of the Gaussian pulse, one can also achieve a phase inversion of the closest peaks, thus providing an additional phase contrast. From the superposition of the selective spin-exchanged SAMPI4 onto the fully excited SAMPI4 spectrum, the 15N sites that are directly adjacent to the selectively excited residues can be easily identified, thereby providing a straightforward method for initiating the assignment process in oriented membrane proteins.

  20. Ion microscopy analysis of the intracellular distribution of {sup 14}C-or {sup 15}N-labelled cytotoxic drugs; Analyse par microscopie ionique de la distribution intracellulaire de drogues cytotoxiques marquees par {sup 14}C ou {sup 15}N

    Energy Technology Data Exchange (ETDEWEB)

    Bisagni, E.; Carrez, D.; Croisy, A.; Favaudon, V.; Guerquin-Kern, J.L.; Nguyen, C.H.; Rivalle, C. [Institut Curie, 91 - Orsay (France); Dennebouy, R.; Slodzian, G. [Paris-11 Univ., 91 - Orsay (France); Galle, P. [Faculte de Medecine, 94 - Creteil (France)

    1994-12-31

    This study has concerned the localization, by the means of ion microscopy of human tumoral cells (in culture), of two synthesized cytostatic compounds: BD40 or Pazelliptine and Intoplicine. These two active compounds (cancer research) have been labelled either with {sup 15}N on the terminal amino dialkyl group of the lateral chain, either, for Intoplicine, with {sup 14}C on the carbon 3 of the aromatic cycle. These two drugs are considered as poisons for topo-isomerases, and they were used to localize these cellular multiplication key-enzymes. Unfortunately, pazelliptine and intoplicine specificity appeared much too low. However, encouraging results were obtained, particularly on {sup 15}N-labelling utilization. 4 figs., 9 refs.

  1. CONTRACT ASSIGNMENT – THEORETICAL ASPECTS

    Directory of Open Access Journals (Sweden)

    Bogdan NAZAT

    2016-12-01

    Full Text Available This project aims to study in detail the theoretical aspects concerning the contract assignment, as provided by the relevant regulation, and the doctrine corresponding to old and current regulations. In this respect, this project aims to give the reader a comprehensive look on the institution in question, the regulation offered by the current Civil Code is reviewed taking into account the national and international doctrine.

  2. Simple approach for the preparation of 15-15N2-enriched water for nitrogen fixation assessments: Evaluation, application and recommendations

    Directory of Open Access Journals (Sweden)

    Isabell eKlawonn

    2015-08-01

    Full Text Available Recent findings revealed that the commonly used 15N2 tracer assay for the determination of dinitrogen (N2 fixation can underestimate the activity of aquatic N2-fixing organisms. Therefore, a modification to the method using pre-prepared 15-15N2-enriched water was proposed. Here, we present a rigorous assessment and outline a simple procedure for the preparation of 15-15N2-enriched water. We recommend to fill sterile-filtered water into serum bottles and to add 15-15N2 gas to the water in amounts exceeding the standard N2 solubility, followed by vigorous agitation (vortex mixing ≥5 min. Optionally, water can be degassed at low-pressure (≥950 mbar for ten minutes prior to the 15-15N2 gas addition to indirectly facilitate the 15-15N2 dissolution. This preparation of 15-15N2-enriched water can be done within one hour using standard laboratory equipment. The final 15N-atom% excess was 5% after replacing 2–5% of the incubation volume with 15-15N2-enriched water. Notably, the addition of 15-15N2-enriched water can alter levels of trace elements in the incubation water due to the contact of 15-15N2-enriched water with glass, plastic and rubber ware during its preparation. In our tests, levels of trace elements (Fe, P, Mn, Mo, Cu, Zn increased by up to 0.1 nmol L-1 in the final incubation volume, which may bias rate measurements in regions where N2 fixation is limited by trace elements. For these regions, we tested an alternative way to enrich water with 15-15N2. The 15-15N2 was injected as a bubble directly to the incubation water, followed by gentle shaking. Immediately thereafter, the bubble was replaced with water to stop the 15-15N2 equilibration. This method achieved a 15N-atom excess of 6.6±1.7% when adding 2 mL 15-15N2 per liter of incubation water. The herein presented methodological tests offer guidelines for the 15N2 tracer assay and thus, are crucial to circumvent methodological draw-backs for future N2 fixation assessments.

  3. Plot-size for {sup 15}N-fertilizer recovery studies by tanzania-grass; Tamanho da parcela para estudos de recuperacao de fertilizante-{sup 15}N por capim-tanzania

    Energy Technology Data Exchange (ETDEWEB)

    Martha Junior, Geraldo Bueno [EMBRAPA Cerrados, Planaltina, DF (Brazil)], e-mail: gbmartha@cpac.embrapa.br; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis], e-mail: pcotrive@cena.usp.br; Corsi, Moacyr [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Zootecnia], e-mail: moa@esalq.usp.br

    2009-07-01

    The understanding of the N dynamics in pasture ecosystems can be improved by studies using the {sup 15}N tracer technique. However, in these experiments it must be ensured that the lateral movement of the labeled fertilizer does not interfere with the results. In this study the plot-size requirements for {sup 15}N-fertilizer recovery experiments with irrigated Panicum maximum cv. Tanzania was determined. Three grazing intensities (light, moderate and intensive grazing) in the winter, spring and summer seasons were considered. A 1 m{sup 2} plot-size, with a grass tussock in the center, was adequate, irrespective of the grazing intensity or season of the year. Increasing the distance from the area fertilized with {sup 15}N negatively affected the N derived from fertilizer (Npfm) recovered in herbage.The lowest decline in Npfm values were observed for moderate and light grazing intensities. This fact might be explained by the vigorous growth characteristics of these plants. Increasing the grazing intensity decreased the tussock mass and, the smaller the tussock mass, the greater was the dependence on fertilizer nitrogen. (author)

  4. A note on ranking assignments using reoptimization

    DEFF Research Database (Denmark)

    Pedersen, Christian Roed; Nielsen, L.R.; Andersen, K.A.

    2005-01-01

    We consider the problem of ranking assignments according to cost in the classical linear assignment problem. An algorithm partitioning the set of possible assignments, as suggested by Murty, is presented where, for each partition, the optimal assignment is calculated using a new reoptimization...

  5. An algorithm for ranking assignments using reoptimization

    DEFF Research Database (Denmark)

    Pedersen, Christian Roed; Nielsen, Lars Relund; Andersen, Kim Allan

    2008-01-01

    We consider the problem of ranking assignments according to cost in the classical linear assignment problem. An algorithm partitioning the set of possible assignments, as suggested by Murty, is presented where, for each partition, the optimal assignment is calculated using a new reoptimization...... technique. Computational results for the new algorithm are presented...

  6. The natural abundance of 15N in litter and soil profiles under six temperate tree species: N cycling depends on tree species traits and site fertility

    DEFF Research Database (Denmark)

    Callesen, Ingeborg; Nilsson, Lars Ola; Schmidt, Inger Kappel;

    2013-01-01

    We investigated the influence of tree species on the natural 15N abundance in forest stands under elevated ambient N deposition.We analysed δ15N in litter, the forest floor and three mineral soil horizons along with ecosystem N status variables at six sites planted three decades ago with five...... European broadleaved tree species and Norway spruce.Litter δ15N and 15N enrichment factor (δ15Nlitter–δ15Nsoil) were positively correlated with N status based on soil and litter N pools, nitrification, subsoil nitrate concentration and forest growth. Tree species differences were also significant...... species leached more nitrate.The δ15N pattern reflected tree species related traits affecting the N cycling as well as site fertility and former land use, and possibly differences in N leaching. The tree species δ15N patterns reflected fractionation caused by uptake of N through mycorrhiza rather than due...

  7. Separation Between d$_{5/2}$ and s$_{1/2}$ Neutron Single Particle Strength in $^{15}$N

    CERN Document Server

    Mertin, C E; Crisp, A M; Keeley, N; Kemper, K W; Momotyuk, O; Roeder, B T; Volya, A

    2014-01-01

    The separation between single particle levels in nuclei plays the dominant role in determining the location of the neutron drip line. The separation also provides a test of current crossed shell model interactions if the experimental data is such that multiple shells are involved. The present work uses the $^{14}$N(d, p)$^{15}$N reaction to extract the 2s$_{1/2}$, and 1d$_{5/2}$ total neutron single particle strengths and then compares these results with a shell model calculation using a p-sd crossed shell interaction to identify the J$^\\pi$ of all levels in $^{15}$N up to 12.8 MeV in excitation.

  8. O potencial da rotulação metabólica de 15N para a pesquisa de esquizofrenia

    Directory of Open Access Journals (Sweden)

    Michaela D. Filiou

    2013-01-01

    Full Text Available Pesquisas em psiquiatria ainda necessitam de estudos não dirigidos por hipóteses para revelar fundamentos neurobiológicos e biomarcadores moleculares para distúrbios psiquiátricos. Metodologias proteômicas disponibilizam uma série de ferramentas para esses fins. Apresentamos o princípio de rotulação metabólica utilizando 15N para proteômica quantitativa e suas aplicações em modelos animais de fenótipos psiquiátricos com um foco particular em esquizofrenia. Exploramos o potencial de rotulação metabólica por 15N em diferentes tipos de experimentos, bem como suas considerações metodológicas.

  9. Determining the source of nitrate pollution in the Niger discontinuous aquifers using the natural {15N }/{14N } ratios

    Science.gov (United States)

    Girard, Pierre; Hillaire-Marcel, Claude

    1997-12-01

    In the semi-arid Niamey area (Niger), more than 10% of the deep wells exploiting the fracture network of the Precambrian aquifer are contaminated by nitrates, with concentrations as high as 10 meq l -1. In order to identify the source(s) of this pollution, nitrate and 15N contents in the polluted wells were monitored over a 20-month period. Potential sources of nitrate contamination were also analyzed for their 15N content. The isotopic compositions of nitrate in polluted waters were > + 12‰ and in rare cases exceeded +17‰. Latrines (˜ + 15‰) may be the major nitrate source for wells showing δ15N values above +15‰. Below this value, waters may be polluted by a combination of nitrates from both latrine and soil sources (˜ + 10‰). In some cases, the soil may account for up to 85% of the groundwater nitrate load. This mode of groundwater pollution is thought to be a consequence of deforestation. Despite their reputation as polluting agents, fertilizers ( +0.5 < δ 15N < + 3.6‰ ) which are used in rice paddies close to the contaminated areas, do not appear to be a significant source of nitrate contamination. Denitrification is probably not a significant process in the study area. Results suggest that nitrate contamination of the aquifer is a consequence of unregulated urbanization (home-made latrines) and deforestation. While latrines are limited to the urban zones, intensive cutting of the forest to meet the city dwellers' wood demand occurs in an ever increasing area around the capital, threatening the local water supply.

  10. Use of a 15N tracer to determine linkages between a mangrove and an upland freshwater swamp

    Science.gov (United States)

    MacKenzie, R. A.; Cormier, N.

    2005-05-01

    Mangrove forests and adjacent upland freshwater swamps are important components of subsistence-based economies of Pacific islands. Mangroves provide valuable firewood (Rhizophora apiculata) and mangrove crabs (Scylla serrata); intact freshwater swamps are often used for agroforestry (e.g., taro cultivation). While these two systems are connected hydrologically via groundwater and surface flows, little information is available on how they may be biogeochemically or ecologically linked. For example, mangrove leaf litter was once thought to be an important food source for resident and transient nekton and invertebrates, but this value may have been overestimated. Instead, nutrients or allochthonous material (e.g., phytoplankton, detritus) delivered via groundwater or surface water from upland freshwater swamps may play a larger role in mangrove food webs. Understanding the linkages between these two ecologically and culturally important ecosystems will help us to understand the potential impacts of hydrological alterations that occur when roads or bridges are constructed through them. We conducted a 15N tracer study in the Yela watershed on the island of Kosrae, Federated States of Micronesia. K15NO3 was continually added at trace levels for 4 weeks to the Yela River in an upland freshwater swamp adjacent to a mangrove forest. Nitrate and ammonium pools, major primary producers, macroinvertebrates, and fish were sampled from stations 5 m upstream (freshwater swamp) and 138, 188, 213, and 313 m downstream (mangrove) from the tracer addition. Samples were collected once a week prior to, during, and after the 15N addition for a total of 6 weeks. Preliminary results revealed no significant enrichment (< 1 ‰) in the 15N isotope composition of either resident shrimp (Macrobrachium sp.) or mudskipper fish (Periophthalmus sp.). However, the 15N signature of ammonium pools was enriched 10-60 ‰ by the end of the third week. These results suggest that the tracer was present

  11. Quantitative study on the fate of residual soil nitrate in winter wheat based on a 15N-labeling method

    Science.gov (United States)

    Zhang, Jing-Ting; Wang, Zhi-Min; Liang, Shuang-Bo; Zhang, Ying-Hua; Lu, Lai-Qing; Wang, Run-Zheng

    2017-01-01

    A considerable amount of surplus nitrogen (N), which primarily takes the form of nitrate, accumulates in the soil profile after harvesting crops from an intensive production system in the North China Plain. The residual soil nitrate (RSN) is a key factor that is included in the N recommendation algorithm. Quantifying the utilization and losses of RSN is a fundamental necessity for optimizing crop N management, improving N use efficiency, and reducing the impact derived from farmland N losses on the environment. In this study, a 15N-labeling method was introduced to study the fate of the RSN quantitatively during the winter wheat growing season by 15N tracer technique combined with a soil column study. A soil column with a 2 m height was vertically divided into 10 20-cm layers, and the RSN in each layer was individually labeled with a 15N tracer before the wheat was sown. The results indicated that approximately 17.68% of the crop N derived from RSN was located in the 0–2 m soil profile prior to wheat sowing. The wheat recovery proportions of RSN at various layers ranged from 0.21% to 33.46%. The percentages that still remained in the soil profile after the wheat harvest ranged from 47.08% to 75.44%, and 19.46–32.64% of the RSN was unaccounted for. Upward and downward movements in the RSN were observed, and the maximum upward and downward distances were 40 cm and 100 cm, respectively. In general, the 15N-labeling method contributes to a deeper understanding of the fates of the RSN. Considering the low crop recovery of the RSN from deep soil layers, water and N saving practices should be adopted during crop production. PMID:28170440

  12. Evaluating mercury biomagnification in fish from a tropical marine environment using stable isotopes (delta13C and delta15N).

    Science.gov (United States)

    Al-Reasi, Hassan A; Ababneh, Fuad A; Lean, David R

    2007-08-01

    Concentrations of total mercury (T-Hg) and methylmercury (MeHg) were measured in zooplankton and 13 fish species from a coastal food web of the Gulf of Oman, an arm of the Arabian Sea between Oman and Iran. Stable isotope ratios (delta13C and delta15N) also were determined to track mercury biomagnification. The average concentration of T-Hg in zooplankton was 21 +/- 8.0 ng g(-1) with MeHg accounting 10% of T-Hg. Total mercury levels in fish species ranged from 3.0 ng g(-1) (Sardinella longiceps) to 760 ng g(-1) (Rhizoprionodon acutus) with relatively lower fraction of MeHg (72%) than that found in other studies. The average trophic difference (Deltadelta13C) between zooplankton and planktivorous fish (Selar crumenopthalmus, Rastrelliger kanagurta, and S. longiceps) was higher (3.4 per thousandth) than expected, suggesting that zooplankton may not be the main diet or direct carbon source for these fish species. However, further sampling would be required to compensate for temporal changes in zooplankton and the influence of their lipid content. Trophic position inferred by delta15N and and slopes of the regression equations (log10[T-Hg] = 0.13[delta15N] - 3.57 and log10[MeHg] = 0.14[delta15N] - 3.90) as estimates of biomagnification indicate that biomagnification of T-Hg and MeHg was lower in this tropical ocean compared to what has been observed in arctic and temperate ecosystems and tropical African lakes. The calculated daily intake of methylmercury in the diet of local people through fish consumption was well below the established World Health Organization (WHO) tolerable daily intake threshold for most of the fish species except Euthynnus affinis, Epinephelus epistictus, R. acutus, and Thunnus tonggol, illustrating safe consumption of the commonly consumed fish species.

  13. Fate of 15N-urea applied to wheat-soybean succession crop Destino de 15N-uréia aplicada em sucessão trigo-soja

    Directory of Open Access Journals (Sweden)

    Antonio Enedi Boaretto

    2004-01-01

    Full Text Available The wheat crop in São Paulo State, Brazil, is fertilized with N, P and K. The rate of applied N (0 to 120 kg.ha-1 depends on the previous grown crop and the irrigation possibility. The response of wheat to rates and time of N application and the fate of N applied to irrigated wheat were studied during two years. Residual N recovery by soybean grown after the wheat was also studied. The maximum grain productivity was obtained with 92 kg.ha-1 of N. The efficiency of 15N-urea utilization ranged from 52% to 85%. The main loss of applied 15N, 5% to 12% occurred as ammonia volatilized from urea applied on soil surface. The N loss by leaching even at the N rate of 135 kg.ha-1, was less than 1% of applied 15N, due to the low amount of rainfall during the wheat grown season and a controlled amount of irrigated water, that were sufficient to moisten only the wheat root zone. The residual 15N after wheat harvest represents around 40% of N applied as urea: 20% in soil, 3% in wheat root system and 16% in the wheat straw. Soybean recovered less than 2% of the 15N applied to wheat at sowing or at tillering stage.No Estado de São Paulo, a cultura do trigo é adubada, além de P e K, com N, cuja dose (0 a 120 kg ha-1 depende do cultivo anterior e da possibilidade de irrigação. A resposta do trigo às doses e épocas de aplicação e o destino do N aplicado foi estudada em dois cultivos de trigo, seguidos pela soja. Também se avaliou a recuperação do N residual pela soja cultivada nas mesmas parcelas após o trigo. A produtividade máxima estimada de grãos seria obtida com a dose de 92 kg.ha-1 de N. A eficiência de absorção 15N-uréia variou de 52% a 85%. A principal perda de N, que variou de 5% a 12%, ocorreu através de volatilização de amônia proveniente da uréia aplicada na superfície do solo. Por lixiviação foi perdido menos que 1% do N aplicado, pois a água da chuva ou da irrigação foi suficiente para molhar somente a camada do solo

  14. Tracing the diet of the monitor lizard Varanus mabitang by stable isotope analyses (δ15N, δ13C)

    Science.gov (United States)

    Struck, Ulrich; Altenbach, Alexander; Gaulke, Maren; Glaw, Frank

    2002-09-01

    In this study, we used analyses of stable isotopes of nitrogen (δ15N) and carbon (δ13C) to determine the trophic ecology of the monitor lizard Varanus mabitang. Stable isotopes from claws, gut contents, and soft tissues were measured from the type specimen. Samples from Varanus olivaceus, Varanus prasinus, Varanus salvator, the herbivorous agamid lizard Hydrosaurus pustulatus, and some plant matter were included for comparison. Our data show a rapid decrease in δ13C (about10‰) from food plants towards gut contents and soft tissues of herbivorous species. For the varanids, we found a significant linear correlation of decreasing δ13C and increasing δ15N from herbivorous towards carnivorous species. In terms of trophic isotope ecology, the type specimen of V. mabitang is a strict herbivore. Thus it differs significantly in its isotopic composition from the morphologically next closest related species V. olivaceus. The most highly carnivorous species is V. salvator, while δ15N values for V. prasinus and V. olivaceus are intermediate. Claws provide very valuable samples for such measurements, because they can be sampled from living animals without harm. Additionally, their range of variability is relatively small in comparison with measurements from soft tissues.

  15. Patterns of δ13 C and δ15 N in wolverine Gulo gulo tissues from the Brooks Range, Alaska

    Institute of Scientific and Technical Information of China (English)

    Fredrik DALERUM; Anders ANGERBJ(O)RN; Kyran KUNKEL; Brad S.SHULTS

    2009-01-01

    Knowledge of carnivore diets is essential to understand hew carnivore populations respond demographically to variations in prey abundance. Analysis of stable isotopes is a useful complement to traditional methods of analyzing carnivore diets. We used data on δ13 C and δ15 N in wolverine tissues to investigate patterns of seasonal and annual diet variation in a wolverine Gulo gulo population in the western Brooks Range, Alaska, USA. The stable isotope ratios in wolverine tissues generally reflected that of terrestrial carnivores, corroborating previous diet studies on wolverines. We also found variation in δ13 C and δ15 N both between muscle samples collected over several years and between tissues with different assimilation rates, even after correcting for isotopic fractionation. This suggests both annual and seasonal diet variation. Our results indicate that data on δ13 C and δ15 N holds promise for qualitative assessments of wolverine diet changes over time. Such temporal variation may be important indicators of ecological responses to environmental perturbations, and we suggest that more refined studies of stable isotopes may be an important tool when studying temporal change in diets of wolverines and similar carnivores.

  16. Affordable uniform isotope labeling with {sup 2}H, {sup 13}C and {sup 15}N in insect cells

    Energy Technology Data Exchange (ETDEWEB)

    Sitarska, Agnieszka; Skora, Lukasz; Klopp, Julia; Roest, Susan; Fernández, César; Shrestha, Binesh; Gossert, Alvar D., E-mail: alvar.gossert@novartis.com [Novartis Institutes for BioMedical Research (Switzerland)

    2015-06-15

    For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80 % can be achieved for {sup 15}N and {sup 13}C with yields comparable to expression in full media. For {sup 2}H,{sup 15}N and {sup 2}H,{sup 13}C,{sup 15}N labeling, incorporation is only slightly lower with 75 and 73 %, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins.

  17. 15N NMR investigation of the covalent binding of reduced TNT amines to soil humic acid, model compounds, and lignocellulose.

    Science.gov (United States)

    Thorn, K A; Kennedy, K R

    2002-09-01

    The five major reductive degradation products of TNT-4ADNT (4-amino-2,6-dinitrotoluene), 2ADNT (2-amino-4,6-dinitrotoluene), 2,4DANT (2,4-diamino-6-nitrotoluene), 2,6DANT (2,6-diamino-4-nitrotoluene), and TAT (2,4,6-triaminotoluene)-labeled with 15N in the amine positions, were reacted with the IHSS soil humic acid and analyzed by 15N NMR spectrometry. In the absence of catalysts, all five amines underwent nucleophilic addition reactions with quinone and other carbonyl groups in the soil humic acid to form both heterocyclic and nonheterocyclic condensation products. Imine formation via 1,2-addition of the amines to quinone groups in the soil humic acid was significant with the diamines and TAT but not the monoamines. Horseradish peroxidase (HRP) catalyzed an increase in the incorporation of all five amines into the humic acid. In the case of the diamines and TAT, HRP also shifted the binding away from heterocyclic condensation product toward imine formation. A comparison of quantitative liquid phase with solid-state CP/MAS 15N NMR indicated that the CP experiment underestimated imine and heterocyclic nitrogens in humic acid, even with contact times optimal for observation of these nitrogens. Covalent binding of the mono- and diamines to 4-methylcatechol, the HRP catalyzed condensation of 4ADNT and 2,4DANT to coniferyl alcohol, and the binding of 2,4DANT to lignocellulose with and without birnessite were also examined.

  18. (13)C and (15)N NMR characterization of amine reactivity and solvent effects in CO2 capture.

    Science.gov (United States)

    Perinu, Cristina; Arstad, Bjørnar; Bouzga, Aud M; Jens, Klaus-J

    2014-08-28

    Factors influencing the reactivity of selected amine absorbents for carbon dioxide (CO2) capture, in terms of the tendency to form amine carbamate, have been studied. Four linear primary alkanolamines at varying chain lengths (MEA, 3A1P, 4A1B , and 5A1P ), two primary amines with different substituents in the β-position to the nitrogen (1A2P and ISOB), a secondary alkanolamine (DEA), and a sterically hindered primary amine (AMP) were investigated. The relationship between the (15)N NMR data of aqueous amines and their ability to form carbamate, as determined at equilibrium by quantitative (13)C NMR experiments, was analyzed, taking into account structural-chemical properties. For all the amines, the (15)N chemical shifts fairly reflected the observed reactivity for carbamate formation. In addition to being a useful tool for the investigation of amine reactivity, (15)N NMR data clearly provided evidence of the importance of solvent effects for the understanding of chemical dynamics in CO2 capture by aqueous amine absorbents.

  19. [Absorption and distribution of nitrogen from 15N labelled urea applied at core-hardening stage in winter jujube].

    Science.gov (United States)

    Zhao, Dengchao; Jiang, Yuanmao; Peng, Futian; Zhang, Jin; Zhang, Xu; Ju, Xiaotang; Zhang, Fusuo

    2006-01-01

    The study with pot experiment showed that at the rapid-swelling stage of winter jujube fruit, the percent of nitrogen derived from fertilizer (Ndff%) was the highest (10.64%) in fine roots, followed by new-growth nutritive organs. The absorbed urea-15N decreased in leaves and deciduous supers, and accumulated preferentially in root systems after harvest. The Ndff% in coarse roots was the highest (3.69%) before budding stage, while that in new-growth organs (new branches, deciduous supers, leaves and flowers) was the highest at full-blooming stage. The urea-15N applied at core-hardening stage mainly allocated in nutritive organs (leaves, deciduous supers, roots) in the first year, with the distribution rate 54.01% in root systems in winter, which was higher than that in branches (45.99%). The 15N stored in main branches changed drastically from post-harvest to budding stage. Main branches could be regarded as the 'target organs' of N storage, while coarse roots were the 'long-term sink' of N storage. The N reserve distributed preferentially in contiguity organs, and the distribution center changed with the growth and development of winter jujube in next spring.

  20. Individual protein balance strongly influences δ15N and δ13C values in Nile tilapia, Oreochromis niloticus

    Science.gov (United States)

    Gaye-Siessegger, Julia; Focken, Ulfert; Abel, Hansjörg; Becker, Klaus

    Although stable isotope ratios in animals have often been used as indicators of the trophic level and for the back-calculation of diets, few experiments have been done under standardized laboratory conditions to investigate factors influencing δ15N and δ13C values. An experiment using Nile tilapia [Oreochromis niloticus (L.)] was therefore carried out to test the effect of different dietary protein contents (35.4, 42.3, and 50.9%) on δ15N and δ13C values of the whole tilapia. The fish were fed the isoenergetic and isolipidic semi-synthetic diets at a relatively low level. δ15N and δ13C values of the lipid-free body did not differ between the fish fed the diets with different protein contents, but the trophic shift for N and C isotopes decreased with increasing protein accretion in the individual fish, for N from 6.5‰ to 4‰ and for C in the lipid-free body from 4‰ to 2.5‰. This is the first study showing the strong influence of the individual protein balance to the degree to which the isotopic signature of dietary protein was modified in tissue protein of fish. The extrapolation of the trophic level or the reconstruction of the diet of an animal from stable isotope ratios without knowledge of the individual physiological condition and the feeding rate may lead to erroneous results.

  1. Differentiation of Pigment in Eggs Using Carbon ((13)C/(12)C) and Nitrogen ((15)N/(14)N) Stable Isotopes.

    Science.gov (United States)

    Sun, Feng M; Shi, Guang Y; Wang, Hui W

    2016-07-01

    Consumers prefer natural and healthy food, but artificial pigments are often abused in egg products. The study aimed at differentiating the origin of pigments in eggs by applying the technique of carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) stable isotope analysis. Five hundred sixty laying hens were randomly distributed into 14 treatments, which were divided into four groups: maize, carophyll red pigment, carophyll yellow pigment, and a mixture of carophyll red and yellow pigments. Eggs were collected and pretreated to determe the values of the Roche Yolk Color Fan (RCF), δ(13)C, and δ(15)N. With increasing maize content, the RCF and δ(13)C values of yolks increased. Moreover, the RCF values in the three pigment groups were significantly influenced by the artificial colors, but δ(13)C values were not significantly different, regardless of the existence of pigment. The δ(15)N values in all treatments did not vary as regularly as the carbon stable isotope. A strong positive correlation was found between RCF and δ(13)C in the maize group, but no such correlation was be observed in the pigment groups. It is concluded that carbon stable isotope ratio analysis (δ(13)C) of the yolk can be used to differentiate the origin of the pigment added to eggs.

  2. 13C and 15N spectral editing inside histidine imidazole ring through solid-state NMR spectroscopy.

    Science.gov (United States)

    Li, Shenhui; Zhou, Lei; Su, Yongchao; Han, Bin; Deng, Feng

    2013-01-01

    Histidine usually exists in three different forms (including biprotonated species, neutral τ and π tautomers) at physiological pH in biological systems. The different protonation and tautomerization states of histidine can be characteristically determined by (13)C and (15)N chemical shifts of imidazole ring. In this work, solid-state NMR techniques were developed for spectral editing of (13)C and (15)N sites in histidine imidazole ring, which provides a benchmark to distinguish the existing forms of histidine. The selections of (13)Cγ, (13)Cδ2, (15)Nδ1, and (15)Nε2 sites were successfully achieved based on one-bond homo- and hetero-nuclear dipole interactions. Moreover, it was demonstrated that (1)H, (13)C, and (15) chemical shifts were roughly linearly correlated with the corresponding atomic charge in histidine imidazole ring by theoretical calculations. Accordingly, the (1)H, (13)C and (15)N chemical shifts variation in different protonation and tautomerization states could be ascribed to the atomic charge change due to proton transfer in biological process.

  3. Individual protein balance strongly influences delta15N and delta13C values in Nile tilapia, Oreochromis niloticus.

    Science.gov (United States)

    Gaye-Siessegger, Julia; Focken, Ulfert; Abel, Hansjörg; Becker, Klaus

    2004-02-01

    Although stable isotope ratios in animals have often been used as indicators of the trophic level and for the back-calculation of diets, few experiments have been done under standardized laboratory conditions to investigate factors influencing delta(15)N and delta(13)C values. An experiment using Nile tilapia [ Oreochromis niloticus (L.)] was therefore carried out to test the effect of different dietary protein contents (35.4, 42.3, and 50.9%) on delta(15)N and delta(13)C values of the whole tilapia. The fish were fed the isoenergetic and isolipidic semi-synthetic diets at a relatively low level. delta(15)N and delta(13)C values of the lipid-free body did not differ between the fish fed the diets with different protein contents, but the trophic shift for N and C isotopes decreased with increasing protein accretion in the individual fish, for N from 6.5 per thousand to 4 per thousand and for C in the lipid-free body from 4 per thousand to 2.5 per thousand. This is the first study showing the strong influence of the individual protein balance to the degree to which the isotopic signature of dietary protein was modified in tissue protein of fish. The extrapolation of the trophic level or the reconstruction of the diet of an animal from stable isotope ratios without knowledge of the individual physiological condition and the feeding rate may lead to erroneous results.

  4. 15N enrichment of ammonium, glutamine-amide and urea, measured via mass isotopomer analysis of hexamethylenetetramine.

    Science.gov (United States)

    Yang, D; Puchowicz, M A; David, F; Powers, L; Halperin, M L; Brunengraber, H

    1999-11-01

    Ammonium is an important intermediate of protein metabolism and is a key component of acid-base balance. Investigations of the metabolism of NH(4)(+) in vivo using isotopic techniques are difficult because of the low concentration of NH(4)(+) in biological fluids and because of frequent artifactual isotopic dilution of the enrichment of NH(4)(+) during the assay. A new gas chromatographic mass spectrometric method was designed to monitor the (15)N enrichment and concentration of NH(4)(+) in vivo. These are both calculated from the mass isotopomer distribution of hexamethylenetetramine (HMT) formed by reacting NH(4)(+) with formaldehyde. The enrichment of NH(4)(+) is amplified four times since the HMT molecule contains four atoms of nitrogen derived from NH(4)(+). This allows the measurement of low (15)N enrichment of NH(4)(+), down to 0.1%. (15)N enrichment of urea and of the amide N of L-glutamine are measured by enzymatic release of NH(4)(+) and conversion of the latter to HMT. These new techniques facilitate in vivo investigations of the metabolism of NH(4)(+) and related compounds.

  5. Nitrogen-containing compounds in two CR2 meteorites: 15N composition, molecular distribution and precursor molecules

    Science.gov (United States)

    Pizzarello, Sandra; Holmes, William

    2009-04-01

    Amino acids, amines and aldehydes were obtained from the water extracts of two CR2 carbonaceous chondrites from Antarctica and analyzed for their molecular and 15N isotopic content. These compounds were found to differ significantly from those of CM chondrites in both overall abundances and molecular distribution. The amino acids suites comprise a preponderant abundance of linear, 2-H amino acids, show rapid non-linear decrease with the compounds' increasing chain length and include protein amino acids never identified in meteorites before, such as threonine, tyrosine and phenylalanine. The presence of tertiary amines as well as a diverse, large abundance of aldehydes and ketones also distinguishes both CR2 organic suites. The δ 15N values determined for CR2 amino acids have a distribution between molecular subgroups that is opposite to the one of their δD values, with 2-H amino acids having higher δ 15N and lower δD values than 2-methyl amino acids, while the opposite is true for 2-methyl amino acids. Based on theoretical data, these isotopic findings would place the formation of the two amino acid groups or their direct precursors at different ISM stages of star formation.

  6. Fertilizer 15N balance in a coffee cropping system: a case study in Brazil Balanço do 15N do fertilizante em uma cultura de café: um estudo de caso no Brasil

    Directory of Open Access Journals (Sweden)

    Tatiele Anete Bergamo Fenilli

    2008-08-01

    Full Text Available Knowledge about the fate of fertilizer nitrogen in agricultural systems is essential for the improvement of management practices in order to maximize nitrogen (N recovery by the crop and reduce N losses from the system to a minimum. This study involves fertilizer management practices using the 15N isotope label applied in a single rate to determine the fertilizer-N balance in a particular soil-coffee-atmosphere system and to deepen the understanding of N plant dynamics. Five replicates consisting of plots of about 120 plants each were randomly defined within a 0.2 ha coffee plantation planted in 2001, in Piracicaba, SP, Brazil. Nine plants of each plot were separated in sub-plots for the 15N balance studies and treated with N rates of 280 and 350 kg ha-1 during 2003/2004 and 2004/2005, respectively, both of them as ammonium sulfate enriched to a 15N abundance of 2.072 atom %. Plant shoots were considered as separate parts: the orthotropic central branch, productive branches, leaves of productive branches, vegetative branches, leaves of vegetative branches and fruit. Litter, consisting of dead leaves accumulated below the plant canopy, was measured by the difference between leaves at harvest and at the beginning of the following flowering. Roots and soil were sampled down to a depth of 1.0 at intervals of 0.2 m. Samples from the isotopic sub-plots were used to evaluate total N and 15N, and plants outside sub-plots were used to evaluate dry matter. Volatilization losses of NH3 were estimated using special collectors. Leaching of fertilizer-N was estimated from deep drainage water fluxes and 15N concentrations of the soil solution at 1 m soil depth. At the end of the 2-year evaluation, the recovery of 15N applied as ammonium sulfate was 19.1 % in aerial plant parts, 9.4 % in the roots, 23.8 % in the litter, 26.3 % in the fruit and 12.6 % remaining in the 0_1.0 m soil profile. Annual leaching and volatilization losses were very small (2.0 % and 0

  7. Configuration-sensitive infrared bands and vibrational assignments of S-alkyldithizones based on isotopic substitutions

    Science.gov (United States)

    Yamada, Osamu; Hiura, Hidehumi; Igarashi, Takashi; Kaneko, Norio; Takahashi, Hiroaki

    By comparison of the infrared spectra of S-methyl-, S-ethyl- and S-isopropyidithizones in the solid state, the configuration-sensitive i.r. bands have been obtained for the trans-syn-s-trans and trans-anti-s-trans configurations with respect to the NN, CN and CN bonds of the formazan skeleton. The vibrational assignment of S-methyldithizone has been made based on the frequency shifts on isotopic substitutions: 1,5- 15N- and 2,4- 15N- substitutions, deuterium substitutions of the methyl group, phenyl groups and NH group, and deuterium substitutions of both phenyl and NH groups.

  8. 15N NMR study of nitrate ion structure and dynamics in hydrotalcite-like compounds

    Science.gov (United States)

    Hou, X.; James, Kirkpatrick R.; Yu, P.; Moore, D.; Kim, Y.

    2000-01-01

    We report here the first nuclear magnetic resonance (NMR) spectroscopic study of the dynamical and structural behavior of nitrate on the surface and in the interlayer of hydrotalcite-like compounds (15NO3--HT). Spectroscopically resolvable surface-absorbed and interlayer NO3- have dramatically different dynamical characteristics. The interlayer nitrate shows a well defined, temperature independent uniaxial chemical shift anisotropy (CS A) powder pattern. It is rigidly held or perhaps undergoes rotation about its threefold axis at all temperatures between -100 ??C and +80 ??C and relative humidities (R.H.) from 0 to 100% at room temperature. For surface nitrate, however, the dynamical behavior depends substantially on temperature and relative humidity. Analysis of the temperature and R.H. dependences of the peak width yields reorieritational frequencies which increase from essentially 0 at -100 ??C to 2.6 ?? 105 Hz at 60 ??C and an activation energy of 12.6 kJ/mol. For example, for samples at R.H. = 33%, the surface nitrate is isotropically mobile at frequencies greater than 105 Hz at room temperature, but it becomes rigid or only rotates on its threefold axis at -100 ??C. For dry samples and samples heated at 200 ??C (R.H. near 0%), the surface nitrate is not isotropically averaged at room temperature. In contrast to our previous results for 35Cl--containing hydrotalcite (35Cl--HT), no NMR detectable structural phase transition is observed for 15NO3--HT. The mobility of interlayer nitrate in HT is intermediate between that of carbonate and chloride.

  9. Redox-controlled backbone dynamics of human cytochrome c revealed by {sup 15}N NMR relaxation measurements

    Energy Technology Data Exchange (ETDEWEB)

    Sakamoto, Koichi [Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810 (Japan); Kamiya, Masakatsu [Graduate School of Life Science, Hokkaido University, Sapporo 060-0810 (Japan); Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810 (Japan); Uchida, Takeshi [Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810 (Japan); Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo 060-0810 (Japan); Kawano, Keiichi [Graduate School of Life Science, Hokkaido University, Sapporo 060-0810 (Japan); Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810 (Japan); Ishimori, Koichiro, E-mail: koichiro@sci.hokudai.ac.jp [Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810 (Japan); Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo 060-0810 (Japan)

    2010-07-23

    Research highlights: {yields} The dynamic parameters for the backbone dynamics in Cyt c were determined. {yields} The backbone mobility of Cyt c is highly restricted due to the covalently bound heme. {yields} The backbone mobility of Cyt c is more restricted upon the oxidation of the heme. {yields} The redox-dependent dynamics are shown in the backbone of Cyt c. {yields} The backbone dynamics of Cyt c would regulate the electron transfer from Cyt c. -- Abstract: Redox-controlled backbone dynamics in cytochrome c (Cyt c) were revealed by 2D {sup 15}N NMR relaxation experiments. {sup 15}N T{sub 1} and T{sub 2} values and {sup 1}H-{sup 15}N NOEs of uniformly {sup 15}N-labeled reduced and oxidized Cyt c were measured, and the generalized order parameters (S{sup 2}), the effective correlation time for internal motion ({tau}{sub e}), the {sup 15}N exchange broadening contributions (R{sub ex}) for each residue, and the overall correlation time ({tau}{sub m}) were estimated by model-free dynamics formalism. These dynamic parameters clearly showed that the backbone dynamics of Cyt c are highly restricted due to the covalently bound heme that functions as the stable hydrophobic core. Upon oxidation of the heme iron in Cyt c, the average S{sup 2} value was increased from 0.88 {+-} 0.01 to 0.92 {+-} 0.01, demonstrating that the mobility of the backbone is further restricted in the oxidized form. Such increases in the S{sup 2} values were more prominent in the loop regions, including amino acid residues near the thioether bonds to the heme moiety and positively charged region around Lys87. Both of the regions are supposed to form the interaction site for cytochrome c oxidase (CcO) and the electron pathway from Cyt c to CcO. The redox-dependent mobility of the backbone in the interaction site for the electron transfer to CcO suggests an electron transfer mechanism regulated by the backbone dynamics in the Cyt c-CcO system.

  10. Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies.

    Directory of Open Access Journals (Sweden)

    Belén Martínez-Alcántara

    Full Text Available Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L. grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95% ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency.

  11. Foliar and fungal 15N:14N ratios reflect development of mycorrhizae and nitrogen supply during primary succession: testing analytical models.

    Science.gov (United States)

    Hobbie, Erik A; Jumpponen, Ari; Trappe, Jim

    2005-12-01

    Nitrogen isotopes (15N/14N ratios, expressed as delta15N values) are useful markers of the mycorrhizal role in plant nitrogen supply because discrimination against 15N during creation of transfer compounds within mycorrhizal fungi decreases the 15N/14N in plants (low delta15N) and increases the 15N/14N of the fungi (high delta15N). Analytical models of 15N distribution would be helpful in interpreting delta15N patterns in fungi and plants. To compare different analytical models, we measured nitrogen isotope patterns in soils, saprotrophic fungi, ectomycorrhizal fungi, and plants with different mycorrhizal habits on a glacier foreland exposed during the last 100 years of glacial retreat and on adjacent non-glaciated terrain. Since plants during early primary succession may have only limited access to propagules of mycorrhizal fungi, we hypothesized that mycorrhizal plants would initially be similar to nonmycorrhizal plants in delta15N and then decrease, if mycorrhizal colonization were an important factor influencing plant delta15N. As hypothesized, plants with different mycorrhizal habits initially showed similar delta15N values (-4 to -6 per thousand relative to the standard of atmospheric N2 at 0 per thousand), corresponding to low mycorrhizal colonization in all plant species and an absence of ectomycorrhizal sporocarps. In later successional stages where ectomycorrhizal sporocarps were present, most ectomycorrhizal and ericoid mycorrhizal plants declined by 5-6 per thousand in delta15N, suggesting transfer of 15N-depleted N from fungi to plants. The values recorded (-8 to -11 per thousand) are among the lowest yet observed in vascular plants. In contrast, the delta15N of nonmycorrhizal plants and arbuscular mycorrhizal plants declined only slightly or not at all. On the forefront, most ectomycorrhizal and saprotrophic fungi were similar in delta15N (-1 to -3 per thousand), but the host-specific ectomycorrhizal fungus Cortinarius tenebricus had values of up to 7

  12. Backbone and side chain NMR assignments of Geobacillus stearothermophilus ZapA allow identification of residues that mediate the interaction of ZapA with FtsZ.

    Science.gov (United States)

    Nogueira, Maria Luiza C; Sforça, Mauricio Luis; Chin, Yanni K-Y; Mobli, Mehdi; Handler, Aaron; Gorbatyuk, Vitaliy Y; Robson, Scott A; King, Glenn F; Gueiros-Filho, Frederico J; Zeri, Ana Carolina de Mattos

    2015-10-01

    Bacterial division begins with the formation of a contractile protein ring at midcell, which constricts the bacterial envelope to generate two daughter cells. The central component of the division ring is FtsZ, a tubulin-like protein capable of self-assembling into filaments which further associate into a higher order structure known as the Z ring. Proteins that bind to FtsZ play a crucial role in the formation and regulation of the Z ring. One such protein is ZapA, a widely conserved 21 kDa homodimeric protein that associates with FtsZ filaments and promotes their bundling. Although ZapA was discovered more than a decade ago, the structural details of its interaction with FtsZ remain unknown. In this work, backbone and side chain NMR assignments for the Geobacillus stearothermophilus ZapA homodimer are described. We titrated FtsZ into (15)N(2)H-ZapA and mapped ZapA residues whose resonances are perturbed upon FtsZ binding. This information provides a structural understanding of the interaction between FtsZ and ZapA.

  13. Nitrogen ({sup 15}N) accumulation in corn grains as affected by source of nitrogen in red latosol;Acumulo de nitrogenio ({sup 15}N) pelos graos de milho em funcao da fonte nitrogenada em latossolo vermelho

    Energy Technology Data Exchange (ETDEWEB)

    Duete, Robson Rui Cotrim, E-mail: rrcduete@oi.com.b [Empresa Baiana de Desenvolvimento Agricola S.A. (EBDA), Cruz das Almas, BA (Brazil); Muraoka, Takashi; Trivelin, Paulo Cesar Ocheuze; Silva, Edson Cabral da, E-mail: muraoka@cena.usp.b, E-mail: pcotrive@cena.usp.b, E-mail: ecsilva@cena.usp.b [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil); Ambrosano, Edmilson Jose, E-mail: ambrosano@aptaregional.sp.gov.b [Agencia Paulista de Tecnologia dos Agronegocios (APTA), Piracicaba, SP (Brazil). Polo Centro Sul

    2009-07-01

    Nitrogen is the most absorbed mineral nutrient by corn crop and most affects grains yield. It is the unique nutrient absorbed by plants as cation (NH{sub 4}{sup +}) or anion (NO{sub 3}{sup -}). The objectives of this work were to investigate the N accumulation by corn grains applied to the soil as NH{sub 4}{sup +} or NO{sub 3}{sup -} in the ammonium nitrate form compared to amidic form of the urea, labeled with {sup 15}N; to determine the corn growth stage with highest fertilizer N utilization by the grains, and to quantify soil nitrogen exported by corn grains. The study was carried out in the Experimental Station of the Regional Pole of the Sao Paulo Northwestern Agribusiness Development (APTA), in Votuporanga, State of Sao Paulo, Brazil, in a Red Latosol. The experimental design was completely randomized blocks, with 13 treatments and four replications, disposed in factorial outline 6x2 + 1 (control, without N application). A nitrogen rate equivalent to 120 kg N ha-1 as urea-{sup 15}N or as ammonium nitrate, labeled in the cation NH{sub 4}{sup +} ({sup 15}NH{sub 4}{sup +}NO{sub 3}{sup -}) or in the anion NO{sub 3}{sup -} (NH{sub 4}{sup +}15N+O{sub 3}{sup -} ), was applied in six fractions of 20 kg N ha-1 each, in different microplots, from seeding to the growth stage 7 (pasty grains). The forms of nitrogen, NH{sub 4}{sup +}-N and N{sub O}{sup 3}--N, were accumulated equitably by corn grains. The corn grains accumulated more N from urea than from ammonium nitrate. The N applied to corn crop at eight expanded leaves stage promoted largest accumulation of this nutrient in the grains. (author)

  14. Assignment Choice, Effort, and Assignment Completion: Does Work Ethic Predict Those Who Choose Higher-Effort Assignments?

    Science.gov (United States)

    Parkhurst, John T.; Fleisher, Matthew S.; Skinner, Christopher H.; Woehr, David J.; Hawthorn-Embree, Meredith L.

    2011-01-01

    After completing the Multidimensional Work-Ethic Profile (MWEP), 98 college students were given a 20-problem math computation assignment and instructed to stop working on the assignment after completing 10 problems. Next, they were allowed to choose to finish either the partially completed assignment that had 10 problems remaining or a new…

  15. Natural (15)N Abundance in Key Amino Acids from Lamb Muscle: Exploring a New Horizon in Diet Authentication and Assessment of Feed Efficiency in Ruminants.

    Science.gov (United States)

    Cantalapiedra-Hijar, Gonzalo; Ortigues-Marty, Isabelle; Schiphorst, Anne-Marie; Robins, Richard J; Tea, Illa; Prache, Sophie

    2016-05-25

    Natural (15)N abundance (δ(15)N) varies between individual amino acids (AAs). We hypothesized that δ(15)N of nontransaminating and essential AAs ("source" AAs, such as phenylalanine) present in animal tissues could be used as a marker of dietary origin, whereas δ(15)N of transaminating AAs ("trophic" AAs, such as glutamic acid) could give more detailed insights into animal feed efficiency. Two diets based on dehydrated Lucerne pellets were tested in growing lambs, which promoted different feed efficiencies. No dietary effects were noted on δ(15)N of any AAs analyzed in lamb muscle. In addition, δ(15)N of phenylalanine was unexpectedly similar to that of glutamic acid, suggesting that δ(15)N of AAs is significantly derived from the metabolism of the rumen microbiota and, thus, are not suited for diet authentication in ruminants. In contrast, the δ(15)N of transaminating AAs facilitates an improved prediction of animal feed efficiency compared to the classical isotopic bulk N analysis.

  16. Quantitative analysis of 15N labeled positional isomers of glutamine and citrulline via electrospray ionization tandem mass spectrometry of their dansyl derivatives

    Science.gov (United States)

    The enteral metabolism of glutamine and citrulline are intertwined because, while glutamine is one of the main fuel sources for the enterocyte, citrulline is one of its products. It has been shown that the administration of 15N labeled glutamine results in the incorporation of the 15N label into cit...

  17. Experimental evidence for diel δ15N-patterns in different tissues, xylem and phloem saps of castor bean (Ricinus communis L.).

    Science.gov (United States)

    Peuke, A D; Gessler, A; Tcherkez, G

    2013-12-01

    Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ(15)N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ(15)N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ(15)N was correlated fairly well between phloem and xylem saps. These variations in δ(15)N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to (15)N-enriched glutamine acting as transport form of N. δ(15)N of the water soluble fraction of roots and leaves partially affected δ(15)N of phloem and xylems saps. δ(15)N patterns are likely the result of a complex set of interactions and N-fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course - a fact that needs to be taken into account when sampling for isotopic studies.

  18. More General Optimal Offset Assignment

    Directory of Open Access Journals (Sweden)

    Sven Mallach

    2015-06-01

    Full Text Available This manuscript presents exact approaches to the general offset assignment problem arising in the address code generation phase of compilers for application-specific processors. First, integer programming models for architecture-dependent and theoretically motivated special cases of the problem are established. Then, these models are extended to provide the first widely applicable formulations for the most general problem setting, supporting processors with several address registers and complex addressing capabilities. Existing heuristics are similarly extended and practical applicability of the proposed methods is demonstrated by experimental evaluation using an established and large benchmark set. The experiments allow us to study the impact of exploiting more complex memory addressing capabilities on the address computation costs of real-world programs. We also show how to integrate operand reordering techniques for commutative instructions into existing solution approaches.

  19. The Random Quadratic Assignment Problem

    Science.gov (United States)

    Paul, Gerald; Shao, Jia; Stanley, H. Eugene

    2011-11-01

    The quadratic assignment problem, QAP, is one of the most difficult of all combinatorial optimization problems. Here, we use an abbreviated application of the statistical mechanics replica method to study the asymptotic behavior of instances in which the entries of at least one of the two matrices that specify the problem are chosen from a random distribution P. Surprisingly, the QAP has not been studied before using the replica method despite the fact that the QAP was first proposed over 50 years ago and the replica method was developed over 30 years ago. We find simple forms for C min and C max , the costs of the minimal and maximum solutions respectively. Notable features of our results are the symmetry of the results for C min and C max and their dependence on P only through its mean and standard deviation, independent of the details of P.

  20. Dynamics of δ(15)N isotopic signatures of different intertidal macroalgal species: Assessment of bioindicators of N sources in coastal areas.

    Science.gov (United States)

    Lemesle, Stéphanie; Erraud, Alexandre; Mussio, Isabelle; Rusig, Anne-Marie; Claquin, Pascal

    2016-09-15

    δ(15)N of annual (Ulva sp., Porphyra sp.) and perennial intertidal seaweed species (Chondrus crispus, Fucus sp.) collected on 17 sampling points along the French coast of the English Channel in 2012 and 2013 were assessed on their suitability as bioindicators of N pollution in coastal areas. A sine function applied for δ(15)N time series data showed for all the species the same seasonal trend with lowest δ(15)N values in April and highest in summer but with no significant interspecific differences of amplitude (α) and phase angle (ϕ). This model provides a useful tool for monitoring the inter-annual changes of N pollution. An interspecific variability of δ(15)N values was observed, probably due to their tolerance to emersion. An in vitro study for comparing the kinetic acquisition of the isotopic signal and N uptake mechanisms of each species underlined the influence of algal physiology on the δ(15)N interspecific variability.

  1. Influence of 15N enrichment on the net isotopic fractionation factor during the reduction of nitrate to nitrous oxide in soil

    DEFF Research Database (Denmark)

    Mathieu, O.; Levegue, J.; Henault, C.

    2007-01-01

    Nitrous oxide, a greenhouse gas, is mainly emitted from soils during the denitrification process. Nitrogen stable-isotope investigations can help to characterise the N(2)O source and N(2)O production mechanisms. The stable-isotope approach is increasingly used with (15)N natural abundance...... or relatively low (15)N enrichment levels and requires a good knowledge of the isotopic fractionation effect inherent to this biological mechanism. This paper reports the measurement of the net and instantaneous isotopic fractionation factor (alpha(i)(s/p)) during the denitrification of NO(3)(-) to N(2)O over...... a range of (15)N substrate enrichments (0.37 to 1.00 atom% (15)N). At natural abundance level, the isotopic fractionation effect reported falls well within the range of data previously observed. For (15)N-enriched substrate, the value of alpha(i)(s/p) was not constant and decreased from 1.024 to 1...

  2. Determination of the delta(15N/14N)of Ammonium (NH4+) in Water: RSIL Lab Code 2898

    Science.gov (United States)

    Hannon, Janet E.; Böhlke, John Karl

    2008-01-01

    The purpose of the technique described by Reston Stable Isotope Laboratory (RSIL) lab code 2898 is to determine the N isotopic composition, delta(15N/14N), abbreviated as d15N, of ammonium (NH4+) in water (freshwater and saline water). The procedure involves converting dissolved NH4+ into NH3 gas by raising the pH of the sample to above 9 with MgO and subsequently trapping the gas quantitatively as (NH4)2SO4 on a glass fiber (GF) filter. The GF filter is saturated with NaHSO4 and pressure sealed between two gas-permeable polypropylene filters. The GF filter 'sandwich' floats on the surface of the water sample in a closed bottle. NH3 diffuses from the water through the polypropylene filter and reacts with NaHSO4, forming (NH4)2SO4 on the GF filter. The GF filter containing (NH4)2SO4 is dried and then combusted with a Carlo Erba NC 2500 elemental analyzer (EA), which is used to convert total nitrogen in a solid sample into N2 gas. The EA is connected to a continuous-flow isotope-ratio mass spectrometer (CF-IRMS), which determines the relative difference in ratios of the amounts of the stable isotopes of nitrogen (15N and 14N) of the product N2 gas and a reference N2 gas. The filters containing the samples are compressed in tin capsules and loaded into a Costech Zero-Blank Autosampler on the EA. Under computer control, samples then are dropped into a heated reaction tube that contains an oxidant, where combustion takes place in a He atmosphere containing an excess of O2 gas. To remove S-O gases produced from the NaHSO4, a plug of Ag-coated Cu wool is inserted at the bottom of the reaction tube. Combustion products are transported by a He carrier through a reduction furnace to remove excess O2, toconvert all nitrogen oxides to N2, and to remove any remaining S-O gases. The gases then pass through a drying tube to remove water. The gas-phase products, mainly N2 and a small amount of background CO2, are separated by a gas chromatograph (GC). The gas is then introduced

  3. Seasonal variation in nitrogen pools and 15N/13C natural abundances in different tissues of grassland plants

    Directory of Open Access Journals (Sweden)

    J. K. Schjoerring

    2011-12-01

    Full Text Available Seasonal changes in nitrogen (N pools, carbon (C content and natural abundance of 13C and 15N in different tissues of ryegrass plants were investigated in two intensively managed grassland fields in order to address their ammonia (NH3 exchange potential. Green leaves generally had the largest total N concentration followed by stems and inflorescences. Senescent leaves had the lowest N concentration, indicating N re-allocation. The seasonal pattern of the Γ value, i.e. the ratio between NH4+ and H+ concentrations, was similar for the various tissues of the ryegrass plants but the magnitude of Γ differed considerably among the different tissues. Green leaves and stems generally had substantially lower Γ values than senescent leaves and litter. Substantial peaks in Γ were observed during spring and summer in response to fertilization and grazing. These peaks were associated with high NH4+ rather than with low H+ concentrations. Peaks in Γ also appeared during the winter, coinciding with increasing δ15N values, indicating absorption of N derived from mineralization of soil organic matter. At the same time, δ13C values were declining, suggesting reduced photosynthesis and capacity for N assimilation. δ15N and δ13C values were more influenced by mean monthly temperature than by the accumulated monthly precipitation. In conclusion, ryegrass plants showed a clear seasonal pattern in N pools. Green leaves and stems of ryegrass plants generally seem to constitute a sink for NH3, while senescent leaves have a large potential for NH3 emission. However, management events such as fertilisation and grazing may create a high NH3 emission potential even in green plant parts. The obtained results provide input for future modelling of plant-atmosphere NH3 exchange.

  4. Unraveling the complexity of protein backbone dynamics with combined (13)C and (15)N solid-state NMR relaxation measurements.

    Science.gov (United States)

    Lamley, Jonathan M; Lougher, Matthew J; Sass, Hans Juergen; Rogowski, Marco; Grzesiek, Stephan; Lewandowski, Józef R

    2015-09-14

    Typically, protein dynamics involve a complex hierarchy of motions occurring on different time scales between conformations separated by a range of different energy barriers. NMR relaxation can in principle provide a site-specific picture of both the time scales and amplitudes of these motions, but independent relaxation rates sensitive to fluctuations in different time scale ranges are required to obtain a faithful representation of the underlying dynamic complexity. This is especially pertinent for relaxation measurements in the solid state, which report on dynamics in a broader window of time scales by more than 3 orders of magnitudes compared to solution NMR relaxation. To aid in unraveling the intricacies of biomolecular dynamics we introduce (13)C spin-lattice relaxation in the rotating frame (R1ρ) as a probe of backbone nanosecond-microsecond motions in proteins in the solid state. We present measurements of (13)C'R1ρ rates in fully protonated crystalline protein GB1 at 600 and 850 MHz (1)H Larmor frequencies and compare them to (13)C'R1, (15)N R1 and R1ρ measured under the same conditions. The addition of carbon relaxation data to the model free analysis of nitrogen relaxation data leads to greatly improved characterization of time scales of protein backbone motions, minimizing the occurrence of fitting artifacts that may be present when (15)N data is used alone. We also discuss how internal motions characterized by different time scales contribute to (15)N and (13)C relaxation rates in the solid state and solution state, leading to fundamental differences between them, as well as phenomena such as underestimation of picosecond-range motions in the solid state and nanosecond-range motions in solution.

  5. PARTITIONING THE RELATIVE INFLUENCE OF SOIL N, MYCORRHIZAE, AND FOLIAR N UPTAKE ON FOLIAR δ15N PATTERNS: CAN WE DETECT FOLIAR UPTAKE OF REACTIVE N?

    Science.gov (United States)

    Vallano, D.; Sparks, J. P.

    2009-12-01

    Vegetation is an important sink for atmospheric reactive N in N-limited systems and may be capable of incorporating reactive N compounds directly into leaves through the foliar uptake pathway. A proxy for atmospheric reactive N entering vegetation would be useful to estimate the impact of direct foliar N uptake on plant metabolism. Natural abundance foliar N isotopic composition (δ15N) is a practical tool for this purpose because plant-available N sources often have different isotopic compositions. Current understanding of foliar δ15N suggests these values primarily represent the integration of soil δ15N, direct foliar N uptake, mycorrhizal fractionation, and within-plant fractionations. Using a potted plant mesocosm system, we estimated the influence of mycorrhizae on foliar δ15N patterns in red maple (Acer rubrum) seedlings along an N deposition gradient in New York State. We found that mycorrhizal associations altered foliar δ15N in red maple seedlings from 0.03 - 1.01‰ across sites. Along the same temporal and spatial scales, we examined the influence of soil δ15N, foliar N uptake, and mycorrhizae on foliar δ15N in adult stands of American beech (Fagus grandifolia), black birch (Betula lenta), red maple (A. rubrum), and red oak (Quercus rubra). Using multiple regression models, atmospheric NO2 concentration explained 0%, 69%, 23%, and 45% of the residual variation in foliar δ15N remaining in American beech, red maple, red oak, and black birch, respectively, after accounting for soil δ15N. Our results suggest that foliar δ15N may be used to estimate pollution-derived atmospheric reactive N entering vegetation via the foliar N uptake pathway.

  6. Compound-specific 15N stable isotope probing of N assimilation by the soil microbial biomass: a new methodological paradigm in soil N cycling

    Science.gov (United States)

    Charteris, A. F.; Knowles, T. D. J.; Michaelides, K.; Evershed, R. P.

    2015-10-01

    A compound-specific nitrogen-15 stable isotope probing (15N-SIP) technique is described which allows investigation of the fate of inorganic- or organic-N amendments to soils. The technique uses gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) to determine the δ15N values of individual amino acids (AAs; determined as N-acetyl, O-isopropyl derivatives) as proxies of biomass protein production. The δ15N values are used together with AA concentrations to quantify N assimilation of 15N-labelled substrates by the soil microbial biomass. The utility of the approach is demonstrated through incubation experiments using inorganic 15N-labelled substrates ammonium (15NH4+) and nitrate (15NO3-) and an organic 15N-labelled substrate, glutamic acid (15N-Glu). Assimilation of all the applied substrates was undetectable based on bulk soil properties, i.e. % total N (% TN), bulk soil N isotope composition and AA concentrations, all of which remained relatively constant throughout the incubation experiments. In contrast, compound-specific AA δ15N values were highly sensitive to N assimilation, providing qualitative and quantitative insights into the cycling and fate of the applied 15N-labelled substrates. The utility of this 15N-AA-SIP technique is considered in relation to other currently available methods for investigating the microbially-mediated assimilation of nitrogenous substrates into the soil organic N pool. This approach will be generally applicable to the study of N cycling in any soil, or indeed, in any complex ecosystem.

  7. Uniform {sup 15}N- and {sup 15}N/{sup 13}C-labeling of proteins in mammalian cells and solution structure of the amino terminal fragment of u-PA

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, A.P.; Petros, A.M.; Meadows, R.P.; Mazar, A.P.; Nettesheim, D.G.; Pederson, T.M.; Fesik, S.W. [Abbott Laboratories, Abbott Park, IL (United States)

    1994-12-01

    Urokinase-type plasminogen activator (u-PA) is a 54-kDa glycoprotein that catalyzes the conversion of plasminogen to plasmin, a broad-specificity protease responsible for the degradation of fibrin clots and extracellular matrix components. The u-PA protein consists of three individual modules: a growth factor domain (GFD), a kringle, and a serine protease domain. The amino terminal fragment (ATF) includes the GFD-responsible for u-PA binding to its receptor-and the kringle domains. This protein was expressed and uniformly {sup 15}N-and {sup 15}N/{sup 13}C-labeled in mammalian cells by methods that will be described. In addition, we present the three-dimensional structure of ATF that was derived from 1299 NOE-derived distance restraints along with the {phi} angle and hydrogen bonding restraints. Although the individual domains in the structures were highly converged, the two domains are structurally independent. The overall structures of the individual domains are very similar to the structures of homologous proteins. However, important structural differences between the growth factor domain of u-PA and other homologous proteins were observed in the region that has been implicated in binding the urokinase receptor. These results may explain, in part, why other growth factors show no appreciable affinity for the urokinase receptor.

  8. 5 CFR 351.705 - Administrative assignment.

    Science.gov (United States)

    2010-01-01

    ... 5 Administrative Personnel 1 2010-01-01 2010-01-01 false Administrative assignment. 351.705 Section 351.705 Administrative Personnel OFFICE OF PERSONNEL MANAGEMENT CIVIL SERVICE REGULATIONS REDUCTION IN FORCE Assignment Rights (Bump and Retreat) § 351.705 Administrative assignment. (a) An...

  9. Competition Based Neural Networks for Assignment Problems

    Institute of Scientific and Technical Information of China (English)

    李涛; LuyuanFang

    1991-01-01

    Competition based neural networks have been used to solve the generalized assignment problem and the quadratic assignment problem.Both problems are very difficult and are ε approximation complete.The neural network approach has yielded highly competitive performance and good performance for the quadratic assignment problem.These neural networks are guaranteed to produce feasible solutions.

  10. A polyhedral approach to quadratic assignment problem

    OpenAIRE

    Köksaldı, Ahmet Sertaç Murat

    1994-01-01

    Ankara : Department of Industrial Engineering and the Institute of Engineering and Sciences of Bilkent University, 1994. Thesis (Master's) -- Bilkent University, 1994. Includes bibliographical references. In this thesis, Quadratic Assignment Problem is considered. Since Quadratic Assignment Problem is JVP-bard, no polynomial time exact solution method exists. Proving optimality of solutions to Quadratic Assignment Problems has been limited to instances of small dimension. In...

  11. The Mechanism Design Approach to Student Assignment

    Science.gov (United States)

    Pathak, Parag A.

    2011-01-01

    The mechanism design approach to student assignment involves the theoretical, empirical, and experimental study of systems used to allocate students into schools around the world. Recent practical experience designing systems for student assignment has raised new theoretical questions for the theory of matching and assignment. This article reviews…

  12. 5 CFR 870.901 - Assignments permitted.

    Science.gov (United States)

    2010-01-01

    ... (CONTINUED) FEDERAL EMPLOYEES' GROUP LIFE INSURANCE PROGRAM Assignments of Life Insurance § 870.901..., and compensationers. (2) An individual may assign ownership of all life insurance under this part... Benefit before assigning the remainder of his/her insurance, the amount of Basic insurance does...

  13. Heteronuclear transverse and longitudinal relaxation in AX4 spin systems: Application to 15N relaxations in 15NH4 +

    OpenAIRE

    Werbeck, Nicolas D; Hansen, D. Flemming

    2014-01-01

    The equations that describe the time-evolution of transverse and longitudinal 15N magnetisations in tetrahedral ammonium ions, 15NH4 +, are derived from the Bloch-Wangsness-Redfield density operator relaxation theory. It is assumed that the relaxation of the spin-states is dominated by (1) the intra-molecular 15N–1H and 1H–1H dipole–dipole interactions and (2) interactions of the ammonium protons with remote spins, which also include the contribution to the relaxations that arise from the exc...

  14. Deep roots of Brassica oleracea have high uptake of 15N-nitrate to 2 meters soil depth

    OpenAIRE

    Kristensen, H.L.; Thorup-Kristensen, K

    2006-01-01

    • Deep roots may be important for uptake of NO3- by annual crops. A field experiment with the deep rooted crop curly kale (Brassica oleracea L. convar. acephala (DC.) Alef. var. sabellica L.) was performed to investigate root distribution and N uptake in deep soil layers. • Root distribution was investigated by use of minirhizotrons and root extractions to 2.4 m depth. The capacity for crop N uptake was studied by deep 15NO3- placement followed by analysis of plant 15N content after 3-30 d...

  15. Determination of the δ15N and δ18O of nitrate in water; RSIL lab code 2900

    Science.gov (United States)

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2900 is to determine the δ15N and δ18O of nitrate (NO3-) in water. The δ15N and δ18O of the dissolved NO3- are analyzed by converting the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of the NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  16. Determination of the δ15N and δ18O of nitrate in solids; RSIL lab code 2897

    Science.gov (United States)

    Coplen, Tyler B.; Qi, Haiping; Revesz, Kinga; Casciotti, Karen; Hannon, Janet E.

    2007-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 2897 is to determine the δ15N and δ18O of nitrate (NO3-) in solids. The NO3- fraction of the nitrogen species is dissolved by water (called leaching) and can be analyzed by the bacterial method covered in RSIL lab code 2900. After leaching, the δ15N and δ18O of the dissolved NO3- is analyzed by conversion of the NO3- to nitrous oxide (N2O), which serves as the analyte for mass spectrometry. A culture of denitrifying bacteria is used in the enzymatic conversion of NO3- to N2O, which follows the pathway shown in equation 1: NO3- → NO2- → NO → 1/2 N2O (1) Because the bacteria Pseudomonas aureofaciens lack N2O reductive activity, the reaction stops at N2O, unlike the typical denitrification reaction that goes to N2. After several hours, the conversion is complete, and the N2O is extracted from the vial, separated from volatile organic vapor and water vapor by an automated -65 °C isopropanol-slush trap, a Nafion drier, a CO2 and water removal unit (Costech #021020 carbon dioxide absorbent with Mg(ClO4)2), and trapped in a small-volume trap immersed in liquid nitrogen with a modified Finnigan MAT (now Thermo Scientific) GasBench 2 introduction system. After the N2O is released, it is further purified by gas chromatography before introduction to the isotope-ratio mass spectrometer (IRMS). The IRMS is a Thermo Scientific Delta V Plus continuous flow IRMS (CF-IRMS). It has a universal triple collector, consisting of two wide cups with a narrow cup in the middle; it is capable of simultaneously measuring mass/charge (m/z) of the N2O molecule 44, 45, and 46. The ion beams from these m/z values are as follows: m/z = 44 = N2O = 14N14N16O; m/z = 45 = N2O = 14N15N16O or 14N14N17O; m/z = 46 = N2O = 14N14N18O. The 17O contributions to the m/z 44 and m/z 45 ion beams are accounted for before δ15N values are reported.

  17. Partitioning Residue-derived and Residue-induced Emissions of N2O Using 15N-labelled Crop Residues

    Science.gov (United States)

    Farrell, R. E.; Carverhill, J.; Lemke, R.; Knight, J. D.

    2014-12-01

    Estimates of N2O emissions in Canada indicate that 17% of all agriculture-based emissions are associated with the decomposition of crop residues. However, research specific to the western Canadian prairies (including Saskatchewan) has shown that the N2O emission factor for N sources in this region typically ranges between 0.2 and 0.6%, which is well below the current IPCC default emission factor of 1.0%. Thus, it stands to reason that emissions from crop residues should also be lower than those calculated using the current IPCC emission factor. Current data indicates that residue decomposition, N mineralization and N2O production are affected by a number of factors such as C:N ratio and chemical composition of the residue, soil type, and soil water content; thus, a bench-scale incubation study was conducted to examine the effects of soil type and water content on N2O emissions associated with the decomposition of different crop residues. The study was carried out using soils from the Black, Dark Brown, Brown, and Gray soil zones and was conducted at both 50% and 70% water-filled pore space (WFPS); the soils were amended with 15N-labeled residues of wheat, pea, canola, and flax, or with an equivalent amount of 15N-labeled urea; 15N2O production was monitored using a Picarro G5101-i isotopic N2O analyzer. Crop residue additions to the soils resulted in both direct and indirect emissions of N2O, with residue derived emissions (RDE; measured as 15N2O) generally exceeding residue-induced emissions (RIE) at 50% WFPS—with RDEs ranging from 42% to 88% (mean = 58%) of the total N2O. Conversely, at 70% WFPS, RDEs were generally lower than RIEs—ranging from 21% to 83% (mean = 48%). Whereas both water content and soil type had an impact on N2O production, there was a clear and consistent trend in the emission factors for the residues; i.e., emissions were always greatest for the canola residue and lowest for the wheat residue and urea fertilizer; and intermediate for pea

  18. Nuclear astrophysics deep underground the case of the 15N(p,γ)16O reaction at LUNA

    CERN Document Server

    Mazzocchi, Chiara

    2010-01-01

    Measuring nuclear reactions of astrophysical interest at the relevant energies is not always possible on the Earth’s surface because of the cosmic-ray background that dominates the spectra. The LUNA collaboration exploits the lowbackground enviroment of Gran Sasso National Laboratory to study these reactions at or close to the Gamow peak. The latest experimental efforts included the measurement of the 15N(p,γ)16O at beam energies between 77 and 350 keV. The status of these measurements is summarised in this contribution.

  19. Nitrogen retention across a gradient of 15N additions to an unpolluted temperate forest soil in Chile

    Science.gov (United States)

    Perakis, Steven S.; Compton, J.E.; Hedin, L.O.

    2005-01-01

    Accelerated nitrogen (N) inputs can drive nonlinear changes in N cycling, retention, and loss in forest ecosystems. Nitrogen processing in soils is critical to understanding these changes, since soils typically are the largest N sink in forests. To elucidate soil mechanisms that underlie shifts in N cycling across a wide gradient of N supply, we added 15NH415NO3 at nine treatment levels ranging in geometric sequence from 0.2 kg to 640 kg NA? ha-1A? yr-1 to an unpolluted old-growth temperate forest in southern Chile. We recovered roughly half of tracers in 0-25 cm of soil, primarily in the surface 10 cm. Low to moderate rates of N supply failed to stimulate N leaching, which suggests that most unrecovered 15N was transferred from soils to unmeasured sinks above ground. However, soil solution losses of nitrate increased sharply at inputs > 160 kg NA? ha-1A? yr-1, corresponding to a threshold of elevated soil N availability and declining 15N retention in soil. Soil organic matter (15N in soils at the highest N inputs and may explain a substantial fraction of the 'missing N' often reported in studies of fates of N inputs to forests. Contrary to expectations, N additions did not stimulate gross N cycling, potential nitrification, or ammonium oxidizer populations. Our results indicate that the nonlinearity in N retention and loss resulted directly from excessive N supply relative to sinks, independent of plant-soil-microbial feedbacks. However, N additions did induce a sharp decrease in microbial biomass C:N that is predicted by N saturation theory, and which could increase long-term N storage in soil organic matter by lowering the critical C:N ratio for net N mineralization. All measured sinks accumulated 15N tracers across the full gradient of N supply, suggesting that short-term nonlinearity in N retention resulted from saturation of uptake kinetics, not uptake capacity, in plant, soil, and microbial pools.

  20. (15)N in tree rings as a bio-indicator of changing nitrogen cycling in tropical forests: an evaluation at three sites using two sampling methods.

    Science.gov (United States)

    van der Sleen, Peter; Vlam, Mart; Groenendijk, Peter; Anten, Niels P R; Bongers, Frans; Bunyavejchewin, Sarayudh; Hietz, Peter; Pons, Thijs L; Zuidema, Pieter A

    2015-01-01

    Anthropogenic nitrogen deposition is currently causing a more than twofold increase of reactive nitrogen input over large areas in the tropics. Elevated (15)N abundance (δ(15)N) in the growth rings of some tropical trees has been hypothesized to reflect an increased leaching of (15)N-depleted nitrate from the soil, following anthropogenic nitrogen deposition over the last decades. To find further evidence for altered nitrogen cycling in tropical forests, we measured long-term δ(15)N values in trees from Bolivia, Cameroon, and Thailand. We used two different sampling methods. In the first, wood samples were taken in a conventional way: from the pith to the bark across the stem of 28 large trees (the "radial" method). In the second, δ(15)N values were compared across a fixed diameter (the "fixed-diameter" method). We sampled 400 trees that differed widely in size, but measured δ(15)N in the stem around the same diameter (20 cm dbh) in all trees. As a result, the growth rings formed around this diameter differed in age and allowed a comparison of δ(15)N values over time with an explicit control for potential size-effects on δ(15)N values. We found a significant increase of tree-ring δ(15)N across the stem radius of large trees from Bolivia and Cameroon, but no change in tree-ring δ(15)N values over time was found in any of the study sites when controlling for tree size. This suggests that radial trends of δ(15)N values within trees reflect tree ontogeny (size development). However, for the trees from Cameroon and Thailand, a low statistical power in the fixed-diameter method prevents to conclude this with high certainty. For the trees from Bolivia, statistical power in the fixed-diameter method was high, showing that the temporal trend in tree-ring δ(15)N values in the radial method is primarily caused by tree ontogeny and unlikely by a change in nitrogen cycling. We therefore stress to account for tree size before tree-ring δ(15)N values can be properly

  1. Integrated assignment and path planning

    Science.gov (United States)

    Murphey, Robert A.

    2005-11-01

    A surge of interest in unmanned systems has exposed many new and challenging research problems across many fields of engineering and mathematics. These systems have the potential of transforming our society by replacing dangerous and dirty jobs with networks of moving machines. This vision is fundamentally separate from the modern view of robotics in that sophisticated behavior is realizable not by increasing individual vehicle complexity, but instead through collaborative teaming that relies on collective perception, abstraction, decision making, and manipulation. Obvious examples where collective robotics will make an impact include planetary exploration, space structure assembly, remote and undersea mining, hazardous material handling and clean-up, and search and rescue. Nonetheless, the phenomenon driving this technology trend is the increasing reliance of the US military on unmanned vehicles, specifically, aircraft. Only a few years ago, following years of resistance to the use of unmanned systems, the military and civilian leadership in the United States reversed itself and have recently demonstrated surprisingly broad acceptance of increasingly pervasive use of unmanned platforms in defense surveillance, and even attack. However, as rapidly as unmanned systems have gained acceptance, the defense research community has discovered the technical pitfalls that lie ahead, especially for operating collective groups of unmanned platforms. A great deal of talent and energy has been devoted to solving these technical problems, which tend to fall into two categories: resource allocation of vehicles to objectives, and path planning of vehicle trajectories. An extensive amount of research has been conducted in each direction, yet, surprisingly, very little work has considered the integrated problem of assignment and path planning. This dissertation presents a framework for studying integrated assignment and path planning and then moves on to suggest an exact

  2. Linking Isotopes and Panmixia: High Within-Colony Variation in Feather δ2H, δ13C, and δ15N across the Range of the American White Pelican.

    Directory of Open Access Journals (Sweden)

    Matthew W Reudink

    Full Text Available Complete panmixia across the entire range of a species is a relatively rare phenomenon; however, this pattern may be found in species that have limited philopatry and frequent dispersal. American white pelicans (Pelecanus erythrorhyncos provide a unique opportunity to examine the role of long-distance dispersal in facilitating gene flow in a species recently reported as panmictic across its broad breeding range. This species is also undergoing a range expansion, with new colonies arising hundreds of kilometers outside previous range boundaries. In this study, we use a multiple stable isotope (δ2H, δ13C, δ15N approach to examine feather isotopic structuring at 19 pelican colonies across North America, with the goal of establishing an isotopic basemap that could be used for assigning individuals at newly established breeding sites to source colonies. Within-colony isotopic variation was extremely high, exceeding 100‰ in δ2H within some colonies (with relatively high variation also observed for δ13C and δ15N. The high degree of within-site variation greatly limited the utility of assignment-based approaches (42% cross-validation success rate; range: 0-90% success. Furthermore, clustering algorithms identified four likely isotopic clusters; however, those clusters were generally unrelated to geographic location. Taken together, the high degree of within-site isotopic variation and lack of geographically-defined isotopic clusters preclude the establishment of an isotopic basemap for American white pelicans, but may indicate that a high incidence of long-distance dispersal is facilitating gene flow, leading to genetic panmixia.

  3. Linking Isotopes and Panmixia: High Within-Colony Variation in Feather δ2H, δ13C, and δ15N across the Range of the American White Pelican

    Science.gov (United States)

    Reudink, Matthew W.; Kyle, Christopher J.; McKellar, Ann E.; Somers, Christopher M.; Reudink, Robyn L. F.; Kyser, T. Kurt; Franks, Samantha E.; Nocera, Joseph J.

    2016-01-01

    Complete panmixia across the entire range of a species is a relatively rare phenomenon; however, this pattern may be found in species that have limited philopatry and frequent dispersal. American white pelicans (Pelecanus erythrorhyncos) provide a unique opportunity to examine the role of long-distance dispersal in facilitating gene flow in a species recently reported as panmictic across its broad breeding range. This species is also undergoing a range expansion, with new colonies arising hundreds of kilometers outside previous range boundaries. In this study, we use a multiple stable isotope (δ2H, δ13C, δ15N) approach to examine feather isotopic structuring at 19 pelican colonies across North America, with the goal of establishing an isotopic basemap that could be used for assigning individuals at newly established breeding sites to source colonies. Within-colony isotopic variation was extremely high, exceeding 100‰ in δ2H within some colonies (with relatively high variation also observed for δ13C and δ15N). The high degree of within-site variation greatly limited the utility of assignment-based approaches (42% cross-validation success rate; range: 0–90% success). Furthermore, clustering algorithms identified four likely isotopic clusters; however, those clusters were generally unrelated to geographic location. Taken together, the high degree of within-site isotopic variation and lack of geographically-defined isotopic clusters preclude the establishment of an isotopic basemap for American white pelicans, but may indicate that a high incidence of long-distance dispersal is facilitating gene flow, leading to genetic panmixia. PMID:26974163

  4. Sequence-specific Assignment of 1H-NMR Resonance and Determination of the Secondary Structure of HWTX-II%虎纹捕鸟蛛毒素HWTX-II的1H-NMR信号归属和二级结构分析

    Institute of Scientific and Technical Information of China (English)

    舒芹; 吕善运; 顾孝诚; 梁宋平

    2001-01-01

    虎纹捕鸟蛛毒素HWTX-II是从虎纹捕鸟蛛Selenocosmia huwena的毒液中分离出的一种新型杀虫肽。应用2D-NMR技术研究该毒素分子的溶液结构特点, 通过分析水及重水DQF-COSY、COSY、TOCSY和NOESY等1H-NMR谱, 识别出HWTX-II全部37个氨基酸残基自旋体系; 通过NOESY谱中的dαN、dαδ、dβN和dNN联系完成了序列专一的谱峰归属, 确认了所有主链质子和除了Lys侧链εNH2质子外的所有侧链质子的化学位移, 为完全解析HWTX-II的溶液三维构象奠定了基础。 并且通过核磁数据分析, 确定HWTX-II的二级结构特点是含有较多的伸展构象, 尤其是C端有一个典型的双股反平行的β折叠(Trp27~Cys29和Cys34~Lys36), 分子中缺乏螺旋结构。 这些二级结构特点与已探明结构的其他蜘蛛毒素的基本相同。%Huwentoxin-II (HWTX-II) is an insecticidal peptide purified from the venom of spider Selenocosmia huwena. The structure of this toxin in solution was investigated using 2D-NMR. The complete sequence-specific assignments of proton resonance in the 1H-NMR spectra of HWTX-II were obtained by analyzing a series of 2D spectrum, including COSY, DQF-COSY, TOCSY and NOESY. All the backbone protons and side chain protons, except εNH2 protons of Lys residues, were identified by dαN, dαδ, dNN and dβN connectivities. The results provide a basis for further determination of the solution conformation of HWTX-II. Furthermore the secondary structure of HWTX-II was determined from NMR data. It contained mainly extended conformation, especially a double-stranded anti-parallel β-sheet with Trp27—Cys29 and Cys34—Lys36 at the C terminal, and it lacked helix. These characters of the secondary structure of HWTX-II were similar to those spider toxins which structure in solution had been reported.

  5. 海南捕鸟蛛毒素-Ⅰ(HNTX-Ⅰ)的1H-NMR信号归属和二级结构分析%SEQUENCE-SPECIFIC ASSIGNMENT OF 1H-NMR RESONANCE AND DETERMINATION OF THE SECONDARY STRUCTURE OF HAINANTOXIN-Ⅰ

    Institute of Scientific and Technical Information of China (English)

    李冬玲; 梁宋平

    2003-01-01

    Hainantoxin-Ⅰ(HNTX-Ⅰ ) is an insect blocker of voltage-gated sodium channel isolated from the venom of the Chinese bird spider Ornithoctonus hainana (early named Selenocosmia hainana). The three-dimensional conformation of HNTX- Ⅰ in aqueous solution was determined using two-dimensional 1H NMR techniques. The complete sequence-specific assignments of proton resonance in the 1H-NMR spectra of HNTX-Ⅰ were obtained by analyzing a series of 2D spectra, including double-quantum-filtered correlation spectroscopy (DQF-COSY), total correlated spectroscopy (TOCSY) and nuclear overhauser effect spectroscopy (NOESY) in H2O and D2O. All the backbone protons and more than 96% of the side-chain protons were identified by dαN, dαN, dNN and dαδ connectivities in NOESY spectrum. Furthermore, it was found that the main element of the secondary structure of HNTX-Ⅰ is a short triple-stranded antiparallel β-sheet with Lys7-Cys9, Tyr20-Asn23 and Trp28-Val31, based on 3JNH-CαH coupling constants, sequential NOE connectivities and slowly exchanging amide protons. These characters of the secondary structure of HNTX-Ⅰ are similar to those of HWTX-Ⅰ, SHL-Ⅰ and HWTX-Ⅳ, which have known solution structures. These results provide a basis for the further determination of the solution conformation of HNTX-Ⅰ.%海南捕鸟蛛毒素-Ⅰ(HNTX-Ⅰ)是从海南捕鸟蛛(Ornithoctonus hainana)的粗毒中纯化的一种新型神经毒素.应用二维1H-NMR技术研究HNTX-Ⅰ的溶液结构特点,通过分析水和重水中的DQF-COSY、TOCSY和NOESY谱,识别出HNTX-Ⅰ全部33个氨基酸残基自旋体系;通过NOESY谱中的dαN、dβN、dNN和dαδ联系完成了序列专一的谱峰归属,从而确认了HNTX-Ⅰ所有的主链质子和大于96%的侧链质子的化学位移.并通过分析3JNH-CαH耦合常数、序列间的NOE联系以及慢氢交换质子等,确定HNTX-Ⅰ的二级结构主要是由三股反平行的β-折迭组成(Lys7-Cys9,Tyr20-Asn23和Trp28-Val31),

  6. The agricultural history of human-nitrogen interactions as recorded in ice core δ15N-NO3-

    Science.gov (United States)

    Felix, J. David; Elliott, Emily M.

    2013-04-01

    The advent and industrialization of the Haber Bosch process in the early twentieth century ushered in a new era of reactive nitrogen distributions on Earth. Since the appearance of the first commercial scale Haber Bosch fertilizer plants, fertilizer application rates have greatly increased in the U.S. While the contributions of fertilizer runoff to eutrophication and anoxic dead zones in coastal regions have been well-documented, the potential influences of increased fertilizer applications on air quality and precipitation chemistry are poorly constrained. Here we combine a 255-year record of precipitation nitrate isotopes preserved in a Greenland ice core, historical reconstructions of fertilizer application rates, and field characterization of the isotopic composition of nitrogen oxides produced biogenically in soils, to provide new constraints on the contributions of biogenic emissions to North American NOx inventories. Our results indicate that increases in twentieth century commercial fertilizer use led to large increases in soil NO, a byproduct released during nitrification and denitrification reactions. These large shifts in soil NO production are evidenced by sharp declines in ice core δ15N-NO3- values. Further, these results suggest that biogenic NOx emissions are underestimated by two to four fold in the U.S. NOx emission inventories used to construct global reactive nitrogen budgets. These results demonstrate that nitrate isotopes in ice cores, coupled with newly constrained δ15N-NOx values for NOx emission sources, provide a novel means for estimating contemporary and historic contributions from individual NOx emission sources to deposition.

  7. Production, Purification, and Characterization of 15N-Labeled DNA Repair Proteins as Internal Standards for Mass Spectrometric Measurements

    Science.gov (United States)

    Jaruga, Pawel; Nelson, Bryant C.; Lowenthal, Mark S.; Jemth, Ann-Sofie; Loseva, Olga; Coskun, Erdem; Helleday, Thomas

    2016-01-01

    Oxidatively induced DNA damage is caused in living organisms by a variety of damaging agents, resulting in the formation of a multiplicity of lesions, which are mutagenic and cytotoxic. Unless repaired by DNA repair mechanisms before DNA replication, DNA lesions can lead to genomic instability, which is one of the hallmarks of cancer. Oxidatively induced DNA damage is mainly repaired by base excision repair pathway with the involvement of a plethora of proteins. Cancer tissues develop greater DNA repair capacity than normal tissues by overexpressing DNA repair proteins. Increased DNA repair in tumors that removes DNA lesions generated by therapeutic agents before they became toxic is a major mechanism in the development of therapy resistance. Evidence suggests that DNA repair capacity may be a predictive biomarker of patient response. Thus, knowledge of DNA–protein expressions in disease-free and cancerous tissues may help predict and guide development of treatments and yield the best therapeutic response. Our laboratory has developed methodologies that use mass spectrometry with isotope dilution for the measurement of expression of DNA repair proteins in human tissues and cultured cells. For this purpose, full-length 15N-labeled analogs of a number of human DNA repair proteins have been produced and purified to be used as internal standards for positive identification and accurate quantification. This chapter describes in detail the protocols of this work. The use of 15N-labeled proteins as internal standards for the measurement of several DNA repair proteins in vivo is also presented. PMID:26791985

  8. Traceability of animal meals in Japanese quail eggs using the technique of 13C e 15N* stable isotopes

    Directory of Open Access Journals (Sweden)

    C Mori

    2013-03-01

    Full Text Available The objective of this study was to trace the inclusion of bovine meat and bone meal (BMBM in the diet of Japanese quails by analyzing eggs and egg fractions (yolk and albumen by the technique of carbon-13 (13C and nitrogen-15 (15N stable isotopes. In the trial, 120 Japanese quails were distributed in six treatments with four replicates of five birds each. The following treatments were applied: feed based on corn and soybean meal, containing graded BMBM inclusions (0, 1, 2, 3, 4 or 5%. After 42 days, 20 eggs per treatment were randomly collected for three consecutive days. Ten eggs were used for yolk and albumen sample collection, and ten for total egg sample collection. It was possible to detect the dietary inclusion of 1% BMBM in the egg and its fractions. Therefore, the technique of isotopes 13C and 15N is able of tracing since 1% inclusion level of BMBM in the diet of Japanese quails in eggs and their fractions.

  9. Determination of trophic relationships within a Bohai Bay food web using stable δ15N and δ13C analysis

    Institute of Scientific and Technical Information of China (English)

    WAN Yi; HU Jianying; AN Lihui; AN Wei; YANG Min; Itoh Mitsuaki; Hattori Tatsuya; TAO Shu

    2005-01-01

    This study measured stable carbon and nitrogen isotope ratios in phytoplankton, zooplankton, five invertebrates species, eight fishes species and three seabirds species collected in Bohai Bay. δ13C ranged from -25.38‰ to -11.08‰ showing a relative low enrichment in the food web from Bohai Bay. The mean δ13C of mullet is higher than that of other organisms, and this might be due to that mullet is migration fish and feeds mainly on inshore sources. δ15N ranged from 4.08‰ to 13.98‰, and showed a step-wise enrichment with trophic level of 3.8‰. The δ15N enrichment factor was used to construct an isotopic food web model to establish trophic relationships within this marine food web. According to this model, exact trophic levels of all organisms were estimated as 1.46-2.10, 1.91-3.32, 2.55-4.23 and 2.98-4.28 for plankton, invertebrates, fishes, and seabirds.

  10. Heteronuclear transverse and longitudinal relaxation in AX4 spin systems: Application to 15N relaxations in 15NH4+

    Science.gov (United States)

    Werbeck, Nicolas D.; Hansen, D. Flemming

    2014-01-01

    The equations that describe the time-evolution of transverse and longitudinal 15N magnetisations in tetrahedral ammonium ions, 15NH4+, are derived from the Bloch-Wangsness-Redfield density operator relaxation theory. It is assumed that the relaxation of the spin-states is dominated by (1) the intra-molecular 15N–1H and 1H–1H dipole–dipole interactions and (2) interactions of the ammonium protons with remote spins, which also include the contribution to the relaxations that arise from the exchange of the ammonium protons with the bulk solvent. The dipole–dipole cross-correlated relaxation mechanisms between each of the 15N–1H and 1H–1H interactions are explicitly taken into account in the derivations. An application to 15N-ammonium bound to a 41 kDa domain of the protein DnaK is presented, where a comparison between experiments and simulations show that the ammonium ion rotates rapidly within its binding site with a local correlation time shorter than approximately 1 ns. The theoretical framework provided here forms the basis for further investigations of dynamics of AX4 spin systems, with ammonium ions in solution and bound to proteins of particular interest. PMID:25128779

  11. Turnover of manure 15N-labelled ammonium during composting and soil application as affected by lime and superphosphate addition

    DEFF Research Database (Denmark)

    Tran, Tien Minh; Luxhøi, Jesper; Jensen, Lars Stoumann

    2012-01-01

    To determine N turnover and losses during aerobic composting of animal manure, a 41-d laboratory study was performed on pig manure composting with three additive treatments (Straw: pig manure + straw only; Lime: pig manure + straw + quick lime; and SSP: pig manure + straw + single superphosphate)...... superphosphate increased the NH4-N content in both. Therefore, superphosphate addition increased the potential fertilizer value of composted pig manure. © Soil Science Society of America.......). The NH4-N pool in the pig manure was initially labeled with 15N to determine the fate of manure NH4-N during composting. The composts were subsequently applied to soil to investigate the effects on soil mineral N and to trace the 15N during 60 d of incubation at 25°C. Of the initial manure 15NH4-N......, approximately 30, 90, and 20% was lost by NH3 volatilization during composting in the Straw, Lime, and SSP treatments, respectively. Concurrently, 62, 16, and 41% of initial 15NH4-N was immobilized in the respective treatments. When the composts were applied to soil, the mineral N in soil with SSP compost...

  12. Revision of the 15N(p,{\\gamma})16O reaction rate and oxygen abundance in H-burning zones

    CERN Document Server

    Caciolli, A; Capogrosso, V; Bemmerer, D; Broggini, C; Corvisiero, P; Costantini, H; Elekes, Z; Formicola, A; Fulop, Zs; Gervino, G; Guglielmetti, A; Gustavino, C; Gyurky, Gy; Imbriani, G; Junker, M; Lemut, A; Marta, M; Menegazzo, R; Palmerini, S; Prati, P; Roca, V; Rolfs, C; Alvarez, C Rossi; Somorjai, E; Straniero, O; Strieder, F; Terrasi, F; Trautvetter, H P; Vomiero, A

    2011-01-01

    The NO cycle takes place in the deepest layer of a H-burning core or shell, when the temperature exceeds T {\\simeq} 30 {\\cdot} 106 K. The O depletion observed in some globular cluster giant stars, always associated with a Na enhancement, may be due to either a deep mixing during the RGB (red giant branch) phase of the star or to the pollution of the primordial gas by an early population of massive AGB (asymptotic giant branch) stars, whose chemical composition was modified by the hot bottom burning. In both cases, the NO cycle is responsible for the O depletion. The activation of this cycle depends on the rate of the 15N(p,{\\gamma})16O reaction. A precise evaluation of this reaction rate at temperatures as low as experienced in H-burning zones in stellar interiors is mandatory to understand the observed O abundances. We present a new measurement of the 15N(p,{\\gamma})16O reaction performed at LUNA covering for the first time the center of mass energy range 70-370 keV, which corresponds to stellar temperatures...

  13. Destino de adubos nitrogenados marcados com 15N em amostras de dois solos da Amazônia central

    Directory of Open Access Journals (Sweden)

    S. S. Alfaia

    1997-09-01

    Full Text Available Foi estudado, em condições de casa de vegetação, em Nancy (França, em 1992, o destino de duas formas de fertilizantes nitrogenados, marcados com 15N, sulfato de amônio e uréia, em amostras do horizonte A de dois principais solos da Amazônia Central, classificados como latossolo amarelo e podzólico vermelho-amarelo. A planta teste foi o "rye-grass" da Itália (Lolium multiflorum L.. Em ambos os solos, a uréia foi mais bem utilizada do que o sulfato de amônio. Entre 60 e 70% do N aplicado como uréia foi absorvido pela planta, enquanto, com a aplicação de sulfato de amônio, esses valores variaram entre 44 e 49%. O balanço do 15N no final do ciclo da cultura mostrou que a imobilização do N nos dois solos foi maior na presença de uréia que na de sulfato de amônio. As perdas, estimadas por diferença, foram mais elevadas no tratamento com sulfato de amônio. Considerando que perdas por lixiviação foram praticamente nulas com a técnica de cultivo utilizada, elas devem ter ocorrido essencialmente por via gasosa.

  14. Accurate determination of order parameters from 1H,15N dipolar couplings in MAS solid-state NMR experiments.

    Science.gov (United States)

    Chevelkov, Veniamin; Fink, Uwe; Reif, Bernd

    2009-10-01

    A reliable site-specific estimate of the individual N-H bond lengths in the protein backbone is the fundamental basis of any relaxation experiment in solution and in the solid-state NMR. The N-H bond length can in principle be influenced by hydrogen bonding, which would result in an increased N-H distance. At the same time, dynamics in the backbone induces a reduction of the experimental dipolar coupling due to motional averaging. We present a 3D dipolar recoupling experiment in which the (1)H,(15)N dipolar coupling is reintroduced in the indirect dimension using phase-inverted CP to eliminate effects from rf inhomogeneity. We find no variation of the N-H dipolar coupling as a function of hydrogen bonding. Instead, variations in the (1)H,(15)N dipolar coupling seem to be due to dynamics of the protein backbone. This is supported by the observed correlation between the H(N)-N dipolar coupling and the amide proton chemical shift. The experiment is demonstrated for a perdeuterated sample of the alpha-spectrin SH3 domain. Perdeuteration is a prerequisite to achieve high accuracy. The average error in the analysis of the H-N dipolar couplings is on the order of +/-370 Hz (+/-0.012 A) and can be as small as 150 Hz, corresponding to a variation of the bond length of +/-0.005 A.

  15. Stereospecific assignments in proteins using exact NOEs

    Energy Technology Data Exchange (ETDEWEB)

    Orts, Julien; Vögeli, Beat; Riek, Roland, E-mail: roland.riek@phys.chem.ethz.ch [Swiss Federal Institute of Technology, Laboratory of Physical Chemistry (Switzerland); Güntert, Peter, E-mail: guentert@em.uni-frankfurt.de [Goethe University Frankfurt am Main, Center for Biomolecular Magnetic Resonance, Institute of Biophysical Chemistry (Germany)

    2013-10-18

    Recently developed methods to measure distances in proteins with high accuracy by “exact” nuclear Overhauser effects (eNOEs) make it possible to determine stereospecific assignments, which are particularly important to fully exploit the accuracy of the eNOE distance measurements. Stereospecific assignments are determined by comparing the eNOE-derived distances to protein structure bundles calculated without stereospecific assignments, or an independently determined crystal structure. The absolute and relative CYANA target function difference upon swapping the stereospecific assignment of a diastereotopic group yields the respective stereospecific assignment. We applied the method to the eNOE data set that has recently been obtained for the third immunoglobulin-binding domain of protein G (GB3). The 884 eNOEs provide relevant data for 47 of the total of 75 diastereotopic groups. Stereospecific assignments could be established for 45 diastereotopic groups (96 %) using the X-ray structure, or for 27 diastereotopic groups (57 %) using structures calculated with the eNOE data set without stereospecific assignments, all of which are in agreement with those determined previously. The latter case is relevant for structure determinations based on eNOEs. The accuracy of the eNOE distance measurements is crucial for making stereospecific assignments because applying the same method to the traditional NOE data set for GB3 with imprecise upper distance bounds yields only 13 correct stereospecific assignments using the X-ray structure or 2 correct stereospecific assignments using NMR structures calculated without stereospecific assignments.

  16. Automatic Assessment of Programming assignment

    Directory of Open Access Journals (Sweden)

    Surendra Gupta

    2012-01-01

    Full Text Available In today’s world study of computer’s language is more important. Effective and good programming skills are need full all computer science students. They can be master in programming, only through intensive exercise practices. Due to day by day increasing number of students in the class, the assessment of programming exercises leads to extensive workload for teacher/instructor, particularly if it has to be carried out manually. In this paper, we propose an automatic assessment system for programming assignments, using verification program with random inputs. One of the most important properties of a program is that, it carries out its intended function. The intended function of a program or part of a program can be verified by using inverse function’s verification program. For checking intended functionality and evaluation of a program, we have used verification program. This assessment system has been tested on basic C programming courses, and results shows that it can work well in basic programming exercises, with some initial promising results

  17. Paramagnetic 13C and 15N NMR analyses of the push and pull effects in cytochrome c peroxidase and Coprinus cinereus peroxidase variants: functional roles of highly conserved amino acids around heme.

    Science.gov (United States)

    Nonaka, Daisuke; Wariishi, Hiroyuki; Welinder, Karen G; Fujii, Hiroshi

    2010-01-12

    Paramagnetic (13)C and (15)N nuclear magnetic resonance (NMR) spectroscopy of heme-bound cyanide ((13)C(15)N) was applied to 11 cytochrome c peroxidase (CcP) and Coprinus cinereus peroxidase (CIP) mutants to investigate contributions to the push and pull effects of conserved amino acids around heme. The (13)C and (15)N NMR data for the distal His and Arg mutants indicated that distal His is the key amino acid residue creating the strong pull effect and that distal Arg assists. The mutation of distal Trp of CcP to Phe, the amino acid at this position in CIP, changed the push and pull effects so they resembled those of CIP, whereas the mutation of distal Phe of CIP to Trp changed this mutant to become CcP-like. The (13)C NMR shifts for the proximal Asp mutants clearly showed that the proximal Asp-His hydrogen bonding strengthens the push effect. However, even in the absence of a hydrogen bond, the push effect of proximal His in peroxidase is significantly stronger than in globins. Comparison of these NMR data with the compound I formation rate constants and crystal structures of these mutants showed that (1) the base catalysis of the distal His is more critical for rapid compound I formation than its acid catalysis, (2) the primary function of the distal Arg is to maintain the distal heme pocket in favor of rapid compound I formation via hydrogen bonding, and (3) the push effect is the major contributor to the differential rates of compound I formation in wild-type peroxidases.

  18. Measurement of (15)N enrichment of glutamine and urea cycle amino acids derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate using liquid chromatography-tandem quadrupole mass spectrometry.

    Science.gov (United States)

    Nakamura, Hidehiro; Karakawa, Sachise; Watanabe, Akiko; Kawamata, Yasuko; Kuwahara, Tomomi; Shimbo, Kazutaka; Sakai, Ryosei

    2015-05-01

    6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is an amino acid-specific derivatizing reagent that has been used for sensitive amino acid quantification by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). In this study, we aimed to evaluate the ability of this method to measure the isotopic enrichment of amino acids and to determine the positional (15)N enrichment of urea cycle amino acids (i.e., arginine, ornithine, and citrulline) and glutamine. The distribution of the M and M+1 isotopomers of each natural AQC-amino acid was nearly identical to the theoretical distribution. The standard deviation of the (M+1)/M ratio for each amino acid in repeated measurements was approximately 0.1%, and the ratios were stable regardless of the injected amounts. Linearity in the measurements of (15)N enrichment was confirmed by measuring a series of (15)N-labeled arginine standards. The positional (15)N enrichment of urea cycle amino acids and glutamine was estimated from the isotopic distribution of unique fragment ions generated at different collision energies. This method was able to identify their positional (15)N enrichment in the plasma of rats fed (15)N-labeled glutamine. These results suggest the utility of LC-MS/MS detection of AQC-amino acids for the measurement of isotopic enrichment in (15)N-labeled amino acids and indicate that this method is useful for the study of nitrogen metabolism in living organisms.

  19. An Investigation of the Partial-Assignment Completion Effect on Students' Assignment Choice Behavior

    Science.gov (United States)

    Hawthorn-Embree, Meredith L.; Skinner, Christopher H.; Parkhurst, John; Conley, Elisha

    2011-01-01

    This study was designed to investigate the partial assignment completion effect. Seventh-grade students were given a math assignment. After working for 5 min, they were interrupted and their partially completed assignments were collected. About 20 min later, students were given their partially completed assignment and a new, control assignment…

  20. Uptake and incorporation of {sup 15}N marked nitrogen oxides by seasonal and woddy plants; Aufnahme und Einbau {sup 15}N-markierter Stickoxide durch krautige und holzige Pflanzen

    Energy Technology Data Exchange (ETDEWEB)

    Eickriede, A.

    1998-07-01

    It the Research Centre Juelich a special system was developed to expose plants under defined climatic conditions to a nitrogen oxide enriched atmosphere. In completition and extension to former studies sunflowers (Helianthus annuus L.), mays (Zea mays L.) and poplar (Populus tremula x P. alba) were exposed to nitrogen oxides. By using {sup 15}N enriched nitrogen oxides ({sup 15}NO{sub 2}, and {sup 15}NO, summarised as {sup 15}NO{sub x}) the uptake and incorporation of {sup 15}NOx in dependance of the exposition concentration (30 to 100 ppb {sup 15}NO, 20 ppb {sup 15}NO{sub 2}), nitrogen nutrition (nitrate or ammonium), and harvesting time were examined. Moreover, the influence of nitrogen oxide exposition on the distribution pattern of {sup 15}NO{sub x} nitrogen within the plants and on the amino acid content of plant leaves was investigated. (orig.)

  1. Nutritional status of sugar cane (planted cane) in {sup 15}N experiments;Estado nutricional da cultura de cana-de-acucar (cana-planta) em experimentos com {sup 15}N{sup (1)}

    Energy Technology Data Exchange (ETDEWEB)

    Faroni, Carlos Eduardo, E-mail: cfaroni@ctc.com.b [Centro de Tecnologia Canavieira (CTC), Piracicaba, SP (Brazil); Trivelin, Paulo Cesar Ocheuze; Franco, Henrique Coutinho Junqueira, E-mail: pcotrive@cena.usp.b, E-mail: hjfranco@cena.usp.b [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil); Vitti, Andre Cesar, E-mail: acvitti@apta.sp.gov.b [Agencia Paulista de Tecnologia dos Agronegocios (APTA), Piracicaba, SP (Brazil). Polo Centro Sul; Otto, Rafael, E-mail: rotto@esalq.usp.b [Universidade de Sao Paulo (USP), Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz; Cantarella, Heitor, E-mail: cantarella@iac.sp.gov.b [Instituto Agronomico de Campinas (IAC), SP (Brazil)

    2009-11-15

    Studies with stable isotopes are becoming more common due to the increased safety of operation and quality and reliability of results. However, the use of microplots is required to decrease the costs of such studies. Since microplots are small compared to regular plot areas, the purpose of this study was to investigate whether nutritional data based on microplot samples can adequately represent the whole area, in a comparison of the nutritional status of microplot sugar cane plants at their maximum development stage with those of the regular plots in experiments with N rates. Three experiments were set up, with three N rates (40, 80, and 120 kg ha{sup -1} N) and a control, with four repetitions, in a randomized complete block design, in the state of Sao Paulo. Microplots of 3 m{sup 2} containing {sup 15}N-fertilizer (5.04% atom {sup 15}N) were included in the main plots formed by 48 lines of sugar cane spaced 1.5 m apart. At the time of maximum development stage, diagnostic leaves were collected in the main and microplots to evaluate the nutritional status of plants by analyzing the total concentration of macro nutrients. There were no differences in N, P, Ca, Mg, and S concentrations in the diagnostic leaves from the main and microplots, so that the latter can be considered representative of the experimental area. Higher nitrogen fertilizer rates induced increased concentrations of not only N, but also of P, Ca, Mg, and S in the diagnostic leaves. (author)

  2. Spatial variations in δ13C and δ15N values of primary consumers in a coastal lagoon

    Science.gov (United States)

    Como, S.; Magni, P.; Van Der Velde, G.; Blok, F. S.; Van De Steeg, M. F. M.

    2012-12-01

    The analysis of the contribution of a food source to a consumer's diet or the trophic position of a consumer is highly sensitive to the variability of the isotopic values used as input data. However, little is known in coastal lagoons about the spatial variations in the isotopic values of primary consumers considered 'end members' in the isotope mixing models for quantifying the diet of secondary consumers or as a baseline for estimating the trophic position of consumers higher up in the food web. We studied the spatial variations in the δ13C and δ15N values of primary consumers and sedimentary organic matter (SOM) within a selected area of the Cabras lagoon (Sardinia, Italy). Our aim was to assess how much of the spatial variation in isotopic values of primary consumers was due to the spatial variability between sites and how much was due to differences in short distances from the shore. Samples were collected at four stations (50-100 m apart) selected randomly at two sites (1.5-2 km apart) chosen randomly at two distances from the shore (i.e. in proximity of the shore -Nearshore - and about 200 m away from the shore -Offshore). The sampling was repeated in March, May and August 2006 using new sites at the two chosen distances from the shore on each date. The isotopic values of size-fractionated seston and macrophytes were also analyzed as a complementary characterization of the study area. While δ15N did not show any spatial variations, the δ13C values of deposit feeders, Alitta (=Neanthes) succinea, Lekanesphaera hookeri, Hydrobia acuta and Gammarus aequicauda, were more depleted Offshore than Nearshore. For these species, there were significant effects of distance or distance × dates in the mean δ13C values, irrespective of the intrinsic variation between sites. SOM showed similar spatial variations in δ13C values, with Nearshore-Offshore differences up to 6‰. This indicates that the spatial isotopic changes are transferred from the food sources to the

  3. δ15N measurement of organic and inorganic substances by EA-IRMS: a speciation-dependent procedure.

    Science.gov (United States)

    Gentile, Natacha; Rossi, Michel J; Delémont, Olivier; Siegwolf, Rolf T W

    2013-01-01

    Little attention has been paid so far to the influence of the chemical nature of the substance when measuring δ(15)N by elemental analysis (EA)-isotope ratio mass spectrometry (IRMS). Although the bulk nitrogen isotope analysis of organic material is not to be questioned, literature from different disciplines using IRMS provides hints that the quantitative conversion of nitrate into nitrogen presents difficulties. We observed abnormal series of δ(15)N values of laboratory standards and nitrates. These unexpected results were shown to be related to the tailing of the nitrogen peak of nitrate-containing compounds. A series of experiments were set up to investigate the cause of this phenomenon, using ammonium nitrate (NH(4)NO(3)) and potassium nitrate (KNO(3)) samples, two organic laboratory standards as well as the international secondary reference materials IAEA-N1, IAEA-N2-two ammonium sulphates [(NH(4))(2)SO(4)]-and IAEA-NO-3, a potassium nitrate. In experiment 1, we used graphite and vanadium pentoxide (V(2)O(5)) as additives to observe if they could enhance the decomposition (combustion) of nitrates. In experiment 2, we tested another elemental analyser configuration including an additional section of reduced copper in order to see whether or not the tailing could originate from an incomplete reduction process. Finally, we modified several parameters of the method and observed their influence on the peak shape, δ(15)N value and nitrogen content in weight percent of nitrogen of the target substances. We found the best results using mere thermal decomposition in helium, under exclusion of any oxygen. We show that the analytical procedure used for organic samples should not be used for nitrates because of their different chemical nature. We present the best performance given one set of sample introduction parameters for the analysis of nitrates, as well as for the ammonium sulphate IAEA-N1 and IAEA-N2 reference materials. We discuss these results considering the

  4. Ecological functions provided by dung beetles are interlinked across space and time: evidence from (15) N isotope tracing.

    Science.gov (United States)

    Nervo, Beatrice; Caprio, Enrico; Celi, Luisella; Lonati, Michele; Lombardi, Giampiero; Falsone, Gloria; Iussig, Gabriele; Palestrini, Claudia; Said-Pullicino, Daniel; Rolando, Antonio

    2017-02-01

    Maintaining multiple ecological functions ("multifunctionality") is crucial to sustain viable ecosystems. To date most studies on biodiversity-ecosystem functioning (BEF) have focused on single or few ecological functions and services. However, there is a critical need to evaluate how species and species assemblages affect multiple processes at the same time, and how these functions are interconnected. Dung beetles represent excellent model organisms because they are key contributors to several ecosystem functions. Using a novel method based on the application of (15) N-enriched dung in a mesocosm field experiment, we assessed the role of dung beetles in regulating multiple aspects of nutrient cycling in alpine pastures over appropriate spatial (up to a soil depth of 20 cm) and temporal (up to 1 yr after dung application) scales. (15) N isotope tracing allowed the evaluation of multiple interrelated ecosystem functions responsible for the cycling of dung-derived nitrogen (DDN) in the soil and vegetation. We also resolved the role of functional group identity and the importance of interactions among co-occurring species for sustaining multiple functions by focusing on two different dung beetle nesting strategies (tunnelers and dwellers). Species interactions were studied by contrasting mixed-species to single-species assemblages, and asking whether the former performed multiple functions better than the latter. Dung beetles influenced at least seven ecological functions by facilitating dung removal, transport of DDN into the soil, microbial ammonification and nitrification processes, uptake of DDN by plants, herbage growth, and changes in botanical composition. Tunnelers and dwellers were found to be similarly efficient for most functions, with differences based on the spatial and temporal scales over which the functions operated. Although mixed-species assemblages seemed to perform better than single-species, this outcome may be dependent on the context. Most

  5. Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy.

    Science.gov (United States)

    Baek, Mi-Hwa; Kamiya, Masakatsu; Kushibiki, Takahiro; Nakazumi, Taichi; Tomisawa, Satoshi; Abe, Chiharu; Kumaki, Yasuhiro; Kikukawa, Takashi; Demura, Makoto; Kawano, Keiichi; Aizawa, Tomoyasu

    2016-04-01

    Antimicrobial peptides (AMPs) are components of the innate immune system and may be potential alternatives to conventional antibiotics because they exhibit broad-spectrum antimicrobial activity. The AMP cecropin P1 (CP1), isolated from nematodes found in the stomachs of pigs, is known to exhibit antimicrobial activity against Gram-negative bacteria. In this study, we investigated the interaction between CP1 and lipopolysaccharide (LPS), which is the main component of the outer membrane of Gram-negative bacteria, using circular dichroism (CD) and nuclear magnetic resonance (NMR). CD results showed that CP1 formed an α-helical structure in a solution containing LPS. For NMR experiments, we expressed (15) N-labeled and (13) C-labeled CP1 in bacterial cells and successfully assigned almost all backbone and side-chain proton resonance peaks of CP1 in water for transferred nuclear Overhauser effect (Tr-NOE) experiments in LPS. We performed (15) N-edited and (13) C-edited Tr-NOE spectroscopy for CP1 bound to LPS. Tr-NOE peaks were observed at the only C-terminal region of CP1 in LPS. The results of structure calculation indicated that the C-terminal region (Lys15-Gly29) formed the well-defined α-helical structure in LPS. Finally, the docking study revealed that Lys15/Lys16 interacted with phosphate at glucosamine I via an electrostatic interaction and that Ile22/Ile26 was in close proximity with the acyl chain of lipid A.

  6. Backbone Assignment of the MALT1 Paracaspase by Solution NMR.

    Science.gov (United States)

    Unnerståle, Sofia; Nowakowski, Michal; Baraznenok, Vera; Stenberg, Gun; Lindberg, Jimmy; Mayzel, Maxim; Orekhov, Vladislav; Agback, Tatiana

    2016-01-01

    Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) is a unique paracaspase protein whose protease activity mediates oncogenic NF-κB signalling in activated B cell-like diffuse large B cell lymphomas (ABC-DLBCLs). ABC-DLBCLs are aggressive lymphomas with high resistance to current chemotherapies. Low survival rate among patients emphasizes the urgent need for alternative treatment options. The characterization of the MALT1 will be an essential tool for developing new target-directed drugs against MALT1 dependent disorders. As the first step in the atomic-level NMR studies of the system, here we report, the (15)N/(13)C/(1)H backbone assignment of the apo form of the MALT1 paracaspase region together with the third immunoglobulin-like (Ig3) domain, 44 kDa, by high resolution NMR. In addition, the non-uniform sampling (NUS) based targeted acquisition procedure is evaluated as a mean of decreasing acquisition and analysis time for larger proteins.

  7. Backbone Assignment of the MALT1 Paracaspase by Solution NMR.

    Directory of Open Access Journals (Sweden)

    Sofia Unnerståle

    Full Text Available Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1 is a unique paracaspase protein whose protease activity mediates oncogenic NF-κB signalling in activated B cell-like diffuse large B cell lymphomas (ABC-DLBCLs. ABC-DLBCLs are aggressive lymphomas with high resistance to current chemotherapies. Low survival rate among patients emphasizes the urgent need for alternative treatment options. The characterization of the MALT1 will be an essential tool for developing new target-directed drugs against MALT1 dependent disorders. As the first step in the atomic-level NMR studies of the system, here we report, the (15N/(13C/(1H backbone assignment of the apo form of the MALT1 paracaspase region together with the third immunoglobulin-like (Ig3 domain, 44 kDa, by high resolution NMR. In addition, the non-uniform sampling (NUS based targeted acquisition procedure is evaluated as a mean of decreasing acquisition and analysis time for larger proteins.

  8. The effect of the recent 17O(p,α14N and 18O(p,α15N fusion cross section measurements in the nucleosynthesis of AGB stars

    Directory of Open Access Journals (Sweden)

    Palmerini S.

    2015-01-01

    Full Text Available The Trojan Horse Method (THM has been used to investigate the low-energy cross sections of the 17O(p,α14N and 18O(p,α15N fusion reactions and to extract the strengths of the resonances that more contribute to the reaction rates at astrophysical energies. Moreover, the strength of the 65 keV resonance in the 17O(p,α14N reaction, measured by means of the THM, has been used to renormalize the corresponding resonance strength in the 17O + p radiative capture channel. Since, proton-induced fusion reactions on 17O and 18O belong to the CNO cycle network for H-burning in stars, the new estimates of the cross sections have been introduced into calculations of Asymptotic giant branch (AGB star nucleosynthesis to determine their impact on astrophysical environments. Results of nucleosynthesis calculations have been compared with geochemical analysis of ”presolar” grains. These solids form in the cold and dusty envelopes that surround AGB stars and once that have been ejected by stellar winds, come to us as inclusions in meteorites providing invaluable benchmarks and constraints for our knowledge of fusion reactions in astrophysical environments.

  9. Mechanism of Solid-State Thermolysis of Ammonia Boraine: 15N NMR Study Using Fast Magic-Angle Spinning and Dynamic Nuclear Polarization

    Energy Technology Data Exchange (ETDEWEB)

    Kobayashi, Takeshi [Ames Laboratory; Gupta, Shalabh [Ames Laboratory; Caporini, Marc A [Bruker BioSpin Corporation; Pecharsky, Vitalij K [Ames Laboratory; Pruski, Marek [Ames Laboratory

    2014-08-28

    The solid-state thermolysis of ammonia borane (NH3BH3, AB) was explored using state-of-the-art 15N solid-state NMR spectroscopy, including 2D indirectly detected 1H{15N} heteronuclear correlation and dynamic nuclear polarization (DNP)-enhanced 15N{1H} cross-polarization experiments as well as 11B NMR. The complementary use of 15N and 11B NMR experiments, supported by density functional theory calculations of the chemical shift tensors, provided insights into the dehydrogenation mechanism of AB—insights that have not been available by 11B NMR alone. Specifically, highly branched polyaminoborane derivatives were shown to form from AB via oligomerization in the “head-to-tail” manner, which then transform directly into hexagonal boron nitride analog through the dehydrocyclization reaction, bypassing the formation of polyiminoborane.

  10. Is delta sup(15)N of sedimentary organic matter a good proxy for paleodenitrification in coastal waters of the eastern Arabian Sea?

    Digital Repository Service at National Institute of Oceanography (India)

    Agnihotri, R.; Naqvi, S.W.A; Kurian, S.; Altabet, M.A; Bratton, J.F.

    Recently published sedimentary records of delta sup(15)N from several coastal areas affected by both natural and anthropogenically produced shallow hypoxia with the objective of testing this as a proxy for denitrification in coastal settings...

  11. Chemical synthesis of glycoproteins with the specific installation of gradient enriched 15N-labeled amino acids for getting insight into glycoprotein behavior.

    Science.gov (United States)

    Kajihara, Yasuhiro; Nguyen, Minh Hien; Izumi, Masayuki; Sato, Hajime; Okamoto, Ryo

    2017-03-09

    We propose a novel partially 15N-labelling method for the amide backbone of a synthetic glycoprotein. By use of a chemical approach utilizing SPPS and NCL, we inserted thirteen 15N-labeled amino acids at specific positions of the protein backbone, while intentionally varying the enrichment of 15N atoms. This idea enables us to discriminate even the same type of amino acid based on the intensities of 1H-15N HSQC signals, thus allowing us to understand the dynamics of the local conformation of a synthetic homogeneous glycoprotein. Results suggested that the attachment of an oligosaccharide of either a bi-antennary complex-type or a high-mannose-type did not disturb protein conformation. However, T1 values suggested that the oligosaccharide influenced dynamics at the local conformation. Temperature-varied CD spectra and T1 values clearly indicated that oligosaccharides appeared to inhibit protein fluctuation or, in other words, stabilize protein structure.

  12. Job assignments, intrinsic motivation and explicit incentives

    OpenAIRE

    Nafziger, Julia

    2008-01-01

    This paper considers the interplay of job assignments with the intrinsic and extrinsic motivation of an agent. Job assignments influence the self confidence of the agent, and thereby his intrinsic motivation. Monetary reward allow the principal to complement intrinsic motivation with extrinsic incentives. The main result is that the principal chooses an inefficient job assignment rule to enhance the agent's intrinsic motivation even though she can motivate him with monetary rewards. This show...

  13. Elucidating the impact of nitrate and labile carbon application on spatial heterogeneity of denitrification by 15N modelling

    Science.gov (United States)

    Cardenas, Laura; Loick, Nadine; Dixon, Liz; Matthews, Peter; Gilsanz, Claudia; Bol, Roland; Lewicka-Szczebak, Dominika; Well, Reinhard

    2016-04-01

    N2O is considered to be an important GHG with soils representing its major source and accounting for approximately 6% of the current global warming and is also implicated in the depletion of stratospheric ozone. The atmospheric N2O concentration has been increasing since the Industrial Revolution making the understanding of its sources and removal processes very important for development of mitigation strategies. Bergstermann et al. (2011) found evidence of the existence of more than one pool of nitrate undergoing denitrification in a silty clay loam arable soil amended with glucose/nitrate solution. The Rayleigh type model was used to simulate d15N of N2O using process rates and associated fractionation factors, but assumptions for some of the model parameters had to be made due to lack of available data. In this study we carried out 2 incubation experiments in order to parameterise the model. To restrict the volume of soil reached by the amendment, we used blocks containing 3 soil cores that were incubated in one vessel to measure emissions of NO, N2O, N2 and CO2 from a clay grassland soil amended with KNO3 (N) and glucose (C) in three treatments: '1C' only 1 core received N and C (the other 2 received water), '3C' 3 cores received N and C, and 'Control' (received water only). The results showed changes in the d15Nbulk trends after day 6 post amendment application, coinciding with the decrease of N2O fluxes. We also report the results in the 15N site preference (SP) and d18O. We will show the results from the model validation based on this data.

  14. Early-stage changes in natural (13)C and (15)N abundance and nutrient dynamics during different litter decomposition.

    Science.gov (United States)

    Gautam, Mukesh Kumar; Lee, Kwang-Sik; Song, Byeong-Yeol; Lee, Dongho; Bong, Yeon-Sik

    2016-05-01

    Decomposition, nutrient, and isotopic (δ(13)C and δ(15)N) dynamics during 1 year were studied for leaf and twig litters of Pinus densiflora, Castanea crenata, Erigeron annuus, and Miscanthus sinensis growing on a highly weathered soil with constrained nutrient supply using litterbags in a cool temperate region of South Korea. Decay constant (k/year) ranged from 0.58 to 1.29/year, and mass loss ranged from 22.36 to 58.43 % among litter types. The results demonstrate that mass loss and nutrient dynamics of decomposing litter were influenced by the seasonality of mineralization and immobilization processes. In general, most nutrients exhibited alternate phases of rapid mineralization followed by gradual immobilization, except K, which was released throughout the field incubation. At the end of study, among all the nutrients only N and P showed net immobilization. Mobility of different nutrients from decomposing litter as the percentage of initial litter nutrient concentration was in the order of K > Mg > Ca > N ≈ P. The δ(13)C (0.32-6.70 ‰) and δ(15)N (0.74-3.90 ‰) values of residual litters showed nonlinear increase and decrease, respectively compared to initial isotopic values during decomposition. Litter of different functional types and chemical quality converged toward a conservative nutrient use strategy through mechanisms of slow decomposition and slow nutrient mobilization. Our results indicate that litter quality and season, are the most important regulators of litter decomposition in these forests. The results revealed significant relationships between litter decomposition rates and N, C:N ratio and P, and seasonality (temperature). These results and the convergence of different litters towards conservative nutrient use in these nutrient constrained ecosystems imply optimization of litter management because litter removal can have cascading effects on litter decomposition and nutrient availability in these systems.

  15. Oxygen determination in materials by {sup 18}O(p,αγ){sup 15}N nuclear reaction

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Sanjiv, E-mail: sanjucccm@rediffmail.com [National Centre for Compositional Characterization of Materials, BARC, ECIL Post, Hyderabad 500062 (India); Sunitha, Y.; Reddy, G.L.N.; Sukumar, A.A.; Ramana, J.V.; Sarkar, A. [National Centre for Compositional Characterization of Materials, BARC, ECIL Post, Hyderabad 500062 (India); Verma, Rakesh [Analytical Chemistry Division, BARC, Mumbai 400085 (India)

    2016-07-01

    The paper presents a proton induced γ-ray emission method based on {sup 18}O(p,αγ){sup 15}N nuclear reaction to determine bulk oxygen in materials. The determination involves the measurement of 5.27 MeV γ-rays emitted following the de-excitation of {sup 15}N nuclei. A description of the energetics of the reaction is given to provide an insight into the origin of 5.27 MeV γ-rays. In addition, thick target γ-ray yields and the limits of detection are measured to ascertain the analytical potential of the reaction. The thick-target γ-ray yields are measured with a high purity germanium detector and a bismuth germanate detector at 0° as well as 90° angles in 3.0–4.2 MeV proton energy region. The best limit of detection of about 1.3 at.% is achieved at 4.2 MeV proton energy for measurements at 0° as well 90° angles with the bismuth germanate detector while the uncertainty in quantitative analysis is <8%. The reaction has a probing depth of several tens of microns. Interferences can arise from fluorine due to the occurrence of {sup 19}F(p,αγ){sup 16}O reaction that emits 6–7 MeV γ-rays. The analytical potential of the methodology is demonstrated by determining oxygen in several oxide as well as non-oxide materials.

  16. Conformation of antifreeze glycoproteins as determined from conformational energy calculations and fully assigned proton NMR spectra

    Energy Technology Data Exchange (ETDEWEB)

    Bush, C.A.; Rao, B.N.N.

    1986-05-01

    The /sup 1/H NMR spectra of AFGP's ranging in molecular weight from 2600 to 30,000 Daltons isolated from several different species of polar fish have been measured. The spectrum of AFGP 1-4 from Pagothenia borchgrevinki with an average of 30 repeating subunits has a single resonance for each proton of the glycotripeptide repeating unit, (ala-(gal-(..beta..-1..-->..3) galNAc-(..cap alpha..--O-)thr-ala)/sub n/. Its /sup 1/H NMR spectrum including resonances of the amide protons has been completely assigned. Coupling constants and nuclear Overhauser enhancements (n.O.e.) between protons on distant residues imply conformational order. The 2600 dalton molecular weight glycopeptides (AFGP-8) have pro in place of ala at certain specific points in the sequence and AFGP-8R of Eleginus gracilis has arg in place of one thr. The resonances of pro and arg were assigned by decoupling. The resonances of the carboxy and amino terminals have distinct chemical shifts and were assigned in AFGP-8 of Boreogadus saida by titration. n.O.e. between ..cap alpha..--protons and amide protons of the adjacent residue (sequential n.O.e.) were used in assignments of additional resonances and to assign the distinctive resonances of thr followed by pro. Conformational energy calculations on the repeating glycotripeptide subunit of AFGP show that the ..cap alpha..--glucosidic linkage has a fixed conformation while the ..beta..--linkage is less rigid. A conformational model for AFGP 1-4, which is based on the calculations has the peptide in an extended left-handed helix with three residues per turn similar to polyproline II. The model is consistent with CD data, amide proton coupling constants, temperature dependence of amide proton chemical shifts.

  17. Are vascular epiphytes nitrogen or phosphorus limited? A study of plant (15) N fractionation and foliar N : P stoichiometry with the tank bromeliad Vriesea sanguinolenta.

    Science.gov (United States)

    Wanek, Wolfgang; Zotz, Gerhard

    2011-10-01

    Although there is unambiguous evidence for vascular epiphytic plants to be limited by insufficient water and nutrient supply under natural conditions, it is an open debate whether they are primarily phosphorus (P) or nitrogen (N) limited. Plant (15) N fractionation and foliar N : P stoichiometry of a tank epiphyte (Vriesea sanguinolenta), and its response to combined N-P fertilization, were studied under semi-natural conditions over 334 d to clarify the type of nutrient limitation. Plants collected in the field and experimental plants with limited nutrient supply showed significant plant (15) N fractionation (mean 5‰) and plant N : P ratios of c. 13.5. Higher relative growth rates and declines in plant (15) N fractionation (0.5‰) and in foliar N : P ratios to 8.5 in the high N-P treatment indicated that these epiphytes were P limited in situ. The critical foliar N : P ratio was 10.4, as derived from the breakpoint in the relationship between plant (15) N fractionation and foliar N : P. We interpret the widespread (15) N depletion of vascular epiphytes relative to their host trees as deriving from (15) N fractionation of epiphytes as a result of P limitation. High foliar N : P ratios (> 12) corroborate widespread P limitation (or co-limitation by N and P) of epiphytic bromeliads and, possibly, other epiphyte species.

  18. NVR-BIP: Nuclear Vector Replacement using Binary Integer Programming for NMR Structure-Based Assignments.

    Science.gov (United States)

    Apaydin, Mehmet Serkan; Çatay, Bülent; Patrick, Nicholas; Donald, Bruce R

    2011-05-01

    Nuclear magnetic resonance (NMR) spectroscopy is an important experimental technique that allows one to study protein structure and dynamics in solution. An important bottleneck in NMR protein structure determination is the assignment of NMR peaks to the corresponding nuclei. Structure-based assignment (SBA) aims to solve this problem with the help of a template protein which is homologous to the target and has applications in the study of structure-activity relationship, protein-protein and protein-ligand interactions. We formulate SBA as a linear assignment problem with additional nuclear overhauser effect constraints, which can be solved within nuclear vector replacement's (NVR) framework (Langmead, C., Yan, A., Lilien, R., Wang, L. and Donald, B. (2003) A Polynomial-Time Nuclear Vector Replacement Algorithm for Automated NMR Resonance Assignments. Proc. the 7th Annual Int. Conf. Research in Computational Molecular Biology (RECOMB), Berlin, Germany, April 10-13, pp. 176-187. ACM Press, New York, NY. J. Comp. Bio., (2004), 11, pp. 277-298; Langmead, C. and Donald, B. (2004) An expectation/maximization nuclear vector replacement algorithm for automated NMR resonance assignments. J. Biomol. NMR, 29, 111-138). Our approach uses NVR's scoring function and data types and also gives the option of using CH and NH residual dipolar coupling (RDCs), instead of NH RDCs which NVR requires. We test our technique on NVR's data set as well as on four new proteins. Our results are comparable to NVR's assignment accuracy on NVR's test set, but higher on novel proteins. Our approach allows partial assignments. It is also complete and can return the optimum as well as near-optimum assignments. Furthermore, it allows us to analyze the information content of each data type and is easily extendable to accept new forms of input data, such as additional RDCs.

  19. Estado nutricional da cultura de cana-de-açúcar (cana-planta em experimentos com 15N Nutritional status of sugar cane (planted cane in 15N experiments

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Faroni

    2009-12-01

    Full Text Available Estudos com isótopos estáveis são cada vez mais comuns pela qualidade e confiabilidade dos resultados obtidos e por não apresentarem risco ao homem e ao ambiente. No entanto, é necessário o uso de microparcelas para diminuir o custo do projeto. Por suas áreas serem muito reduzidas em relação à parcela onde está inserida, o objetivo deste trabalho foi comparar o estado nutricional de plantas das microparcelas com as do restante das parcelas experimentais, na fase de máximo desenvolvimento da cultura da cana-de-açúcar, em experimentos com doses de N. Foram instalados três experimentos, no Estado de São Paulo, com três doses de N e uma testemunha, com quatro repetições, em delineamento em blocos completos casualizados. Nas parcelas, de 48 linhas de 15 m cada, foram instaladas microparcelas de 3 m² com fertilizante-15N. Na época de máximo desenvolvimento da cultura foram coletadas folhas-diagnóstico para a avaliação de seu estado nutricional em todas as parcelas e microparcelas. Nessas folhas foram determinados os teores de macronutrientes, e os resultados foram submetidos à análise de variância. Foram comparados, em cada área experimental e de forma conjunta, os resultados das parcelas com os das microparcelas quanto aos teores foliares dos nutrientes e, quando significativo o efeito das doses de N na variação desses, foram feitas regressões. Não houve diferença entre os teores de N, P, Ca, Mg e S das folhas-diagnóstico entre as parcelas e as microparcelas, as quais foram consideradas representativas da área experimental. A adubação nitrogenada aumentou os teores dos nutrientes nas folhas-diagnóstico, o que deve ser considerado na avaliação nutricional da cultura quando este tipo de experimento for realizado.Studies with stable isotopes are becoming more common due to the increased safety of operation and quality and reliability of results. However, the use of microplots is required to decrease the costs of

  20. 33 CFR 401.61 - Assigned frequencies.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Assigned frequencies. 401.61 Section 401.61 Navigation and Navigable Waters SAINT LAWRENCE SEAWAY DEVELOPMENT CORPORATION, DEPARTMENT... frequencies. The Seaway stations operate on the following assigned VHF frequencies: 156.8 MHz—(channel...

  1. Optimization of Rated Officer Staff Assignments

    Science.gov (United States)

    2007-06-01

    determinant +1, -1 or 0, which holds for the assignment problem ( Bazaraa et al., 1990). The Unimodularity Theorem states for an integer matrix A with...Prentice-Hall, 1993. 2. Air Force Instruction 36-2110, Assignments, 20 April 2005. 3. Bazaraa , Mokhtar S., Jarvis, John J. and Sherali, Hanif D

  2. A Role-Based Fuzzy Assignment Model

    Institute of Scientific and Technical Information of China (English)

    ZUO Bao-he; FENG Shan

    2002-01-01

    It's very important to dynamically assign the tasks to corresponding actors in workflow management system, especially in complex applications. This improves the flexibility of workflow systems.In this paper, a role-based workflow model with fuzzy optimized intelligent assignment is proposed and applied in the investment management system. A groupware-based software model is also proposed.

  3. Stress Assignment in Reading Italian Polysyllabic Pseudowords

    Science.gov (United States)

    Sulpizio, Simone; Arduino, Lisa S.; Paizi, Despina; Burani, Cristina

    2013-01-01

    In 4 naming experiments we investigated how Italian readers assign stress to pseudowords. We assessed whether participants assign stress following distributional information such as stress neighborhood (the proportion and number of existent words sharing orthographic ending and stress pattern) and whether such distributional information affects…

  4. Effects of Individualized Assignments on Biology Achievement.

    Science.gov (United States)

    Kremer, Philip L.

    1983-01-01

    Compared detailed (favoring field dependence and induction) and nondetailed (favoring field dependence and deduction) assignments on biology achievement of grade 10 male students (N=95) over a seven-month period. Detailed assignments, employing pictorial and verbal block diagrams and high structure, significantly enhanced learning among some…

  5. Online Discussion Assignments Improve Students' Class Preparation

    Science.gov (United States)

    Lineweaver, Tara T.

    2010-01-01

    To increase the number of students who read the text before class and to promote student interaction centering on text material, I developed an online discussion assignment as a required component of a cognitive psychology course. Across 2 studies, this assignment had a limited effect on examination performance, but students completing online…

  6. 48 CFR 208.7002 - Assignment authority.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Assignment authority. 208..., DEPARTMENT OF DEFENSE ACQUISITION PLANNING REQUIRED SOURCES OF SUPPLIES AND SERVICES Coordinated Acquisition 208.7002 Assignment authority. (a) Under the DoD Coordinated Acquisition Program,...

  7. 7 CFR 601.1 - Functions assigned.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 6 2010-01-01 2010-01-01 false Functions assigned. 601.1 Section 601.1 Agriculture... AGRICULTURE GENERAL FUNCTIONS § 601.1 Functions assigned. The Natural Resources Conservation Service (NRCS) is... conserve, improve, and sustain natural resources. The following functions support the mission. (a)...

  8. A Model of Hierarchical Key Assignment Scheme

    Institute of Scientific and Technical Information of China (English)

    ZHANG Zhigang; ZHAO Jing; XU Maozhi

    2006-01-01

    A model of the hierarchical key assignment scheme is approached in this paper, which can be used with any cryptography algorithm. Besides, the optimal dynamic control property of a hierarchical key assignment scheme will be defined in this paper. Also, our scheme model will meet this property.

  9. Detecting Plagiarism in MS Access Assignments

    Science.gov (United States)

    Singh, Anil

    2013-01-01

    Assurance of individual effort from students in computer-based assignments is a challenge. Due to digitization, students can easily use a copy of their friend's work and submit it as their own. Plagiarism in assignments puts students who cheat at par with those who work honestly and this compromises the learning evaluation process. Using a…

  10. Nitrogen-15 labeling of Crotalaria juncea green manure Marcação do adubo verde Crotalaria juncea com 15N

    Directory of Open Access Journals (Sweden)

    Edmilson José Ambrosano

    2003-02-01

    Full Text Available Most studies dealing with the utilization of 15N labeled plant material do not present details about the labeling technique. This is especially relevant for legume species since biological nitrogen fixation difficults plant enrichment. A technique was developed for labeling leguminous plant tissue with 15N to obtain labeled material for nitrogen dynamics studies. Sun hemp (Crotalaria juncea L. was grown on a Paleudalf, under field conditions. An amount of 58.32 g of urea with 70.57 ± 0.04 atom % 15N was sprayed three times on plants grown on eight 6-m²-plots. The labelled material presented 2.412 atom % 15N in a total dry matter equivalent to 9 Mg ha-1 This degree of enrichment enables the use of the green manure in pot or field experiments requiring 15N-labeled material.A grande maioria dos estudos com a utilização de material vegetal marcado com o isótopo 15N não apresentam detalhes tão importantes sobre como foram obtidos esses materiais. Em se tratando de marcação de leguminosas as dificuldades em se obter material marcado com 15N são ainda maiores pelo fato de serem plantas fixadoras de nitrogênio. Isso posto foi estabelecida uma técnica de marcação de leguminosas com nitrogênio (15N, com o objetivo de obter material vegetal marcado isotopicamente para estudos de dinâmica do nitrogênio. Cultivou-se a leguminosa crotalária júncea (Crotalaria juncea L., em Argissolo Vermelho Amarelo distrófico, em campo. Ao se aplicarem via foliar 58,32 gramas de uréia em oito canteiros experimentais, (uréia com 70,57 ± 0,04% de átomos de 15N parceladas em três vezes, obteve-se material vegetal marcado seco que continha 2,412 % em átomos de 15N em uma massa seca equivalente a 9 Mg ha-1. Essa marcação permite o uso dessa massa vegetal em estudos de dinâmica de nitrogênio.

  11. Preferential flow, nitrogen transformations and 15N balance under urine-affected areas of irrigated and non-irrigated clover-based pastures

    Science.gov (United States)

    Pakro, Naser; Dillon, Peter

    1995-12-01

    Urine-affected areas can lead to considerable losses of N by leaching, ammonia volatilisation and denitrification from dairy pastures in the southeast of South Australia. Potable groundwater supplies are considered to have become contaminated by nitrate as a result of leaching from these leguminous pastures. Dairy cow urine, labelled with 15N urea, was applied to micro-plots and mini-lysimeters installed in two adjacent irrigated (white clover-rye grass) and non-irrigated (subterranean clover-annual grasses) paddocks of a dairy farm on four occasions representing different seasonal conditions. These experiments allowed measurement of nitrogen transformations, recovery of 15N in the pasture and soil, and leaching below various depths. Gaseous losses were calculated from the nitrogen balance. The results of the four experiments showed that within a day of urine application up to 40% of the applied urinary-N was leached below a depth of 150 mm as a result of macropore flow in the irrigated paddock, and up to 24% in the non-irrigated one. After application to the irrigated paddock 17% of the urinary-N moved immediately below 300 mm but only 2% below the 450-mm depth. The urinary-N remaining in the soil was converted from urea to ammonium within a day regardless of season. Within the first 7 days of application six times more nitrate was produced in summer than in winter. This has obvious implications for leaching potential. Leaching of 15N from the top 150 mm of soil, following urine applications in all seasons, was between 41% and 62% of the applied 15N in the irrigated paddock and 25-51% in the non-irrigated paddock. However, leaching losses measured at depths of 300 or 450 mm were smaller by a factor of 2-4. The leaching loss of 15N applied in spring in both paddocks was 41% below 150 mm and 12% below 450 mm. Recovery of 15N from the soil-plant system in the 450-nm deep lysimeters was ˜60% of that applied. Estimated ammonia was ˜9% of applied 15N with no paddock

  12. Airport Gate Assignment: New Model and Implementation

    CERN Document Server

    Li, Chendong

    2008-01-01

    Airport gate assignment is of great importance in airport operations. In this paper, we study the Airport Gate Assignment Problem (AGAP), propose a new model and implement the model with Optimization Programming language (OPL). With the objective to minimize the number of conflicts of any two adjacent aircrafts assigned to the same gate, we build a mathematical model with logical constraints and the binary constraints, which can provide an efficient evaluation criterion for the Airlines to estimate the current gate assignment. To illustrate the feasibility of the model we construct experiments with the data obtained from Continental Airlines, Houston Gorge Bush Intercontinental Airport IAH, which indicate that our model is both energetic and effective. Moreover, we interpret experimental results, which further demonstrate that our proposed model can provide a powerful tool for airline companies to estimate the efficiency of their current work of gate assignment.

  13. Photothermal resonance

    DEFF Research Database (Denmark)

    2014-01-01

    The present invention relates to a method for detecting photo-thermal absorbance of a material utilising a mechanically temperature sensitive resonator (20) and a sample being arrange in thermal communication with the temperature sensitive resonator. The present invention further relates to an ap......The present invention relates to a method for detecting photo-thermal absorbance of a material utilising a mechanically temperature sensitive resonator (20) and a sample being arrange in thermal communication with the temperature sensitive resonator. The present invention further relates...... to an apparatus for detecting photo-thermal absorbance of a sample....