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Sample records for 13c 15n nmr

  1. Biosynthetic uniform 13C,15N-labelling of zervamicin IIB. Complete 13C and 15N NMR assignment.

    PubMed

    Ovchinnikova, Tatyana V; Shenkarev, Zakhar O; Yakimenko, Zoya A; Svishcheva, Natalia V; Tagaev, Andrey A; Skladnev, Dmitry A; Arseniev, Alexander S

    2003-01-01

    Zervamicin IIB is a member of the alpha-aminoisobutyric acid containing peptaibol antibiotics. A new procedure for the biosynthetic preparation of the uniformly 13C- and 15N-enriched peptaibol is described This compound was isolated from the biomass of the fungus-producer Emericellopsis salmosynnemata strain 336 IMI 58330 obtained upon cultivation in the totally 13C, 15N-labelled complete medium. To prepare such a medium the autolysed biomass and the exopolysaccharides of the obligate methylotrophic bacterium Methylobacillus flagellatus KT were used. This microorganism was grown in totally 13C, 15N-labelled minimal medium containing 13C-methanol and 15N-ammonium chloride as the only carbon and nitrogen sources. Preliminary NMR spectroscopic analysis indicated a high extent of isotope incorporation (> 90%) and led to the complete 13C- and 15N-NMR assignment including the stereospecific assignment of Aib residues methyl groups. The observed pattern of the structurally important secondary chemical shifts of 1H(alpha), 13C=O and 13C(alpha) agrees well with the previously determined structure of zervamicin IIB in methanol solution. PMID:14658801

  2. 15N and13C NMR investigation of hydroxylamine-derivatized humic substances

    USGS Publications Warehouse

    Thorn, K.A.; Arterburn, J.B.; Mikita, M.A.

    1992-01-01

    Five fulvic and humic acid samples of diverse origins were derivatized with 15N-labeled hydroxylamine and analyzed by liquid-phase 15N NMR spectrometry. The 15N NMR spectra indicated that hydroxylamine reacted similarly with all samples and could discriminate among carbonyl functional groups. Oximes were the major derivatives; resonances attributable to hydroxamic acids, the reaction products of hydroxylamine with esters, and resonances attributable to the tautomeric equilibrium position between the nitrosophenol and monoxime derivatives of quinones, the first direct spectroscopic evidence for quinones, also were evident. The 15N NMR spectra also suggested the presence of nitriles, oxazoles, oxazolines, isocyanides, amides, and lactams, which may all be explained in terms of Beckmann reactions of the initial oxime derivatives. INEPT and ACOUSTIC 15N NMR spectra provided complementary information on the derivatized samples. 13C NMR spectra of derivatized samples indicated that the ketone/quinone functionality is incompletely derivatized with hydroxylamine. ?? 1991 American Chemical Society.

  3. 1H, 13C and 15N NMR assignments of phenazopyridine derivatives.

    PubMed

    Burgueño-Tapia, Eleuterio; Mora-Pérez, Yolanda; Morales-Ríos, Martha S; Joseph-Nathan, Pedro

    2005-03-01

    Phenazopyridine hydrochloride (1), a drug in clinical use for many decades, and some derivatives were studied by one- and two-dimensional (1)H, (13)C and (15)N NMR methodology. The assignments, combined with DFT calculations, reveal that the preferred protonation site of the drug is the pyridine ring nitrogen atom. The chemoselective acetylation of phenazopyridine (2) and its influence on the polarization of the azo nitrogen atoms were evidenced by the (15)N NMR spectra. Molecular calculations of the phenazopyridines 2-4 show that the pyridine and phenyl groups are oriented in an antiperiplanar conformation with intramolecular hydrogen bonding between the N-b atom and the C-2 amino group preserving the E-azo stereochemistry. PMID:15625718

  4. Preparation of 13C and 15N labelled RNAs for heteronuclear multi-dimensional NMR studies.

    PubMed

    Nikonowicz, E P; Sirr, A; Legault, P; Jucker, F M; Baer, L M; Pardi, A

    1992-09-11

    A procedure is described for the efficient preparation of isotopically enriched RNAs of defined sequence. Uniformly labelled nucleotide 5'triphosphates (NTPs) were prepared from E.coli grown on 13C and/or 15N isotopically enriched media. These procedures routinely yield 180 mumoles of labelled NTPs per gram of 13C enriched glucose. The labelled NTPs were then used to synthesize RNA oligomers by in vitro transcription. Several 13C and/or 15N labelled RNAs have been synthesized for the sequence r(GGCGCUUGCGUC). Under conditions of high salt or low salt, this RNA forms either a symmetrical duplex with two U.U base pairs or a hairpin containing a CUUG loop respectively. These procedures were used to synthesize uniformly labelled RNAs and a RNA labelled only on the G and C residues. The ability to generate milligram quantities of isotopically labelled RNAs allows application of multi-dimensional heteronuclear magnetic resonance experiments that enormously simplify the resonance assignment and solution structure determination of RNAs. Examples of several such heteronuclear NMR experiments are shown. PMID:1383927

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

    PubMed

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

    2015-06-01

    (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. PMID:25843843

  6. Application of unsymmetrical indirect covariance NMR methods to the computation of the (13)C <--> (15)N HSQC-IMPEACH and (13)C <--> (15)N HMBC-IMPEACH correlation spectra.

    PubMed

    Martin, Gary E; Hilton, Bruce D; Irish, Patrick A; Blinov, Kirill A; Williams, Antony J

    2007-10-01

    Utilization of long-range (1)H--(15)N heteronuclear chemical shift correlation has continually grown in importance since the first applications were reported in 1995. More recently, indirect covariance NMR methods have been introduced followed by the development of unsymmetrical indirect covariance processing methods. The latter technique has been shown to allow the calculation of hyphenated 2D NMR data matrices from more readily acquired nonhyphenated 2D NMR spectra. We recently reported the use of unsymmetrical indirect covariance processing to combine (1)H--(13)C GHSQC and (1)H--(15)N GHMBC long-range spectra to yield a (13)C--(15)N HSQC-HMBC chemical shift correlation spectrum that could not be acquired in a reasonable period of time without resorting to (15)N-labeled molecules. We now report the unsymmetrical indirect covariance processing of (1)H--(13)C GHMBC and (1)H--(15)N IMPEACH spectra to afford a (13)C--(15)N HMBC-IMPEACH spectrum that has the potential to span as many as six to eight bonds. Correlations for carbon resonances long-range coupled to a protonated carbon in the (1)H--(13)C HMBC spectrum are transferred via the long-range (1)H--(15)N coupling pathway in the (1)H--(15)N IMPEACH spectrum to afford a much broader range of correlation possibilities in the (13)C--(15)N HMBC-IMPEACH correlation spectrum. The indole alkaloid vincamine is used as a model compound to illustrate the application of the method. PMID:17729230

  7. (1)H, (13)C and (15)N NMR assignments of a calcium-binding protein from Entamoeba histolytica.

    PubMed

    Verma, Deepshikha; Bhattacharya, Alok; Chary, Kandala V R

    2016-04-01

    We report almost complete sequence specific (1)H, (13)C and (15)N NMR assignments of a 150-residue long calmodulin-like calcium-binding protein from Entamoeba histolytica (EhCaBP6), as a prelude to its structural and functional characterization. PMID:26377206

  8. Bonding in hard and elastic amorphous carbon nitride films investigated using 15N, 13C, and 1H NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Gammon, W. J.; Hoatson, G. L.; Holloway, B. C.; Vold, R. L.; Reilly, A. C.

    2003-11-01

    The nitrogen bonding in hard and elastic amorphous carbon nitride (a-CNx) films is examined with 15N, 13C, and 1H nuclear magnetic resonance (NMR) spectroscopy. Films were deposited by dc magnetron sputtering, in a pure nitrogen discharge on Si(001) substrates at 300 °C. Nanoindentation tests revealed an elastic recovery of 80%, a hardness of 5 GPa, and an elastic modulus of 47 GPa. The NMR results show that nitrogen bonding in this material is consistent with sp2 hybridized nitrogen incorporated in an aromatic carbon environment. The data also indicate that the a-CNx prepared for this study has very low hydrogen content and is hydrophilic. Specifically, analysis of 15N and 13C cross polarization magic angle spinning and 1H NMR experiments suggests that water preferentially protonates nitrogen sites.

  9. Quantitation of metabolic compartmentation in hyperammonemic brain by natural abundance 13C-NMR detection of 13C-15N coupling patterns and isotopic shifts.

    PubMed

    Lapidot, A; Gopher, A

    1997-02-01

    In the present study, the removal of cerebral ammonia by glutamine synthetase (GS) and by reductive amination of 2-oxoglutarate by glutamate dehydrogenase in the presence of an amino donor group, was determined in hyperammonemic rabbit brains. The 15N enrichments of brain metabolite alpha-amino and amide positions of glutamine, glutamate, and alanine were determined by the indirect detection of 15N-labeled compounds of the 13C-15N spin coupling patterns of natural abundance 13C-NMR spectra. The 13C-NMR spectra of brain extracts were obtained from rabbits infused with 15NH4Cl with or without intraperitoneal infusion of the GS inhibitor, L-methionine DL-sulfoximine, in a reasonable acquisition time period. When 15NH4Cl was infused, [5-15N]glutamine and [2-15N]glutamine concentrations reached 5.2 mumol/100 mg protein and 3.6 mumol/100 mg protein, respectively, which indicates the relatively high activity of reductive amination of 2-oxoglutarate in the glutamate dehydrogenase reaction. The low concentration of [2-15N]glutamate, which is about 30% of that of [2-15N]glutamine obtained in this study, suggests that very little glutamine serves as a precursor of neuronal glutamate. When GS was inhibited by L-methionine DL-sulfoximine, a flux of 15NH4+ via the residual activity of GS was accompanied by an apparent increase of [2-15N]glutamate and [15N]alanine concentrations (2.9 mumol/100 mg protein and 1.8 mumol/100 mg protein, respectively). These findings and those obtained from 13C-13C isotopomer analysis (Lapidot and Gopher, 1994b) suggest that astrocytic 2-oxoglutarate is partially utilized (together with an amino group donor) as a precursor for neuronal glutamate in the hyperammonemic brain when GS is inhibited. This process can partly replace GS activity in metabolizing ammonia in the hyperammonemic rabbit brain. PMID:9057821

  10. /sup 15/N and /sup 13/C NMR determination of methionine metabolism in developing soybean cotyledons

    SciTech Connect

    Coker, G.T. III; Garbow, J.R.; Schaefer, J.

    1987-03-01

    The metabolism of D- and L-methionine by immature cotyledons of soybean (Glycine max, L. cv Elf) grown in culture has been investigated using solid-state /sup 13/C and /sup 15/N nuclear magnetic resonance. D-Methionine is taken up by the cotyledons and converted to an amide, most likely by N-malonylation. About 16% of the L-methionine taken up is incorporated intact into protein, and 25% remains as soluble methionine. Almost two-thirds of the L-methionine that enters the cotyledons is degraded. The largest percentage of this is used in transmethylation of the carboxyl groups of pectin. Methionine is not extensively converted to polyamines. The authors attribute the stimulation of growth of the cotyledons by exogenous methionine to the bypassing of a rate-limiting methyl-transfer step in the synthesis of methionine itself, and subsequently of pectins and proteins.

  11. Simple, efficient protocol for enzymatic synthesis of uniformly 13C, 15N-labeled DNA for heteronuclear NMR studies.

    PubMed Central

    Masse, J E; Bortmann, P; Dieckmann, T; Feigon, J

    1998-01-01

    The use of uniformly 13C,15N-labeled RNA has greatly facilitated structural studies of RNA oligonucleotides by NMR. Application of similar methodologies for the study of DNA has been limited, primarily due to the lack of adequate methods for sample preparation. Methods for both chemical and enzymatic synthesis of DNA oligonucleotides uniformly labeled with 13C and/or 15N have been published, but have not yet been widely used. We have developed a modified procedure for preparing uniformly 13C,15N-labeled DNA based on enzymatic synthesis using Taq DNA polymerase. The highly efficient protocol results in quantitative polymerization of the template and approximately 80% incorporation of the labeled dNTPs. Procedures for avoiding non-templated addition of nucleotides or for their removal are given. The method has been used to synthesize several DNA oligonucleotides, including two complementary 15 base strands, a 32 base DNA oligonucleotide that folds to form an intramolecular triplex and a 12 base oligonucleotide that dimerizes and folds to form a quadruplex. Heteronuclear NMR spectra of the samples illustrate the quality of the labeled DNA obtained by these procedures. PMID:9592146

  12. Differentiation of histidine tautomeric states using (15)N selectively filtered (13)C solid-state NMR spectroscopy.

    PubMed

    Miao, Yimin; Cross, Timothy A; Fu, Riqiang

    2014-08-01

    The histidine imidazole ring in proteins usually contains a mixture of three possible tautomeric states (two neutral - τ and π states and a charged state) at physiological pHs. Differentiating the tautomeric states is critical for understanding how the histidine residue participates in many structurally and functionally important proteins. In this work, one dimensional (15)N selectively filtered (13)C solid-state NMR spectroscopy is proposed to differentiate histidine tautomeric states and to identify all (13)C resonances of the individual imidazole rings in a mixture of tautomeric states. When (15)N selective 180° pulses are applied to the protonated or non-protonated nitrogen region, the (13)C sites that are bonded to the non-protonated or protonated nitrogen sites can be identified, respectively. A sample of (13)C, (15)N labeled histidine powder lyophilized from a solution at pH 6.3 has been used to illustrate the usefulness of this scheme by uniquely assigning resonances of the neutral τ and charged states from the mixture. PMID:25026459

  13. Differentiation of Histidine Tautomeric States using 15N Selectively Filtered 13C Solid-State NMR Spectroscopy

    PubMed Central

    Miao, Yimin; Cross, Timothy A.; Fu, Riqiang

    2014-01-01

    The histidine imidazole ring in proteins usually contains a mixture of three possible tautomeric states (two neutral - τ and π states and a charged state) at physiological pHs. Differentiating the tautomeric states is critical for understanding how the histidine residue participates in many structurally and functionally important proteins. In this work, one dimensional 15N selectively filtered 13C solid-state NMR spectroscopy is proposed to differentiate histidine tautomeric states and to identify all 13C resonances of the individual imidazole rings in a mixture of tautomeric states. When 15N selective 180° pulses are applied to the protonated or non-protonated nitrogen region, the 13C sites that are bonded to the non-protonated or protonated nitrogen sites can be identified, respectively. A sample of 13C,15N labeled histidine powder lyophilized from a solution at pH 6.3 has been used to illustrate the usefulness of this scheme by uniquely assigning resonances of the neutral τ and charged states from the mixture. PMID:25026459

  14. Differentiation of histidine tautomeric states using 15N selectively filtered 13C solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Miao, Yimin; Cross, Timothy A.; Fu, Riqiang

    2014-08-01

    The histidine imidazole ring in proteins usually contains a mixture of three possible tautomeric states (two neutral - τ and π states and a charged state) at physiological pHs. Differentiating the tautomeric states is critical for understanding how the histidine residue participates in many structurally and functionally important proteins. In this work, one dimensional 15N selectively filtered 13C solid-state NMR spectroscopy is proposed to differentiate histidine tautomeric states and to identify all 13C resonances of the individual imidazole rings in a mixture of tautomeric states. When 15N selective 180° pulses are applied to the protonated or non-protonated nitrogen region, the 13C sites that are bonded to the non-protonated or protonated nitrogen sites can be identified, respectively. A sample of 13C, 15N labeled histidine powder lyophilized from a solution at pH 6.3 has been used to illustrate the usefulness of this scheme by uniquely assigning resonances of the neutral τ and charged states from the mixture.

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

    PubMed

    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. PMID:19019638

  16. A New Tool for NMR Crystallography: Complete (13)C/(15)N Assignment of Organic Molecules at Natural Isotopic Abundance Using DNP-Enhanced Solid-State NMR.

    PubMed

    Märker, Katharina; Pingret, Morgane; Mouesca, Jean-Marie; Gasparutto, Didier; Hediger, Sabine; De Paëpe, Gaël

    2015-11-01

    NMR crystallography of organic molecules at natural isotopic abundance (NA) strongly relies on the comparison of assigned experimental and computed NMR chemical shifts. However, a broad applicability of this approach is often hampered by the still limited (1)H resolution and/or difficulties in assigning (13)C and (15)N resonances without the use of structure-based chemical shift calculations. As shown here, such difficulties can be overcome by (13)C-(13)C and for the first time (15)N-(13)C correlation experiments, recorded with the help of dynamic nuclear polarization. We present the complete de novo (13)C and (15)N resonance assignment at NA of a self-assembled 2'-deoxyguanosine derivative presenting two different molecules in the asymmetric crystallographic unit cell. This de novo assignment method is exclusively based on aforementioned correlation spectra and is an important addition to the NMR crystallography approach, rendering firstly (1)H assignment straightforward, and being secondly a prerequisite for distance measurements with solid-state NMR. PMID:26485326

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

    USGS Publications Warehouse

    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

  18. 1H, 13C, 15N and 195Pt NMR studies of Au(III) and Pt(II) chloride organometallics with 2-phenylpyridine.

    PubMed

    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. PMID:19691018

  19. Simultaneous cross polarization to 13C and 15N with 1H detection at 60 kHz MAS solid-state NMR

    NASA Astrophysics Data System (ADS)

    Das, Bibhuti B.; Opella, Stanley J.

    2016-01-01

    We describe high resolution MAS solid-state NMR experiments that utilize 1H detection with 60 kHz magic angle spinning; simultaneous cross-polarization from 1H to 15N and 13C nuclei; bidirectional cross-polarization between 13C and 15N nuclei; detection of both amide nitrogen and aliphatic carbon 1H; and measurement of both 13C and 15N chemical shifts through multi-dimensional correlation experiments. Three-dimensional experiments correlate amide 1H and alpha 1H selectively with 13C or 15N nuclei in a polypeptide chain. Two separate three-dimensional spectra correlating 1Hα/13Cα/1HN and 1HN/15N/1Hα are recorded simultaneously in a single experiment, demonstrating that a twofold savings in experimental time is potentially achievable. Spectral editing using bidirectional coherence transfer pathways enables simultaneous magnetization transfers between 15N, 13Cα(i) and 13C‧(i-1), facilitating intra- and inter-residue correlations for sequential resonance assignment. Non-uniform sampling is integrated into the experiments, further reducing the length of experimental time.

  20. Multinuclear 1H, 13C and 15N NMR study of some substituted 2-amino-4-nitropyridines and their N-oxides

    NASA Astrophysics Data System (ADS)

    Laihia, K.; Kolehmainen, E.; Kauppinen, R.; Lorenc, J.; Puszko, A.

    2002-05-01

    1H, 13C and 15N NMR chemical shift assignments based on pulsed field gradient selected PFG 1H,X (X= 13C and 15N) HMQC and HMBC experiments are reported for three 4-nitropyridine N-oxides and four 4-nitropyridines. It was found that an ortho effect of a methyl group inhibits the deshielding effect of the 4-nitro group and that this effect and the so-called back donation is influenced by electronegativity and position of substituents in the multisubstituted pyridine N-oxides. The shielding effect of N-oxide group is most pronounced in the 15N NMR chemical shifts of the studied compounds. This effect is further modified by methylamino, methylnitramino, 5- or 3-methyl and 4-nitro groups. Among them the 4-nitro group exerts the highest influence on the shielding effect of the N-oxide functionality. Experimental 1H, 13C and 15N NMR chemical shifts and GIAO/DFT theoretical calculations are consistent with each other and supported by the reactivity on nucleophilic substitution, the UV spectral and the dipole moment data.

  1. Global Fold of Human Cannabinoid Type 2 Receptor Probed by Solid-State 13C-, 15N-MAS NMR and Molecular Dynamics Simulations

    PubMed Central

    Kimura, Tomohiro; Vukoti, Krishna; Lynch, Diane L.; Hurst, Dow P.; Grossfield, Alan; Pitman, Michael C.; Reggio, Patricia H.; Yeliseev, Alexei A.; Gawrisch, Klaus

    2014-01-01

    The global fold of human cannabinoid type 2 (CB2) receptor in the agonist-bound active state in lipid bilayers was investigated by solid-state 13C- and 15N magic-angle spinning (MAS) NMR, in combination with chemical-shift prediction from a structural model of the receptor obtained by microsecond-long molecular dynamics (MD) simulations. Uniformly 13C-, and 15N-labeled CB2 receptor was expressed in milligram quantities by bacterial fermentation, purified, and functionally reconstituted into liposomes. 13C MAS NMR spectra were recorded without sensitivity enhancement for direct comparison of Cα, Cβ, and C=O bands of superimposed resonances with predictions from protein structures generated by MD. The experimental NMR spectra matched the calculated spectra reasonably well indicating agreement of the global fold of the protein between experiment and simulations. In particular, the 13C chemical shift distribution of Cα resonances was shown to be very sensitive to both the primary amino acid sequence and the secondary structure of CB2. Thus the shape of the Cα band can be used as an indicator of CB2 global fold. The prediction from MD simulations indicated that upon receptor activation a rather limited number of amino acid residues, mainly located in the extracellular loop 2 and the second half of intracellular loop 3, change their chemical shifts significantly (≥1.5 ppm for carbons and ≥5.0 ppm for nitrogens). Simulated two-dimensional 13Cα(i)-13C=O(i) and 13C=O(i)-15NH(i+1) dipolar-interaction correlation spectra provide guidance for selective amino-acid labeling and signal assignment schemes to study the molecular mechanism of activation of CB2 by solid-state MAS NMR. PMID:23999926

  2. HCN, a triple-resonance NMR technique for selective observation of histidine and tryptophan side chains in 13C/15N-labeled proteins.

    PubMed

    Sudmeier, J L; Ash, E L; Günther, U L; Luo, X; Bullock, P A; Bachovchin, W W

    1996-12-01

    HCN, a new 3D NMR technique for stepwise coherence transfer from 1H to 13C to 15N and reverse through direct spin couplings 1JCH and 1JCN, is presented as a method for detection and assignment of histidine and tryptophan side-chain 1H, 13C, and 15N resonances in uniformly 13C/15N-labeled proteins. Product-operator calculations of cross-peak volumes vs adjustable delay tau 3 were employed for determination of optimal tau 3. For the phosphatidylinositol 3-kinase (PI3K SH3 domain, MW = 9.6 kD) at pH 6, H(C)N, the 1H/15N projection, produced observable cross peaks within 20 min. and was completely selective for the single tryptophan and single histidine. The 3D HCN experiment yielded well-defined cross peaks in 20 h for the 13C/15N-labeled origin-specific DNA binding domain from simian virus 40 T-antigen (T-ag-OBD131-259, MW = 15.4 kD) at pH 5.5. Resonances from all six histidines in T-ag-OBD were observed, and 11 of the 12 1H and 13C chemical shifts and 10 of the 12 15N chemical shifts were determined. The 13C dimension proved essential in assignment of the multiply overlapping 1H and 15N resonances. From the spectra recorded at a single pH, three of the imidazoles were essentially neutral and the other three were partially protonated (22-37%). HCN yielded strong cross peaks after 18 h on a 2.0 mM sample of phenylmethanesulfonyl fluoride (PMSF)-inhibited alpha-lytic protease (MW = 19.8 kD) at pH 4.4. No spectra have been obtained, however, of native or boronic acid-inhibited alpha-lytic protease after 18 h at various temperatures ranging from 5 to 55 degrees C, probably due to efficient relaxation of active-site imidazole 1H and/or 15N nuclei. PMID:8995843

  3. HCN, A Triple-Resonance NMR Technique for Selective Observation of Histidine and Tryptophan Side Chains in 13C/ 15N-Labeled Proteins

    NASA Astrophysics Data System (ADS)

    Sudmeier, James L.; Ash, Elissa L.; Günther, Ulrich L.; Luo, Xuelian; Bullock, Peter A.; Bachovchin, William W.

    1996-12-01

    HCN, a new 3D NMR technique for stepwise coherence transfer from1H to13C to15N and reverse through direct spin couplings1JCHand1JCN, is presented as a method for detection and assignment of histidine and tryptophan side-chain1H,13C, and15N resonances in uniformly13C/15N-labeled proteins. Product-operator calculations of cross-peak volumes vs adjustable delay τ3were employed for determination of optimal τ3. For the phosphatidylinositol 3-kinase (PI3K SH3 domain, MW = 9.6 kD) at pH 6, H(C)N, the1H/15N projection, produced observable cross peaks within 20 min. and was completely selective for the single tryptophan and single histidine. The 3D HCN experiment yielded well-defined cross peaks in 20 h for the13C/15N-labeled origin-specific DNA binding domain from simian virus 40 T-antigen (T-ag-OBD131-259, MW = 15.4 kD) at pH 5.5. Resonances from all six histidines in T-ag-OBD were observed, and 11 of the 121H and13C chemical shifts and 10 of the 1215N chemical shifts were determined. The13C dimension proved essential in assignment of the multiply overlapping1H and15N resonances. From the spectra recorded at a single pH, three of the imidazoles were essentially neutral and the other three were partially protonated (22-37%). HCN yielded strong cross peaks after 18 h on a 2.0 mMsample of phenylmethanesulfonyl fluoride (PMSF)-inhibited α-lytic protease (MW = 19.8 kD) at pH 4.4. No spectra have been obtained, however, of native or boronic acid-inhibited α-lytic protease after 18 h at various temperatures ranging from 5 to 55°C, probably due to efficient relaxation of active-site imidazole1H and/or15N nuclei.

  4. NMR profiling of biomolecules at natural abundance using 2D 1H-15N and 1H-13C multiplicity-separated (MS) HSQC spectra

    NASA Astrophysics Data System (ADS)

    Chen, Kang; Freedberg, Darón I.; Keire, David A.

    2015-02-01

    2D NMR 1H-X (X = 15N or 13C) HSQC spectra contain cross-peaks for all XHn moieties. Multiplicity-edited1H-13C HSQC pulse sequences generate opposite signs between peaks of CH2 and CH/CH3 at a cost of lower signal-to-noise due to the 13C T2 relaxation during an additional 1/1JCH period. Such CHn-editing experiments are useful in assignment of chemical shifts and have been successfully applied to small molecules and small proteins (e.g. ubiquitin) dissolved in deuterated solvents where, generally, peak overlap is minimal. By contrast, for larger biomolecules, peak overlap in 2D HSQC spectra is unavoidable and peaks with opposite phases cancel each other out in the edited spectra. However, there is an increasing need for using NMR to profile biomolecules at natural abundance dissolved in water (e.g., protein therapeutics) where NMR experiments beyond 2D are impractical. Therefore, the existing 2D multiplicity-edited HSQC methods must be improved to acquire data on nuclei other than 13C (i.e.15N), to resolve more peaks, to reduce T2 losses and to accommodate water suppression approaches. To meet these needs, a multiplicity-separated1H-X HSQC (MS-HSQC) experiment was developed and tested on 500 and 700 MHz NMR spectrometers equipped with room temperature probes using RNase A (14 kDa) and retroviral capsid (26 kDa) proteins dissolved in 95% H2O/5% D2O. In this pulse sequence, the 1/1JXH editing-period is incorporated into the semi-constant time (semi-CT) X resonance chemical shift evolution period, which increases sensitivity, and importantly, the sum and the difference of the interleaved 1JXH-active and the 1JXH-inactive HSQC experiments yield two separate spectra for XH2 and XH/XH3. Furthermore we demonstrate improved water suppression using triple xyz-gradients instead of the more widely used z-gradient only water-suppression approach.

  5. Oligomeric complexes of some heteroaromatic ligands and aromatic diamines with rhodium and molybdenum tetracarboxylates: 13C and 15N CPMAS NMR and density functional theory studies.

    PubMed

    Leniak, Arkadiusz; Kamieński, Bohdan; Jaźwiński, Jarosław

    2015-05-01

    Seven new oligomeric complexes of 4,4'-bipyridine; 3,3'-bipyridine; benzene-1,4-diamine; benzene-1,3-diamine; benzene-1,2-diamine; and benzidine with rhodium tetraacetate, as well as 4,4'-bipyridine with molybdenum tetraacetate, have been obtained and investigated by elemental analysis and solid-state nuclear magnetic resonance spectroscopy, (13)C and (15)N CPMAS NMR. The known complexes of pyrazine with rhodium tetrabenzoate, benzoquinone with rhodium tetrapivalate, 4,4'-bipyridine with molybdenum tetrakistrifluoroacetate and the 1 : 1 complex of 2,2'-bipyridine with rhodium tetraacetate exhibiting axial-equatorial ligation mode have been obtained as well for comparison purposes. Elemental analysis revealed 1 : 1 complex stoichiometry of all complexes. The (15)N CPMAS NMR spectra of all new complexes consist of one narrow signal, indicating regular uniform structures. Benzidine forms a heterogeneous material, probably containing linear oligomers and products of further reactions. The complexes were characterized by the parameter complexation shift Δδ (Δδ = δcomplex  - δligand). This parameter ranged from around -40 to -90 ppm in the case of heteroaromatic ligands, from around -12 to -22 ppm for diamines and from -16 to -31 ppm for the complexes of molybdenum tetracarboxylates with 4,4'-bipyridine. The experimental results have been supported by a density functional theory computation of (15)N NMR chemical shifts and complexation shifts at the non-relativistic Becke, three-parameter, Perdew-Wang 91/[6-311++G(2d,p), Stuttgart] and GGA-PBE/QZ4P levels of theory and at the relativistic scalar and spin-orbit zeroth order regular approximation/GGA-PBE/QZ4P level of theory. Nucleus-independent chemical shifts have been calculated for the selected compounds. PMID:25614975

  6. Amino-acid selective experiments on uniformly 13C and 15N labeled proteins by MAS NMR: Filtering of lysines and arginines

    NASA Astrophysics Data System (ADS)

    Jehle, Stefan; Rehbein, Kristina; Diehl, Anne; van Rossum, Barth-Jan

    2006-12-01

    Amino-acid selective magic-angle spinning (MAS) NMR experiments can aid the assignment of ambiguous cross-peaks in crowded spectra of solid proteins. In particular for larger proteins, data analysis can be hindered by severe resonance overlap. In such cases, filtering techniques may provide a good alternative to site-specific spin-labeling to obtain unambiguous assignments that can serve as starting points in the assignment procedure. In this paper we present a simple pulse sequence that allows selective excitation of arginine and lysine residues. To achieve this, we make use of a combination of specific cross-polarization for selective excitation [M. Baldus, A.T. Petkova, J. Herzfeld, R.G. Griffin, Cross polarization in the tilted frame: assignment and spectral simplification in heteronuclear spin systems, Mol. Phys. 95 (1998) 1197-1207.] and spin diffusion for transfer along the amino-acid side-chain. The selectivity of the filter is demonstrated with the excitation of lysine and arginine side-chain resonances in a uniformly 13C and 15N labeled protein preparation of the α-spectrin SH3 domain. It is shown that the filter can be applied as a building block in a 13C- 13C lysine-only correlation experiment.

  7. 1H, 15N and 13C assignment of the amyloidogenic protein medin using fast-pulsing NMR techniques.

    PubMed

    Davies, H A; Phelan, M M; Madine, J

    2016-04-01

    Thirty-one proteins are known to form extracellular fibrillar amyloid in humans. Molecular information about many of these proteins in their monomeric, intermediate or fibrillar form and how they aggregate and interact to form the insoluble fibrils is sparse. This is because amyloid proteins are notoriously difficult to study in their soluble forms, due to their inherent propensity to aggregate. Using recent developments in fast NMR techniques, band-selective excitation short transient and band-selective optimized flip-angle short-transient heteronuclear multiple quantum coherence we have been able to assign a 5 kDa full-length amyloidogenic protein called medin. Medin is the key protein component of the most common form of localised amyloid with a proposed role in aortic aneurysm and dissection. This assignment will now enable the study of the early interactions that could influence initiation and progression of medin aggregation. The chemical shifts have been deposited in the BioMagRes-Bank accession Nos. 25399 and 26576. PMID:26377205

  8. Restraints on backbone conformations in solid state NMR studies of uniformly labeled proteins from quantitative amide 15N–15N and carbonyl 13C–13C dipolar recoupling data

    PubMed Central

    Hu, Kan-Nian; Qiang, Wei; Bermejo, Guillermo A.; Schwieters, Charles D.; Tycko, Robert

    2013-01-01

    Recent structural studies of uniformly 15N, 13C-labeled proteins by solid state nuclear magnetic resonance (NMR) rely principally on two sources of structural restraints: (i) restraints on backbone conformation from isotropic 15N and 13C chemical shifts, based on empirical correlations between chemical shifts and backbone torsion angles; (ii) restraints on inter-residue proximities from qualitative measurements of internuclear dipole–dipole couplings, detected as the presence or absence of inter-residue crosspeaks in multidimensional spectra. We show that site-specific dipole–dipole couplings among 15N-labeled backbone amide sites and among 13C-labeled backbone carbonyl sites can be measured quantitatively in uniformly-labeled proteins, using dipolar recoupling techniques that we call 15N-BARE and 13C-BARE (BAckbone REcoupling), and that the resulting data represent a new source of restraints on backbone conformation. 15N-BARE and 13C-BARE data can be incorporated into structural modeling calculations as potential energy surfaces, which are derived from comparisons between experimental 15N and 13C signal decay curves, extracted from crosspeak intensities in series of two-dimensional spectra, with numerical simulations of the 15N-BARE and 13C-BARE measurements. We demonstrate this approach through experiments on microcrystalline, uniformly 15N, 13C-labeled protein GB1. Results for GB1 show that 15N-BARE and 13C-BARE restraints are complementary to restraints from chemical shifts and inter-residue crosspeaks, improving both the precision and the accuracy of calculated structures. PMID:22449573

  9. (1)H NMR assignment corrections and (1)H, (13)C, (15)N NMR coordination shifts structural correlations in Fe(II), Ru(II) and Os(II) cationic complexes with 2,2'-bipyridine and 1,10-phenanthroline.

    PubMed

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

    2010-06-01

    (1)H, (13)C and (15)N NMR studies of iron(II), ruthenium(II) and osmium(II) tris-chelated cationic complexes with 2,2'-bipyridine and 1,10-phenanthroline of the general formula [M(LL)(3)](2+) (M = Fe, Ru, Os; LL = bpy, phen) were performed. Inconsistent literature (1)H signal assignments were corrected. Significant shielding of nitrogen-adjacent protons [H(6) in bpy, H(2) in phen] and metal-bonded nitrogens was observed, being enhanced in the series Ru(II) --> Os(II) --> Fe(II) for (1)H, Fe(II) --> Ru(II) --> Os(II) for (15)N and bpy --> phen for both nuclei. The carbons are deshielded, the effect increasing in the order Ru(II) --> Os(II) --> Fe(II). PMID:20474023

  10. Structural correlations for (1)H, (13)C and (15)N NMR coordination shifts in Au(III), Pd(II) and Pt(II) chloride complexes with lutidines and collidine.

    PubMed

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

    2010-06-01

    (1)H, (13)C and (15)N NMR studies of gold(III), palladium(II) and platinum(II) chloride complexes with dimethylpyridines (lutidines: 2,3-lutidine, 2,3lut; 2,4-lutidine, 2,4lut; 3,5-lutidine, 3,5lut; 2,6-lutidine, 2,6lut) and 2,4,6-trimethylpyridine (2,4,6-collidine, 2,4,6col) having general formulae [AuLCl(3)], trans-[PdL(2)Cl(2)] and trans-/cis-[PtL(2)Cl(2)] were performed and the respective chemical shifts (delta(1H), delta(13C), delta(15N)) reported. The deshielding of protons and carbons, as well as the shielding of nitrogens was observed. The (1)H, (13)C and (15)N NMR coordination shifts (Delta(1H) (coord), Delta(13C) (coord), Delta(15N) (coord); Delta(coord) = delta(complex) - delta(ligand)) were discussed in relation to some structural features of the title complexes, such as the type of the central atom [Au(III), Pd(II), Pt(II)], geometry (trans- or cis-), metal-nitrogen bond lengths and the position of both methyl groups in the pyridine ring system. PMID:20474019

  11. Study of stereospecificity of 1H, 13C, 15N and 77Se shielding constants in the configurational isomers of the selenophene-2-carbaldehyde azine by NMR spectroscopy and MP2-GIAO calculations.

    PubMed

    Afonin, Andrei V; Pavlov, Dmitry V; Albanov, Alexander I; Levanova, Ekaterina P; Levkovskaya, Galina G

    2011-11-01

    In the (1)H and (13)C NMR spectra of selenophene-2-carbaldehyde azine, the (1)H-5, (13)C-3 and (13)C-5 signals of the selenophene ring are shifted to higher frequencies, whereas those of the (1)H-1, (13)C-1, (13)C-2 and (13)C-4 are shifted to lower frequencies on going from the EE to ZZ isomer or from the E moiety to the Z moiety of EZ isomer. The (15)N chemical shift is significantly larger in the EE isomer relative to the ZZ isomer and in the E moiety relative to the Z moiety of EZ isomer. A very pronounced difference (60-65 mg/g) between the (77)Se resonance positions is revealed in the studied azine isomers, the (77)Se peak being shifted to higher frequencies in the ZZ isomer and in the Z moiety of EZ isomer. The trends in the changes of the measured chemical shifts are reasonably reproduced by the GIAO calculations at the MP2 level of the (1)H, (13)C, (15)N and (77)Se shielding constants in the energy-favorable conformation with the syn orientation of both selenophene rings relative to the C = N groups. The NBO analysis suggests that such an arrangement of the selenophene rings may take place because of a higher energy of some intramolecular interactions. PMID:22002712

  12. Secondary Structure, Backbone Dynamics, and Structural Topology of Phospholamban and Its Phosphorylated and Arg9Cys-Mutated Forms in Phospholipid Bilayers Utilizing 13C and 15N Solid-State NMR Spectroscopy

    PubMed Central

    2015-01-01

    Phospholamban (PLB) is a membrane protein that regulates heart muscle relaxation rates via interactions with the sarcoplasmic reticulum Ca2+ ATPase (SERCA). When PLB is phosphorylated or Arg9Cys (R9C) is mutated, inhibition of SERCA is relieved. 13C and 15N solid-state NMR spectroscopy is utilized to investigate conformational changes of PLB upon phosphorylation and R9C mutation. 13C=O NMR spectra of the cytoplasmic domain reveal two α-helical structural components with population changes upon phosphorylation and R9C mutation. The appearance of an unstructured component is observed on domain Ib. 15N NMR spectra indicate an increase in backbone dynamics of the cytoplasmic domain. Wild-type PLB (WT-PLB), Ser16-phosphorylated PLB (P-PLB), and R9C-mutated PLB (R9C-PLB) all have a very dynamic domain Ib, and the transmembrane domain has an immobile component. 15N NMR spectra indicate that the cytoplasmic domain of R9C-PLB adopts an orientation similar to P-PLB and shifts away from the membrane surface. Domain Ib (Leu28) of P-PLB and R9C-PLB loses the alignment. The R9C-PLB adopts a conformation similar to P-PLB with a population shift to a more extended and disordered state. The NMR data suggest the more extended and disordered forms of PLB may relate to inhibition relief. PMID:24511878

  13. Experimental and quantum-chemical studies of 1H, 13C and 15N NMR coordination shifts in Au(III), Pd(II) and Pt(II) chloride complexes with picolines.

    PubMed

    Pazderski, Leszek; Tousek, Jaromír; Sitkowski, Jerzy; Malináková, Katerina; Kozerski, Lech; Szłyk, Edward

    2009-03-01

    (1)H, (13)C and (15)N NMR studies of gold(III), palladium(II) and platinum(II) chloride complexes with picolines, [Au(PIC)Cl(3)], trans-[Pd(PIC)(2)Cl(2)], trans/cis-[Pt(PIC)(2)Cl(2)] and [Pt(PIC)(4)]Cl(2), were performed. After complexation, the (1)H and (13)C signals were shifted to higher frequency, whereas the (15)N ones to lower (by ca 80-110 ppm), with respect to the free ligands. The (15)N shielding phenomenon was enhanced in the series [Au(PIC)Cl(3)] < trans-[Pd(PIC)(2)Cl(2)] < cis-[Pt(PIC)(2)Cl(2)] < trans-[Pt(PIC)(2)Cl(2)]; it increased following the Pd(II) --> Pt(II) replacement, but decreased upon the trans --> cis-transition. Experimental (1)H, (13)C and (15)N NMR chemical shifts were compared to those quantum-chemically calculated by B3LYP/LanL2DZ + 6-31G**//B3LYP/LanL2DZ + 6-31G*. PMID:19097135

  14. Structure and reactivity of lithium amides. /sup 6/Li, /sup 13/C, and /sup 15/N NMR spectroscopic studies and colligative measurements of lithium diphenylamide and lithium diphenylamide-lithium bromide complex solvated by tetrahydrofuran

    SciTech Connect

    DePue, J.S.; Collum, D.B.

    1988-08-03

    /sup 6/Li, /sup 13/C, and /sup 15/N NMR spectroscopic studies of lithium diphenylamide in THF/hydrocarbon solutions (THF = tetrahydrofuran) detected two different species. /sup 6/Li and /sup 15/N NMR spectroscopic studies of (/sup 6/Li, /sup 15/N)lithium diphenylamide showed the species observed at low THF concentrations to be a cyclic oligomer. Structural analogies provided strong support for a dimer while colligative measurements at 0/degrees/C indicated the dimer to be di- or trisolvated. On the basis of the observed mass action effects, the species appearing at intermediate THF concentrations is assigned as a contact or solvent-separated ion-paired monomer. Lithium diphenylamide forms a 1:1 adduct with lithium bromide at low THF concentrations. A combination of /sup 6/Li-/sup 15/N double labeling studies and colligative measurements supports a trisolvated cyclic mixed dimer structure. Although detailed spectroscopic studies at elevated THF concentrations were precluded by high fluctionality, the similarity of the /sup 13/C chemical shifts of lithium diphenylamide in the presence and absence of lithium bromide provide indirect evidence that the mixed dimer undergoes a THF concentration dependent dissociation to the monomeric amide and free lithium bromide. 24 references, 9 figures, 2 tables.

  15. Backbone and Ile-δ1, Leu, Val Methyl 1H, 13C and 15N NMR chemical shift assignments for human interferon-stimulated gene 15 protein

    SciTech Connect

    Yin, Cuifeng; Aramini, James M.; Ma, LiChung; Cort, John R.; Swapna, G.V.T.; Krug, R. M.; Montelione, Gaetano

    2011-10-01

    Human interferon-stimulated gene 15 protein (ISG15), also called ubiquitin cross-reactive protein (UCRP), is the first identified ubiquitin-like protein containing two ubiquitin-like domains fused in tandem. The active form of ISG15 is conjugated to target proteins via the C-terminal glycine residue through an isopeptide bond in a manner similar to ubiquitin. The biological role of ISG15 is strongly associated with the modulation of cell immune function, and there is mounting evidence suggesting that many viral pathogens evade the host innate immune response by interfering with ISG15 conjugation to both host and viral proteins in a variety of ways. Here we report nearly complete backbone 1HN, 15N, 13CO, and 13Ca, as well as side chain 13Cb, methyl (Ile-d1, Leu, Val), amide (Asn, Gln), and indole NH (Trp) NMR resonance assignments for the 157-residue human ISG15 protein. These resonance assignments provide the basis for future structural and functional solution NMR studies of the biologically important human ISG15 protein.

  16. 1H, 13C and 15N NMR coordination shifts in gold(III), cobalt(III), rhodium(III) chloride complexes with pyridine, 2,2'-bipyridine and 1,10-phenanthroline.

    PubMed

    Pazderski, Leszek; Tousek, Jaromír; Sitkowski, Jerzy; Kozerski, Lech; Marek, Radek; Szłyk, Edward

    2007-01-01

    Au(III), Co(III) and Rh(III) chloride complexes with pyridine (py), 2,2'-bipyridine (bpy) and 1,10-phenanthroline (phen) of the general formulae [M1LCl3], trans-[M2L4Cl2]+, mer-[M2L3Cl3], [M1(LL)Cl2]+, cis-[M2(LL)2Cl2]+, where M1=Au; M2=Co, Rh; L=py; LL=bpy, phen, were studied by 1H--13C HMBC and 1H--15N HMQC/HSQC. The 1H, 13C and 15N coordination shifts (the latter from ca-78 to ca-107 ppm) are discussed in relation to the type of metal, electron configuration, coordination sphere geometry and the type of ligand. The 13C and 15N chemical shifts were also calculated by quantum-chemical NMR methods, which reproduced well the experimental tendencies concerning the coordination sphere geometry and the ligand type. PMID:17048265

  17. Phosphorus-31, sup 15 N, and sup 13 C NMR of glyphosate: Comparison of pH titrations to the herbicidal dead-end complex with 5-enolpyruvoylshikimate-3-phosphate synthase

    SciTech Connect

    Castellino, S.; Leo, G.C.; Sammons, R.D.; Sikorski, J.A. )

    1989-05-02

    The herbicidal dead-end ternary complex (E{sup S3P}{sub Glyph}) of glyphosate (N-(phosphonomethyl)glycine) with 5-enolpyruvoylshikimate-3-phosphate synthase (EPSPS) and the substrate shikimate 3-phosphate (S3P) has been characterized by {sup 31}P, {sup 15}N, and {sup 13}C NMR. The NMR spectra of EPSPS-bound glyphosate show unique chemical shifts ({delta}) for each of the three nuclei. By {sup 31}P NMR, glyphosate in the dead-end complex is a distinct species 3.5 ppm downfield from free glyphosate. The {sup 13}C signal of glyphosate in the dead-end complex is shifted 4 ppm downfield from that of free glyphosate. The {sup 15}N signal for glyphosate (99%) in the dead-end complex is 5 ppm further downfield than that of any free zwitterionic species and 10 ppm downfield from that of the average free species at pH 10.1. The structures of each ionic state of glyphosate are modeled with force field calculations by using MacroModel. A correlation is made for the {sup 31}P {delta} and the C-P-O bond angle, and the {sup 13}C and {sup 15}N {delta} values are postulated to be related to C-C-O and C-N-C bond angles, respectively. The downfield {sup 31}P chemical shift perturbation for S3P in the EPSPS binary complex is consistent with ionization of the 3-phosphate of S3P upon binding. Comparison with the S3P {sup 31}P {delta} vs pH titration curve specifies predominantly the dianion of the 3-phosphate in the E{sup S3P} binary complex, while the E{sup S3P}{sub Glyph} complex indicates net protonation at the 3-phosphate. Chemical shift perturbations of this latter type may be explained by changes in the O-P-O bond angle.

  18. Experimental and quantum-chemical studies of 1H, 13C and 15N NMR coordination shifts in Pd(II) and Pt(II) chloride complexes with quinoline, isoquinoline, and 2,2'-biquinoline.

    PubMed

    Pazderski, Leszek; Tousek, Jaromír; Sitkowski, Jerzy; Kozerski, Lech; Szłyk, Edward

    2007-12-01

    1H, 13C, and 15N NMR studies of platinide(II) (M=Pd, Pt) chloride complexes with quinolines (L=quinoline-quin, or isoquinoline-isoquin; LL=2,2'-biquinoline-bquin), having the general formulae trans-/cis-[ML2Cl2] and [M(LL)Cl2], were performed and the respective chemical shifts (delta1H, delta13C, delta15N) reported. 1H coordination shifts of various signs and magnitudes (Delta1Hcoord=delta1Hcomplex-delta1Hligand) are discussed in relation to the changes of diamagnetic contribution to the relevant 1H shielding constants. The comparison to the literature data for similar complexes containing auxiliary ligands other than chlorides exhibited a large dependence of delta1H parameters on electron density variations and ring-current effects (inductive and anisotropic phenomena). The influence of deviations from planarity, concerning either MN2Cl2 chromophores or azine ring systems, revealed by the known X-ray structures of [Pd(bquin)Cl2] and [Pt(bquin)Cl2], is discussed in respect to 1H NMR spectra. 15N coordination shifts (Delta15Ncoord=delta15Ncomplex-delta15Nligand) of ca. 78-100 ppm (to lower frequency) are attributed mainly to the decrease of the absolute value of paramagnetic contribution in the relevant 15N shielding constants, this phenomenon being noticeably dependent on the type of a platinide metal and coordination sphere geometry. The absolute magnitude of Delta15Ncoord parameter increased by ca 15 ppm upon Pd(II)-->Pt(II) replacement but decreased by ca. 15 ppm following trans-->cis transition. Experimental 1H, 13C, 15N NMR chemical shifts are compared to those quantum-chemically calculated by B3LYP/LanL2DZ+6-31G**//B3LYP/LanL2DZ+6-31G*, both in vacuo and in CHCl3 or DMF solution. PMID:18044805

  19. QUANTITATIVE 15N NMR SPECTROSCOPY

    EPA Science Inventory

    Line intensities in 15N NMR spectra are strongly influenced by spin-lattice and spin-spin relaxation times, relaxation mechanisms and experimental conditions. Special care has to be taken in using 15N spectra for quantitative purposes. Quantitative aspects are discussed for the 1...

  20. 1H, 13C, 195Pt and 15N NMR structural correlations in Pd(II) and Pt(II) chloride complexes with various alkyl and aryl derivatives of 2,2'-bipyridine and 1,10-phenanthroline.

    PubMed

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

    2011-02-01

    (1)H, (13)C, (195)Pt and (15)N NMR studies of platinide(II) (M = Pd, Pt) chloride complexes with such alkyl and aryl derivatives of 2,2'-bipyridine and 1,10-phenanthroline as LL = 6,6'-dimethyl-bpy, 5,5'-dimethyl-bpy, 4,4'-di-tert-butyl-bpy, 2,9-dimethyl-phen, 2,9-dimethyl-4,7-diphenyl-phen, 3,4,7,8-tetramethyl-phen, having the general [M(LL)Cl(2)] formula were performed and the respective chemical shifts (δ(1H), δ(13C), δ(195Pt), δ(15N)) reported. (1)H high-frequency coordination shifts (Δ(coord)(1H) = δ(complex)(1H)-δ(ligand)(1H)) mostly pronounced for nitrogen-adjacent protons and methyl groups in the nearest adjacency of nitrogen, as well as (15)N low-frequency coordination shifts (Δ(coord)(15H) = δ(complex)(15H)-δ(ligand)(15H)) were discussed in relation to the molecular structures. PMID:21254225

  1. Measuring (13)C/(15)N chemical shift anisotropy in [(13)C,(15)N] uniformly enriched proteins using CSA amplification.

    PubMed

    Hung, Ivan; Ge, Yuwei; Liu, Xiaoli; Liu, Mali; Li, Conggang; Gan, Zhehong

    2015-11-01

    Extended chemical shift anisotropy amplification (xCSA) is applied for measuring (13)C/(15)N chemical shift anisotropy (CSA) of uniformly labeled proteins under magic-angle spinning (MAS). The amplification sequence consists of a sequence of π-pulses that repetitively interrupt MAS averaging of the CSA interaction. The timing of the pulses is designed to generate amplified spinning sideband manifolds which can be fitted to extract CSA parameters. The (13)C/(13)C homonuclear dipolar interactions are not affected by the π-pulses due to the bilinear nature of the spin operators and are averaged by MAS in the xCSA experiment. These features make the constant evolution-time experiment suitable for measuring CSA of uniformly labeled samples. The incorporation of xCSA with multi-dimensional (13)C/(15)N correlation is demonstrated with a GB1 protein sample as a model system for measuring (13)C/(15)N CSA of all backbone (15)NH, (13)CA and (13)CO sites. PMID:26404770

  2. 1H, 13C and 15N NMR assignments of the E. coli peptide deformylase in complex with a natural inhibitor called actinonin.

    PubMed

    Larue, Valéry; Seijo, Bili; Tisne, Carine; Dardel, Frédéric

    2009-06-01

    In eubacteria, the formyl group of nascent polypeptides is removed by peptide deformylase protein (PDF). This is the reason why PDF has received special attention in the course of the search for new antibacterial agents. We observed by NMR that actinonin, a natural inhibitor, induced drastic changes in the HSQC spectrum of E. coli PDF. We report here the complete NMR chemical shift assignments of PDF resonances bound to actinonin. PMID:19636969

  3. 1H, 13C, 15N NMR coordination shifts in Fe(II), Ru(II) and Os(II) cationic complexes with 2,2':6',2″-terpyridine.

    PubMed

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

    2011-05-01

    (1)H, (13)C and (15)N NMR studies of iron(II), ruthenium(II) and osmium(II) bis-chelated cationic complexes with 2,2':6',2″-terpyridine ([M(terpy)(2) ](2+) ; M = Fe, Ru, Os) were performed. Significant shielding of nitrogen-adjacent H(6) and deshielding of H(3'), H(4') protons were observed, both effects being mostly expressed for Fe(II) compounds. The metal-bonded nitrogens were shielded, this effect being much larger for the outer N(1), N(1″) than the inner N(1') atoms, and enhanced in the Fe(II) → Ru(II) → Os(II) series. PMID:21491480

  4. (1)H-, (13)C- and (15)N-NMR assignment of the N-terminal domain of human cerebral dopamine neurotrophic factor (CDNF).

    PubMed

    Latgé, Cristiane; Cabral, Kátia M S; Almeida, Marcius S; Foguel, Débora

    2013-04-01

    Parkinson's disease (PD) is a neurodegenerative disorder that is caused by the death of midbrain dopaminergic neurons. Current therapies for PD do not halt the neurodegeneration nor repair the affected neurons. Therefore, search for novel neurotrophic factors (NTF) for midbrain dopaminergic neurons, which could be used in novel therapeutic approaches, is highly wanted. In 2007, a potent NTF for dopaminergic neurons was described as the conserved dopamine neurotrophic factor (CDNF). Single doses of this protein protect and restore dopaminergic neurons in experimental models of PD. CDNF has two domains; an N-terminal saposin-like domain, which may bind to membranes; and a presumably intrinsically unstructured C-terminal which contains an internal cysteine bridge in a CXXC motif similar to that of thiol/disulphide oxidoreductases and isomerases, and may thus reduce the endoplasmic reticulum stress caused by incorrectly folded proteins. We show for the first time the nuclear magnetic resonance assignment of N-terminal domain of recombinant CDNF (residues 1-105) by solution 2D and 3D NMR spectroscopy. We were able to obtain a nearly complete resonance assignment, which is the first step toward the solution structure determination of this neurotrophic factor. PMID:22528768

  5. 1H, 13C and 15N resonance assignments of URNdesign, a computationally redesigned RRM protein

    SciTech Connect

    Dobson, Neil; Dantas, Gautam; Varani, Gabriele

    2005-10-01

    Protein design represents one of the great challenges of computational structural biology. The ability to successfully design new proteins would allow us to generate new reagents and enzymes, while at the same time providing us with an understanding of the principles of protein stability. Here we report 1H, 15N and 13C resonance assignments of a redesigned U1A protein, URNdesign. U1A has been studied extensively by our group and hence was chosen as a design target. For the assignments we sued 2D and 3D heteronuclearNMR experiments with uniformly 13C, 15N-labeled URNdesign. The assignments for the backbone NH, CO,Ca and Cb nuclei are 94%complete. Sidechain 1Hand13C, aromatic andQ/NNH2 resonances are essentially complete with guanidinium and K NH3 residues unassigned. BMRB deposit with accession number 6493

  6. 13C NMR Metabolomics: INADEQUATE Network Analysis

    PubMed Central

    Clendinen, Chaevien S.; Pasquel, Christian; Ajredini, Ramadan; Edison, Arthur S.

    2015-01-01

    The many advantages of 13C NMR are often overshadowed by its intrinsically low sensitivity. Given that carbon makes up the backbone of most biologically relevant molecules, 13C NMR offers a straightforward measurement of these compounds. Two-dimensional 13C-13C correlation experiments like INADEQUATE (incredible natural abundance double quantum transfer experiment) are ideal for the structural elucidation of natural products and have great but untapped potential for metabolomics analysis. We demonstrate a new and semi-automated approach called INETA (INADEQUATE network analysis) for the untargeted analysis of INADEQUATE datasets using an in silico INADEQUATE database. We demonstrate this approach using isotopically labeled Caenorhabditis elegans mixtures. PMID:25932900

  7. 1H, 13C, 15N backbone and side chain NMR resonance assignments for the N-terminal RNA recognition motif of the HvGR-RBP1 protein involved in the regulation of barley (Hordeum vulgare L.) senescence

    PubMed Central

    Mason, Katelyn E.; Tripet, Brian P.; Parrott, David; Fischer, Andreas M.; Copié, Valérie

    2013-01-01

    Leaf senescence is an important process in the developmental life of all plant species. Senescence efficiency influences important agricultural traits such as grain protein content and plant growth, which are often limited by nitrogen use. Little is known about the molecular mechanisms regulating this highly orchestrated process. To enhance our understanding of leaf senescence and its regulation, we have undertaken the structural and functional characterization of previously unknown proteins that are involved in the control of senescence in barley (Hordeum vulgare L.). Previous microarray analysis highlighted several barley genes whose transcripts are differentially expressed during senescence, including a specific gene which is greater than 40 fold up-regulated in the flag leaves of early- as compared to late-senescing near-isogenic barley lines at 14 and 21 days past flowering (anthesis). From inspection of its amino acid sequence, this gene is predicted to encode a glycine-rich RNA-binding protein herein referred to as HvGR-RBP1. HvGR-RBP1 has been expressed as a recombinant protein in E. coli, and preliminary NMR data analysis has revealed that its glycine-rich C-terminal region [residues: 93–162] is structurally disordered whereas its N-terminal region [residues: 1–92] forms a well-folded domain. Herein, we report the complete 1H, 13C, and 15N resonance assignments of backbone and sidechain atoms, and the secondary structural topology of the N-terminal RNA Recognition Motif (RRM) domain of HvGR-RBP1, as a first step to unraveling its structural and functional role in the regulation of barley leaf senescence. PMID:23417794

  8. Study of conformations and hydrogen bonds in the configurational isomers of pyrrole-2-carbaldehyde oxime by 1H, 13C and 15N NMR spectroscopy combined with MP2 and DFT calculations and NBO analysis.

    PubMed

    Afonin, Andrei V; Ushakov, Igor A; Pavlov, Dmitry V; Ivanov, Andrei V; Mikhaleva, Al'bina I

    2010-09-01

    The (1)H, (13)C and (15)N NMR studies have shown that the E and Z isomers of pyrrole-2-carbaldehyde oxime adopt preferable conformation with the syn orientation of the oxime group with respect to the pyrrole ring. The syn conformation of E and Z isomers of pyrrole-2-carbaldehyde oxime is stabilized by the N-H...N and N-H...O intramolecular hydrogen bonds, respectively. The N-H...N hydrogen bond in the E isomer causes the high-frequency shift of the bridge proton signal by about 1 ppm and increase the (1)J(N, H) coupling by approximately 3 Hz. The bridge proton shows further deshielding and higher increase of the (1)J(N, H) coupling constant due to the strengthening of the N-H...O hydrogen bond in the Z isomer. The MP2 calculations indicate that the syn conformation of E and Z isomers is by approximately 3.5 kcal/mol energetically less favorable than the anti conformation. The calculations of (1)H shielding and (1)J(N, H) coupling in the syn and anti conformations allow the contribution to these constants from the N-H...N and N-H...O hydrogen bondings to be estimated. The NBO analysis suggests that the N-H...N hydrogen bond in the E isomer is a pure electrostatic interaction while the charge transfer from the oxygen lone pair to the antibonding orbital of the N-H bond through the N-H...O hydrogen bond occurs in the Z isomer. PMID:20623827

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

    NASA Astrophysics Data System (ADS)

    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.

  10. Site-Selective Synthesis of (15)N- and (13)C-Enriched Flavin Mononucleotide Coenzyme Isotopologues.

    PubMed

    Neti, Syam Sundar; Poulter, C Dale

    2016-06-17

    Flavin mononucleotide (FMN) is a coenzyme for numerous proteins involved in key cellular and physiological processes. Isotopically labeled flavin is a powerful tool for studying the structure and mechanism of flavoenzyme-catalyzed reactions by a variety of techniques, including NMR, IR, Raman, and mass spectrometry. In this report, we describe the preparation of labeled FMN isotopologues enriched with (15)N and (13)C isotopes at various sites in the pyrazine and pyrimidine rings of the isoalloxazine core of the cofactor from readily available precursors by a five-step chemo-enzymatic synthesis. PMID:27176708

  11. Optimization of amino acid type-specific 13C and 15N labeling for the backbone assignment of membrane proteins by solution- and solid-state NMR with the UPLABEL algorithm.

    PubMed

    Hefke, Frederik; Bagaria, Anurag; Reckel, Sina; Ullrich, Sandra Johanna; Dötsch, Volker; Glaubitz, Clemens; Güntert, Peter

    2011-02-01

    We present a computational method for finding optimal labeling patterns for the backbone assignment of membrane proteins and other large proteins that cannot be assigned by conventional strategies. Following the approach of Kainosho and Tsuji (Biochemistry 21:6273-6279 (1982)), types of amino acids are labeled with (13)C or/and (15)N such that cross peaks between (13)CO(i - 1) and (15)NH(i) result only for pairs of sequentially adjacent amino acids of which the first is labeled with (13)C and the second with (15)N. In this way, unambiguous sequence-specific assignments can be obtained for unique pairs of amino acids that occur exactly once in the sequence of the protein. To be practical, it is crucial to limit the number of differently labeled protein samples that have to be prepared while obtaining an optimal extent of labeled unique amino acid pairs. Our computer algorithm UPLABEL for optimal unique pair labeling, implemented in the program CYANA and in a standalone program, and also available through a web portal, uses combinatorial optimization to find for a given amino acid sequence labeling patterns that maximize the number of unique pair assignments with a minimal number of differently labeled protein samples. Various auxiliary conditions, including labeled amino acid availability and price, previously known partial assignments, and sequence regions of particular interest can be taken into account when determining optimal amino acid type-specific labeling patterns. The method is illustrated for the assignment of the human G-protein coupled receptor bradykinin B2 (B(2)R) and applied as a starting point for the backbone assignment of the membrane protein proteorhodopsin. PMID:21170670

  12. Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction

    DOEpatents

    Chen, Xian; Gupta, Goutam; Bradbury, E. Morton

    2001-01-01

    Preparation of .sup.13 C/.sup.15 N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (.sup.13 C/.sup.15 N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly .sup.13 C/.sup.15 N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly .sup.13 C/.sup.15 N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.

  13. Determination of methyl 13C-15N dipolar couplings in peptides and proteins by three-dimensional and four-dimensional magic-angle spinning solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Helmus, Jonathan J.; Nadaud, Philippe S.; Höfer, Nicole; Jaroniec, Christopher P.

    2008-02-01

    We describe three- and four-dimensional semiconstant-time transferred echo double resonance (SCT-TEDOR) magic-angle spinning solid-state nuclear magnetic resonance (NMR) experiments for the simultaneous measurement of multiple long-range N15-Cmethyl13 dipolar couplings in uniformly C13, N15-enriched peptides and proteins with high resolution and sensitivity. The methods take advantage of C13 spin topologies characteristic of the side-chain methyl groups in amino acids alanine, isoleucine, leucine, methionine, threonine, and valine to encode up to three distinct frequencies (N15-Cmethyl13 dipolar coupling, N15 chemical shift, and Cmethyl13 chemical shift) within a single SCT evolution period of initial duration ˜1/JCC1 (where JCC1≈35Hz, is the one-bond Cmethyl13-C13 J-coupling) while concurrently suppressing the modulation of NMR coherences due to C13-C13 and N15-C13 J-couplings and transverse relaxation. The SCT-TEDOR schemes offer several important advantages over previous methods of this type. First, significant (approximately twofold to threefold) gains in experimental sensitivity can be realized for weak N15-Cmethyl13 dipolar couplings (corresponding to structurally interesting, ˜3.5Å or longer, distances) and typical Cmethyl13 transverse relaxation rates. Second, the entire SCT evolution period can be used for Cmethyl13 and/or N15 frequency encoding, leading to increased spectral resolution with minimal additional coherence decay. Third, the experiments are inherently "methyl selective," which results in simplified NMR spectra and obviates the use of frequency-selective pulses or other spectral filtering techniques. Finally, the N15-C13 cross-peak buildup trajectories are purely dipolar in nature (i.e., not influenced by J-couplings or relaxation), which enables the straightforward extraction of N15-Cmethyl13 distances using an analytical model. The SCT-TEDOR experiments are demonstrated on a uniformly C13, N15-labeled peptide, N-acetyl-valine, and a 56

  14. Rapid and accurate calculation of protein 1H, 13C and 15N chemical shifts.

    PubMed

    Neal, Stephen; Nip, Alex M; Zhang, Haiyan; Wishart, David S

    2003-07-01

    A computer program (SHIFTX) is described which rapidly and accurately calculates the diamagnetic 1H, 13C and 15N chemical shifts of both backbone and sidechain atoms in proteins. The program uses a hybrid predictive approach that employs pre-calculated, empirically derived chemical shift hypersurfaces in combination with classical or semi-classical equations (for ring current, electric field, hydrogen bond and solvent effects) to calculate 1H, 13C and 15N chemical shifts from atomic coordinates. The chemical shift hypersurfaces capture dihedral angle, sidechain orientation, secondary structure and nearest neighbor effects that cannot easily be translated to analytical formulae or predicted via classical means. The chemical shift hypersurfaces were generated using a database of IUPAC-referenced protein chemical shifts--RefDB (Zhang et al., 2003), and a corresponding set of high resolution (<2.1 A) X-ray structures. Data mining techniques were used to extract the largest pairwise contributors (from a list of approximately 20 derived geometric, sequential and structural parameters) to generate the necessary hypersurfaces. SHIFTX is rapid (<1 CPU second for a complete shift calculation of 100 residues) and accurate. Overall, the program was able to attain a correlation coefficient (r) between observed and calculated shifts of 0.911 (1Halpha), 0.980 (13Calpha), 0.996 (13Cbeta), 0.863 (13CO), 0.909 (15N), 0.741 (1HN), and 0.907 (sidechain 1H) with RMS errors of 0.23, 0.98, 1.10, 1.16, 2.43, 0.49, and 0.30 ppm, respectively on test data sets. We further show that the agreement between observed and SHIFTX calculated chemical shifts can be an extremely sensitive measure of the quality of protein structures. Our results suggest that if NMR-derived structures could be refined using heteronuclear chemical shifts calculated by SHIFTX, their precision could approach that of the highest resolution X-ray structures. SHIFTX is freely available as a web server at http

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

    SciTech Connect

    Ashburn, D.A.; Garcia, K.; Hanners, J.L.; Silks, L.A. III; Unkefer, C.J.

    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.

  16. 15N chemical shift referencing in solid state NMR.

    PubMed

    Bertani, Philippe; Raya, Jésus; Bechinger, Burkhard

    2014-01-01

    Solid-state NMR spectroscopy has much advanced during the last decade and provides a multitude of data that can be used for high-resolution structure determination of biomolecules, polymers, inorganic compounds or macromolecules. In some cases the chemical shift referencing has become a limiting factor to the precision of the structure calculations and we have therefore evaluated a number of methods used in proton-decoupled (15)N solid-state NMR spectroscopy. For (13)C solid-state NMR spectroscopy adamantane is generally accepted as an external standard, but to calibrate the (15)N chemical shift scale several standards are in use. As a consequence the published chemical shift values exhibit considerable differences (up to 22 ppm). In this paper we report the (15)N chemical shift of several commonly used references compounds in order to allow for comparison and recalibration of published data and future work. We show that (15)NH4Cl in its powdered form (at 39.3 ppm with respect to liquid NH3) is a suitable external reference as it produces narrow lines when compared to other reference compounds and at the same time allows for the set-up of cross-polarization NMR experiments. The compound is suitable to calibrate magic angle spinning and static NMR experiments. Finally the temperature variation of (15)NH4Cl chemical shift is reported. PMID:24746715

  17. Experimental and quantum-chemical studies of 1H, 13C and 15N NMR coordination shifts in Pd(II) and Pt(II) chloride complexes with methyl and phenyl derivatives of 2,2'-bipyridine and 1,10-phenanthroline.

    PubMed

    Pazderski, Leszek; Tousek, Jaromír; Sitkowski, Jerzy; Kozerski, Lech; Szłyk, Edward

    2007-12-01

    1H, 13C and 15N NMR studies of platinide(II) (M=Pd, Pt) chloride complexes with methyl and phenyl derivatives of 2,2'-bipyridine and 1,10-phenanthroline [LL=4,4'-dimethyl-2,2'-bipyridine (dmbpy); 4,4'-diphenyl-2,2'-bipyridine (dpbpy); 4,7-dimethyl-1,10-phenanthroline (dmphen); 4,7-diphenyl-1,10-phenanthroline (dpphen)] having a general [M(LL)Cl2] formula were performed and the respective chemical shifts (delta1H, delta13C, delta15N) reported. 1H high-frequency coordination shifts (Delta1Hcoord=delta1Hcomplex-delta1Hligand) were discussed in relation to the changes of diamagnetic contribution in the relevant 1H shielding constants. The comparison to literature data for similar [M(LL)(XX)], [M(LL)X2] and [M(LL)XY] coordination or organometallic compounds containing various auxiliary ligands revealed a large dependence of delta1H parameters on inductive and anisotropic effects. 15N low-frequency coordination shifts (Delta15Ncoord=delta 15Ncomplex-delta15Nligand) of ca 88-96 ppm for M=Pd and ca 103-111 ppm for M=Pt were attributed to both the decrease of the absolute value of paramagnetic contribution and the increase of the diamagnetic term in the expression for 15N shielding constants. The absolute magnitude of Delta15Ncoord parameter increased by ca 15 ppm upon Pd(II)-->Pt(II) transition and by ca 6-7 ppm following dmbpy-->dmphen or dpbpy-->dpphen ligand replacement; variations between analogous complexes containing methyl and phenyl ligands (dmbpy vs dpbpy; dmphen vs dpphen) did not exceed+/-1.5 ppm. Experimental 1H, 13C, 15N NMR chemical shifts were compared to those quantum-chemically calculated by B3LYP/LanL2DZ+6-31G**//B3LYP/LanL2DZ+6-31G*, both in vacuo and in DMSO or DMF solution. PMID:18044804

  18. Vertical δ13C and δ15N changes during pedogenesis

    NASA Astrophysics Data System (ADS)

    Brunn, Melanie; Spielvogel, Sandra; Wells, Andrew; Condron, Leo; Oelmann, Yvonne

    2015-04-01

    The natural abundance of soil organic matter (SOM) stable C and N isotope ratios are subjected to vertical changes throughout the soil profile. This vertical distribution is a widely reported phenomenon across varieties of ecosystems and constitutes important insights of soil carbon cycling. In most ecosystems, SOM becomes enriched in heavy isotopes by several per mill in the first few centimeters of the topsoil. The enrichment of 13C in SOM with soil depth is attributed to biological and physical-chemical processes in soil e.g., plant physiological impacts, microbial decomposition, sorption and transport processes. Such vertical trends in 13C and 15N abundance have rarely been related to SOM composition during pedogenesis. The aims of our study were to investigate short and long-term δ13C and δ15N depth changes and their interrelations under progressing pedogenesis and ecosystem development. We sampled soils across the well studied fordune progradation Haast-chronosequence, a dune ridge system under super-humid climate at the West Coast of New Zealand's South Island (43° 53' S, 169° 3' E). Soils from 11 sites with five replicates each covered a time span of around 2870 yr of soil development (from Arenosol to Podzol). Vertical changes of δ13C and δ15N values of SOM were investigated in the organic layers and in 1-cm depth intervals of the upper 10 cm of the mineral soil. With increasing soil depth SOM became enriched in δ13C by 1.9 ± SE 0.1 o and in δ15N by 6.0 ± 0.4 ‰˙Litter δ13C values slightly decreased with increasing soil age (r = -0.61; p = 0.00) likely due to less efficient assimilation linked to nutrient limitations. Fractionation processes during mycorrhizal transfer appeared to affect δ15N values in the litter. We found a strong decrease of δ15N in the early succession stages ≤ 300 yr B.P. (r = -0.95; p = 0.00). Positive relations of vertical 13C and 15N enrichment with soil age might be related to decomposition and appeared to be

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

    PubMed

    Wiedemann, Christoph; Goradia, Nishit; Häfner, Sabine; Herbst, Christian; Görlach, Matthias; Ohlenschläger, Oliver; Ramachandran, Ramadurai

    2015-10-01

    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 ((13)C, (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 (1) J CαN and (2) J CαN couplings to transfer the (15)N x magnetisation from amino acid residue 'i' to adjacent residues via the application of a band-selective (15)N-(13)C(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. PMID:26282620

  20. Isotopic variability of cave bears (δ15N, δ13C) across Europe during MIS 3

    NASA Astrophysics Data System (ADS)

    Krajcarz, Magdalena; Pacher, Martina; Krajcarz, Maciej T.; Laughlan, Lana; Rabeder, Gernot; Sabol, Martin; Wojtal, Piotr; Bocherens, Hervé

    2016-01-01

    Collagen, the organic fraction of bone, records the isotopic parameters of consumed food for carbon (δ13C) and nitrogen (δ15N). This relationship of isotopic signature between diet and tissue is an important tool for the study of dietary preferences of modern and fossil animal species. Since the first information on the isotopic signature of cave bear was reported, numerous data from Europe have become available. The goal of this work is to track the geographical variation of cave bear collagen isotopic values in Europe during Marine Isotopic Stage 3 (about 60,000-25,000 yr BP). In this study the results of new δ13C and δ15N isotopic analyses of cave bear collagen from four Central-Eastern European sites are presented, as well as a review of all published isotopic data for cave bears of the same period. The main conclusion is a lack of geographical East-West pattern in the variations of δ13C and δ15N values of cave bear collagen. Moreover, no relationship was found between cave bear taxonomy and isotopic composition. The cave bears from Central-Eastern Europe exhibit δ13C and δ15N values near the average of the range of Central, Western and Southern European cave bears. Despite the fact that most cave bear sites follow an altitudinal gradient, separate groups of sites exhibit shift in absolute values of δ13C, what disturbs an altitude-related isotopic pattern. The most distinct groups are: high Alpine sites situated over 1500 m a.s.l. - in terms of δ13C; and two Romanian sites Peştera cu Oase and Urşilor - in case of δ15N. Although the cave bear isotopic signature is driven by altitude, the altitudinal adjustment of isotopic data is not enough to explain the isotopic dissimilarity of these cave bears. The unusually high δ15N signature of mentioned Romanian sites is an isolated case in Europe. Cave bears from relatively closely situated Central-Eastern European sites and other Romanian sites are more similar to Western European than to Romanian

  1. Functional groups identified by solid state 13C NMR spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal manure is generally high in organic matter intensity so it is well suitable for 13C nuclear magnetic resonance (NMR) analysis. Solid-state 13C NMR techniques used in characterizing organic matter and its components include, but are not limited to, cross-polarization /magic angle spinning (CP...

  2. Food webs in Mongolian grasslands: the analysis of 13C and 15N natural abundances.

    PubMed

    Kohzu, Ayato; Iwata, T; Kato, M; Nishikawa, J; Wada, Eitaro; Amartuvshin, N; Namkhaidorj, B; Fujita, N

    2009-09-01

    Overgrazing often lowers species richness and productivity of grassland communities. For Mongolian grassland ecosystems, a lack of detailed information about food-web structures makes it difficult to predict the effects of overgrazing on species diversity and community composition. We analysed the delta13C and delta15N signatures of herbaceous plants, arthropods (grouped by feeding habit), wild and domestic mammals, and humans in central Mongolia to understand the predominant food-web pathways in this grassland ecosystem. The delta13C and delta15N values of mammals showed little variation within species, but varied considerably with slope position for arthropods. The apparent isotopic discrimination between body tissue and hair of mammals was estimated as 2.0 per thousand for delta13C and 2.1 per thousand for delta15N, which was large enough to cause overestimation of the trophic level of mammals if not taken into account when using hair samples to measure isotopic enrichment. PMID:19507080

  3. Development of LC-13C NMR

    NASA Technical Reports Server (NTRS)

    Dorn, H. C.; Wang, J. S.; Glass, T. E.

    1986-01-01

    This study involves the development of C-13 nuclear resonance as an on-line detector for liquid chromatography (LC-C-13 NMR) for the chemical characterization of aviation fuels. The initial focus of this study was the development of a high sensitivity flow C-13 NMR probe. Since C-13 NMR sensitivity is of paramount concern, considerable effort during the first year was directed at new NMR probe designs. In particular, various toroid coil designs were examined. In addition, corresponding shim coils for correcting the main magnetic field (B sub 0) homogeneity were examined. Based on these initial probe design studies, an LC-C-13 NMR probe was built and flow C-13 NMR data was obtained for a limited number of samples.

  4. Whole-core analysis by sup 13 C NMR

    SciTech Connect

    Vinegar, H.J.; Tutunjian, P.N. ); Edelstein, W.A.; Roemer, P.B. )

    1991-06-01

    This paper reports on a whole-core nuclear magnetic resonance (NMR) system that was used to obtain natural abundance {sup 13}C spectra. The system enables rapid, nondestructive measurements of bulk volume of movable oil, aliphatic/aromatic ratio, oil viscosity, and organic vs. carbonate carbon. {sup 13}C NMR can be used in cores where the {sup 1}H NMR spectrum is too broad to resolve oil and water resonances separately. A 5 1/4-in. {sup 13}C/{sup 1}H NMR coil was installed on a General Electric (GE) CSI-2T NMR imager/spectrometer. With a 4-in.-OD whole core, good {sup 13}C signal/noise ratio (SNR) is obtained within minutes, while {sup 1}H spectra are obtained in seconds. NMR measurements have been made of the {sup 13}C and {sup 1}H density of crude oils with a wide range of API gravities. For light- and medium-gravity oils, the {sup 13}C and {sup 1}H signal per unit volume is constant within about 3.5%. For heavy crudes, the {sup 13}C and {sup 1}H density measured by NMR is reduced by the shortening of spin-spin relaxation time. {sup 13}C and {sup 1}H NMR spin-lattice relaxation times were measured on a suite of Cannon viscosity standards, crude oils (4 to 60{degrees} API), and alkanes (C{sub 5} through C{sub 16}) with viscosities at 77{degrees}F ranging from 0.5 cp to 2.5 {times} 10{sup 7} cp. The {sup 13}C and {sup 1}H relaxation times show a similar correlation with viscosity from which oil viscosity can be estimated accurately for viscosities up to 100 cp. The {sup 13}C surface relaxation rate for oils on water-wet rocks is very low. Nonproton decoupled {sup 13}C NMR is shown to be insensitive to kerogen; thus, {sup 13}C NMR measures only the movable hydrocarbon content of the cores. In carbonates, the {sup 13}C spectrum also contains a carbonate powder pattern useful in quantifying inorganic carbon and distinguishing organic from carbonate carbon.

  5. Benchmark fragment-based (1)H, (13)C, (15)N and (17)O chemical shift predictions in molecular crystals.

    PubMed

    Hartman, Joshua D; Kudla, Ryan A; Day, Graeme M; Mueller, Leonard J; Beran, Gregory J O

    2016-08-21

    The performance of fragment-based ab initio(1)H, (13)C, (15)N and (17)O chemical shift predictions is assessed against experimental NMR chemical shift data in four benchmark sets of molecular crystals. Employing a variety of commonly used density functionals (PBE0, B3LYP, TPSSh, OPBE, PBE, TPSS), we explore the relative performance of cluster, two-body fragment, and combined cluster/fragment models. The hybrid density functionals (PBE0, B3LYP and TPSSh) generally out-perform their generalized gradient approximation (GGA)-based counterparts. (1)H, (13)C, (15)N, and (17)O isotropic chemical shifts can be predicted with root-mean-square errors of 0.3, 1.5, 4.2, and 9.8 ppm, respectively, using a computationally inexpensive electrostatically embedded two-body PBE0 fragment model. Oxygen chemical shieldings prove particularly sensitive to local many-body effects, and using a combined cluster/fragment model instead of the simple two-body fragment model decreases the root-mean-square errors to 7.6 ppm. These fragment-based model errors compare favorably with GIPAW PBE ones of 0.4, 2.2, 5.4, and 7.2 ppm for the same (1)H, (13)C, (15)N, and (17)O test sets. Using these benchmark calculations, a set of recommended linear regression parameters for mapping between calculated chemical shieldings and observed chemical shifts are provided and their robustness assessed using statistical cross-validation. We demonstrate the utility of these approaches and the reported scaling parameters on applications to 9-tert-butyl anthracene, several histidine co-crystals, benzoic acid and the C-nitrosoarene SnCl2(CH3)2(NODMA)2. PMID:27431490

  6. Determination of 15N/14N and 13C/12C in Solid and Aqueous Cyanides

    USGS Publications Warehouse

    Johnson, C.A.

    1996-01-01

    The stable isotopic compositions of nitrogen and carbon in cyanide compounds can be determined by combusting aliquots in sealed tubes to form N2 gas and CO2 gas and analyzing the gases by mass spectrometry. Free cyanide (CN-aq + HCNaq) in simple solutions can also be analyzed by first precipitating the cyanide as copper(II) ferrocyanide and then combusting the precipitate. Reproducibility is ??0.5??? or better for both ??15N and ??13C. If empirical corrections are made on the basis of carbon yields, the reproducibility of ??13C can be improved to ??0.2???. The analytical methods described herein are sufficiently accurate and precise to apply stable isotope techniques to problems of cyanide degradation in natural waters and industrial process solutions.

  7. Acid treatment biasing to C/N, δ13C and δ15N of organic matter: A Molecular insight

    NASA Astrophysics Data System (ADS)

    Brodie, C.; Casford, J.; Leng, M. J.; Zong, Y.

    2011-12-01

    It is known that acid treatment methods employed to remove inorganic carbon (IC) from sample material prior to analysis for C/N, δ13C and δ15N cause non-linear, unpredictable biasing to the organic matter (OM) fraction [1, 2, 3]. Consequently, measured C/N, δ13C and δ15N have an uncertainty much greater than instrument precision with biases for C/N reported in the range of 1 - 100, for δ13C in the range of 0.2 - 6.8 % and for δ15N in the range of 0.2 - 1.5 % [1, 2, 3], in both modern and palaeo environmental materials. Brodie et al [3] extended this investigation to a down-core lake sedimentary archive (Lake Tianyang, South China) and noted the potential for this biasing to preclude "common" interpretations of the data (e.g., C/N values as an OM provenance tool; δ13C as a proxy for changes in C3 and C4 vegetation). It is evident that the size of biasing between sample horizons varies considerably implying a differential relative reaction to acid treatment down-core (i.e., as the type, relative amount and physical state of organic components changes. We now investigate this biasing at the molecular level by employing 13C-NMR and GC-IRMS techniques on a suite of modern and palaeo environmental materials and on a lake sedimentary archive. This will provide an important insight into the effect of acid treatment on organic compounds (i.e. removal from the sample, breakdown of compounds and partial removal) and associated isotopic fractionation. From an improved understanding of the type of compounds most susceptible to alteration/removal during the acid treatment processes it will be possible to consider refinements to the acid pre-treatment process and provide information on the relative down-core changes in those compounds susceptible to change (which we may be able to glean environmental information from). [1] Brodie, C.R., Leng, M.J., Casford, J.S.L., Kendrick, C.K., Lloyd, J.M., Zong, Y.Q., and Bird, M.I. (2011) Evidence for bias in C and N concentrations

  8. Study of molecular interactions with 13C DNP-NMR

    NASA Astrophysics Data System (ADS)

    Lerche, Mathilde H.; Meier, Sebastian; Jensen, Pernille R.; Baumann, Herbert; Petersen, Bent O.; Karlsson, Magnus; Duus, Jens Ø.; Ardenkjær-Larsen, Jan H.

    2010-03-01

    NMR spectroscopy is an established, versatile technique for the detection of molecular interactions, even when these interactions are weak. Signal enhancement by several orders of magnitude through dynamic nuclear polarization alleviates several practical limitations of NMR-based interaction studies. This enhanced non-equilibrium polarization contributes sensitivity for the detection of molecular interactions in a single NMR transient. We show that direct 13C NMR ligand binding studies at natural isotopic abundance of 13C gets feasible in this way. Resultant screens are easy to interpret and can be performed at 13C concentrations below μM. In addition to such ligand-detected studies of molecular interaction, ligand binding can be assessed and quantified with enzymatic assays that employ hyperpolarized substrates at varying enzyme inhibitor concentrations. The physical labeling of nuclear spins by hyperpolarization thus provides the opportunity to devise fast novel in vitro experiments with low material requirement and without the need for synthetic modifications of target or ligands.

  9. (1)H, (13)C and (15)N backbone assignment of the EC-1 domain of human E-cadherin.

    PubMed

    Prasasty, Vivitri D; Krause, Mary E; Tambunan, Usman S F; Anbanandam, Asokan; Laurence, Jennifer S; Siahaan, Teruna J

    2015-04-01

    The Extracellular 1 (EC1) domain of E-cadherin has been shown to be important for cadherin-cadherin homophilic interactions. Cadherins are responsible for calcium-mediated cell-cell adhesion located at the adherens junction of the biological barriers (i.e., intestinal mucosa and the blood-brain barrier (BBB)). Cadherin peptides can modulate cadherin interactions to improve drug delivery through the BBB. However, the mechanism of modulating the E-cadherin interactions by cadherin peptides has not been fully elucidated. To provide a basis for subsequent examination of the structure and peptide-binding properties of the EC1 domain of human E-cadherin using solution NMR spectroscopy, the (1)H, (13)C and (15)N backbone resonance of the uniformly labeled-EC1 were assigned and the secondary structure was determined based on the chemical shift values. These resonance assignments are essential for assessing protein-ligand interactions and are reported here. PMID:24510398

  10. High resolution (13)C MRI with hyperpolarized urea: in vivo T(2) mapping and (15)N labeling effects.

    PubMed

    Reed, Galen D; von Morze, Cornelius; Bok, Robert; Koelsch, Bertram L; Van Criekinge, Mark; Smith, Kenneth J; Hong Shang; Larson, Peder E Z; Kurhanewicz, John; Vigneron, Daniel B

    2014-02-01

    (13)C steady state free precession (SSFP) magnetic resonance imaging and effective spin-spin relaxation time (T2) mapping were performed using hyperpolarized [(13)C] urea and [(13) C,(15)N2] urea injected intravenously in rats. (15)N labeling gave large T2 increases both in solution and in vivo due to the elimination of a strong scalar relaxation pathway. The T2 increase was pronounced in the kidney, with [(13) C,(15) N2] urea giving T2 values of 6.3±1.3 s in the cortex and medulla, and 11±2 s in the renal pelvis. The measured T2 in the aorta was 1.3±0.3 s. [(13)C] urea showed shortened T2 values in the kidney of 0.23±0.03 s compared to 0.28±0.03 s measured in the aorta. The enhanced T2 of [(13)C,(15)N2] urea was utilized to generate large signal enhancement by SSFP acquisitions with flip angles approaching the fully refocused regime. Projection images at 0.94 mm in-plane resolution were acquired with both urea isotopes, with [(13)C,(15) N2] urea giving a greater than four-fold increase in signal-to-noise ratio over [(13)C] urea. PMID:24235273

  11. Regional, seasonal and interspecific variation in 15N and 13C in sympatric mouse lemurs

    NASA Astrophysics Data System (ADS)

    Rakotondranary, S. Jacques; Struck, Ulrich; Knoblauch, Christian; Ganzhorn, Jörg U.

    2011-11-01

    Madagascar provides some of the rare examples where two or more primate species of the same genus and with seemingly identical niche requirements occur in sympatry. If congeneric primate species co-occur in other parts of the world, they differ in size in a way that is consistent with Hutchinson's rule for coexisting species, or they occupy different ecological niches. In some areas of Madagascar, mouse lemurs do not follow these "rules" and thus seem to violate one of the principles of community ecology. In order to understand the mechanisms that allow coexistence of sympatric congeneric species without obvious niche differentiation, we studied food composition of two identical sized omnivorous mouse lemur species, Microcebus griseorufus and M. murinus with the help of stable isotope analyses ( δ 15N and δ 13C). The two species are closely related sister species. During the rich season, when food seems abundant, the two species do not differ in their nitrogen isotope composition, indicating that the two species occupy the same trophic level. But they differ in their δ 13C values, indicating that M. griseorufus feeds more on C4 and CAM (Crassulacean-acid-metabolism) plants than M. murinus. During the lean season, M. murinus has lower δ 15N values, indicating that the two species feed at different trophic levels during times of food shortage. Hybrids between the two species showed intermediate food composition. The results reflect subtle differences in foraging or metabolic adaptations that are difficult to quantify by traditional observations but that represent possibilities to allow coexistence of species.

  12. Intraskeletal isotopic compositions (δ(13) C, δ(15) N) of bone collagen: nonpathological and pathological variation.

    PubMed

    Olsen, Karyn C; White, Christine D; Longstaffe, Fred J; von Heyking, Kristin; McGlynn, George; Grupe, Gisela; Rühli, Frank J

    2014-04-01

    Paleodiet research traditionally interprets differences in collagen isotopic compositions (δ(13) C, δ(15) N) as indicators of dietary distinction even though physiological processes likely play some role in creating variation. This research investigates the degree to which bone collagen δ(13) C and δ(15) N values normally vary within the skeleton and examines the influence of several diseases common to ancient populations on these isotopic compositions. The samples derive from two medieval German cemeteries and one Swiss reference collection and include examples of metabolic disease (rickets/osteomalacia), degenerative joint disease (osteoarthritis), trauma (fracture), infection (osteomyelitis), and inflammation (periostitis). A separate subset of visibly nonpathological skeletal elements from the German collections established normal intraindividual variation. For each disease type, tests compared bone lesion samples to those near and distant to the lesions sites. Results show that normal (nonpathological) skeletons exhibit limited intraskeletal variation in carbon- and nitrogen-isotope ratios, suggesting that sampling of distinct elements is appropriate for paleodiet studies. In contrast, individuals with osteomyelitis, healed fractures, and osteoarthritis exhibit significant intraskeletal differences in isotope values, depending on whether one is comparing lesions to near or to distant sites. Skeletons with periostitis result in significant intraskeletal differences in nitrogen isotope values only, while those with rickets/osteomalacia do not exhibit significant intraskeletal differences. Based on these results, we suggest that paleodiet researchers avoid sampling collagen at or close to lesion sites because the isotope values may be reflecting both altered metabolic processes and differences in diet relative to others in the population. PMID:24374993

  13. Afforestation impacts microbial biomass and its natural (13)C and (15)N abundance in soil aggregates in central China.

    PubMed

    Wu, Junjun; Zhang, Qian; Yang, Fan; Lei, Yao; Zhang, Quanfa; Cheng, Xiaoli

    2016-10-15

    We investigated soil microbial biomass and its natural abundance of δ(13)C and δ(15)N in aggregates (>2000μm, 250-2000μm, 53-250μm and <53μm) of afforested (implementing woodland and shrubland plantations) soils, adjacent croplands and open area (i.e., control) in the Danjiangkou Reservoir area of central China. The afforested soils averaged higher microbial biomass carbon (MBC) and nitrogen (MBN) levels in all aggregates than in open area and cropland, with higher microbial biomass in micro-aggregates (<250μm) than in macro-aggregates (>2000μm). The δ(13)C of soil microbial biomass was more enriched in woodland soils than in other land use types, while δ(15)N of soil microbial biomass was more enriched compared with that of organic soil in all land use types. The δ(13)C and δ(15)N of microbial biomass were positively correlated with the δ(13)C and δ(15)N of organic soil across aggregates and land use types, whereas the (13)C and (15)N enrichment of microbial biomass exhibited linear decreases with the corresponding C:N ratio of organic soil. Our results suggest that shifts in the natural (13)C and (15)N abundance of microbial biomass reflect changes in the stabilization and turnover of soil organic matter (SOM) and thereby imply that afforestation can greatly impact SOM accumulation over the long-term. PMID:27285796

  14. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis.

    PubMed

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian; Ulrich, Anne S

    2015-06-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly (13)C/(15)N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive (13)C/(15)N-labeled amino acids. The most cost-effective production of (13)C/(15)N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% (13)C-glycerol and 0.5% (15)N-ammonium sulfate, supplemented with only 0.025% of (13)C/(15)N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state. PMID:25795666

  15. Fermentation and Cost-Effective 13C/15N Labeling of the Nonribosomal Peptide Gramicidin S for Nuclear Magnetic Resonance Structure Analysis

    PubMed Central

    Berditsch, Marina; Afonin, Sergii; Steineker, Anna; Orel, Nataliia; Jakovkin, Igor; Weber, Christian

    2015-01-01

    Gramicidin S (GS) is a nonribosomally synthesized decapeptide from Aneurinibacillus migulanus. Its pronounced antibiotic activity is attributed to amphiphilic structure and enables GS interaction with bacterial membranes. Despite its medical use for over 70 years, the peptide-lipid interactions of GS and its molecular mechanism of action are still not fully understood. Therefore, a comprehensive structural analysis of isotope-labeled GS needs to be performed in its biologically relevant membrane-bound state, using advanced solid-state nuclear magnetic resonance (NMR) spectroscopy. Here, we describe an efficient method for producing the uniformly 13C/15N-labeled peptide in a minimal medium supplemented by selected amino acids. As GS is an intracellular product of A. migulanus, we characterized the producer strain DSM 5759 (rough-convex phenotype) and examined its biosynthetic activity in terms of absolute and biomass-dependent peptide accumulation. We found that the addition of either arginine or ornithine increases the yield only at very high supplementing concentrations (1% and 0.4%, respectively) of these expensive 13C/15N-labeled amino acids. The most cost-effective production of 13C/15N-GS, giving up to 90 mg per gram of dry cell weight, was achieved in a minimal medium containing 1% 13C-glycerol and 0.5% 15N-ammonium sulfate, supplemented with only 0.025% of 13C/15N-phenylalanine. The 100% efficiency of labeling is corroborated by mass spectrometry and preliminary solid-state NMR structure analysis of the labeled peptide in the membrane-bound state. PMID:25795666

  16. Stellar Origins of Extremely 13C- and 15N-enriched Presolar SiC Grains: Novae or Supernovae?

    NASA Astrophysics Data System (ADS)

    Liu, Nan; Nittler, Larry R.; O'D. Alexander, Conel M.; Wang, Jianhua; Pignatari, Marco; José, Jordi; Nguyen, Ann

    2016-04-01

    Extreme excesses of 13C (12C/13C < 10) and 15N (14N/15N < 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 13C- and 15N-enriched presolar SiC grains (12C/13C < 16 and 14N/15N < ˜100) from Murchison, and their correlated Mg-Al, S, and Ca-Ti isotope data when available. These grains are enriched in 13C and 15N, but with quite diverse Si isotopic signatures. Four grains with 29,30Si 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 30Si excesses and 29Si depletions show lower-than-solar 34S/32S 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 13C enriched, but have a range of higher 14N/15N. We found that 15N-enriched AB grains (˜50 < 14N/15N < ˜100) have distinctive isotopic signatures compared to putative nova grains, such as higher 14N/15N, lower 26Al/27Al, and lack of 30Si excess, indicating weaker proton-capture nucleosynthetic environments.

  17. Effect of body size and body mass on δ 13 C and δ 15 N in coastal fishes and cephalopods

    NASA Astrophysics Data System (ADS)

    Vinagre, C.; Máguas, C.; Cabral, H. N.; Costa, M. J.

    2011-11-01

    Carbon and nitrogen isotopes have been widely used in the investigation of trophic relations, energy pathways, trophic levels and migrations, under the assumption that δ 13C is independent of body size and that variation in δ 15N occurs exclusively due to ontogenetic changes in diet and not body size increase per se. However, several studies have shown that these assumptions are uncertain. Data from food-webs containing an important number of species lack theoretical support on these assumptions because very few species have been tested for δ 13C and δ 15N variation in captivity. However, if sampling comprises a wide range of body sizes from various species, the variation of δ 13C and δ 15N with body size can be investigated. While correlation between body size and δ 13C and δ 15N can be due to ontogenetic diet shifts, stability in such values throughout the size spectrum can be considered an indication that δ 13C and δ 15N in muscle tissues of such species is independent of body size within that size range, and thus the basic assumptions can be applied in the interpretation of such food webs. The present study investigated the variation in muscle δ 13C and δ 15N with body size and body mass of coastal fishes and cephalopods. It was concluded that muscle δ 13C and δ 15N did not vary with body size or mass for all bony fishes with only one exception, the dragonet Callionymus lyra. Muscle δ 13C and δ 15N also did not vary with body size or mass in cartilaginous fishes and cephalopods, meaning that body size/mass per se have no effect on δ 13C or δ 15N, for most species analysed and within the size ranges sampled. The assumption that δ 13C is independent of body size and that variation in δ 15N is not affected by body size increase per se was upheld for most organisms and can be applied to the coastal food web studied taking into account that C. lyra is an exception.

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

    SciTech Connect

    Strzelecka, T.; Gronenborn, A.M.; Clore, G.M.

    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.

  19. A 13C-NMR study of azacryptand complexes.

    PubMed

    Wild, Aljoscha A C; Fennell, Kevin; Morgan, Grace G; Hewage, Chandralal M; Malthouse, J Paul G

    2014-09-28

    An azacryptand has been solubilised in aqueous media containing 50% (v/v) dimethyl sulphoxide. (13)C-NMR has been used to determine how the azacryptand is affected by zinc binding at pH 10. Using (13)C-NMR and (13)C-enriched bicarbonate we have been able to observe the formation of 4 different carbamate derivatives of the azacryptand at pH 10. The azacryptand was shown to solubilise zinc or cadmium at alkaline pHs. Two moles of zinc are bound per mole of azacryptand and this complex binds 1 mole of carbonate. By replacing the zinc with cadmium-113 we have shown that the (13)C-NMR signal of the (13)C-enriched carbon of the bound carbonate is split into two triplets at 2.2 °C. This shows that two cadmium complexes are formed and in each of these complexes the carbonate group is bound by two magnetically equivalent metal ions. It also demonstrates that these cadmium complexes are not in fast exchange. From temperature studies we show that in the zinc complexes both complexes are in fast exchange with each other but are in slow exchange with free bicarbonate. HOESY is used to determine the position of the carbonate carbon in the complex. The solution and crystal structures of the zinc-carbonate-azacryptand complexes are compared. PMID:25091182

  20. 13C NMR spectroscopy applications to brain energy metabolism

    PubMed Central

    Rodrigues, Tiago B.; Valette, Julien; Bouzier-Sore, Anne-Karine

    2013-01-01

    13C nuclear magnetic resonance (NMR) spectroscopy is the method of choice for studying brain metabolism. Indeed, the most convincing data obtained to decipher metabolic exchanges between neurons and astrocytes have been obtained using this technique, thus illustrating its power. It may be difficult for non-specialists, however, to grasp thefull implication of data presented in articles written by spectroscopists. The aim of the review is, therefore, to provide a fundamental understanding of this topic to facilitate the non-specialists in their reading of this literature. In the first part of this review, we present the metabolic fate of 13C-labeled substrates in the brain in a detailed way, including an overview of some general neurochemical principles. We also address and compare the various spectroscopic strategies that can be used to study brain metabolism. Then, we provide an overview of the 13C NMR experiments performed to analyze both intracellular and intercellular metabolic fluxes. More particularly, the role of lactate as a potential energy substrate for neurons is discussed in the light of 13C NMR data. Finally, new perspectives and applications offered by 13C hyperpolarization are described. PMID:24367329

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

    NASA Astrophysics Data System (ADS)

    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

  2. Two new organic reference materials for δ13C and δ15N measurements and a new value for the δ13C of NBS 22 oil

    USGS Publications Warehouse

    Qi, Haiping; Coplen, Tyler B.; Geilmann, Heike; Brand, Willi A.; Bohlke, John Karl

    2003-01-01

    Analytical grade L-glutamic acid is chemically stable and has a C/N mole ratio of 5, which is close to that of many of natural biological materials, such as blood and animal tissue. Two L-glutamic acid reference materials with substantially different 13C and 15N abundances have been prepared for use as organic reference materials for C and N isotopic measurements. USGS40 is analytical grade L-glutamic acid and has a δ13C value of −26.24‰ relative to VPDB and a δ15N value of −4.52‰ relative to N2 in air. USGS41 was prepared by dissolving analytical grade L-glutamic acid with L-glutamic acid enriched in 13C and 15N. USGS41 has a δ13C value of +37.76‰ and a δ15N value of +47.57‰. The δ13C and δ15N values of both materials were measured against the international reference materials NBS 19 calcium carbonate (δ13C = +1.95‰), L-SVEC lithium carbonate (δ13C = −46.48‰), IAEA-N-1 ammonium sulfate (δ15N = 0.43‰), and USGS32 potassium nitrate (δ15N = 180‰) by on-line combustion continuous-flow and off-line dual-inlet isotope-ratio mass spectrometry. Both USGS40 and USGS41 are isotopically homogeneous; reproducibility of δ13C is better than 0.13‰, and that of δ15N is better than 0.13‰ in 100-μg amounts. These two isotopic reference materials can be used for (i) calibrating local laboratory reference materials, and (ii) quantifying drift with time, mass-dependent fractionations, and isotope-ratio-scale contraction in the isotopic analysis of various biological materials. Isotopic results presented in this paper yield a δ13C value for NBS 22 oil of −29.91‰, in contrast to the commonly accepted value of −29.78‰ for which off-line blank corrections probably have not been quantified satisfactorily.

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

    PubMed Central

    Yang, Yang; Siegwolf, Rolf T. W.; Körner, Christian

    2015-01-01

    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. Two thousand five hundred meter 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. PMID:26097487

  4. Choice of dietary protein of vegetarians and omnivores is reflected in their hair protein 13C and 15N abundance.

    PubMed

    Petzke, Klaus J; Boeing, Heiner; Metges, Cornelia C

    2005-01-01

    Stable isotopic (15N, 13C) composition of tissues depends on isotopic pattern of food sources. We investigated whether the isotopic compositions of human hair protein and amino acids reflect the habitual dietary protein intake. Hair samples were analyzed from 100 omnivores (selected randomly out of the 1987-1988 German nutrition survey VERA), and from 15 ovo-lacto-vegetarians (OLV), and from 6 vegans recruited separately. Hair bulk and amino acid specific isotopic compositions were analyzed by isotope-ratio mass spectrometry (EA/IRMS and GC/C/IRMS, respectively) and the results were correlated with data of the 7 day dietary records. Hair bulk 15N and 13C abundances clearly reflect the particular eating habits. Vegans can be distinguished from OLV and both are significantly distinct from omnivores in both 15N and 13C abundances. 15N and 13C abundances rose with a higher proportion of animal to total protein intake (PAPI). Individual proportions of animal protein consumption (IPAP) were calculated using isotopic abundances and a linear regression model using animal protein consumption data of vegans (PAPI = 0) and omnivores (mean PAPI = 0.639). IPAP values positively correlated with the intake of protein, meat, meat products, and animal protein. Distinct patterns for hair amino acid specific 15N and 13C abundances were measured but with lower resolution between food preference groups compared with bulk values. In conclusion, hair 13C and 15N values both reflected the extent of animal protein consumption. Bulk isotopic abundance of hair can be tested for future use in the validation of dietary assessment methods. PMID:15880664

  5. Variation in hair δ13C and δ15N values in long-tailed macaques (Macaca fascicularis) from Singapore

    USGS Publications Warehouse

    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.

  6. Spatial Patterns of Plant δ13C and δ15N Along a Topoedaphic Gradient in a Subtropical Savanna Landscape

    NASA Astrophysics Data System (ADS)

    Bai, E.; Boutton, T. W.; Liu, F.; Wu, B.; Archer, S. R.

    2005-12-01

    δ13C and δ15N values of plants are powerful tools in physiological ecology, ecosystem science, and global biogeochemistry, yet we know relatively little about their variation and controls at the landscape scale. In this study, we investigated landscape-scale spatial variations in the foliar isotopic composition of 3 woody plant species across a 308 m topoedaphic gradient, along which soil texture and plant resources (water and nitrogen availability) varied from upland (86 m) to lowland (84 m) portions of the landscape. The study was conducted in a subtropical savanna at the La Copita Research Area, approximately 60 km west of Corpus Christi, TX. Foliar δ13C, δ15N, leaf nitrogen concentration ([N]), and specific leaf area (SLA) were measured on all individuals of Prosopis glandulosa, Condalia hookeri, and Zanthoxylum fagara present within a belt transect 308 m long x 12 m wide. Soil texture, available soil moisture, and total N were measured at 1 m intervals along the center-line of the belt transect. Clay content, available soil moisture, and soil total N were all negatively correlated with elevation along the transect. Leaf δ13C and δ15N values for all 3 species increased by 1-4 o/oo with decreasing elevation along the transect. Contrary to theory and previous studies, δ13C values were highest where soil water was most available, suggesting that some other variable could be overriding or interacting with water availability. Foliar [N] appeared to exert the strongest control over landscape-level variation, and was positively correlated with δ13C of all species (R 2 = 0.58, p<0.0001). Since leaf [N] is positively related to photosynthetic capacity, plants with high [N] are likely to have low Ci/Ca ratios and therefore higher δ13C values. δ15N values of Zanthoxylum and Condalia were positively correlated with leaf [N] and soil water availability; however, these relationships were absent for Prosopis, an N-fixing tree legume. We speculate that the

  7. Development of a 13C-optimized 1.5-mm high temperature superconducting NMR probe

    NASA Astrophysics Data System (ADS)

    Ramaswamy, Vijaykumar; Hooker, Jerris W.; Withers, Richard S.; Nast, Robert E.; Brey, William W.; Edison, Arthur S.

    2013-10-01

    We report a 1.5-mm NMR probe based on high temperature superconductors operating at 14.1 T optimized for 13C detection. The probe has a total sample volume of about 35 microliters (μL) with an active volume of 20 μL and provides exceptional mass sensitivity for 13C detection. The probe also has excellent 1H sensitivity and employs a 2H lock; 15N irradiation capability can be added in the future. The coils are cooled to about 20 K using a standard Agilent cryogenic refrigeration system, and the sample temperature is regulated near room temperature. The coil design considerations are discussed in detail. This probe is ideal for directly detected 13C NMR experiments for natural products chemistry and metabolomics applications, for which 35 μL is an optimal sample volume. The outstanding 13C sensitivity of this probe allowed us to directly determine the 13C connectivity on 1.1 mg of natural abundance histidine using an INADEQUATE experiment. We demonstrated the utility of this probe for 13C-based metabolomics using a synthetic mixture of common natural abundance metabolites whose concentrations ranged from 1 to 5 mM (40-200 nmol).

  8. Development of a 13C-Optimized 1.5-mm High Temperature Superconducting NMR Probe

    PubMed Central

    Ramaswamy, Vijaykumar; Hooker, Jerris W.; Withers, Richard S.; Nast, Robert E.; Brey, William W.; Edison, Arthur S.

    2013-01-01

    We report a 1.5-mm NMR probe based on high temperature superconductors operating at 14.1 T optimized for 13C detection. The probe has a total sample volume of about 35 microliters (μL) with an active volume of 20 μL and provides exceptional mass sensitivity for 13C detection. The probe also has excellent 1H sensitivity and employs a 2H channel lock; 15N irradiation capability can be added in the future. The coils are cooled to about 20 K using a standard Agilent cryogenic refrigeration system, and the sample temperature is regulated near room temperature. The coil design considerations are discussed in detail. This probe is ideal for directly detected 13C NMR experiments for natural products chemistry and metabolomics applications, for which 35 μL is an optimal sample volume. The outstanding 13C sensitivity of this probe allowed us to directly determine the 13C connectivity on 1.1 mg of natural abundance histidine using an INADEQUATE experiment. We demonstrated the utility of this probe for 13C-based metabolomics using a synthetic mixture of common natural abundance metabolites whose concentrations ranged from 1 to 5 mM (40 to 200 nmol). PMID:23969086

  9. Differentiation of Pigment in Eggs Using Carbon ((13)C/(12)C) and Nitrogen ((15)N/(14)N) Stable Isotopes.

    PubMed

    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. PMID:27302905

  10. 1H, 13C and 15N Backbone Assignment of the EC-1 Domain of Human E-Cadherin

    PubMed Central

    Prasasty, Vivitri D.; Krause, Mary E.; Tambunan, Usman S. F.; Anbanandam, Asokan; Laurence, Jennifer S.; Siahaan, Teruna J.

    2014-01-01

    The EC1 domain of E-cadherin has been shown to be important for cadherin-cadherin homophilic interactions. Cadherins are responsible for calcium-mediated cell-cell adhesion located at the adherens junction of the biological barriers (i.e., intestinal mucosa and the blood-brain barrier (BBB). Cadherin peptides can modulate cadherin interactions to improve drug delivery through the blood-brain barriers (BBB). However, the mechanism of modulating the E-cadherin interactions by cadherin peptides has not been fully elucidated. To provide a basis for subsequent examination of the structure and peptide-binding properties of the EC1 domain of human E-cadherin using solution NMR spectroscopy, the 1H, 13C and 15N backbone resonance of the uniformly labeled-EC1 were assigned and the secondary structure was determined based on the chemical shift values. These resonance assignments are essential for assessing protein-ligand interactions and are reported here. PMID:24510398

  11. Stable isotope (13C, 15N and 34S) analysis of the hair of modern humans and their domestic animals.

    PubMed

    Bol, Roland; Pflieger, Christian

    2002-01-01

    Relationships between dietary status and recent migration were examined by delta(13)C, delta(15)N and delta(34)S analysis of hair samples from 43 modern humans living in a rural community in SW England. The isotopic content of 38 'local' hair samples was compared with that of five recently arrived individuals (from Canada, Chile, Germany and the USA). Hair samples from domestic animals (i.e. mainly cats, dogs, cows and horses) were analysed to examine the difference in delta(13)C, delta(15)N and delta(34)S values between herbivores and carnivores. Generally, modern human hair data from the triple stable isotope (delta(13)C, delta(15)N and delta(34)S) provided enough information to confirm the dietary status and origin of the individual subjects. The dietary intake was generally reflected in the animal hair delta(15)N and delta(13)C values, i.e. highest in the carnivores (cats). However, a non-local origin of food sources given to domesticated omnivores (i.e. dogs) was suggested by their hair delta(34)S values. PMID:12442295

  12. Backbone and sidechain 1H, 15N and 13C assignments of the KSR1 CA1 domain

    PubMed Central

    Koveal, Dorothy; Pinheiro, Anderson S.; Peti, Wolfgang; Page, Rebecca

    2014-01-01

    The backbone and side chain resonance assignments of the murine KSR1 CA1 domain have been determined based on triple-resonance experiments using uniformly [13C, 15N]-labeled protein. This assignment is the first step towards the determination of the three-dimensional structure of the unique KSR1 CA1 domain. PMID:20737253

  13. Elucidating the trophodynamics of four coral reef fishes of the Solomon Islands using δ15N and δ13C

    NASA Astrophysics Data System (ADS)

    Greenwood, N. D. W.; Sweeting, C. J.; Polunin, N. V. C.

    2010-09-01

    Size-related diet shifts are important characteristics of fish trophodynamics. Here, body size-related changes in muscle δ15N and δ13C of four coral reef fishes, Acanthurus nigrofuscus (herbivore), Chaetodon lunulatus (corallivore) , Chromis xanthura (planktivore) and Plectropomus leopardus (piscivore) were investigated at two locations in the Solomon Islands. All four species occupied distinct isotopic niches and the concurrent δ13C' values of C. xanthura and P. leopardus suggested a common planktonic production source. Size-related shifts in δ15N, and thus trophic level, were observed in C. xanthura, C. lunulatus and P. leopardus, and these trends varied between location, indicating spatial differences in trophic ecology. A literature review of tropical fishes revealed that positive δ15N-size trends are common while negative δ15N-size trends are rare. Size-δ15N trends fall into approximately equal groups representing size-based feeding within a food chain, and that associated with a basal resource shift and occurs in conjunction with changes in production source, indicated by δ13C. The review also revealed large scale differences in isotope-size trends and this, combined with small scale location differences noted earlier, highlights a high degree of plasticity in the reef fishes studied. This suggests that trophic size analysis of reef fishes would provide a productive avenue to identify species potentially vulnerable to reef impacts as a result of constrained trophic behaviour.

  14. (15)N NMR studies of a nitrile-modified nucleoside.

    PubMed

    Gillies, Anne T; Gai, Xin Sonia; Buckwalter, Beth L; Fenlon, Edward E; Brewer, Scott H

    2010-12-30

    Nitrile-modified molecules have proven to be excellent probes of local environments in biomolecules via both vibrational and fluorescence spectroscopy. The utility of the nitrile group as a spectroscopic probe has been expanded here to (15)N NMR spectroscopy by selective (15)N incorporation. The (15)N NMR chemical shift (δ((15)N)) of the (15)N-labeled 5-cyano-2'-deoxyuridine (C(15)NdU, 1a) was found to change from 153.47 to 143.80 ppm in going from THF-d(8) to D(2)O. A 0.81 ppm downfield shift was measured upon formation of a hydrogen-bond-mediated heterodimer between 2,6-diheptanamidopyridine and a silyl ether analogue of 1a in chloroform, and the small intrinsic temperature dependence of δ((15)N) of C(15)NdU was measured as a 0.38 ppm downfield shift from 298 to 338 K. The experiments were complemented with density functional theory calculations exploring the effect of solvation on the (15)N NMR chemical shift. PMID:21126044

  15. Organic vs. Conventional Grassland Management: Do 15N and 13C Isotopic Signatures of Hay and Soil Samples Differ?

    PubMed Central

    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 δ15N and δ13C isotopic composition of soil and hay samples of 21 organic and 34 conventional grasslands in two German regions. We also used Δδ15N15N plant - δ15N 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 δ13C in hay and δ15N in both soil and hay between management types, but showed that δ13C abundances were significantly lower in soil of organic compared to conventional grasslands. Δδ15N values implied that management types did not substantially differ in nitrogen cycling. Only δ13C 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 used in practice

  16. State-of-the-Art Direct 13C and Indirect 1H-[13C] NMR Spectroscopy In Vivo

    PubMed Central

    de Graaf, Robin A.; Rothman, Douglas L.; Behar, Kevin L.

    2013-01-01

    Carbon-13 NMR spectroscopy in combination with 13C-labeled substrate infusion is a powerful technique to measure a large number of metabolic fluxes non-invasively in vivo. It has been used to quantify glycogen synthesis rates, establish quantitative relationships between energy metabolism and neurotransmission and evaluate the importance of different substrates. All measurements can, in principle, be performed through direct 13C NMR detection or via indirect 1H-[13C] NMR detection of the protons attached to 13C nuclei. The choice for detection scheme and pulse sequence depends on the magnetic field strength, whereas substrate selection depends on the metabolic pathways that are studied. 13C NMR spectroscopy remains a challenging technique that requires several non-standard hardware modifications, infusion of 13C-labeled substrates and sophisticated processing and metabolic modeling. Here the various aspects of direct 13C and indirect 1H-[13C] NMR are reviewed with the aim of providing a practical guide. PMID:21919099

  17. Accurate measurements of {sup 13}C-{sup 13}C distances in uniformly {sup 13}C-labeled proteins using multi-dimensional four-oscillating field solid-state NMR spectroscopy

    SciTech Connect

    Straasø, Lasse Arnt; Nielsen, Jakob Toudahl; Bjerring, Morten; Nielsen, Niels Chr.; Khaneja, Navin

    2014-09-21

    Application of sets of {sup 13}C-{sup 13}C internuclear distance restraints constitutes a typical key element in determining the structure of peptides and proteins by magic-angle-spinning solid-state NMR spectroscopy. Accurate measurements of the structurally highly important {sup 13}C-{sup 13}C distances in uniformly {sup 13}C-labeled peptides and proteins, however, pose a big challenge due to the problem of dipolar truncation. Here, we present novel two-dimensional (2D) solid-state NMR experiments capable of extracting distances between carbonyl ({sup 13}C′) and aliphatic ({sup 13}C{sub aliphatic}) spins with high accuracy. The method is based on an improved version of the four-oscillating field (FOLD) technique [L. A. Straasø, M. Bjerring, N. Khaneja, and N. C. Nielsen, J. Chem. Phys. 130, 225103 (2009)] which circumvents the problem of dipolar truncation, thereby offering a base for accurate extraction of internuclear distances in many-spin systems. The ability to extract reliable accurate distances is demonstrated using one- and two-dimensional variants of the FOLD experiment on uniformly {sup 13}C,{sup 15}N-labeled-L-isoleucine. In a more challenging biological application, FOLD 2D experiments are used to determine a large number of {sup 13}C′-{sup 13}C{sub aliphatic} distances in amyloid fibrils formed by the SNNFGAILSS fibrillating core of the human islet amyloid polypeptide with uniform {sup 13}C,{sup 15}N-labeling on the FGAIL fragment.

  18. Identification of novel hydrazine metabolites by 15N-NMR.

    PubMed

    Preece, N E; Nicholson, J K; Timbrell, J A

    1991-05-01

    15N-NMR has been used to study the metabolism of hydrazine in rats in vivo. Single doses of [15N2]hydrazine (2.0 mmol/kg: 98.6% g atom) were administered to rats and urine collected for 24 hr over ice. A number of metabolites were detected by 15N-NMR analysis of lyophilized urine. Ammonia was detected as a singlet at 0 ppm and unchanged [15N2]hydrazine was present in the urine detectable as a singlet at 32 ppm. Peaks were observed at 107 and 110 ppm which were identified as being due to the hydrazido nitrogen of acetylhydrazine and diacetylhydrazine, respectively. A resonance at 85 ppm was ascribed to carbazic acid, resulting from reaction of hydrazine with carbon dioxide. A singlet detected at 316 ppm was thought to be due to the hydrazono nitrogen of the pyruvate hydrazone. The resonance at 56 ppm was assigned to 15N-enriched urea, this together with the presence of ammonia indicates that the N-N bond of hydrazine is cleaved in vivo, possibly by N-oxidation, and the resultant ammonia is incorporated into urea. A doublet centred at 150 ppm and a singlet at 294 ppm were assigned to a metabolite which results from cyclization of the 2-oxoglutarate hydrazone. Therefore 15N-NMR spectroscopic analysis of urine has yielded significant new information on the metabolism of hydrazine. PMID:2018564

  19. Accurate quantitative 13C NMR spectroscopy: repeatability over time of site-specific 13C isotope ratio determination.

    PubMed

    Caytan, Elsa; Botosoa, Eliot P; Silvestre, Virginie; Robins, Richard J; Akoka, Serge; Remaud, Gérald S

    2007-11-01

    The stability over time (repeatability) for the determination of site-specific 13C/12C ratios at natural abundance by quantitative 13C NMR spectroscopy has been tested on three probes: enriched bilabeled [1,2-13C2]ethanol; ethanol at natural abundance; and vanillin at natural abundance. It is shown in all three cases that the standard deviation for a series of measurements taken every 2-3 months over periods between 9 and 13 months is equal to or smaller than the standard deviation calculated from 5-10 replicate measurements made on a single sample. The precision which can be achieved using the present analytical 13C NMR protocol is higher than the prerequisite value of 1-2 per thousand for the determination of site-specific 13C/12C ratios at natural abundance (13C-SNIF-NMR). Hence, this technique permits the discrimination of very small variations in 13C/12C ratios between carbon positions, as found in biogenic natural products. This observed stability over time in 13C NMR spectroscopy indicates that further improvements in precision will depend primarily on improved signal-to-noise ratio. PMID:17900175

  20. Carbon-rich Presolar Grains from Massive Stars: Subsolar 12C/13C and 14N/15N Ratios and the Mystery of 15N

    NASA Astrophysics Data System (ADS)

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

    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 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 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 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 puzzle of the Ne-E(L) component in LD graphite grains. This scenario is attractive for the SiC grains of type AB with 14N/15N 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 14N and 15N in the Galaxy, helping to produce the 14N/15N ratio in the solar system.

  1. Stereospecificity of (1) H, (13) C and (15) N shielding constants in the isomers of methylglyoxal bisdimethylhydrazone: problem with configurational assignment based on (1) H chemical shifts.

    PubMed

    Afonin, Andrei V; Pavlov, Dmitry V; Ushakov, Igor A; Keiko, Natalia A

    2012-07-01

    In the (13) C NMR spectra of methylglyoxal bisdimethylhydrazone, the (13) C-5 signal is shifted to higher frequencies, while the (13) C-6 signal is shifted to lower frequencies on going from the EE to ZE isomer following the trend found previously. Surprisingly, the (1) H-6 chemical shift and (1) J(C-6,H-6) coupling constant are noticeably larger in the ZE isomer than in the EE isomer, although the configuration around the -CH═N- bond does not change. This paradox can be rationalized by the C-H⋯N intramolecular hydrogen bond in the ZE isomer, which is found from the quantum-chemical calculations including Bader's quantum theory of atoms in molecules analysis. This hydrogen bond results in the increase of δ((1) H-6) and (1) J(C-6,H-6) parameters. The effect of the C-H⋯N hydrogen bond on the (1) H shielding and one-bond (13) C-(1) H coupling complicates the configurational assignment of the considered compound because of these spectral parameters. The (1) H, (13) C and (15) N chemical shifts of the 2- and 8-(CH(3) )(2) N groups attached to the -C(CH(3) )═N- and -CH═N- moieties, respectively, reveal pronounced difference. The ab initio calculations show that the 8-(CH(3) )(2) N group conjugate effectively with the π-framework, and the 2-(CH(3) )(2) N group twisted out from the plane of the backbone and loses conjugation. As a result, the degree of charge transfer from the N-2- and N-8- nitrogen lone pairs to the π-framework varies, which affects the (1) H, (13) C and (15) N shieldings. PMID:22615146

  2. Isotope ratio mass spectrometry: delta13C and delta15 N analysis for tracing the origin of illicit drugs.

    PubMed

    Galimov, E M; Sevastyanov, V S; Kulbachevskaya, E V; Golyavin, A A

    2005-01-01

    Gas chromatography/combustion/mass spectrometry (GC-C-MS) and elemental analysis/mass spectrometry (EA-MS) techniques are proposed to estimate delta(13)C and delta(15)N values in heroin, morphine, cocaine and hemp leaves, for the purposes of tracing the geographical origins of seized drugs. The values of isotope ratios for pure drugs and drugs with impurities were compared. It was demonstrated that large samples (up to 3 x 10(-6) g C) were combusted completely, so that the results obtained were valid. The data are considered to be an essential supplement to a wide-scale database designed specifically for the delta(13)C and delta(15)N values of drugs. The potential forensic and academic significance of the results is discussed. PMID:15832288

  3. Secondary structure and (1)H, (13)C, (15)N resonance assignments of the endosomal sorting protein sorting nexin 3.

    PubMed

    Overduin, Michael; Rajesh, Sandya; Gruenberg, Jean; Lenoir, Marc

    2015-10-01

    Sorting nexin 3 (SNX3) belongs to a sub-family of sorting nexins that primarily contain a single Phox homology domain capable of binding phosphoinositides and membranes. We report the complete (1)H, (13)C and (15)N resonance assignments of the full-length human SNX3 protein and identification of its secondary structure elements, revealing a canonical fold and unstructured termini. PMID:25893673

  4. Strategy for Enhancement of (13)C-Photo-CIDNP NMR Spectra by Exploiting Fractional (13)C-Labeling of Tryptophan.

    PubMed

    Eisenreich, Wolfgang; Joshi, Monika; Illarionov, Boris; Kacprzak, Sylwia; Lukaschek, Michail; Kothe, Gerd; Budisa, Nediljko; Fischer, Markus; Bacher, Adelbert; Weber, Stefan

    2015-10-29

    The photo-CIDNP effect has proven to be useful to strongly enhance NMR signals of photochemically active proteins simply by irradiation with light. The evolving characteristic patterns of enhanced absorptive and emissive NMR lines can be exploited to elucidate the photochemistry and photophysics of light-driven protein reactions. In particular, by the assignment of (13)C NMR resonances, redox-active amino acids may be identified and thereby electron-transfer pathways unraveled, in favorable cases, even with (13)C at natural abundance. If signal enhancement is weak, uniform (13)C isotope labeling is traditionally applied to increase the signal strength of protein (13)C NMR. However, this typically leads to cross relaxation, which transfers light-induced nuclear-spin polarization to adjacent (13)C nuclei, thereby preventing an unambiguous analysis of the photo-CIDNP effect. In this contribution, two isotope labeling strategies are presented; one leads to specific but ubiquitous (13)C labeling in tryptophan, and the other is based on fractional isotope labeling affording sets of isotopologs with low probability of next-neighbor isotope accumulation within individual tryptophan molecules. Consequently, cross relaxation is largely avoided while the signal enhancement by (13)C enrichment is preserved. This results in significantly simplified polarization patterns that are easier to analyze with respect to the generation of light-generated nuclear-spin polarization. PMID:26244593

  5. Tracing the diet of the monitor lizard Varanus mabitang by stable isotope analyses (δ15N, δ13C)

    NASA Astrophysics Data System (ADS)

    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.

  6. Individual protein balance strongly influences δ15N and δ13C values in Nile tilapia, Oreochromis niloticus

    NASA Astrophysics Data System (ADS)

    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.

  7. Seasonal δ13C and δ15N isoscapes of fish populations along a continental shelf trophic gradient

    NASA Astrophysics Data System (ADS)

    Radabaugh, Kara R.; Hollander, David J.; Peebles, Ernst B.

    2013-10-01

    The West Florida Shelf, located in the eastern Gulf of Mexico, transitions from a eutrophic ecosystem dominated by the Mississippi River plume to mesotrophic and oligotrophic ecosystems off the coast of peninsular Florida. Three extensive trawl surveys in this region were used to acquire samples of fish muscle, benthic algae from sea urchin stomach contents, and filtered particulate organic matter (POM) to create δ13C and δ15N isoscapes. Muscle δ15N from three widely distributed fish species, Synodus foetens (inshore lizardfish), Calamus proridens (littlehead porgy), and Syacium papillosum (dusky flounder), exhibited strong longitudinal correlations (Pearson‧s r=-0.67 to -0.90, p<0.001) that coincided with the principal trophic gradient, whereas δ13C values of fish muscle and benthic algae were correlated with depth (Pearson‧s r=-0.34 to -0.73, p<0.05). Correlations between isotopic values and surface concentrations of chlorophyll and particulate organic carbon (POC) imply linkages between the isotopic baseline and transitions from eutrophic to oligotrophic waters. The δ13C depth gradient and the δ15N longitudinal gradient were consistent between seasons and years, providing a foundation for future stable isotope studies of animal migration in the Gulf of Mexico.

  8. Retrospective characterization of ontogenetic shifts in killer whale diets via δ13C and δ15N analysis of teeth

    USGS Publications Warehouse

    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

  9. Retrospective characterization of ontogenetic shifts in killer whale diets via δ13C and δ15N analysis of teeth

    USGS Publications Warehouse

    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 ??13C and ??15N 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 ??15N 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 ??15N and ??13C values throughout the remainder of their lives, whereas ??15N values of most (n = 11) increased by ???1.5%, suggestive of an ontogenetic increase in trophic level. Significant differences in mean ??13C and ??15N 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. ?? Inter-Research 2009.

  10. Histidine side-chain dynamics and protonation monitored by 13C CPMG NMR relaxation dispersion.

    PubMed

    Hass, Mathias A S; Yilmaz, Ali; Christensen, Hans E M; Led, Jens J

    2009-08-01

    The use of 13C NMR relaxation dispersion experiments to monitor micro-millisecond fluctuations in the protonation states of histidine residues in proteins is investigated. To illustrate the approach, measurements on three specifically 13C labeled histidine residues in plastocyanin (PCu) from Anabaena variabilis (A.v.) are presented. Significant Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion is observed for 13C(epsilon1) nuclei in the histidine imidazole rings of A.v. PCu. The chemical shift changes obtained from the CPMG dispersion data are in good agreement with those obtained from the chemical shift titration experiments, and the CPMG derived exchange rates agree with those obtained previously from 15N backbone relaxation measurements. Compared to measurements of backbone nuclei, 13C(epsilon1) dispersion provides a more direct method to monitor interchanging protonation states or other kinds of conformational changes of histidine side chains or their environment. Advantages and shortcomings of using the 13C(epsilon1) dispersion experiments in combination with chemical shift titration experiments to obtain information on exchange dynamics of the histidine side chains are discussed. PMID:19533375

  11. Insight on RDX degradation mechanism by Rhodococcus strains using 13C and 15N kinetic isotope effects.

    PubMed

    Bernstein, Anat; Ronen, Zeev; Gelman, Faina

    2013-01-01

    The explosive Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is known to be degraded aerobically by various isolates of the Rhodococcus species, with denitration being the key step, mediated by Cytochrome P450. Our study aimed at gaining insight into the RDX degradation mechanism by Rhodococcus species and comparing isotope effects associated with RDX degradation by distinct Rhodococcus strains. For these purposes, enrichment in (13)C and (15)N isotopes throughout RDX denitration was studied for three distinct Rhodococcus strains, isolated from soil and groundwater in an RDX-contaminated site. The observable (15)N enrichment throughout the reaction, together with minor (13)C enrichment, suggests that N-N bond cleavage is likely to be the key rate-limiting step in the reaction. The similarity in the kinetic (15)N isotope effect between the three tested strains suggests that either isotope-masking effects are negligible, or are of a similar extent for all tested strains. The lack of variability in the kinetic (15)N isotope effect allows the interpretation of environmental studies with greater confidence. PMID:23215036

  12. Dipolar-coupling-mediated total correlation spectroscopy in solid-state 13C NMR: Selection of individual 13C- 13C dipolar interactions

    NASA Astrophysics Data System (ADS)

    Spano, Justin; Wi, Sungsool

    2010-06-01

    Herein is described a useful approach in solid-state NMR, for selecting homonuclear 13C- 13C spin pairs in a multiple- 13C homonuclear dipolar coupled spin system. This method builds upon the zero-quantum (ZQ) dipolar recoupling method introduced by Levitt and coworkers (Marin-Montesinos et al., 2006 [30]) by extending the originally introduced one-dimensional (1D) experiment into a two-dimensional (2D) method with selective irradiation scheme, while moving the 13C- 13C mixing scheme from the transverse to the longitudinal mode, together with a dramatic improvement in the proton decoupling efficiency. Selective spin-pair recoupling experiments incorporating Gaussian and cosine-modulated Gaussian pulses for inverting specific spins were performed, demonstrating the ability to detect informative, simplified/individualized, long-range 13C- 13C homonuclear dipolar coupling interactions more accurately by removing less informative, stronger, short-range 13C- 13C interactions from 2D correlation spectra. The capability of this new approach was demonstrated experimentally on uniformly 13C-labeled Glutamine and a tripeptide sample, GAL.

  13. Endogenous and environmental factors influence the dietary fractionation of 13C and 15N in hissing cockroaches Gromphadorhina portentosa.

    PubMed

    McCue, Marshall D

    2008-01-01

    Since DeNiro and Epstein's discovery that the (13)C and (15)N isotopic signatures of animals approximate those of their respective diets, the measurement of stable isotope signatures has become an important tool for ecologists studying the diets of wild animals. This study used Madagascar hissing cockroaches (Gromphadorhina portentosa) to examine several preexisting hypotheses about the relationship between the isotopic composition of an animal and its diet. Contrary to my predictions, the results revealed that the tissues of adult cockroaches raised for two generations on a diet of known isotopic composition did not demonstrate enrichment of heavy stable isotopes. Moreover, the (15)N signatures of cockroaches were neither influenced by periods of rapid growth (i.e., 300-fold increase in dry body mass over 120 d) nor by imposed periods of starvation lasting up to 80 d. The offspring born to mothers raised on known diets were enriched in (15)N. Diet-switching experiments showed that turnover times of (13)C were highly correlated with age and ranged from 9 to 10 d to 60 to 75 d in subadults and adults, respectively. Adults subjected to diet switches differed from the subadults in that the adults achieved equilibrated isotopic signatures that were shifted approximately 1.0 per thousand toward their respective original diets. Lipid fractions of adult cockroaches averaged 2.9 per thousand more depleted in (13)C than in lipid-free fractions, but no changes in (13)C were observed in aging adults. Exposure to reduced ambient temperature from 33 degrees C to 23 degrees C over 120 d did not influence isotopic signatures of tissues. Overall, the results of this study reveal that different endogenous and exogenous factors can influence the isotopic signatures of cockroaches. These findings reinforce the need to conduct controlled studies to further examine environmental factors that influence the relationships between the isotopic signatures of animals and their diets. PMID

  14. Biosynthetic preparation of L-(/sup 13/C)- and (/sup 15/N)glutamate by Brevibacterium flavum

    SciTech Connect

    Walker, T.E.; London, R.E.

    1987-01-01

    The biosynthesis of isotopically labeled L-glutamic acid by the microorganism Brevibacterium flavum was studied with a variety of carbon-13-enriched precursors. The purpose of this study was twofold: (i) to develop techniques for the efficient preparation of labeled L-glutamate with a variety of useful labeling patterns which can be used for other metabolic studies, and (ii) to better understand the metabolic events leading to label scrambling in these strains. B. flavum, which is used commercially for the production of monosodium glutamate, has the capability of utilizing glucose or acetate as a sole carbon source, and important criterion from the standpoint of developing labeling strategies. Unfortunately, singly labeled glucose precursors lead to excessive isotopic dilution which reduces their usefulness. Studies with (3-/sup 13/C)pyruvate indicate that this problem can in principle be overcome by using labeled three-carbon precursors; however, conditions could not be found which would lead to an acceptable yield of isotopically labeled L-glutamate. In contrast, (1-/sup 13/C)- or (2-/sup 13/C)acetate provides relatively inexpensive, readily available precursors for the production of selectively labeled, high enriched L-glutamate. The preparation of L-(/sup 15/N)glutamate from (/sup 15/N)ammonium sulfate was carried out and is a very effective labeling strategy. Analysis of the isotopic distribution in labeled glutamate provides details about the metabolic pathways in these interesting organisms.

  15. 13C and 15N fractionation of CH4/N2 mixtures during photochemical aerosol formation: Relevance to Titan

    NASA Astrophysics Data System (ADS)

    Sebree, Joshua A.; Stern, Jennifer C.; Mandt, Kathleen E.; Domagal-Goldman, Shawn D.; Trainer, Melissa G.

    2016-05-01

    The ratios of the stable isotopes that comprise each chemical species in Titan's atmosphere provide critical information towards understanding the processes taking place within its modern and ancient atmosphere. Several stable isotope pairs, including 12C/13C and 14N/15N, have been measured in situ or probed spectroscopically by Cassini-borne instruments, space telescopes, or through ground-based observations. Current attempts to model the observed isotope ratios incorporate fractionation resulting from atmospheric diffusion, hydrodynamic escape, and primary photochemical processes. However, the effect of a potentially critical pathway for isotopic fractionation - organic aerosol formation and subsequent deposition onto the surface of Titan - has not been considered due to insufficient data regarding fractionation during aerosol formation. To better understand the nature of this process, we have conducted a laboratory study to measure the isotopic fractionation associated with the formation of Titan aerosol analogs, commonly referred to as 'tholins', via far-UV irradiation of several methane (CH4) and dinitrogen (N2) mixtures. Analysis of the δ13C and δ15N isotopic signatures of the photochemical aerosol products using an isotope ratio mass spectrometer (IRMS) show that fractionation direction and magnitude are dependent on the initial bulk composition of the gas mixture. In general, the aerosols showed enrichment in 13C and 14N, and the observed fractionation trends can provide insight into the chemical mechanisms controlling photochemical aerosol formation.

  16. Validating the Incorporation of 13C and 15N in a Shorebird That Consumes an Isotopically Distinct Chemosymbiotic Bivalve

    PubMed Central

    van Gils, Jan A.; Ahmedou Salem, Mohamed Vall

    2015-01-01

    The wealth of field studies using stable isotopes to make inferences about animal diets require controlled validation experiments to make proper interpretations. Despite several pleas in the literature for such experiments, validation studies are still lagging behind, notably in consumers dwelling in chemosynthesis-based ecosystems. In this paper we present such a validation experiment for the incorporation of 13C and 15N in the blood plasma of a medium-sized shorebird, the red knot (Calidris canutus canutus), consuming a chemosymbiotic lucinid bivalve (Loripes lucinalis). Because this bivalve forms a symbiosis with chemoautotrophic sulphide-oxidizing bacteria living inside its gill, the bivalve is isotopically distinct from ‘normal’ bivalves whose food has a photosynthetic basis. Here we experimentally tested the hypothesis that isotope discrimination and incorporation dynamics are different when consuming such chemosynthesis-based prey. The experiment showed that neither the isotopic discrimination factor, nor isotopic turnover time, differed between birds consuming the chemosymbiotic lucinid and a control group consuming a photosynthesis-based bivalve. This was true for 13C as well as for 15N. However, in both groups the 15N discrimination factor was much higher than expected, which probably had to do with the birds losing body mass over the course of the experiment. PMID:26458005

  17. Validating the Incorporation of 13C and 15N in a Shorebird That Consumes an Isotopically Distinct Chemosymbiotic Bivalve.

    PubMed

    van Gils, Jan A; Ahmedou Salem, Mohamed Vall

    2015-01-01

    The wealth of field studies using stable isotopes to make inferences about animal diets require controlled validation experiments to make proper interpretations. Despite several pleas in the literature for such experiments, validation studies are still lagging behind, notably in consumers dwelling in chemosynthesis-based ecosystems. In this paper we present such a validation experiment for the incorporation of 13C and 15N in the blood plasma of a medium-sized shorebird, the red knot (Calidris canutus canutus), consuming a chemosymbiotic lucinid bivalve (Loripes lucinalis). Because this bivalve forms a symbiosis with chemoautotrophic sulphide-oxidizing bacteria living inside its gill, the bivalve is isotopically distinct from 'normal' bivalves whose food has a photosynthetic basis. Here we experimentally tested the hypothesis that isotope discrimination and incorporation dynamics are different when consuming such chemosynthesis-based prey. The experiment showed that neither the isotopic discrimination factor, nor isotopic turnover time, differed between birds consuming the chemosymbiotic lucinid and a control group consuming a photosynthesis-based bivalve. This was true for 13C as well as for 15N. However, in both groups the 15N discrimination factor was much higher than expected, which probably had to do with the birds losing body mass over the course of the experiment. PMID:26458005

  18. Theoretical and experimental study of 15N NMR protonation shifts.

    PubMed

    Semenov, Valentin A; Samultsev, Dmitry O; Krivdin, Leonid B

    2015-06-01

    A combined theoretical and experimental study revealed that the nature of the upfield (shielding) protonation effect in 15N NMR originates in the change of the contribution of the sp(2)-hybridized nitrogen lone pair on protonation resulting in a marked shielding of nitrogen of about 100 ppm. On the contrary, for amine-type nitrogen, protonation of the nitrogen lone pair results in the deshielding protonation effect of about 25 ppm, so that the total deshielding protonation effect of about 10 ppm is due to the interplay of the contributions of adjacent natural bond orbitals. A versatile computational scheme for the calculation of 15N NMR chemical shifts of protonated nitrogen species and their neutral precursors is proposed at the density functional theory level taking into account solvent effects within the supermolecule solvation model. PMID:25891386

  19. The Titan 14N/ 15N and 12C/ 13C isotopic ratios in HCN from Cassini/CIRS

    NASA Astrophysics Data System (ADS)

    Vinatier, Sandrine; Bézard, Bruno; Nixon, Conor A.

    2007-11-01

    We report the detection of H 13CN and HC 15N in mid-infrared spectra recorded by the Composite Infrared Spectrometer (CIRS) aboard Cassini, along with the determination of the 12C/ 13C and 14N/ 15N isotopic ratios. We analyzed two sets of limb spectra recorded near 13-15° S (Tb flyby) and 83° N (T4 flyby) at 0.5 cm -1 resolution. The spectral range 1210-1310 cm -1 was used to retrieve the temperature profile in the range 145-490 km at 13° S and 165-300 km at 83° N. These two temperature profiles were then incorporated in the atmospheric model to retrieve the abundance profile of H 12C 14N, H 13CN and HC 15N from their bands at 713, 706 and 711 cm -1, respectively. The HCN abundance profile was retrieved in the range 90-460 km at 15° S and 165-305 km at 83° N. There is no evidence for vertical variations of the isotopic ratios. Constraining the isotopic abundance profiles to be proportional to the HCN one, we find C12/C13=89-18+22 at 15° S, and 68-12+16 at 83° N, two values that are statistically consistent. A combination of these results yields a 12C/ 13C value equal to 75±12. This global result, as well as the 15° S one, envelop the value in Titan's methane ( 82.3±1) [Niemann, H.B., and 17 colleagues, 2005. Nature 438, 779-784] measured at 10° S and is slightly lower than the terrestrial inorganic standard value (89). The 14N/ 15N isotopic ratio is found equal to 56-13+16 at 15° S and 56-9+10 at 83° N. Combining the two values yields 14N/ 15N = 56 ± 8, which corresponds to an enrichment in 15N of about 4.9 compared with the terrestrial ratio. These results agree with the values obtained from previous ground-based millimeter observations [Hidayat, T., Marten, A., Bézard, B., Gautier, D., Owen, T., Matthews, H.E., Paubert, G., 1997. Icarus 126, 170-182; Marten, A., Hidayat, T., Biraud, Y., Moreno, R., 2002. Icarus 158, 532-544]. The 15N/ 14N ratio found in HCN is ˜3 times higher than in N 2 [Niemann, H.B., and 17 colleagues, 2005. Nature 438, 779

  20. Improved mass analysis of oligoribonucleotides by 13C, 15N double depletion and electrospray ionization FT-ICR mass spectrometry.

    PubMed

    Xiong, Ying; Schroeder, Kersten; Greenbaum, Nancy L; Hendrickson, Christopher L; Marshall, Alan G

    2004-03-15

    13C, 15N doubly depleted 32-ribonucleotide was synthesized enzymatically by in vitro transcription from nucleoside triphosphates isolated from E. coli grown in a minimal medium containing 12C, 14N-enriched glucose and ammonium sulfate. Following purification and desalting by reversed-phase HPLC, buffer exchange with Microcon YM-3, and ethanol precipitation, electrospray ionization Fourier transform ion cyclotron resonance mass spectra revealed greatly enhanced abundance of monoisotopic ions (by a factor of approximately 100) and a narrower isotopic distribution with higher signal-to-noise ratio. The abrupt onset and high magnitude of the monoisotopic species promise to facilitate accurate mass measurement of RNA's. PMID:15018587

  1. Stable isotope-enhanced two- and three-dimensional diffusion ordered 13C NMR spectroscopy (SIE-DOSY 13C NMR)

    NASA Astrophysics Data System (ADS)

    Vermillion, Karl; Price, Neil P. J.

    2009-06-01

    The feasibility of obtaining high quality homonuclear or heteronuclear diffusion-ordered 13C NMR data is shown to be greatly improved by using 13C isotopically-enriched samples. Stable isotope-enhanced diffusion ordered (SIE-DOSY) 13C NMR has been applied to 13C-enriched carbohydrates, and has been used to determine diffusion coefficients for pentose and hexose monosaccharides, and a disaccharide and trisaccharide. These 2D spectra were obtained with as little as 8 min of acquisition time. Fully resolved 3D DOSY-HMQC NMR spectra of [U- 13C]xylose, [U- 13C]glucose, and [1- 13C gal]lactose were obtained in 5 h. Sample derivatization with [ carbonyl- 13C]acetate (peracetylation) extends the usefulness of the technique to included non-labeled sugars; the 13C-carbonyl - carbohydrate ring proton 1H- 13C correlations also provide additional structural information, as shown for the 3-D DOSY-HMQC analysis of a mixture of maltotriose and lactose per-[ carbonyl- 13C]acetates.

  2. Evaluating mercury biomagnification in fish from a tropical marine environment using stable isotopes (delta13C and delta15N).

    PubMed

    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. PMID:17702328

  3. Stable isotope analysis (δ13C and δ15N) of soil nematodes from four feeding groups

    PubMed Central

    Griffiths, Bryan; Dyckmans, Jens; Schmidt, Olaf

    2016-01-01

    Soil nematode feeding groups are a long-established trophic categorisation largely based on morphology and are used in ecological indices to monitor and analyse the biological state of soils. Stable isotope ratio analysis (13C/12C and 15N/14N, expressed as δ13C and δ15N) has provided verification of, and novel insights into, the feeding ecology of soil animals such as earthworms and mites. However, isotopic studies of soil nematodes have been limited to date as conventional stable isotope ratio analysis needs impractically large numbers of nematodes (up to 1,000) to achieve required minimum sample weights (typically >100 µg C and N). Here, micro-sample near-conventional elemental analysis–isotopic ratio mass spectrometry (μEA–IRMS) of C and N using microgram samples (typically 20 µg dry weight), was employed to compare the trophic position of selected soil nematode taxa from four feeding groups: predators (Anatonchus and Mononchus), bacterial feeders (Plectus and Rhabditis), omnivores (Aporcelaimidae and Qudsianematidae) and plant feeder (Rotylenchus). Free-living nematodes were collected from conventionally and organically managed arable soils. As few as 15 nematodes, for omnivores and predators, were sufficient to reach the 20 µg dry weight target. There was no significant difference in δ15N (p = 0.290) or δ13C (p = 0.706) between conventional and organic agronomic treatments but, within treatments, there was a significant difference in N and C stable isotope ratios between the plant feeder, Rotylenchus (δ15N = 1.08 to 3.22 mUr‰, δ13C = –29.58 to –27.87 mUr) and all other groups. There was an average difference of 9.62 mUr in δ15N between the plant feeder and the predator group (δ15N = 9.89 to 12.79 mUr, δ13C = –27.04 to –25.51 mUr). Isotopic niche widths were calculated as Bayesian derived standard ellipse areas and were smallest for the plant feeder (1.37 mUr2) and the predators (1.73 mUr2), but largest for omnivores (3.83 mUr2

  4. 1H, 15N and 13C resonance assignments of light organ-associated fatty acid-binding protein of Taiwanese fireflies.

    PubMed

    Tseng, Kai-Li; Lee, Yi-Zong; Chen, Yun-Ru; Lyu, Ping-Chiang

    2016-04-01

    Fatty acid-binding proteins (FABPs) are a family of proteins that modulate the transfer of various fatty acids in the cytosol and constitute a significant portion in many energy-consuming cells. The ligand binding properties and specific functions of a particular type of FABP seem to be diverse and depend on the respective binding cavity as well as the cell type from which this protein is derived. Previously, a novel FABP (lcFABP; lc: Luciola cerata) was identified in the light organ of Taiwanese fireflies. The lcFABP was proved to possess fatty acids binding capabilities, especially for fatty acids of length C14-C18. However, the structural details are unknown, and the structure-function relationship has remained to be further investigated. In this study, we finished the (1)H, (15)N and (13)C chemical shift assignments of (15)N/(13)C-enriched lcFABP by solution NMR spectroscopy. In addition, the secondary structure distribution was revealed based on the backbone N, H, Cα, Hα, C and side chain Cβ assignments. These results can provide the basis for further structural exploration of lcFABP. PMID:26373428

  5. δ13C and δ15N values in scales of Micropterus salmoides largemouth bass as a freshwater environmental indicator.

    PubMed

    Inamura, O; Zhang, J; Minagawa, M

    2012-01-15

    We have investigated the effectiveness of using the Micropterus salmoides largemouth bass, which is a top predator found throughout the world, as the index of a hydrosphere environment and its food chain. To this end, we used stable carbon and nitrogen isotope analysis (SIA). Largemouth bass were collected from eight dam reservoirs and two ponds in Toyama Prefecture, Japan. Toyama is located in central Japan and features a variety of distinct geographical environments, a result of the 3000-m elevation that changes over short distances, and abundant water systems. The mean δ(13)C and δ(15)N values for the lipid-extracted muscle of largemouth bass from all sampling locations showed large variability, but there were only small standard deviations at each sampling location. The isotope ratios for largemouth bass express the characteristics of each investigated hydrosphere environment and food chain. A very high correlation (δ(13)C: Y(scale) = 0.96 X(muscle) + 1.58, R(2) = 0.98, δ(15)N: Y(scale) = 0.92 X(muscle) - 1.15, R(2) = 0.95) of SIA values was found between largemouth bass scales and lipid-extracted muscles, which suggests that the more easily analyzed scales are useful as SIA samples for the monitoring and comparison of hydrosphere environments throughout the world. PMID:22215573

  6. Characterising ontogenetic niche shifts in Nile crocodile using stable isotope (δ13C, δ15N) analyses of scute keratin.

    PubMed

    Radloff, Frans G T; Hobson, Keith A; Leslie, Alison J

    2012-09-01

    Nile crocodiles undergo a three to five order of magnitude increase in body size during their lifespan. This shift coincides with a change in resource and habitat use which influences the strength, type and symmetry of interactions with other species. Identifying size-specific crocodile groups displaying similar traits is important for conservation planning. Here, we illustrate how stable carbon (δ(13) C) and nitrogen (δ(15) N) isotope analysis of scute keratin, together with breakpoint modelling analysis can be used to characterise ontogenetic niche shifts. Using a sample set of 238 crocodiles from the Okavango Delta, Botswana (35-463 cm total length), we found prominent size-related changes in the scute keratin δ(13) C and δ(15) N profiles close to 40 and 119 cm snout-vent length. The first shift corroborated the findings of a traditional stomach-content study conducted on the same population at the same time, and the second conformed to known crocodile ecology. This approach can be used as a first approximation to identify size-specific groups within crocodile populations, and these can then be investigated further using isotopic or other methods. PMID:22462522

  7. Comparison of the solution and crystal structures of staphylococcal nuclease with /sup 13/C and /sup 15/N chemical shifts used as structural fingerprints

    SciTech Connect

    Cole, H.B.R.; Sparks, S.W.; Torchia, D.A.

    1988-09-01

    The authors report high-resolution /sup 13/C and /sup 15/N NMR spectra of crystalline staphylococcal nuclease (Nase) complexed to thymidine 3',5'-diphosphate and Ca/sup 2+/. High sensitivity and resolution are obtained by applying solid-state NMR techniques-high power proton decoupling and cross-polarization magic angle sample spinning (CPMASS)-to protein samples that have been efficiently synthesized and labeled by an overproducing strain of Escherichia coli. A comparison of CPMASS and solution spectra of Nase labeled with either (methyl-/sup 13/C)methionine or (/sup 15/)valine shows that the chemical shifts in the crystalline and solution states are virtually identical. This result is strong evidence that the protein conformations in the solution and crystalline states are nearly the same. Because of the close correspondence of the crystal and solution chemical shifts, sequential assignments obtained in solution apply to the crystal spectra. It should therefore be possible to study the molecular structure and dynamics of many sequentially assigned atomic sites in Nase crystals. Similar experiments are applicable to the growing number of proteins that can be obtained from efficient expression systems.

  8. Stable isotope-enhanced two- and three-dimensional diffusion ordered 13C-NMR spectroscopy (SIE-DOSY 13C-NMR)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stable Isotope-Enhanced Diffusion Ordered (SIE-DOSY) 13C-NMR has been applied to 13C-enriched carbohydrates and has been used to determine diffusion coefficients for pentose and hexose monosaccharides, a disaccharide and a trisaccharide. These 2D spectra were obtained with as little as 8 min of acq...

  9. 1H, 13C, and 15N resonance assignments for the protein coded by gene locus BB0938 of Bordetella bronchiseptica

    SciTech Connect

    Rossi, Paolo; Ramelot, Theresa A.; Xiao, Rong; Ho, Chi K.; Ma, LiChung; Acton, Thomas; Kennedy, Michael A.; Montelione, Gaetano

    2005-11-01

    The product of gene locus BB0938 from Bordetella bronchiseptica (Swiss-Prot ID: Q7WNU7-BORBR; NESG target ID: BoR11; Wunderlich et al., 2004; Pfam ID: PF03476) is a 128-residue protein of unknown function. This broadly conserved protein family is found in eubacteria and eukaryotes. Using triple resonance NMR techniques, we have determined 98% of backbone and 94% of side chain 1H, 13C, and 15N resonance assignments. The chemical shift and 3J(HN?Ha) scalar coupling data reveal a b topology with a seven-residue helical insert, ??????????. BMRB deposit with accession number 6693. Reference: Wunderlich et al. (2004) Proteins, 56, 181?187.

  10. 1H, 13C, and 15N resonance assignments for Escherichia coli ytfP, a member of the broadly conserved UPF0131 protein domain family

    SciTech Connect

    Aramini, James M.; Swapna, G.V.T.; Huang, Yuanpeng; Rajan, Paranji K.; Xiao, Rong; Shastry, Ritu; Acton, Thomas; Cort, John R.; Kennedy, Michael A.; Montelione, Gaetano

    2005-11-01

    Protein ytfP from Escherichia coli (Swiss-Prot ID: YTFP-ECOLI; NESG target ID: ER111; Wunderlich et al., 2004) is a 113-residue member of the UPF0131 protein family (Pfam ID: PF03674) of unknown function. This domain family is found in organisms from all three kingdoms, archaea, eubacteria and eukaryotes. Using triple resonance NMR techniques, we have determined 97% of backbone and 91% of side chain 1H, 13C, and 15N resonance assignments. The chemical shift and 3J(HN?Ha) scalar coupling data reveal a mixed a/b topology,????????. BMRB deposit with Accession No. 6448. Reference: Wunderlich et al. (2004) Proteins, 56, 181?187.

  11. 15N NMR chemical shifts in papaverine decomposition products

    NASA Astrophysics Data System (ADS)

    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.

  12. (13)C NMR assignments of regenerated cellulose from solid-state 2D NMR spectroscopy.

    PubMed

    Idström, Alexander; Schantz, Staffan; Sundberg, Johan; Chmelka, Bradley F; Gatenholm, Paul; Nordstierna, Lars

    2016-10-20

    From the assignment of the solid-state (13)C NMR signals in the C4 region, distinct types of crystalline cellulose, cellulose at crystalline surfaces, and disordered cellulose can be identified and quantified. For regenerated cellulose, complete (13)C assignments of the other carbon regions have not previously been attainable, due to signal overlap. In this study, two-dimensional (2D) NMR correlation methods were used to resolve and assign (13)C signals for all carbon atoms in regenerated cellulose. (13)C-enriched bacterial nanocellulose was biosynthesized, dissolved, and coagulated as highly crystalline cellulose II. Specifically, four distinct (13)C signals were observed corresponding to conformationally different anhydroglucose units: two signals assigned to crystalline moieties and two signals assigned to non-crystalline species. The C1, C4 and C6 regions for cellulose II were fully examined by global spectral deconvolution, which yielded qualitative trends of the relative populations of the different cellulose moieties, as a function of wetting and drying treatments. PMID:27474592

  13. Multiple regression models of δ13C and δ15N for fish populations in the eastern Gulf of Mexico

    NASA Astrophysics Data System (ADS)

    Radabaugh, Kara R.; Peebles, Ernst B.

    2014-08-01

    Multiple regression models were created to explain spatial and temporal variation in the δ13C and δ15N values of fish populations on the West Florida Shelf (eastern Gulf of Mexico, USA). Extensive trawl surveys from three time periods were used to acquire muscle samples from seven groundfish species. Isotopic variation (δ13Cvar and δ15Nvar) was calculated as the deviation from the isotopic mean of each fish species. Static spatial data and dynamic water quality parameters were used to create models predicting δ13Cvar and δ15Nvar in three fish species that were caught in the summers of 2009 and 2010. Additional data sets were then used to determine the accuracy of the models for predicting isotopic variation (1) in a different time period (fall 2010) and (2) among four entirely different fish species that were collected during summer 2009. The δ15Nvar model was relatively stable and could be applied to different time periods and species with similar accuracy (mean absolute errors 0.31-0.33‰). The δ13Cvar model had a lower predictive capability and mean absolute errors ranged from 0.42 to 0.48‰. δ15N trends are likely linked to gradients in nitrogen fixation and Mississippi River influence on the West Florida Shelf, while δ13C trends may be linked to changes in algal species, photosynthetic fractionation, and abundance of benthic vs. planktonic basal resources. These models of isotopic variability may be useful for future stable isotope investigations of trophic level, basal resource use, and animal migration on the West Florida Shelf.

  14. Early-stage changes in natural (13)C and (15)N abundance and nutrient dynamics during different litter decomposition.

    PubMed

    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. PMID:26915037

  15. Performance evaluation of quantitative adiabatic (13)C NMR pulse sequences for site-specific isotopic measurements.

    PubMed

    Thibaudeau, Christophe; Remaud, Gérald; Silvestre, Virginie; Akoka, Serge

    2010-07-01

    (2)H/(1)H and (13)C/(12)C site-specific isotope ratios determined by NMR spectroscopy may be used to discriminate pharmaceutically active ingredients based on the synthetic process used in production. Extending the Site-specific Natural Isotope Fractionation NMR (SNIF-NMR) method to (13)C is highly beneficial for complex organic molecules when measurements of (2)H/(1)H ratios lead to poorly defined molecular fingerprints. The current NMR methodology to determine (13)C/(12)C site-specific isotope ratios suffers from poor sensitivity and long experimental times. In this work, several NMR pulse sequences based on polarization transfer were evaluated and optimized to measure precise quantitative (13)C NMR spectra within a short time. Adiabatic 180 degrees (1)H and (13)C pulses were incorporated into distortionless enhancement by polarization transfer (DEPT) and refocused insensitive nuclei enhanced by polarization transfer (INEPT) to minimize the influence of 180 degrees pulse imperfections and of off-resonance effects on the precision of the measured (13)C peak areas. The adiabatic DEPT sequence was applied to draw up a precise site-specific (13)C isotope profile of ibuprofen. A modified heteronuclear cross-polarization (HCP) experiment featuring (1)H and (13)C spin-locks with adiabatic 180 degrees pulses is also introduced. This sequence enables efficient magnetization transfer across a wide (13)C frequency range although not enough for an application in quantitative (13)C isotopic analysis. PMID:20527737

  16. A highly sensitive assay for xanthine oxidoreductase activity using a combination of [(13) C2 ,(15) N2 ]xanthine and liquid chromatography/triple quadrupole mass spectrometry.

    PubMed

    Murase, Takayo; Oka, Mitsuru; Nampei, Mai; Miyachi, Atsushi; Nakamura, Takashi

    2016-05-15

    In this study, we developed a highly sensitive assay for xanthine oxidoreductase (XOR) activity utilizing a combination of [(13) C2 ,(15) N2 ]xanthine and liquid chromatography (LC)/triple quadrupole mass spectrometry (TQMS). In this assay, the amount of [(13) C2 ,(15) N2 ]uric acid (UA) produced by XOR was determined by using LC/TQMS. For this assay, we synthesized [(13) C2 ,(15) N2 ]xanthine as a substrate, [(13) C2 ,(15) N2 ]UA as an analytical standard, and [(13) C3 ,(15) N3 ]UA as an internal standard. The [(13) C2 ,(15) N2 ]UA calibration curve obtained using LC/TQMS under the selected reaction monitoring mode was evaluated, and the results indicated good linearity (R(2)  = 0.998, weighting of 1/x(2) ) in the range of 20 to 4000 nM. As a model reaction of less active samples, the XOR activity of serial-diluted mouse plasma was measured. Thereby, the XOR activity of the 1024-fold-diluted mouse plasma was 4.49 ± 0.44 pmol/100 μL/h (mean ± standard deviation, n = 3). This value is comparable to the predicted XOR activity value of healthy human plasma. Hence, this combination method may be used to obtain high-sensitivity measurements required for XOR activity analysis on various organs or human plasma. PMID:27006202

  17. 1H, 13C and 15N nuclear magnetic resonance coordination shifts in Au(III), Pd(II) and Pt(II) chloride complexes with phenylpyridines.

    PubMed

    Pazderski, Leszek; Tousek, Jaromír; Sitkowski, Jerzy; Kozerski, Lech; Szłyk, Edward

    2009-08-01

    1H, 13C and 15N nuclear magnetic resonance studies of gold(III), palladium(II) and platinum(II) chloride complexes with phenylpyridines (PPY: 4-phenylpyridine, 4ppy; 3-phenylpyridine, 3ppy; and 2-phenylpyridine, 2ppy) having the general formulae [Au(PPY)Cl3], trans-/cis-[Pd(PPY)2Cl2] and trans-/cis-[Pt(PPY)2Cl2] were performed and the respective chemical shifts (delta1H, delta13C and delta15N) reported. 1H, 13C and 15N coordination shifts (i.e. differences between chemical shifts of the same atom in the complex and ligand molecules: Delta(coord)(1H) = delta(complex)(1H)-delta(ligand)(1H), Delta(coord)(13C) = delta(complex)(13C)-delta(ligand)(13C), Delta(coord)(15N) = delta(complex)(15N)-delta(ligand)(15N)) were discussed in relation to the type of the central atom (Au(III), Pd(II) and Pt(II)), geometry (trans-/cis-) and the position of a phenyl group in the pyridine ring system. PMID:19472306

  18. Rotary resonance recoupling of 13C- 1H dipolar interactions in magic angle spinning 13C NMR of dynamic solids

    NASA Astrophysics Data System (ADS)

    Kitchin, Simon J.; Harris, Kenneth D. M.; Aliev, Abil E.; Apperley, David C.

    2000-06-01

    Rotary resonance recoupling of heteronuclear 13C- 1H dipolar interactions in magic angle spinning solid state 13C NMR spectra (recorded under conditions of 1H decoupling at frequency ν1 and magic angle spinning at frequency νr) has been studied for three examples of molecular solids (adamantane, ferrocene and hexamethylbenzene) in which substantial molecular motion is known to occur. It is shown that when rotary resonance conditions are satisfied (i.e. ν1/νr= n, for n=1 or 2), the recoupling can lead to motionally averaged Pake-like powder patterns from which information on 13C- 1H internuclear distances and/or molecular motion can be derived.

  19. Backbone dynamics of barstar: a (15)N NMR relaxation study.

    PubMed

    Sahu, S C; Bhuyan, A K; Majumdar, A; Udgaonkar, J B

    2000-12-01

    Backbone dynamics of uniformly (15)N-labeled barstar have been studied at 32 degrees C, pH 6.7, by using (15)N relaxation data obtained from proton-detected 2D (1)H-(15)N NMR spectroscopy. (15)N spin-lattice relaxation rate constants (R(1)), spin-spin relaxation rate constants (R(2)), and steady-state heteronuclear (1)H-(15)N NOEs have been determined for 69 of the 86 (excluding two prolines and the N-terminal residue) backbone amide (15)N at a magnetic field strength of 14.1 Tesla. The primary relaxation data have been analyzed by using the model-free formalism of molecular dynamics, using both isotropic and axially symmetric diffusion of the molecule, to determine the overall rotational correlation time (tau(m)), the generalized order parameter (S(2)), the effective correlation time for internal motions (tau(e)), and NH exchange broadening contributions (R(ex)) for each residue. As per the axially symmetric diffusion, the ratio of diffusion rates about the unique and perpendicular axes (D( parallel)/D( perpendicular)) is 0.82 +/- 0.03. The two results have only marginal differences. The relaxation data have also been used to map reduced spectral densities for the NH vectors of these residues at three frequencies: 0, omega(H), and omega(N), where omega(H),(N) are proton and nitrogen Larmor frequencies. The value of tau(m) obtained from model-free analysis of the relaxation data is 5.2 ns. The reduced spectral density analysis, however, yields a value of 5.7 ns. The tau(m) determined here is different from that calculated previously from time-resolved fluorescence data (4.1 ns). The order parameter ranges from 0.68 to 0.98, with an average value of 0.85 +/- 0.02. A comparison of the order parameters with the X-ray B-factors for the backbone nitrogens of wild-type barstar does not show any considerable correlation. Model-free analysis of the relaxation data for seven residues required the inclusion of an exchange broadening term, the magnitude of which ranges from 2

  20. Comparing isotope signatures of prey fish: does gut removal affect δ13C or δ15N?

    USGS Publications Warehouse

    Chipps, Steven R.; Fincel, Mark J.; VanDeHey, Justin A.; Wuestewald, Andrew

    2011-01-01

    Stable isotope analysis is a quick and inexpensive method to monitor the effects of food web changes on aquatic communities. Traditionally, whole specimens have been used when determining isotope composition of prey fish or age-0 recreational fishes. However, gut contents of prey fish could potentially alter isotope composition of the specimen, especially when recent foraging has taken place or when the gut contains non-assimilated material that would normally pass through fishes undigested. To assess the impacts of gut content on prey fish isotope signatures, we examined the differences in isotopic variation of five prey fish species using whole fish, whole fish with the gut contents removed, and dorsal muscle only. We found significant differences in both δ15N and δ13C between the three tissue treatments. In most cases, muscle tissue was enriched compared to whole specimens or gut-removed specimens. Moreover, differences in mean δ15N within a species were up to 2‰ among treatments. This would result in a change of over half a trophic position (TP) based on a 3.4‰ increase per trophic level. However, there were no apparent relationships between tissue isotope values in fish with increased gut fullness (more prey tissue present). We suggest that muscle tissue should be used as the standard tissue for determining isotope composition of prey fish or age-0 recreational fishes, especially when determining enrichment for mixing models, calculating TP, or constructing aquatic food webs.

  1. Mangrove isotopic (δ15N and δ13C) fractionation across a nitrogen vs. phosphorus limitation gradient

    USGS Publications Warehouse

    Mckee, Karen L.; Feller, Ilka C.; Popp, Marianne; Wanek, Wolfgang

    2002-01-01

    Mangrove islands in Belize are characterized by a unique switching from nitrogen (N) to phosphorus (P) limitation to tree growth from shoreline to interior. Fertilization has previously shown that Rhizophora mangle (red mangrove) fringe trees (5–6 m tall) growing along the shoreline are N limited; dwarf trees (!1.5 m tall) in the forestinterior are P limited; and transition trees (2–4 m tall) are co-limited by both N and P.  Growth patterns paralleled a landward decrease in soil flushing by tides and an increase in bioavailable N, but P availability remained consistently low across the gradient. Stable isotopic composition was measured in R. mangle leaves to aid in explaining this nutrient switching pattern and growth variation. Along control transects, leaf !15N decreased from "0.10‰ (fringe) to #5.38‰ (dwarf). The !15N of N-fertilized trees also varied spatially, but the values were consistently more negative (by $3‰) compared to control trees. Spatial variation in !15N values disappeared when the trees were fertilized with P, and values averaged "0.12‰, similar to that in control fringe trees. Neither variation in source inputs nor microbial fractionation could fully account for the observed patterns in !15N. The results instead suggest that the lower !15N values in transition and dwarf control trees were due to plant fractionation as a consequence of slower growth and lower N demand. P fertilization increased N demand and decreased fractionation. Although leaf !13C was unaffected by fertilization, values increased from fringe (#28.6‰) to transition (#27.9‰) to dwarf (#26.4‰) zones, indicating spatial variation in environmental stresses affecting stomatal conductance or carboxylation. The results thus suggest an interaction of external supply, internal demand, and plant ability to acquire nutrients under different hydro-edaphic conditions that vary across this tree-height gradient. The findings not only aid in understanding

  2. Near-complete 1H, 13C, 15N resonance assignments of dimethylsulfoxide-denatured TGFBIp FAS1-4 A546T.

    PubMed

    Kulminskaya, Natalia V; Yoshimura, Yuichi; Runager, Kasper; Sørensen, Charlotte S; Bjerring, Morten; Andreasen, Maria; Otzen, Daniel E; Enghild, Jan J; Nielsen, Niels Chr; Mulder, Frans A A

    2016-04-01

    The transforming growth factor beta induced protein (TGFBIp) is a major protein component of the human cornea. Mutations occurring in TGFBIp may cause corneal dystrophies, which ultimately lead to loss of vision. The majority of the disease-causing mutations are located in the C-terminal domain of TGFBIp, referred as the fourth fascilin-1 (FAS1-4) domain. In the present study the FAS1-4 Ala546Thr, a mutation that causes lattice corneal dystrophy, was investigated in dimethylsulfoxide using liquid-state NMR spectroscopy, to enable H/D exchange strategies for identification of the core formed in mature fibrils. Isotope-labeled fibrillated FAS1-4 A546T was dissolved in a ternary mixture 95/4/1 v/v/v% dimethylsulfoxide/water/trifluoroacetic acid, to obtain and assign a reference 2D (1)H-(15)N HSQC spectrum for the H/D exchange analysis. Here, we report the near-complete assignments of backbone and aliphatic side chain (1)H, (13)C and (15)N resonances for unfolded FAS1-4 A546T at 25 °C. PMID:26275916

  3. 1H, 13C and 15N resonance assignments and secondary structure analysis of CmPI-II, a serine protease inhibitor isolated from marine snail Cenchritis muricatus.

    PubMed

    Cabrera-Muñoz, Aymara; Rojas, Laritza; Alonso-del-Rivero Antigua, Maday; Pires, José Ricardo

    2016-04-01

    A protease inhibitor (CmPI-II) (UNIPROT: IPK2_CENMR) from the marine mollusc Cenchritis muricatus, has been isolated and characterized. It is the first member of a new group (group 3) of non-classical Kazal-type inhibitors. CmPI-II is a tight-binding inhibitor of serine proteases: trypsin, human neutrophil elastase (HNE), subtilisin A and pancreatic elastase. This specificity is exceptional in the members of Kazal-type inhibitor family. Several models of three-dimensional structure of CmPI-II have been constructed by homology with other inhibitors of the family but its structure has not yet been solved experimentally. Here we report the (1)H, (15)N and (13)C chemical shift assignments of CmPI-II as basis for NMR structure determination and interaction studies. Secondary structure analyses deduced from the NMR chemical shift data have identified three β-strands β1: residues 14-19, β2: 23-35 and β3: 43-45 and one helix α1: 28-37 arranged in the sequential order β1-β2-α1-β3. These secondary structure elements suggest that CmPI-II adopts the typical scaffold of a Kazal-type inhibitor. PMID:26547437

  4. An economic approach to efficient isotope labeling in insect cells using homemade 15N-, 13C- and 2H-labeled yeast extracts.

    PubMed

    Opitz, Christian; Isogai, Shin; Grzesiek, Stephan

    2015-07-01

    Heterologous expression of proteins in insect cells is frequently used for crystallographic structural studies due to the high yields even for challenging proteins requiring the eukaryotic protein processing capabilities of the host. However for NMR studies, the need for isotope labeling poses extreme challenges in eukaryotic hosts. Here, we describe a robust method to achieve uniform protein (15)N and (13)C labeling of up to 90 % in baculovirus-infected insect cells. The approach is based on the production of labeled yeast extract, which is subsequently supplemented to insect cell growth media. The method also allows deuteration at levels of >60 % without decrease in expression yield. The economic implementation of the labeling procedures into a standard structural biology laboratory environment is described in a step-by-step protocol. Applications are demonstrated for a variety of NMR experiments using the Abelson kinase domain, GFP, and the beta-1 adrenergic receptor as examples. Deuterated expression of the latter provides spectra of very high quality of a eukaryotic G-protein coupled receptor. PMID:26070442

  5. Investigations into the \\delta 15N and \\delta 13C of POM, Primary Producers, and Filter Feeders in the Florida Keys Coral Reef Tract

    NASA Astrophysics Data System (ADS)

    Lamb, K. A.; Swart, P. K.

    2004-12-01

    In order to examine the possible influences of anthropogenic wastes on the Florida Keys coral reef tract, water column particulate organic matter \\(POM\\), primary producers, and filter feeders were sampled from within the coastal waters of the reef tract, and analyzed for \\delta 15N and \\delta 13C isotopic composition. The POM was found to have a mean \\delta 15N value of +4.13\\permil (\\pm 1.03\\permil\\) and a mean \\delta 13C value of -19.93\\permil (\\pm 0.58\\permil). The \\delta 13C values for POM across the entire reef tract were heavier inshore \\(avg \\delta 13C = -18.29\\permil \\(\\pm 1.04\\permil\\)\\) and became lighter offshore \\(avg \\delta 13C = -21.41\\permil \\(\\pm 0.89\\permil\\)\\), which is consistent with influences from benthic algae/seagrass closer to shore in contrast to open-marine influences offshore. The \\delta 15N values for POM were neither temporally nor spatially interrelated, as the data showed no clear trend moving in either the inshore or offshore direction. Instead, our \\delta 15N values for POM reflect a combined influence from nitrogen fixation \\(\\approx0\\permil\\), zooplankton and other suspended particles, and not anthropogenic wastes \\(>+10\\permil\\), as other authors claim. The mean isotopic composition of various primary producers suggests a nitrogen fixation source of nutrients to the reef system (Halimeda sp. (\\delta 15N = 2.43\\permil; \\delta 13C = -18.04\\permill), Dictyota sp. (\\delta 15N = +2.86\\permil; \\delta 13C = -16.30\\permil), Thalassia sp. (\\delta 15N = 2.39\\permil; \\delta 13C = -10.32\\permil)), while filter feeders were found to have a mean isotopic composition more similar to POM values (sponge \\delta 15N = 4.95\\permil; \\delta 13C = -18.02\\permil). Our data simply do not appear to show evidence of anthropogenic influence on the Florida Keys reef tract. Further more, we suggest that \\delta 15N values of +4\\permil, which have recently been suggested to indicate anthropogenic

  6. Similarities and differences in 13C and 15N stable isotope ratios in two non-lethal tissue types from shovelnose sturgeon Scaphirhynchus platorynchus (Rafinesque, 1820)

    USGS Publications Warehouse

    DeVries, R. J.; Schramm, Harold L., Jr.

    2015-01-01

    We tested the hypothesis that δ13C and δ15N signatures of pectoral spines would provide measures of δ13C and δ15N similar to those obtained from fin clips for adult shovelnose sturgeon Scaphirhynchus platorynchus. Thirty-two shovelnose sturgeon (fork length [FL] = 500–724 mm) were sampled from the lower Mississippi River, USA on 23 February 2013. Isotopic relationships between the two tissue types were analyzed using mixed model analysis of covariance. Tissue types differed significantly for both δ13C (P < 0.01; spine: mean = −23.83, SD = 0.62; fin clip: mean = −25.74, SD = 0.97) and δ15N (P = 0.01; spine: mean = 17.01, SD = 0.51; fin clip: mean = 17.19, SD = 0.62). Neither FL nor the FL × tissue type interaction had significant (P > 0.05) effects on δ13C. Fin clip δ13C values were highly variable and weakly correlated (r = 0.16, P = 0.40) with those from pectoral spines. We found a significant FL-tissue type interaction for δ15N, reflecting increasing δ15N with FL for spines and decreasing δ15N with FL for fin clips. These results indicate that spines are not a substitute for fin clip tissue for measuring δ13C and δ15N for shovelnose sturgeon in the lower Mississippi River, but the two tissues have different turnover rates they may provide complementary information for assessing trophic position at different time scales.

  7. Interresidue carbonyl-carbonyl polarization transfer experiments in uniformly 13C, 15N-labeled peptides and proteins

    NASA Astrophysics Data System (ADS)

    Janik, Rafal; Ritz, Emily; Gravelle, Andrew; Shi, Lichi; Peng, Xiaohu; Ladizhansky, Vladimir

    2010-03-01

    In this work, we demonstrate that Homonuclear Rotary Resonance Recoupling (HORROR) can be used to reintroduce carbonyl-carbonyl interresidue dipolar interactions and to achieve efficient polarization transfer between carbonyl atoms in uniformly 13C, 15N-labeled peptides and proteins. We show that the HORROR condition is anisotropically broadened and overall shifted to higher radio frequency intensities because of the CSA effects. These effects are analyzed theoretically using Average Hamiltonian Theory. At spinning frequencies used in this study, 22 kHz, this broadening is experimentally found to be on the order of a kilohertz at a proton field of 600 MHz. To match HORROR condition over all powder orientations, variable amplitude radio frequency (RF) fields are required, and efficient direct transfers on the order of 20-30% can be straightforwardly established. Two- and three-dimensional chemical shift correlation experiments establishing long-range interresidue connectivities (e.g., (N[i]-CO[i - 2])) are demonstrated on the model peptide N-acetyl-valine-leucine, and on the third immunoglobulin binding domain of protein G. Possible future developments are discussed.

  8. Interresidue carbonyl-carbonyl polarization transfer experiments in uniformly 13C,15N-labeled peptides and proteins.

    PubMed

    Janik, Rafal; Ritz, Emily; Gravelle, Andrew; Shi, Lichi; Peng, Xiaohu; Ladizhansky, Vladimir

    2010-03-01

    In this work, we demonstrate that Homonuclear Rotary Resonance Recoupling (HORROR) can be used to reintroduce carbonyl-carbonyl interresidue dipolar interactions and to achieve efficient polarization transfer between carbonyl atoms in uniformly (13)C,(15)N-labeled peptides and proteins. We show that the HORROR condition is anisotropically broadened and overall shifted to higher radio frequency intensities because of the CSA effects. These effects are analyzed theoretically using Average Hamiltonian Theory. At spinning frequencies used in this study, 22kHz, this broadening is experimentally found to be on the order of a kilohertz at a proton field of 600MHz. To match HORROR condition over all powder orientations, variable amplitude radio frequency (RF) fields are required, and efficient direct transfers on the order of 20-30% can be straightforwardly established. Two- and three-dimensional chemical shift correlation experiments establishing long-range interresidue connectivities (e.g., (N[i]-CO[i-2])) are demonstrated on the model peptide N-acetyl-valine-leucine, and on the third immunoglobulin binding domain of protein G. Possible future developments are discussed. PMID:20060344

  9. 13C- and 15N-Labeling Strategies Combined with Mass Spectrometry Comprehensively Quantify Phospholipid Dynamics in C. elegans

    PubMed Central

    Drechsler, Robin; Gafken, Philip R.; Olsen, Carissa Perez

    2015-01-01

    Membranes define cellular and organelle boundaries, a function that is critical to all living systems. Like other biomolecules, membrane lipids are dynamically maintained, but current methods are extremely limited for monitoring lipid dynamics in living animals. We developed novel strategies in C. elegans combining 13C and 15N stable isotopes with mass spectrometry to directly quantify the replenishment rates of the individual fatty acids and intact phospholipids of the membrane. Using multiple measurements of phospholipid dynamics, we found that the phospholipid pools are replaced rapidly and at rates nearly double the turnover measured for neutral lipid populations. In fact, our analysis shows that the majority of membrane lipids are replaced each day. Furthermore, we found that stearoyl-CoA desaturases (SCDs), critical enzymes in polyunsaturated fatty acid production, play an unexpected role in influencing the overall rates of membrane maintenance as SCD depletion affected the turnover of nearly all membrane lipids. Additionally, the compromised membrane maintenance as defined by LC-MS/MS with SCD RNAi resulted in active phospholipid remodeling that we predict is critical to alleviate the impact of reduced membrane maintenance in these animals. Not only have these combined methodologies identified new facets of the impact of SCDs on the membrane, but they also have great potential to reveal many undiscovered regulators of phospholipid metabolism. PMID:26528916

  10. Application of (13)C and (15)N stable isotope probing to characterize RDX degrading microbial communities under different electron-accepting conditions.

    PubMed

    Cho, Kun-Ching; Lee, Do Gyun; Fuller, Mark E; Hatzinger, Paul B; Condee, Charles W; Chu, Kung-Hui

    2015-10-30

    This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or its metabolites as carbon and/or nitrogen sources under different electron-accepting conditions using (13)C and (15)N stable isotope probing (SIP). Mesocosms were constructed using groundwater and aquifer solids from an RDX-contaminated aquifer. The mesocosms received succinate as a carbon source and one of four electron acceptors (nitrate, manganese(IV), iron(III), or sulfate) or no additional electron acceptor (to stimulate methanogenesis). When RDX degradation was observed, subsamples from each mesocosm were removed and amended with (13)C3- or ring-(15)N3-, nitro-(15)N3-, or fully-labeled (15)N6-RDX, followed by additional incubation and isolation of labeled nucleic acids. A total of fifteen 16S rRNA sequences, clustering in α- and γ-Proteobacteria, Clostridia, and Actinobacteria, were detected in the (13)C-DNA fractions. A total of twenty seven sequences were derived from different (15)N-DNA fractions, with the sequences clustered in α- and γ-Proteobacteria, and Clostridia. Interestingly, sequences identified as Desulfosporosinus sp. (in the Clostridia) were not only observed to incorporate the labeled (13)C or (15)N from labeled RDX, but also were detected under each of the different electron-accepting conditions. The data suggest that (13)C- and (15)N-SIP can be used to characterize microbial communities involved in RDX biodegradation, and that the dominant pathway of RDX biodegradation may differ under different electron-accepting conditions. PMID:25935409

  11. δ13C and δ15N of moss Haplocladium microphyllum (Hedw.) Broth. for indicating growing environment variation and canopy retention on atmospheric nitrogen deposition

    NASA Astrophysics Data System (ADS)

    Liu, Xue-Yan; Xiao, Hua-Yun; Liu, Cong-Qiang; Li, You-Yi

    Mosses have been recognized as a useful tool for biomonitoring atmospheric deposition and assessing regional environment. This study was carried on whether the same moss growing in areas with identical regional atmospheric deposition while under different growing environments would have the same indicating signals. Similar variations in mean δ13C and δ15N signatures were found between mosses collected from five habitats, with an increasing sequence from mosses under canopies to epilithic mosses, indicating that habitats were potentially regulating δ13C and δ15N values of mosses. Dryer habitats (lower water availability) and input of more aerosol N were the main reasons for higher δ13C and δ15N values of mosses at open sites (especially for epilithic species), while more negative values of mosses under canopies were attributed to their wetter habitats and less uptake of aerosol N. Additionally, δ15N values not δ13C varied linearly with canopy thickness from -7.84‰ (1 m) to -4.71±0.7‰ (4 m), suggesting δ15N was more sensitive for indicating canopy retention. Consequently, isotopic data of mosses under different environments could not be compared for atmospheric deposition research with each other even collected at the same site. Moss δ13C and δ15N were affected not only by regional atmospheric N sources but also by their growing environments. δ15N of epilithic Haplocladium microphyllum at open sites can be taken as confident bio-indicator of atmospheric N deposition, which would deepen the application of stable nitrogen isotope of bryophytes in atmosphere-plant system study.

  12. Complementary constraints from carbon (13C) and nitrogen (15N) isotopes on the glacial ocean's soft-tissue biological pump

    NASA Astrophysics Data System (ADS)

    Schmittner, A.; Somes, C. J.

    2016-06-01

    A three-dimensional, process-based model of the ocean's carbon and nitrogen cycles, including 13C and 15N isotopes, is used to explore effects of idealized changes in the soft-tissue biological pump. Results are presented from one preindustrial control run (piCtrl) and six simulations of the Last Glacial Maximum (LGM) with increasing values of the spatially constant maximum phytoplankton growth rate μmax, which accelerates biological nutrient utilization mimicking iron fertilization. The default LGM simulation, without increasing μmax and with a shallower and weaker Atlantic Meridional Overturning Circulation and increased sea ice cover, leads to 280 Pg more respired organic carbon (Corg) storage in the deep ocean with respect to piCtrl. Dissolved oxygen concentrations in the colder glacial thermocline increase, which reduces water column denitrification and, with delay, nitrogen fixation, thus increasing the ocean's fixed nitrogen inventory and decreasing δ15NNO3 almost everywhere. This simulation already fits sediment reconstructions of carbon and nitrogen isotopes relatively well, but it overestimates deep ocean δ13CDIC and underestimates δ15NNO3 at high latitudes. Increasing μmax enhances Corg and lowers deep ocean δ13CDIC, improving the agreement with sediment data. In the model's Antarctic and North Pacific Oceans modest increases in μmax result in higher δ15NNO3 due to enhanced local nutrient utilization, improving the agreement with reconstructions there. Models with moderately increased μmax fit both isotope data best, whereas large increases in nutrient utilization are inconsistent with nitrogen isotopes although they still fit the carbon isotopes reasonably well. The best fitting models reproduce major features of the glacial δ13CDIC, δ15N, and oxygen reconstructions while simulating increased Corg by 510-670 Pg compared with the preindustrial ocean. These results are consistent with the idea that the soft-tissue pump was more efficient

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

    PubMed

    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. PMID:25451865

  14. 1H-13C/1H-15N Heteronuclear Dipolar Recoupling by R-Symmetry Sequences Under Fast Magic Angle Spinning for Dynamics Analysis of Biological and Organic Solids

    PubMed Central

    Hou, Guangjin; Byeon, In-Ja L.; Ahn, Jinwoo; Gronenborn, Angela M.; Polenova, Tatyana

    2011-01-01

    Fast magic angle spinning (MAS) NMR spectroscopy is becoming increasingly important in structural and dynamics studies of biological systems and inorganic materials. Superior spectral resolution due to the efficient averaging of the dipolar couplings can be attained at MAS frequencies of 40 kHz and higher with appropriate decoupling techniques, while proton detection gives rise to significant sensitivity gains, therefore making fast MAS conditions advantageous across the board compared with the conventional slow- and moderate-MAS approaches. At the same time, many of the dipolar recoupling approaches that currently constitute the basis for structural and dynamics studies of solid materials and that are designed for MAS frequencies of 20 kHz and below, fail above 30 kHz. In this report, we present an approach for 1H-13C/1H-15N heteronuclear dipolar recoupling under fast MAS conditions using R-type symmetry sequences, which is suitable even for fully protonated systems. A series of rotor-synchronized R-type symmetry pulse schemes are explored for the determination of structure and dynamics in biological and organic systems. The investigations of the performance of the various RNnv-symmetry sequences at the MAS frequency of 40 kHz experimentally and by numerical simulations on [U-13C,15N]-alanine and [U-13C,15N]-N-acetyl-valine, revealed excellent performance for sequences with high symmetry number ratio (N/2n > 2.5). Further applications of this approach are presented for two proteins, sparsely 13C/uniformly 15N enriched CAP-Gly domain of dynactin and U-13C,15N-Tyr enriched C-terminal domain of HIV-1 CA protein. 2D and 3D R1632-based DIPSHIFT experiments carried out at the MAS frequency of 40 kHz, yielded site-specific 1H-13C/1H-15N heteronuclear dipolar coupling constants for CAP-Gly and CTD CA, reporting on the dynamic behavior of these proteins on time scales of nano- to microseconds. The R-symmetry based dipolar recoupling under fast MAS is expected to find

  15. Food partitioning of leaf-eating mangrove crabs ( Sesarminae): Experimental and stable isotope ( 13C and 15N) evidence

    NASA Astrophysics Data System (ADS)

    Kristensen, Ditte K.; Kristensen, Erik; Mangion, Perrine

    2010-05-01

    The feasibility of mangrove leaves as a full diet for sesarmid crabs has been questioned for decades. Since these leaves are nitrogen-poor, sesarmids probably obtain nitrogen from other sources to sustain growth. The aim of this study was to assess the food partitioning of the sesarmid species Neoepisesarma versicolor with emphasis on nitrogen allocation. The preference for animal tissue when crabs were pre-fed diets of different nitrogen content was determined in the laboratory. Furthermore, the possible in situ diet composition of N. versicolor was established from carbon and nitrogen stable isotope signature ( δ13C and δ15N) of freshly caught individuals and their potential food sources, using a concentration-dependent mixing model. N. versicolor showed significantly higher feeding preferences for fish meat when pre-fed leaf material without than with access to meat, indicating that this crab species can meet its nitrogen demand by ingesting animal tissue. The stable isotope mixing model based on in situ materials suggests that the diet of N. versicolor consists of ˜60% leaves in terms of biomass, leaving ˜40% for other sources such as animal tissue and benthic microorganisms. The biomass contribution from animal tissues, in form of e.g. other crustaceans and fish carcasses, was found to account for ˜15%. Despite the relative low biomass fraction, animal food sources may contribute with up to half of the nitrogen in the diet of N. versicolor. The quantity of ingested sediment most likely exceeds that of animal tissues. However, due to the low concentration of assimilable microalgae and other microorganism, we propose that sediment associated sources are less important as a nitrogen source for N. versicolor than hitherto presumed.

  16. 13C NMR spectroscopy of methane adsorbed in SAPO-11 molecular sieve

    NASA Astrophysics Data System (ADS)

    Koskela, Tuomas; Ylihautala, Mika; Vaara, Juha; Jokisaari, Jukka

    1996-10-01

    Static 13C and 13C-{ 1H} NMR spectra of carbon-13 enriched methane ( 13CH 4) adsorbed into SAPO-11 molecular sieve were recorded at variable temperatures. Moreover, the corresponding MAS NMR spectra were measured. These experiments reveal a temperature-dependent, anisotropic and asymmetric 13C nuclear shielding tensor. Ab initio model calculations of methane in the field of a positive point charge suggest that the deformation of the shielding tensor may be related to the interaction between the methane molecule and the charge-compensating protons. A comparison with existing Xe data is made.

  17. Sequence-specific (1)H, (15)N, and (13)C resonance assignments of the autophagy-related protein LC3C.

    PubMed

    Krichel, Carsten; Weiergräber, Oliver H; Pavlidou, Marina; Mohrlüder, Jeannine; Schwarten, Melanie; Willbold, Dieter; Neudecker, Philipp

    2016-04-01

    Autophagy is a versatile catabolic pathway for lysosomal degradation of cytoplasmic material. While the phenomenological and molecular characteristics of autophagic non-selective (bulk) decomposition have been investigated for decades, the focus of interest is increasingly shifting towards the selective mechanisms of autophagy. Both, selective as well as bulk autophagy critically depend on ubiquitin-like modifiers belonging to the Atg8 (autophagy-related 8) protein family. During evolution, Atg8 has diversified into eight different human genes. While all human homologues participate in the formation of autophagosomal membrane compartments, microtubule-associated protein light chain 3C (LC3C) additionally plays a unique role in selective autophagic clearance of intracellular pathogens (xenophagy), which relies on specific protein-protein recognition events mediated by conserved motifs. The sequence-specific (1)H, (15)N, and (13)C resonance assignments presented here form the stepping stone to investigate the high-resolution structure and dynamics of LC3C and to delineate LC3C's complex network of molecular interactions with the autophagic machinery by NMR spectroscopy. PMID:26280529

  18. Backbone 1H, 13C and 15N resonance assignments of the 39 kDa staphylococcal hemoglobin receptor IsdH.

    PubMed

    Spirig, Thomas; Clubb, Robert T

    2012-10-01

    During infections Stahpylococcus aureus preferentially uses heme as an iron source, which it captures from human hemoglobin using the Iron regulated surface determinant (Isd) system. On the cell surface two related staphylococcal surface receptors called IsdH and IsdB bind to hemoglobin and extract its heme. Both receptors contain multiple NEAr iron Transporter (NEAT) domains that either bind to hemoglobin, or to heme. All previous structural studies have investigated individual NEAT domains and have not explored how the domains might interact with one another to synergistically extract heme from hemoglobin. Here, we report the near complete (1)H, (13)C and (15)N backbone resonance assignments of a bi-domain unit from IsdH that contains the N2 and N3 NEAT domains, which bind to hemoglobin and heme, respectively (IsdH(N2N3), residues 326-660, 39 kDa). The assigned backbone resonances lay the foundation for future NMR studies that will explore the molecular basis of IsdH function. PMID:22101872

  19. Design and operation of a continuous 13C and 15N labeling chamber for uniform or differential, metabolic and structural, plant tissue isotope labeling

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tracing heavy stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2 fluxes and soil organic matter formation to small-scale stable-isotope biomarker probing. Coupling multiple stable isotopes such as 13C with 15N, 18O o...

  20. An efficient NMR method for the characterisation of 14N sites through indirect 13C detection

    PubMed Central

    Jarvis, James A.; Haies, Ibraheem M.

    2013-01-01

    Nitrogen is one of the most abundant elements and plays a key role in the chemistry of biological systems. Despite its widespread distribution, the study of the naturally occurring isotope of nitrogen, 14N (99.6%), has been relatively limited as it is a spin-1 nucleus that typically exhibits a large quadrupolar interaction. Accordingly, most studies of nitrogen sites in biomolecules have been performed on samples enriched with 15N, limiting the application of NMR to samples which can be isotopically enriched. This precludes the analysis of naturally occurring samples and results in the loss of the wealth of structural and dynamic information that the quadrupolar interaction can provide. Recently, several experimental approaches have been developed to characterize 14N sites through their interaction with neighboring ‘spy’ nuclei. Here we describe a novel version of these experiments whereby coherence between the 14N site and the spy nucleus is mediated by the application of a moderate rf field to the 14N. The resulting 13C/14N spectra show good sensitivity on natural abundance and labeled materials; whilst the 14N lineshapes permit the quantitative analysis of the quadrupolar interaction. PMID:23589073

  1. Selective {sup 2}H and {sup 13}C labeling in NMR analysis of solution protein structure and dynamics

    SciTech Connect

    LeMaster, D.M.

    1994-12-01

    Preparation of samples bearing combined isotope enrichment patterns has played a central role in the recent advances in NMR analysis of proteins in solution. In particular, uniform {sup 13}C, {sup 15}N enrichment has made it possible to apply heteronuclear multidimensional correlation experiments for the mainchain assignments of proteins larger than 30 KDa. In contrast, selective labeling approaches can offer advantages in terms of the directedness of the information provided, such as chirality and residue type assignments, as well as through enhancements in resolution and sensitivity that result from editing the spectral complexity, the relaxation pathways and the scalar coupling networks. In addition, the combination of selective {sup 13}C and {sup 2}H enrichment can greatly facilitate the determination of heteronuclear relaxation behavior.

  2. Production and NMR signal optimization of hyperpolarized 13C-labeled amino acids

    NASA Astrophysics Data System (ADS)

    Parish, Christopher; Niedbalski, Peter; Ferguson, Sarah; Kiswandhi, Andhika; Lumata, Lloyd

    Amino acids are targeted nutrients for consumption by cancers to sustain their rapid growth and proliferation. 13C-enriched amino acids are important metabolic tracers for cancer diagnostics using nuclear magnetic resonance (NMR) spectroscopy. Despite this diagnostic potential, 13C NMR of amino acids however is hampered by the inherently low NMR sensitivity of the 13C nuclei. In this work, we have employed a physics technique known as dynamic nuclear polarization (DNP) to enhance the NMR signals of 13C-enriched amino acids. DNP works by transferring the high polarization of electrons to the nuclear spins via microwave irradiation at low temperature and high magnetic field. Using a fast dissolution method in which the frozen polarized samples are dissolved rapidly with superheated water, injectable solutions of 13C-amino acids with highly enhanced NMR signals (by at least 5,000-fold) were produced at room temperature. Factors that affect the NMR signal enhancement levels such as the choice of free radical polarizing agents and sample preparation will be discussed along with the thermal mixing physics model of DNP. The authors would like to acknowledge the support by US Dept of Defense Award No. W81XWH-14-1-0048 and Robert A. Welch Foundation Grant No. AT-1877.

  3. The magnitude of spatial and temporal variation in δ15N and δ13C differs between taxonomic groups: Implications for food web studies

    NASA Astrophysics Data System (ADS)

    Hyndes, Glenn A.; Hanson, Christine E.; Vanderklift, Mathew A.

    2013-03-01

    Understanding variability in stable isotope abundance is essential for effective hypothesis testing and evaluating food sources, trophic levels and food web structure. The magnitude and sources of variability are likely to differ among taxonomic and functional groups. We aimed to quantify variability of δ13C and δ15N for 16 species representing seven distinct taxonomic groups of benthic invertebrates and autotrophs in a marine ecosystem. We quantified the magnitude of variability among individuals or shoots separated by metres, among eight sites separated by kilometres, and between two survey occasions separated by months. δ13C varied by as much as 7‰ for primary producers, 4‰ for consumers, while δ15N varied by as much as 9‰ and 2‰ respectively. Variation in δ15N of seagrass was largely accounted for by differences among sites, while variation in δ13C was mainly attributable to shoots collected a few metres apart. Compared to seagrasses, variation in macroalgae was mainly explained by differences between the two survey occasions for δ15N and among individuals collected a few metres apart for δ13C. Variation was generally lower for consumers and typically explained by differences among individuals for δ15N but displayed inconsistent patterns for δ13C. Dual isotope Bayesian mixing models showed that the potential contributions of food sources for herbivorous consumers varied among sites and between survey occasions, and also that there was high variability or uncertainty in the contributions of sources within sites. The relative consistency in the main sources of variation among broad taxonomic groups in autotrophs suggests that aspects of physiology that are phylogenetically conserved might be important influences on variation in natural abundances of stable isotopes. In comparison, the sources of variability were less consistent within and among broad consumer groups, suggesting complex interactions between consumers and their food sources.

  4. Accessible NMR Experiments Studying the Hydrodynamics of [subscript 15]N-Enriched Ubiquitin at Low Fields

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

    We have recently developed and implemented two experiments in biomolecular NMR for an undergraduate-level biophysical chemistry laboratory with commercially available [subscript 15]N-enriched human ubiquitin. These experiments take advantage of [subscript 15]N direct detection of the NMR signal. The first experiment develops skills in acquiring…

  5. Accessible NMR Experiments Studying the Hydrodynamics of [superscript 15]N-Enriched Ubiquitin at Low Fields

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

    We have recently developed and implemented two experiments in biomolecular NMR for an undergraduate-level biophysical chemistry laboratory with commercially available [superscript 15]N-enriched human ubiquitin. These experiments take advantage of [superscript 15]N direct detection of the NMR signal. The first experiment develops skills in…

  6. Investigating brain metabolism at high fields using localized 13C NMR spectroscopy without 1H decoupling.

    PubMed

    Deelchand, Dinesh Kumar; Uğurbil, Kâmil; Henry, Pierre-Gilles

    2006-02-01

    Most in vivo 13C NMR spectroscopy studies in the brain have been performed using 1H decoupling during acquisition. Decoupling imposes significant constraints on the experimental setup (particularly for human studies at high magnetic field) in order to stay within safety limits for power deposition. We show here that incorporation of the 13C label from 13C-labeled glucose into brain amino acids can be monitored accurately using localized 13C NMR spectroscopy without the application of 1H decoupling. Using LCModel quantification with prior knowledge of one-bond and multiple-bond J(CH) coupling constants, the uncertainty on metabolites concentrations was only 35% to 91% higher (depending on the carbon resonance of interest) in undecoupled spectra compared to decoupled spectra in the rat brain at 9.4 Tesla. Although less sensitive, 13C NMR without decoupling dramatically reduces experimental constraints on coil setup and pulse sequence design required to keep power deposition within safety guidelines. This opens the prospect of safely measuring 13C NMR spectra in humans at varied brain locations (not only the occipital lobe) and at very high magnetic fields above 4 Tesla. PMID:16345037

  7. Comprehensive signal assignment of 13C-labeled lignocellulose using multidimensional solution NMR and 13C chemical shift comparison with solid-state NMR.

    PubMed

    Komatsu, Takanori; Kikuchi, Jun

    2013-09-17

    A multidimensional solution NMR method has been developed using various pulse programs including HCCH-COSY and (13)C-HSQC-NOESY for the structural characterization of commercially available (13)C labeled lignocellulose from potatoes (Solanum tuberosum L.), chicory (Cichorium intybus), and corn (Zea mays). This new method allowed for 119 of the signals in the (13)C-HSQC spectrum of lignocelluloses to be assigned and was successfully used to characterize the structures of lignocellulose samples from three plants in terms of their xylan and xyloglucan structures, which are the major hemicelluloses in angiosperm. Furthermore, this new method provided greater insight into fine structures of lignin by providing a high resolution to the aromatic signals of the β-aryl ether and resinol moieties, as well as the diastereomeric signals of the β-aryl ether. Finally, the (13)C chemical shifts assigned in this study were compared with those from solid-state NMR and indicated the presence of heterogeneous dynamics in the polysaccharides where rigid cellulose and mobile hemicelluloses moieties existed together. PMID:24010724

  8. Effects of climate on deer bone δ15N and δ13C: Lack of precipitation effects on δ15N for animals consuming low amounts of C 4 plants

    NASA Astrophysics Data System (ADS)

    Cormie, A. B.; Schwarcz, H. P.

    1996-11-01

    We have examined the relationship of bone collagen δ15N and δ13C to climatic variables, humidity, temperature, and amount of precipitation using fifty-nine specimens of North American white-tailed deer ( Odocoileus virginianus) from forty-six different locations. In previous studies of African mammals there was a significant correlation between bone collagen δ15N and local amount of precipitation. Results presented here similarly show an increase in δ15N with decreasing amount of precipitation but only for 25% of the animals, namely those consuming more than 10% C 4 plants. These animals also exhibited a significant correlation between δ13C and temperature which mirrors previous observations for grasses suggesting that these deer consume grasses during times of population and nutrient stress. In contrast, even in dry areas containing high proportions of C 4 grasses, the majority of the deer had consumed low amounts of C 4 plants and these deer did not have δ15N which correlate with amount of precipitation. Only when deer deviated from their normal feeding pattern by consuming C 4 plants or grasses did their δ15N correlate with amount of rainfall. For these animals, consumption of C 4 plants or grasses may signal conditions of water and nutrient stress. An increase in δ15N of bone collagen may result from combined effects from excretion of concentrated urine (to conserve water) and increased internal recycling of nitrogen (to conserve nitrogen).

  9. 2H-DNP-enhanced 2H–13C solid-state NMR correlation spectroscopy

    PubMed Central

    Maly, Thorsten; Andreas, Loren B.; Smith, Albert A.

    2015-01-01

    Perdeuteration of biological macromolecules for magic angle spinning solid-state NMR spectroscopy can yield high-resolution 2H–13C correlation spectra and the method is therefore of great interest for the structural biology community. Here we demonstrate that the combination of sample deuteration and dynamic nuclear polarization yields resolved 2H–13C correlation spectra with a signal enhancement of ε ≥ 700 compared to a spectrum recorded with microwaves off and otherwise identical conditions. To our knowledge, this is the first time that 2H-DNP has been employed to enhance MAS-NMR spectra of a biologically relevant system. The DNP process is studied using several polarizing agents and the technique is applied to obtain 2H–13C correlation spectra of U-[2H, 13C] proline. PMID:20458422

  10. Comparative effect of salinity on growth, grain yield, water use efficiency, δ(13)C and δ(15)N of landraces and improved durum wheat varieties.

    PubMed

    Chamekh, Zoubeir; Ayadi, Sawsen; Karmous, Chahine; Trifa, Youssef; Amara, Hajer; Boudabbous, Khaoula; Yousfi, Salima; Serret, Maria Dolors; Araus, José Luis

    2016-10-01

    Supplemental irrigation with low-quality water will be paramount in Mediterranean agriculture in the future, where durum wheat is a major crop. Breeding for salinity tolerance may contribute towards improving resilience to irrigation with brackish water. However, identification of appropriate phenotyping traits remains a bottleneck in breeding. A set of 25 genotypes, including 19 landraces and 6 improved varieties most cultivated in Tunisia, were grown in the field and irrigated with brackish water (6, 13 and 18dSm(-1)). Improved genotypes exhibited higher grain yield (GY) and water use efficiency at the crop level (WUEyield or 'water productivity'), shorter days to flowering (DTF), lower N concentration (N) and carbon isotope composition (δ(13)C) in mature kernels and lower nitrogen isotope composition (δ(15)N) in the flag leaf compared with landraces. GY was negatively correlated with DTF and the δ(13)C and N of mature kernels and was positively correlated with the δ(15)N of the flag leaf. Moreover, δ(13)C of mature kernels was negatively correlated with WUEyield. The results highlight the importance of shorter phenology together with photosynthetic resilience to salt-induced water stress (lower δ(13)C) and nitrogen metabolism (higher N and δ(15)N) for assessing genotypic performance to salinity. PMID:27593462

  11. Tracking spatial distribution of human-derived wastewater from Davis Station, East Antarctica, using δ15N and δ13C stable isotopes.

    PubMed

    Corbett, Patricia A; King, Catherine K; Mondon, Julie A

    2015-01-15

    Stable isotope ratios, δ15N and δ13C were effectively used to determine the geographical dispersion of human derived sewage from Davis Station, East Antarctica, using Antarctic rock cod (Trematomus bernacchii). Fish within 0-4 km downstream of the outfall exhibited higher δ15N and δ13C values relative to reference sites. Nitrogen in particular showed a stepped decrease in δ15N with increasing distance from the discharge point by 1-2‰. Stable isotopes were better able to detect the extent of wastewater contamination than other techniques including faecal coliform and sterol measures. Uptake and assimilation of δ15N and δ13C up to 4 km from the outfall adds to growing evidence indicating the current level of wastewater treatment at Davis Station is not sufficient to avoid impact to the surrounding environment. Isotopic assimilation in T. bernacchii is a viable biomarker for investigation of initial sewage exposure and longer term monitoring in the future. PMID:25487089

  12. Millimeter-Wave Observations of Circumstellar 14N/15N and 12C/13C Ratios: New Insights into J-Type Stars

    NASA Astrophysics Data System (ADS)

    Adande, Gilles; Ziurys, Lucy M.; Woolf, Neville

    2016-06-01

    Measurements of 14N/15N and 12C/13C isotopic ratios have been conducted towards circumstellar envelopes of a sample of evolved stars using the J = 3→2 rotational transitions of the isotopologues of HCN, observed with the Submillimeter Telescope (SMT) of the Arizona Radio Observatory (ARO). Towards the J-type stars Y CVn and RY Dra, where 12C/13C ~ 3, the 14N/15N ratios were found to be 120-180 and 225, respectively. The 14N/15N ratio is thus anomalously low relative to interstellar values and a factor ~100 lower than equilibrium values predicted from the CNO cycle. Combining these results with previous chemical and isotopic prior observations of these stars, we conclude that two anomalous behaviors are likely to have occurred in Y CVn and RY Dra. First, the stellar envelope failed to participate in the normal mixing seen in low mass red giants, in which C and then O are substantially converted to N. Secondly, both the carbon enrichment and anomalous isotopic composition of both 13C and15N could have been caused by a plume of hot gas, hydrogen poor but enriched in 12C, from a helium flash mixing into the envelope.

  13. Assessing waterbird habitat use in coastal evaporative systems using stable isotopes (δ 13C, δ 15N and δD) as environmental tracers

    NASA Astrophysics Data System (ADS)

    Ramírez, Francisco; Abdennadher, Aida; Sanpera, Carola; Jover, Lluís; Wassenaar, Leonard I.; Hobson, Keith A.

    2011-04-01

    Isotopic patterns of biota across salinity gradients in man-made evaporative systems could assist in determining the use of these habitats by animals. Here we report δ 13C, δ 15N and δD measurements of a euryhaline fish, the Mediterranean toothcarp ( Aphanius fasciatus), inhabiting a range of salinities in the Thyna saltworks near Sfax (Tunisia). The contribution of these salinity niches to egg formation of two typically piscivorous bird species breeding in the area and feeding within saltworks, Little Tern ( Sternula albifrons) and Little Egret ( Egretta garzetta), was inferred trough a triple-isotope (δ 13C, δ 15N and δD) Bayesian mixing model. Isotopic trends for fish δ 15N and δD across the salinity gradient followed the equations: δ 15N = e (1.1 + 47.68/Salinity) and δD = -175.74 + Salinity + Salinity 2; whereas fish δ 13C increased as salinity rose (δ 13C = -10.83 + 0.02·Salinity), after a sudden drop in fish isotopic values for salinities >60 (Practical Salinity Scale) (average fish δ 13C for salinities <60 = -5.92‰). Both bird species fed largely on low hypersalinity ponds (salinity = 43; average contribution = 37% and 22% for Little Egrets and Little Terns, respectively), although the use of intermediate hypersalinities (salinities 63 and 70) by Little Terns also occurred (16% and 21%, respectively). Isotopic patterns across salinity gradients allow the use of isotopic measurements to inform studies of habitat occupancy within evaporative systems and provide further insights into how wildlife communities interact with them.

  14. Bomb-pulse 14C analysis combined with 13C and 15N measurements in blood serum from residents of Malmö, Sweden.

    PubMed

    Georgiadou, Elisavet; Stenström, Kristina Eriksson; Uvo, Cintia Bertacchi; Nilsson, Peter; Skog, Göran; Mattsson, Sören

    2013-05-01

    The (14)C content of 60 human blood serum samples from residents of Malmö (Sweden) in 1978, obtained from a biobank, has been measured to estimate the accuracy of (14)C bomb-pulse dating. The difference between the date estimated using the Calibomb software and sampling date varied between -3 ± 0.4 and +0.2 ± 0.5 years. The average age deviation of all samples was -1.5 ± 0.7 years, with the delay between production and consumption of foodstuffs being probably the dominating cause. The potential influence of food habits on the (14)C date has been evaluated using stable isotope δ(13)C and δ(15)N analysis and information about the dietary habits of the investigated individuals. Although the group consisting of lacto-ovo vegetarians and vegans (pooled group) was not completely separated from the omnivores in a stable isotopic trophic level diagram, this analysis proved to add valuable information on probable dietary habits. The age deviation of the sampling date from the respective Calibomb date was found strongly correlated with the δ(13)C values, probably due to influence from marine diet components. For the omnivore individuals, there were indications of seasonal effects on δ(13)C and the age deviation. No significant correlation was found between the age deviation and the δ(15)N values of any dietary group. No influence of sex or year of birth was found on neither the (14)C nor the δ(13)C and δ(15)N values of the serum samples. The data were also divided into two groups (omnivores and pooled group), based on the level of δ(15)N in the samples. The consumption of high δ(15)N-valued fish and birds can be responsible for this clustering. PMID:23358598

  15. Magnetic susceptibility effects on 13C MAS NMR spectra of carbon materials and graphite.

    PubMed

    Freita, J C; Emmerich, F G; Cernicchiaro, G R; Sampaio, L C; Bonagamba, T J

    2001-01-01

    13C high-resolution solid-state nuclear magnetic resonance (NMR) was employed to study carbon materials prepared through the thermal decomposition of four different organic precursors (rice hulls, endocarp of babassu coconut, peat, and PVC). For heat treatment temperatures (HTTs) above about 600 C, all materials presented 13C NMR spectra composed of a unique resonance line associated with carbon atoms in aromatic planes. With increasing HTT a continuous broadening of this resonance and a diamagnetic shift in its central frequency were verified for all samples. The evolution of the magnitude and anisotropy of the magnetic susceptibility of the heat-treated carbon samples with HTT explains well these findings. It is shown that these results are better understood when a comparison is made with the features of the 13C NMR spectrum of polycrystalline graphite, for which the magnetic susceptibility effect is also present and is much more pronounced. PMID:11529420

  16. A 13C-NMR study of exopolysaccharide synthesis in Rhizobium meliloti Su47 strain

    NASA Astrophysics Data System (ADS)

    Tavernier, P.; Portais, J.-C.; Besson, I.; Courtois, J.; Courtois, B.; Barbotin, J.-N.

    1998-02-01

    Metabolic pathways implied in the synthesis of succinoglycan produced by the Su47 strain of R. meliloti were evaluated by 13C-NMR spectroscopy after incubation with [1{-}13C] or [2{-}13C] glucose. The biosynthesis of this polymer by R. meliloti from glucose occurred by a direct polymerisation of the introduced glucose and by the pentose phosphate pathway. Les voies métaboliques impliquées dans la synthèse du succinoglycane produit par la souche Su47 de R. meliloti ont été évaluées par la spectroscopie de RMN du carbone 13 après incubation des cellules avec du [1{-}13C] ou [2{-}13C] glucose. La biosynthèse de ce polymère à partir du glucose se produit par polymérisation directe du glucose et par la voie des pentoses phosphate.

  17. Spectroscopic labeling of A, S/T in the 1H- 15N HSQC spectrum of uniformly ( 15N- 13C) labeled proteins

    NASA Astrophysics Data System (ADS)

    Chugh, Jeetender; Hosur, Ramakrishna V.

    2008-10-01

    A new triple resonance two-dimensional experiment, termed (HC)NH, has been described to generate specific labels on the peaks of alanines and serines/threonines, separately, in the 1H- 15N HSQC spectrum of a protein. The performance of the pulse sequence has been demonstrated with a 151 residue protein. The method permits the investigation of local environments around those specific residues without actually having to obtain complete resonance assignments for the entire protein. With this one can envisage use of the technique for studying large protein systems from different points of view.

  18. In vivo 31P and multilabel 13C NMR measurements for evaluation of plant metabolic pathways.

    PubMed

    Rijhwani, S K; Ho, C H; Shanks, J V

    1999-01-01

    Reliable measurements of intracellular metabolites are useful for effective plant metabolic engineering. This study explored the application of in situ 31P and 13C NMR spectroscopy for long-term measurements of intracellular pH and concentrations of several metabolites in glycolysis, glucan synthesis, and central carbon metabolic pathways in plant tissues. An NMR perfusion reactor system was designed to allow Catharanthus roseus hairy root cultures to grow for 3-6 weeks, during which time NMR spectroscopy was performed. Constant cytoplasmic pH (7.40+/-0.06), observed during the entire experiment, indicated adequate oxygenation. 13C NMR spectroscopy was performed on hairy root cultures grown in solutions containing 1-13C-, 2-13C-, and 3-13C-labeled glucose in separate experiments and the flow of label was monitored. Activities of pentose phosphate pathways, nonphotosynthetic CO2 fixation, and glucan synthesis pathways were evident from the experimental results. Scrambling of label in glucans also indicated recycling of triose phosphate and their subsequent conversion to hexose phosphates. PMID:10935751

  19. Food web implications of delta13C and delta15N variability over 370 km of the regulated Colorado River USA.

    PubMed

    Shannon, J P; Blinn, D W; Haden, G A; Benenati, E P; Wilson, K P

    2001-01-01

    Dual stable isotope analysis in the regulated Colorado River through Grand Canyon National Park, USA, revealed a food web that varied spatially through this arid biome. Down-river enrichment of delta13C data was detected across three trophic levels resulting in shifted food webs. Humpack chub delta13C and delta15N values from muscle plugs and fin clips did not differ significantly. Humpback chub and rainbow trout trophic position is positively correlated with standard length indicating an increase in piscivory by larger fishes. Recovery of the aquatic community from impoundment by Glen Canyon Dam and collecting refinements for stable isotope analysis within large rivers are discussed. PMID:11924849

  20. 13C-NMR study of labeled vinyl groups in paramagnetic myoglobin derivatives.

    PubMed

    Sankar, S S; La Mar, G N; Smith, K M; Fujinari, E M

    1987-04-01

    The 13C-NMR spectra of high-spin met-aquo myoglobin, spin-equilibrium met-azido myoglobin, low-spin met-cyano myoglobin, deoxy myoglobin and carbonmonoxy myoglobin from sperm whale reconstituted with hemin 13C enriched at both vinyl alpha or beta positions have been recorded. In all cases the labeled vinyl 13C signals are clearly resolved and useful spectra could be obtained within approx. 15 minutes. The decoupling of multiplet structure due to attached proton(s) has led to the specific assignment of vinyl 13C alpha signals in all paramagnetic derivatives and the 13C beta signals in met-cyano myoglobin. In all other cases, the collapse of the proton multiplet structure as a function of 1H decoupling frequency has located, but not assigned, the attached 1H resonance positions which are obscured by the intense diamagnetic envelope in the 1H-NMR spectrum. The resulting vinyl 13C hyperfine shifts follow Curie behavior, and the patterns closely resemble those in the appropriate model complexes in the same oxidation/spin/ligation state, except that the protein exhibits more in-plane asymmetry. The hyperfine shift patterns are indicative of dominant pi contact shifts for all ferric complexes. Deoxy myoglobin vinyl 13C and 1H contact shifts provide little evidence for pi bonding. PMID:3828362

  1. Mid-late Holocene North Pacific trade winds as inferred from deep-sea coral skeletal δ15N and δ13C biogeochemistry

    NASA Astrophysics Data System (ADS)

    Guilderson, T. P.; Mccarthy, M. D.; Dunbar, R. B.; Englebrecht, A.; Roark, E.

    2013-12-01

    δ15N and δ13C data obtained from samples of proteinaceous deep-sea corals collected from the North Pacific Subtropical Gyre (Hawai'ian archipelago) and the central equatorial Pacific (Line Islands) document multi-decadal to century scale variability in the isotopic composition of surface-produced particulate organic matter exported to the deep sea. Comparison of the δ13C data, where Line Island samples are 0.6‰ more positive than the Hawai'ian samples, support the contention that the North Pacific Subtropical Gyre is more efficient than the tropical upwelling system at trapping and/or recycling nutrients within the mixed layer. δ15N values from the Line Island samples are also more positive than those from the central gyre, and within the Hawai'ian samples there is a gradient with more positive δ15N values in samples from the main Hawai'ian Islands versus French Frigate Shoals in the Northwestern Hawai'ian Islands. The gradient in the Hawai'ian samples likely reflects the relative importance of algal acquisition of metabolic N via dissolved seawater nitrate uptake versus nitrogen fixation. The Hawai'ian sample set also exhibits a strong decrease in δ15N values from the mid-Holocene to present. We hypothesize that this decrease is most likely the result of decreasing tradewinds, and possibly a commensurate decrease in entrainment of more positive δ15N-NO3 subthermocline water masses.

  2. Dynamic nuclear polarization-enhanced 13C NMR spectroscopy of static biological solids

    PubMed Central

    Potapov, Alexey; Yau, Wai-Ming; Tycko, Robert

    2013-01-01

    We explore the possibility of using dynamic nuclear polarization (DNP) to enhance signals in structural studies of biological solids by solid state NMR without sample spinning. Specifically, we use 2D 13C-13C exchange spectroscopy to probe the peptide backbone torsion angles (ϕ,ψ) in a series of selectively 13C-labeled 40-residue β-amyloid (Aβ1–40) samples, in both fibrillar and non-fibrillar states. Experiments are carried out at 9.39 T and 8 K, using a static double-resonance NMR probe and low-power microwave irradiation at 264 GHz. In frozen solutions of Aβ1–40 fibrils doped with DOTOPA-TEMPO, we observe DNP signal enhancement factors of 16–21. We show that the orientation- and frequency-dependent spin polarization exchange between sequential backbone carbonyl 13C labels can be simulated accurately using a simple expression for the exchange rate, after experimentally determined homogeneous 13C lineshapes are incorporated in the simulations. The experimental 2D 13C-13C exchange spectra place constraints on the ϕ and ψ angles between the two carbonyl labels. Although the data are not sufficient to determine ϕ and ψ uniquely, the data do provide non-trivial constraints that could be included in structure calculations. With DNP at low temperatures, 2D 13C-13C exchange spectra can be obtained from a 3.5 mg sample of Aβ1–40 fibrils in 4 hr or less, despite the broad 13C chemical shift anisotropy line shapes that are observed in static samples. PMID:23562665

  3. Dynamic nuclear polarization-enhanced 13C NMR spectroscopy of static biological solids

    NASA Astrophysics Data System (ADS)

    Potapov, Alexey; Yau, Wai-Ming; Tycko, Robert

    2013-06-01

    We explore the possibility of using dynamic nuclear polarization (DNP) to enhance signals in structural studies of biological solids by solid state NMR without sample spinning. Specifically, we use 2D 13C-13C exchange spectroscopy to probe the peptide backbone torsion angles (ϕ, ψ) in a series of selectively 13C-labeled 40-residue β-amyloid (Aβ1-40) samples, in both fibrillar and non-fibrillar states. Experiments are carried out at 9.39 T and 8 K, using a static double-resonance NMR probe and low-power microwave irradiation at 264 GHz. In frozen solutions of Aβ1-40 fibrils doped with DOTOPA-TEMPO, we observe DNP signal enhancement factors of 16-21. We show that the orientation- and frequency-dependent spin polarization exchange between sequential backbone carbonyl 13C labels can be simulated accurately using a simple expression for the exchange rate, after experimentally determined homogeneous 13C lineshapes are incorporated in the simulations. The experimental 2D 13C-13C exchange spectra place constraints on the ϕ and ψ angles between the two carbonyl labels. Although the data are not sufficient to determine ϕ and ψ uniquely, the data do provide non-trivial constraints that could be included in structure calculations. With DNP at low temperatures, 2D 13C-13C exchange spectra can be obtained from a 3.5 mg sample of Aβ1-40 fibrils in 4 h or less, despite the broad 13C chemical shift anisotropy line shapes that are observed in static samples.

  4. Benchmarks for the 13C NMR chemical shielding tensors in peptides in the solid state

    NASA Astrophysics Data System (ADS)

    Czernek, Jiří; Pawlak, Tomasz; Potrzebowski, Marek J.

    2012-02-01

    The benchmark set is proposed, which comprises 126 principal elements of chemical shielding tensors, and the respective isotropic chemical shielding values, of all 42 13C nuclei in crystalline Tyr-D-Ala-Phe and Tyr-Ala-Phe tripeptides with known, but highly dissimilar structures. These data are obtained by both the NMR measurements and the density functional theory in the pseudopotential plane-wave scheme. Using the CASTEP program, several computational strategies are employed, for which the level of agreement between calculations and experiment is established. This set is mainly intended for the validation of methods capable of predicting the 13C NMR parameters in solid-state systems.

  5. 13C-NMR spectra and contact time experiment for Skjervatjern fulvic and humic acids

    USGS Publications Warehouse

    Malcolm, R.L.

    1992-01-01

    The T(CP) and T(1p) time constants for Skjervatjern fulvic and humic acids were determined to be short with T(CP) values ranging from 0.14 ms to 0.53 ms and T(1p) values ranging from 3.3 ms to 5.9 ms. T(CP) or T(1p) time constants at a contact time of 1 ms are favorable for quantification of 13C-NMR spectra. Because of the short T(CP) values, correction factors for signal intensity for various regions of the 13C-NMR spectra would be necessary at contact times greater than 1.1 ms or less than 0.9 ms. T(CP) and T(1p) values have a limited non-homogeneity within Skjervatjern fulvic and humic acids. A pulse delay or repeat time of 700 ms is more than adequate for quantification of these 13C-NMR spectra. Paramagnetic effects in these humic substances are precluded due to low inorganic ash contents, low contents of Fe, Mn, and Co, and low organic free-radical contents. The observed T(CP) values suggest that all the carbon types in Skjervatjern fulvic and humic acids are fully cross-polarized before significant proton relaxation occurs. The 13C-NMR spectra for Skjervatjern fulvic acid is similar to most aquatic fulvic acids as it is predominantly aliphatic, low in aromaticity (fa1 = 24), low in phenolic content, high in carboxyl content, and has no resolution of a methoxyl peak. The 13C-NMR spectra for Skjervatjern humic acid is also similar to most other aquatic humic acids in that it is also predominantly aliphatic, high in aromaticity (fa1 = 38), moderate in phenolic content, moderate in carboxyl content, and has a clear resolution of a methoxyl carbon region. After the consideration of the necessary 13C-NMR experimental conditions, these spectra are considered to be quantitative. With careful consideration of the previously determined 13C-NMR experimental conditions, quantitative spectra can be obtained for humic substances in the future from the HUMEX site. Possible changes in humic substances due to acidification should be determined from 13C-NMR data.

  6. In vivo 13C NMR metabolite profiling: potential for understanding and assessing conifer seed quality.

    PubMed

    Terskikh, Victor V; Feurtado, J Allan; Borchardt, Shane; Giblin, Michael; Abrams, Suzanne R; Kermode, Allison R

    2005-08-01

    High-resolution 13C MAS NMR spectroscopy was used to profile a range of primary and secondary metabolites in vivo in intact whole seeds of eight different conifer species native to North America, including six of the Pinaceae family and two of the Cupressaceae family. In vivo 13C NMR provided information on the total seed oil content and fatty acid composition of the major storage lipids in a non-destructive manner. In addition, a number of monoterpenes were identified in the 13C NMR spectra of conifer seeds containing oleoresin; these compounds showed marked variability in individual seeds of Pacific silver fir within the same seed lot. In imbibed conifer seeds, the 13C NMR spectra showed the presence of considerable amounts of dissolved sucrose presumed to play a protective role in the desiccation-tolerance of seeds. The free amino acids arginine and asparagine, generated as a result of storage protein mobilization, were detected in vivo during seed germination and early seedling growth. The potential for NMR to profile metabolites in a non-destructive manner in single conifer seeds and seed populations is discussed. It is a powerful tool to evaluate seed quality because of its ability to assess reserve accumulation during seed development or at seed maturity; it can also be used to monitor reserve mobilization, which is critical for seedling emergence. PMID:15996983

  7. Combining combing and secondary ion mass spectrometry to study DNA on chips using (13)C and (15)N labeling.

    PubMed

    Cabin-Flaman, Armelle; Monnier, Anne-Francoise; Coffinier, Yannick; Audinot, Jean-Nicolas; Gibouin, David; Wirtz, Tom; Boukherroub, Rabah; Migeon, Henri-Noël; Bensimon, Aaron; Jannière, Laurent; Ripoll, Camille; Norris, Victor

    2016-01-01

    Dynamic secondary ion mass spectrometry ( D-SIMS) imaging of combed DNA - the combing, imaging by SIMS or CIS method - has been developed previously using a standard NanoSIMS 50 to reveal, on the 50 nm scale, individual DNA fibers labeled with different, non-radioactive isotopes in vivo and to quantify these isotopes. This makes CIS especially suitable for determining the times, places and rates of DNA synthesis as well as the detection of the fine-scale re-arrangements of DNA and of molecules associated with combed DNA fibers. Here, we show how CIS may be extended to (13)C-labeling via the detection and quantification of the (13)C (14)N (-) recombinant ion and the use of the (13)C: (12)C ratio, we discuss how CIS might permit three successive labels, and we suggest ideas that might be explored using CIS. PMID:27429742

  8. Combining combing and secondary ion mass spectrometry to study DNA on chips using 13C and 15N labeling

    PubMed Central

    Cabin-Flaman, Armelle; Monnier, Anne-Francoise; Coffinier, Yannick; Audinot, Jean-Nicolas; Gibouin, David; Wirtz, Tom; Boukherroub, Rabah; Migeon, Henri-Noël; Bensimon, Aaron; Jannière, Laurent; Ripoll, Camille; Norris, Victor

    2016-01-01

    Dynamic secondary ion mass spectrometry ( D-SIMS) imaging of combed DNA – the combing, imaging by SIMS or CIS method – has been developed previously using a standard NanoSIMS 50 to reveal, on the 50 nm scale, individual DNA fibers labeled with different, non-radioactive isotopes in vivo and to quantify these isotopes. This makes CIS especially suitable for determining the times, places and rates of DNA synthesis as well as the detection of the fine-scale re-arrangements of DNA and of molecules associated with combed DNA fibers. Here, we show how CIS may be extended to 13C-labeling via the detection and quantification of the 13C 14N - recombinant ion and the use of the 13C: 12C ratio, we discuss how CIS might permit three successive labels, and we suggest ideas that might be explored using CIS. PMID:27429742

  9. Vibrational spectra and structure of RDX and its 13C- and 15N-labeled derivatives: a theoretical and experimental study.

    PubMed

    Infante-Castillo, Ricardo; Pacheco-Londoño, Leonardo; Hernández-Rivera, Samuel P

    2010-07-01

    Unambiguous vibrational band assignments have been made to cyclic nitramine hexahydro-1,3,5-trinitro-s-triazine, commonly known as the alpha-phase of RDX or alpha-RDX, with the use of (13)C and (15)N (on ring) enriched isotopic RDX analogues. Vibrational spectra were collected using Raman and IR spectroscopy in solid state and ab initio normal mode calculations were performed using density functional theory (DFT) and a 6-311G++** basis set. The calculated isotopic frequency shifts, induced by (13)C and (15)N labeling, are in very good accordance with measures ones. The changes in vibrational modes associated with the isotopic substitutions are well modeled by the calculation and previous assignments of the vibrational spectra have been revised, especially where the exact nature of the vibrational modes had been either vague or contradictory. PMID:20381411

  10. Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling

    PubMed Central

    Soong, Jennifer L; Reuss, Dan; Pinney, Colin; Boyack, Ty; Haddix, Michelle L; Stewart, Catherine E; Cotrufo, M. Francesca

    2014-01-01

    Tracing rare stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2 fluxes and soil organic matter formation to small-scale stable-isotope biomarker probing. Coupling multiple stable isotopes such as 13C with 15N, 18O or 2H has the potential to reveal even more information about complex stoichiometric relationships during biogeochemical transformations. Isotope labeled plant material has been used in various studies of litter decomposition and soil organic matter formation1-4. From these and other studies, however, it has become apparent that structural components of plant material behave differently than metabolic components (i.e. leachable low molecular weight compounds) in terms of microbial utilization and long-term carbon storage5-7. The ability to study structural and metabolic components separately provides a powerful new tool for advancing the forefront of ecosystem biogeochemical studies. Here we describe a method for producing 13C and 15N labeled plant material that is either uniformly labeled throughout the plant or differentially labeled in structural and metabolic plant components. Here, we present the construction and operation of a continuous 13C and 15N labeling chamber that can be modified to meet various research needs. Uniformly labeled plant material is produced by continuous labeling from seedling to harvest, while differential labeling is achieved by removing the growing plants from the chamber weeks prior to harvest. Representative results from growing Andropogon gerardii Kaw demonstrate the system's ability to efficiently label plant material at the targeted levels. Through this method we have produced plant material with a 4.4 atom%13C and 6.7 atom%15N uniform plant label, or material that is differentially labeled by up to 1.29 atom%13C and 0.56 atom%15N in its metabolic and structural components (hot water extractable and hot water residual components

  11. Topological Constraints on Chain-Folding Structure of Semicrystalline Polymer as Studied by 13C-13C Double Quantum NMR

    NASA Astrophysics Data System (ADS)

    Hong, Youlee; Miyoshi, Toshikazu

    Chain-folding process is a prominent feature of long polymer chains during crystallization. Over the last half century, much effort has been paid to reveal the chain trajectory. Even though various chain-folding models as well as theories of crystallization at molecule levels have been proposed, they could be not reconciled due to the limited experimental evidences. Recent development of double quantum NMR with selective isotope labeling identified the chain-trajectory of 13C labeled isotactic poly(1-butene). The systematic experiments covered a wide range of parameters, i.e. kinetics, concentration, and molecular weight (Mw) . It was demonstrated that i) adjacent re-entry site was invariant as a function of crystallization temperature (Tc) , concentration, andMw, ii) long-range order of adjacent re-entry sequence is independence of kinetics at a given concentration while it decreased with increasing the polymer concentration at a given Tc due to the increased interruption between the chains, and iii) high Mw chains led to the multilayer folded structures in single crystals, but the melt state induced the identical short adjacent sequences of long and short polymer over a wide range of Tc due to the entanglements. The behaviors indicated that the topological restriction plays significant roles in the chain-folding process rather than the kinetics. The proposed framework to control the chain-folding structure presents a new perspective into the chain organization by either the intra- or inter-chain interaction. National Science Foundation Grants DMR-1105829 and 1408855.

  12. δ 13C and δ 15N Values of Soil Organic Matter Over Drought and Non-drought Affected Elevation Gradients in Ethiopia: Calibrating for Environmental Reconstruction

    NASA Astrophysics Data System (ADS)

    Terwilliger, V. J.; Eshetu, Z.; Colman, A. S.; Fogel, M.

    2004-12-01

    Portions of Ethiopia today are experiencing increasing temperatures and drought frequencies. The longest known hominid record is in Ethiopia's Awash Basin. Reconstructing past environments in Ethiopia may, therefore, contribute both to understanding present day and past consequences of climate change. Studies suggest that at least 7000 years of environmental reconstruction may be possible from isotopic analyses of organic matter in some Ethiopian paleosols. We have measured δ 13C and δ 15N of organic matter from modern soils in Ethiopia to explore the climatic dependence of these signals and thus to determine the maximum resolution of climatic reconstruction possible by bulk isotopic analyses of soil organic matter (SOM). Surface soil samples were taken at elevations from 350 - 3500 m in drought affected regions and from 1050 - 3100 m in regions with no history of drought. Collections were made at an altitude resolution of better than 150 m. Deeper soil samples (max. 27 m) were also obtained at 22 elevations in sites of the Awash Basin that had already been studied using other paleoenvironmental proxies. Soils were sampled in grassland, shrubland, forest, and grass/sedge wetland. The δ 15N values of SOM decreased significantly with increase in elevation and were sensitive to both overlaying vegetation type and drought proclivity. Our results support hypotheses that δ 15N values vary with total nitrogen pools in soils which, in turn vary with humidity and associated microbial influences. The δ 13C values of SOM had a quadratic relationship to elevation that most likely reflected the relative compositions of C3 and C4 biomass in overlying vegetation. Exposure to drought could not be detected by δ 13C values. At sites in the Hadar region where depth profile measurements were made to 27 m, δ 13C values decreased with depth. This result conforms to inferences from other proxy that a cooler, wetter climate previously existed. Nitrogen contents of soils below 3 m

  13. Sequence-specific 1H, 15N and 13C resonance assignments of the 23.7-kDa homodimeric toxin CcdB from Vibrio fischeri.

    PubMed

    Respondek, Michal; Buts, Lieven; De Jonge, Natalie; Haesaerts, Sarah; Loris, Remy; Van Melderen, Laurence; Wyns, Lode; Zangger, Klaus

    2009-06-01

    CcdB is the toxic component of a bacterial toxin-antitoxin system. It inhibits DNA gyrase (a type II topoisomerase), and its toxicity can be neutralized by binding of its antitoxin CcdA. Here we report the sequential backbone and sidechain (1)H, (15)N and (13)C resonance assignments of CcdB(Vfi) from the marine bacterium Vibrio fischeri. The BMRB accession number is 16135. PMID:19636967

  14. Stable isotope ((13)C/(12)C and (15)N/(14)N) composition of the woolly rhinoceros Coelodonta antiquitatis horn suggests seasonal changes in the diet.

    PubMed

    Tiunov, Alexei V; Kirillova, Irina V

    2010-11-15

    The extinct woolly rhinoceros Coelodonta antiquitatis is a prominent member of the Mammuthus-Coelodonta faunal complex, but its biology is poorly known, partly because very few specimens with well-preserved soft tissues have been discovered to date. However, the permafrost-preserved horns of the woolly rhinoceros are recording structures which contain isotopic records of the diet, environmental conditions and physiological status of the animal during most of its life. In this study we report the first data on the pattern of carbon ((13)C/(12)C) and nitrogen ((15)N/(14)N) isotopic composition along the nasal horn of woolly rhinoceros. We found systematic variations in δ(13)C and δ(15)N values associated with morphologically expressed transverse banding of the horn. The comparative analysis of isotopic variation in keratinous tissues of extant and extinct herbivores suggests that the oscillation in isotopic composition of the horn was induced by seasonal changes in the diet. Although the compiled evidence is in part contradictory, we suggest that more positive δ(13)C and δ(15)N values associated with dark-colored and less dense zones of the horn indicate a summer diet. More dense and light-colored zones of the horn have lower δ(13)C and δ(15)N values possibly indicating a larger proportion of woody and shrub vegetation in the winter diet. The validity of these conclusions has to be proven in further investigations, but our data underline the potential of isotopic analysis for studies on diet and habitat use by extinct members of Pleistocene fauna. PMID:20941761

  15. Importance of bacterivory and preferential selection toward diatoms in larvae of Crepidula fornicata (L.) assessed by a dual stable isotope (13C, 15N) labeling approach

    NASA Astrophysics Data System (ADS)

    Leroy, Fanny; Riera, Pascal; Jeanthon, Christian; Edmond, Frédérique; Leroux, Cédric; Comtet, Thierry

    2012-05-01

    In Europe, the gastropod Crepidula fornicata is an invasive species characterized by a long reproductive period (from February to November). Thus, its larvae are exposed to variations in available food sources (in terms of quantity and quality). We aimed to investigate if bacteria could contribute to larval food both in presence or absence of phytoplankton, and to compare these results to seasonal variations of bacteria and phytoplankton abundances at a coastal site in the English Channel. First, ingestion of fluorescent beads of 0.5 to 2 μm diameter, showed that larvae were able to ingest particles of typical bacterial size. Then we used a dual stable isotope labeling approach which consisted in labeling a bacterial pelagic community with 15N and a diatom (Chaetoceros gracilis) culture with 13C, and supplying larvae with 15N-labeled bacteria, 13C-labeled diatoms, and both labeled sources. This technique has, to our knowledge, never been applied to invertebrate larvae. After 24 h of experiment, larvae were significantly enriched in all treatments: + 21.5‰ (∆δ13C) when supplied with diatoms, + 1364‰ (∆δ15N) when supplied with bacteria, and + 24‰ (∆δ13C) and + 135‰ (∆δ15N) when supplied with the two mixed sources. These results indicated that bacteria can contribute to the larval nutrition in C. fornicata, even in the presence of phytoplankton. Our results however suggested that larvae of C. fornicata preferentially used diatoms and showed that the supply of free bacteria did not alter the uptake of diatoms. Considering the seasonal variations of bacteria and phytoplankton abundances at the study site, these results suggested that bacteria may constitute a complementary resource for the larvae of C. fornicata when phytoplankton is abundant and may become a substitute resource when phytoplankton is less available. This approach offers promising perspectives to trace food sources and assess nitrogen and carbon fluxes between planktotrophic larvae

  16. Investigating Past Ocean Ecosystem Variability with δ13C and δ15N Records in Long Lived Deep Sea Proteinaceous Corals from the Central Equatorial Pacific

    NASA Astrophysics Data System (ADS)

    Miles, K. W.; Glynn, D. S.; McMahon, K.; McCarthy, M. D.

    2014-12-01

    The Central Tropical Pacific Ocean may experience large, abrupt climate variations on a century to millennial time scale that greatly impact ocean ecosystem structure and function, including primary productivity, plankton community composition, and biogeochemical cycling. Stable carbon and nitrogen isotope analyses of exported production are valuable proxies for past ocean plankton community composition, trophic structure, and nutrient utilization. However, very few long-term, high-resolution records exist from the tropics, which can establish how millennial climate variability is inherently linked to tropical ecosystem dynamics. Sediment cores are valuable archives of past ocean ecosystem dynamics, but have limited temporal resolution. In contrast, deep-sea proteinaceous corals act as living sediment traps due to direct incorporation of sinking organic matter into their organic skeletons. Since colonies can live for thousands of years, proteinaceous coral archives can significantly extend the length and resolution of δ13C and δ15N records in the open ocean. Here we present three δ13C and δ15N records obtained from the coral species Kulamanamana haumeaae that provide multi-decadal scale resolution δ13C and δ15N records, for the first time extending up to ~2600 years into the past. We observed large-scale oscillations in δ13C and δ15N values on century to millennial time scales that correspond to known major climate events, including the Medieval Climate Anomaly, the Little Ice Age, and the onset of the Industrial Revolution. These records will provide new insights into the variations in plankton community structure and nutrient dynamics in the equatorial pacific linked to long-term climate oscillations.

  17. Measuring Level Alignment at the Metal–Molecule Interface by In Situ Electrochemical 13C NMR

    SciTech Connect

    Li, Ying; Zelakiewicz, Brian S.; Allison, Thomas C.; Tong, Yu ye J.

    2015-03-16

    A new technique to measure energy-level alignment at a metal–molecule interface between the Fermi level of the metal and the frontier orbitals of the molecule is proposed and experimentally demonstrated. The method, which combines the electrochemistry of organo-ligand-stabilized Au nanoparticles with 13C NMR spectroscopy (i.e. in situ electrochemical NMR), enables measuring both occupied and unoccupied states.

  18. Carbon flow and trophic structure of an Antarctic coastal benthic community as determined by δ 13C and δ 15N

    NASA Astrophysics Data System (ADS)

    Gillies, C. L.; Stark, J. S.; Johnstone, G. J.; Smith, S. D. A.

    2012-01-01

    Stable isotopes of carbon and nitrogen were used to determine the different carbon pathways and trophic assemblages amongst coastal benthic fauna of the Windmill Islands, East Antarctica. Macroalgae, pelagic POM, sediment POM and sea ice POM had well-separated δ 13C signatures, which ranged from -36.75‰ for the red alga Phyllophora antarctica, to -10.35‰ for sea ice POM. Consumers were also well separated by δ 13C, ranging from -21.42‰ for the holothurian Staurocucumis sp. up to -7.47‰ for the urchin Sterechinus neumayeri. Analysis of δ 13C and δ 15N revealed distinct groups for suspension feeders, grazer/herbivores and deposit feeders, whilst predators and predator/scavengers showed less grouping. Consumers spanned a δ 15N range of 8.71‰, equivalent to four trophic levels, although δ 15N ratios amongst consumers were continuous, rather than grouped into discrete trophic levels. The study has built a trophic model for the Windmill Islands and summarises three main carbon pathways utilised by the benthos: (1) pelagic POM; (2) macroalgae/epiphytic/benthic diatoms and (3) sediment POM/benthic diatoms. The movement of carbon within the coastal benthic community of the Windmill Islands is considered complex, and stable isotopes of carbon and nitrogen were valuable tools in determining specific feeding guilds and in tracing carbon flow, particularly amongst lower-order consumers.

  19. Trace elements and stable isotopes (delta13C and delta15N) in shallow and deep-water organisms from the East China Sea.

    PubMed

    Asante, Kwadwo Ansong; Agusa, Tetsuro; Mochizuki, Hiroko; Ramu, Karri; Inoue, Suguru; Kubodera, Tsunemi; Takahashi, Shin; Subramanian, Annamalai; Tanabe, Shinsuke

    2008-12-01

    Trace elements (22) and stable isotope ratios (delta15N and delta13C) were analyzed in marine organisms from shallow (SW) and deep-water (DW) of the East China Sea to understand biomagnification and prey source of trace elements. In the benthic marine organisms from DW, delta15N values were negatively correlated with Ba, Cu, Ag, Mo, Sr, As, and Co concentrations. This may be due to the specific accumulation in lower trophic animals and/or the biodilution through the food web in DW. Relationships between delta15N and concentrations of Co, Cr, Bi, and Tl in fish and Ag, Bi, V, Hg, and Tl in crustaceans showed positive correlations, suggesting that trophic position was affecting the concentrations of those elements in phyla, with higher trophic animals retaining higher concentrations than the lower trophic animals. Positive correlations between delta13C and Rb were observed in marine organisms. Therefore, Rb may be a possible substitute of delta13C as tracer of prey source in the East China Sea although further investigation is required. PMID:18583004

  20. Multi-isotope ((15)N, (18)O and (13)C) indicators of sources and fate of nitrate in the upper stream of Chaobai River, Beijing, China.

    PubMed

    Li, Cai; Jiang, Yongbin; Guo, Xinyue; Cao, Yang; Ji, Hongbing

    2014-11-01

    Dual isotopes of nitrate ((15)N and (18)O) and carbon isotopes of dissolved inorganic carbon ((13)C) together with water chemistry were used to identify the sources and fate of nitrate in the upper stream of Chaobai River, north China. The results show that NO3(-) concentrations ranges from 0.03 mmol L(-1) to 0.80 mmol L(-1). Sampling sites from watershed with dominant forest land had higher NO3(-) concentrations and lower δ(15)N-NO3(-) (<10‰) in the wet season than in the dry season, while those from watershed with more anthropogenic activities had lower NO3(-) concentrations and higher δ(15)N-NO3(-) (>10‰) in the wet season. Compositions of isotopes and chemistry indicated that NO3(-) originated mainly from soil N, sewage and livestock wastes and atmospheric nitrogen. Furthermore, the mixing model suggested that soil N was the major NO3(-) source in the wet season, while the sewage and livestock wastes contributed the most in the dry season. Compared to rivers, the Miyun Reservoir had a higher contribution of atmospheric N and the N input from the upper rivers exerted significant influence over the reservoir. Mineralization and nitrification played an important role in N biogeochemistry based on the isotopes ((15)N and (18)O and (13)C) and chemical data. There appeared to be no significant denitrification in the watershed according to the three isotopes and chemical ions. The combined use of (15)N, (18)O and (13)C proved to be useful for further identification of the sources and fate of nitrate in watersheds with dominant forest land in the wet season. PMID:25283837

  1. 15N and 1H NMR spectroscopy of the catalytic histidine in chloromethyl ketone-inhibited complexes of serine proteases.

    PubMed

    Tsilikounas, E; Rao, T; Gutheil, W G; Bachovchin, W W

    1996-02-20

    The hemiketal hydroxyl groups in chloromethyl ketone (cmk) complexes of trypsin and chymotrypsin have been reported to ionize to the oxyanion with pK(a) values 2-4 pK(a) units below expectations for such a functional group on the basis of the behavior of the hemiketal carbon atom in 13C NMR spectra [Finucane, M. D., & Malthouse, J. P. G. (1992) Biochem. J. 286, 889-900]. The low pK(a) indicates the enzymes selectively stabilize the oxyanion form of the bound inhibitor, and therefore that cmk complexes may be good models of enzyme-mediated transition-state stabilization. However, the 13C NMR studies could not rule out His57 as the titrating group. Here we report the behavior of the ring 15N atoms of His57 in the Ala-Ala-Pro-Val-cmk complex of alpha-lytic protease. Both N(delta 1) and N(epsilon 2) of His57 respond to an ionization with a pK(a) of approximately 7.5, but His57 itself does not titrate as N(epsilon 2) remains alkylated and N(delta 1) remains bonded to a proton over the entire pH range. The species titrating with a pK(a) of approximately 7.5 must therefore be the hemiketal hydroxyl. The results also show that the 1H NMR signal from the proton in the Asp-His hydrogen bond behaves in a characteristic manner in cmk complexes and can be used diagnostically to confirm that His57 does not titrate and to measure the pK(a) of the hemiketal hydroxyl in cmk-protease complexes without resorting to 15N-labeling. We have used the behavior of this signal to directly confirm that His57 does not titrate in the trypsin and chymotrypsin complexes that were the subjects of the original 13C NMR studies. PMID:8652587

  2. Characterizing biomass fast pyrolysis oils by 13C-NMR and chemometric analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several biomass fast pyrolysis oils were characterized by 13C and DEPT NMR analysis to determine chemical functional group compositions as related to their energy content. Pyrolysis oils were produced from a variety of feedstocks including energy crops, woods, animal wastes and oil seed presscakes,...

  3. 1H and 13C Solid-state NMR of Gossypium barbadense (Pima) Cotton

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in G. barbadense (Pima) cotton were investigated by 1H and 13C solid-state NMR techniques. 1H spin diffusion results from a Goldman-Shen experiment indicate that the water is multilayered. 1H MAS experiments pro...

  4. 1H and 13C Solid-state NMR of G. barbadense (Pima) Cotton

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in G. barbadense (Pima) cotton were investigated with 1H and 13C solid-state NMR techniques. 1H spin diffusion results from a Goldman-Shen experiment indicate that the water is multilayered. 1H MAS experiment...

  5. Trophic ecology of the rocky shore community associated with the Ascophyllum nodosum zone (Roscoff, France): A δ 13C vs δ 15N investigation

    NASA Astrophysics Data System (ADS)

    Riera, Pascal; Escaravage, Carole; Leroux, Cédric

    2009-01-01

    This study aimed to characterize the structure and functioning of the benthic food web associated with the Ascophyllum nodosum zone of the rocky shore of Roscoff by using δ 13C and δ 15N. Several characteristics of the trophic ecology of the invertebrates associated with this mid-littoral habitat and which belong to different functional groups (e.g., grazers, filter-feeders, predators and omnivores) were highlighted. In particular, the filter feeder species (including mostly sponges) used macroalgae-derived organic matter as a substantial food requirement. The results also pointed out an important stable isotopes variability for strict coexisting primary consumers which: (1) is directly related to the high δ 15N range of the food sources; (2) makes it impossible to establish a unique trophic level scale based on δ 15N values, as previously done in coastal environments; and (3) points out the existence of major co-occurring trophic pathways which characterise the Ascophyllum nodosum habitat.

  6. Nicotine, acetanilide and urea multi-level 2H-, 13C- and 15N-abundance reference materials for continuous-flow isotope ratio mass spectrometry.

    PubMed

    Schimmelmann, Arndt; Albertino, Andrea; Sauer, Peter E; Qi, Haiping; Molinie, Roland; Mesnard, François

    2009-11-01

    Accurate determinations of stable isotope ratios require a calibration using at least two reference materials with different isotopic compositions to anchor the isotopic scale and compensate for differences in machine slope. Ideally, the delta values of these reference materials should bracket the isotopic range of samples with unknown delta values. While the practice of analyzing two isotopically distinct reference materials is common for water (VSMOW-SLAP) and carbonates (NBS 19 and L-SVEC), the lack of widely available organic reference materials with distinct isotopic composition has hindered the practice when analyzing organic materials by elemental analysis/isotope ratio mass spectrometry (EA-IRMS). At present only L-glutamic acids USGS40 and USGS41 satisfy these requirements for delta13C and delta15N, with the limitation that L-glutamic acid is not suitable for analysis by gas chromatography (GC). We describe the development and quality testing of (i) four nicotine laboratory reference materials for on-line (i.e. continuous flow) hydrogen reductive gas chromatography-isotope ratio mass-spectrometry (GC-IRMS), (ii) five nicotines for oxidative C, N gas chromatography-combustion-isotope ratio mass-spectrometry (GC-C-IRMS, or GC-IRMS), and (iii) also three acetanilide and three urea reference materials for on-line oxidative EA-IRMS for C and N. Isotopic off-line calibration against international stable isotope measurement standards at Indiana University adhered to the 'principle of identical treatment'. The new reference materials cover the following isotopic ranges: delta2H(nicotine) -162 to -45 per thousand, delta13C(nicotine) -30.05 to +7.72 per thousand, delta15N(nicotine) -6.03 to +33.62 per thousand; delta15N(acetanilide) +1.18 to +40.57 per thousand; delta13C(urea) -34.13 to +11.71 per thousand, delta15N(urea) +0.26 to +40.61 per thousand (recommended delta values refer to calibration with NBS 19, L-SVEC, IAEA-N-1, and IAEA-N-2). Nicotines fill a gap as

  7. DFT study of zigzag (n, 0) single-walled carbon nanotubes: (13)C NMR chemical shifts.

    PubMed

    Kupka, Teobald; Stachów, Michal; Stobiński, Leszek; Kaminský, Jakub

    2016-06-01

    (13)C NMR chemical shifts of selected finite-size models of pristine zigzag single walled carbon nanotubes (SWCNTs) with a diameter of ∼0.4-0.8nm and length up to 2.2nm were studied theoretically. Results for finite SWCNTs models containing 1, 4 and 10 adjacent bamboo-type units were compared with data obtained for infinite tubes in order to estimate the reliability of small finite models in predicting magnetic properties of real-size nanotubes and to assess their tube-length dependence. SWCNTs were fully optimized using unrestricted density functional theory (DFT-UB3LYP/6-31G*). Cyclacenes, as the shortest models of open-ended zigzag SWCNTs, with systematically varying diameter were calculated as well. GIAO NMR calculations on the SWCNT and cyclacene models were performed using the BHandH density functional combined with relatively small STO-3Gmag basis set, developed by Leszczyński and coworkers for accurate description of magnetic properties. Regular changes of carbon (13)C chemical shifts along the tube axis of real size (6, 0) and (9, 0) zigzag carbon nanotubes were shown. The (13)C NMR shifts according to increasing diameter calculated for zigzag (n, 0, n=5-10) cyclacenes followed the trends observed for zigzag (n, 0) SWCNTs. The results for 4-units long SWCNTs match reasonably well with the data obtained for infinite zigzag (n, 0) SWCNTs, especially to those with bigger diameter (n=8-15). The presence of rim hydrogens obviously affects theoretical (13)C chemical shieldings and shifts in cyclacenes and thus cyclacenes can provide only approximate estimation of (13)C NMR parameters of real-size SWCNTs. The NMR properties predicted for the longest 10-units long models of SWCNTs reliably correspond to results obtained for infinite nanotubes. They were thus able to accurately predict also recently reported experimental chemical shift of chiral (6, 5) SWCNT. PMID:27155813

  8. Characterisation of the 1H and 13C NMR spectra of methylcitric acid

    NASA Astrophysics Data System (ADS)

    Krawczyk, Hanna; Martyniuk, Tomasz

    2007-06-01

    Methylcitric acid (MCA) was synthesised in Reformatsky reaction (2 RS, 3 RS stereoisomers) and in the nucleophilic addition (2 RS, 3 SR stereoisomers). The stereoselectivity of these reactions was analysed. 1H and 13C NMR spectra of diastereoisomers of methylcitric acid were recorded and interpreted. The values of 1H chemical shifts and 1H- 1H coupling constants were analysed. Proton-decoupled high-resolution 13C NMR spectra of MCA diastereoisomers were measured in a series of dilute water solutions of various acidities. These data may provide a basis for unequivocal determination of the presence of MCA in the urine samples of patients' suffering from propionic acidemia, methylmalonic aciduria, or holocarboxylase synthetase deficiency. NMR spectroscopy enables determination of MCA diastereoisomers in body fluids and can be a complementary and useful diagnostic tool.

  9. Effect of salinity and water stress during the reproductive stage on growth, ion concentrations, Delta 13C, and delta 15N of durum wheat and related amphiploids.

    PubMed

    Yousfi, Salima; Serret, Maria Dolores; Voltas, Jordi; Araus, José Luis

    2010-08-01

    The physiological performance of durum wheat and two related amphiploids was studied during the reproductive stage under different combinations of salinity and irrigation. One triticale, one tritordeum, and four durum wheat genotypes were grown in pots in the absence of stress until heading, when six different treatments were imposed progressively. Treatments resulted from the combination of two irrigation regimes (100% and 35% of container water capacity) with three levels of water salinity (1.8, 12, and 17 dS m(-1)), and were maintained for nearly 3 weeks. Gas exchange and chlorophyll fluorescence and content were measured prior to harvest; afterwards shoot biomass and height were recorded, and Delta(13)C, delta(15)N, and the concentration of nitrogen (N), phosphorus, and several ions (K(+), Na(+), Ca(2+), Mg(2+)) were analysed in shoot material. Compared with control conditions (full irrigation with Hoagland normal) all other treatments inhibited photosynthesis through stomatal closure, accelerated senescence, and decreased biomass. Full irrigation with 12 dS m(-1) outperformed other stress treatments in terms of biomass production and physiological performance. Biomass correlated positively with N and delta(15)N, and negatively with Na(+) across genotypes and fully irrigated treatments, while relationships across deficit irrigation conditions were weaker or absent. Delta(13)C did not correlate with biomass across treatments, but it was the best trait correlating with phenotypic differences in biomass within treatments. Tritordeum produced more biomass than durum wheat in all treatments. Its low Delta(13)C and high K(+)/Na(+) ratio, together with a high potential growth, may underlie this finding. Mechanisms relating delta(15)N and Delta(13)C to biomass are discussed. PMID:20660293

  10. The structural properties of the transmembrane segment of the integral membrane protein phospholamban utilizing (13)C CPMAS, (2)H, and REDOR solid-state NMR spectroscopy.

    PubMed

    Karp, Ethan S; Tiburu, Elvis K; Abu-Baker, Shadi; Lorigan, Gary A

    2006-06-01

    Solid-state NMR spectroscopic techniques were used to investigate the secondary structure of the transmembrane peptide phospholamban (TM-PLB), a sarcoplasmic Ca(2+) regulator. (13)C cross-polarization magic angle spinning spectra of (13)C carbonyl-labeled Leu39 of TM-PLB exhibited two peaks in a pure 1-palmitoyl-2-oleoyl-phosphocholine (POPC) bilayer, each due to a different structural conformation of phospholamban as characterized by the corresponding (13)C chemical shift. The addition of a negatively charged phospholipid (1-palmitoyl-2-oleoylphosphatidylglycerol (POPG)) to the POPC bilayer stabilized TM-PLB to an alpha-helical conformation as monitored by an enhancement of the alpha-helical carbonyl (13)C resonance in the corresponding NMR spectrum. (13)C-(15)N REDOR solid-state NMR spectroscopic experiments revealed the distance between the (13)C carbonyl carbon of Leu39 and the (15)N amide nitrogen of Leu42 to be 4.2+/-0.2A indicating an alpha-helical conformation of TM-PLB with a slight deviation from an ideal 3.6 amino acid per turn helix. Finally, the quadrupolar splittings of three (2)H labeled leucines (Leu28, Leu39, and Leu51) incorporated in mechanically aligned DOPE/DOPC bilayers yielded an 11 degrees +/-5 degrees tilt of TM-PLB with respect to the bilayer normal. In addition to elucidating valuable TM-PLB secondary structure information, the solid-state NMR spectroscopic data indicates that the type of phospholipids and the water content play a crucial role in the secondary structure and folding of TM-PLB in a phospholipid bilayer. PMID:16839519

  11. Food web structure of the epibenthic and infaunal invertebrates on the Catalan slope (NW Mediterranean): Evidence from δ 13C and δ 15N analysis

    NASA Astrophysics Data System (ADS)

    Fanelli, E.; Papiol, V.; Cartes, J. E.; Rumolo, P.; Brunet, C.; Sprovieri, M.

    2011-01-01

    The food-web structure of the epibenthic and infaunal invertebrates on the continental slope of the Catalan Sea (Balearic basin, NW Mediterranean) was investigated using carbon and nitrogen stable isotopes on a total of 34 species, and HPLC pigment analyses for three key species. Samples were collected close to Barcelona (NE Iberian Peninsula), between 650 and 800 m depth and between February 2007 and February 2008. Mean δ 13C values ranged from -21.0‰ (small Calocaris macandreae and Amphipholis squamata) to -14.5‰ ( Sipunculus norvegicus). Values of δ 15N ranged from 4.0‰ ( A. squamata) to 12.1‰ ( Molpadia musculus). The stable isotope ratios of benthic fauna displayed a continuum of values (e.g. δ 15N range of 8‰), confirming a wide spectrum of feeding strategies (from active suspension feeders to predators) and complex food webs. According to the available information on diets of benthic fauna, the lowest values were found for surface deposit feeders (small C. macandrae and the two ophiuroids A. squamata and Amphiura chiajei) and active suspension feeders ( Abra longicallus and Scalpellum scalpellum) feeding on different sizes of particulate organic matter (POM), among which small particles may exhibit lower δ 15N. High annual mean δ 15N values were found among sub-surface deposit feeders, exploiting refractory or frequently recycled organic matter that is enriched in δ 15N. Carnivorous polychaetes ( Nephtys spp., Oenonidae and Polynoidae) and large decapods ( Geryon longipes and Paromola cuvieri) also displayed high δ 15N values. δ 13C ranges were particularly wide among surface deposit feeders (ranging from -21.0‰ to -16.4‰), suggesting exploitation of POM of both terrigenous and oceanic origins. Correlation between δ 13C and δ 15N was generally weak, indicating multiple carbon sources, likely due to the consumption of different kinds of sinking particles (e.g. marine snow, phytodetritus, etc.), sedimented and frequently recycled POM

  12. Natural abundances of 15N and 13C in leaves of some N2-fixing and non-N2-fixing trees and shrubs in Syria.

    PubMed

    Kurdali, F; Al-Shamma'a, M

    2009-09-01

    A survey study was conducted on man-made plantations located at two different areas in the arid region of Syria to determine the variations in natural abundances of the (15)N and (13)C isotopes in leaves of several woody legume and non-legume species, and to better understand the consequence of such variations on nitrogen fixation and carbon assimilation. In the first study area (non-saline soil), the delta(15)N values in four legume species (Acacia cyanophylla,-1.73 per thousand Acacia farnesiana,-0.55 per thousand Prosopis juliflora,-1.64 per thousand; and Medicago arborea,+1.6 \\textperthousand) and one actinorhizal plant (Elaeagnus angustifolia,-0.46 to-2.1 per thousand) were found to be close to that of the atmospheric value pointing to a major contribution of N(2) fixing in these species; whereas, delta(15)N values of the non-fixing plant species were highly positive. delta(13)C per thousand; in leaves of the C3 plants were found to be affected by plant species, ranging from a minimum of-28.67 per thousand; to a maximum of-23 per thousand. However, they were relatively similar within each plant species although they were grown at different sites. In the second study area (salt affected soil), a higher carbon discrimination value (Delta(13)C per thousand) was exhibited by P. juliflora, indicating that the latter is a salt tolerant species; however, its delta(15)N was highly positive (+7.03 per thousand) suggesting a negligible contribution of the fixed N(2). Hence, it was concluded that the enhancement of N(2) fixation might be achieved by selection of salt-tolerant Rhizobium strains. PMID:20183233

  13. Millennial scale oscillations in bulk δ15N and δ13C over the Mid- to Late Holocene seen in proteinaceous corals from the North Pacific Subtropical Gyre

    NASA Astrophysics Data System (ADS)

    Glynn, D. S.; Mccarthy, M. D.; McMahon, K.; Guilderson, T. P.

    2014-12-01

    The North Pacific Subtropical Gyre (NPSG) is the largest continuous ecosystem on this planet and is an important regulator of biogeochemical cycling and carbon sequestration. With evidence of its expansion in a warming climate, it is necessary to develop a more complete understanding of the variability in productivity and nutrient dynamics in this important ecosystem through time. We constructed a long-term, high resolution record of bulk record of stable nitrogen (δ15N) and carbon isotopes (δ13C) from multiple proteinaceous deep sea corals around Hawaii extending back ~5300 years with few gaps. Our data confirms the decreasing trend in δ15N since the Little Ice Age (1850s), which matches previously published results in part attributed to anthropogenic climate change (e.g. Sherwood et al. 2014). However, while the rate of change since the Little Ice Age (δ15N declines ~1‰ over ~150yrs) remains by far the most rapid throughout the longer record, there also appear to be longer-term (near-millennial scale) climatic oscillations of even greater magnitude (δ15N shifts ~1.5-2‰ over ~1000yrs). After removal of the Seuss Effect, δ13C values also declined ~1.5‰ since the Little Ice Age. Furthermore, there also appear to be oscillations in δ13C of ~1-2‰ over millennial timescales. These results reveal the existence of previously unrecognized long-term oscillations in NPSG biogeochemical cycles, which are likely linked to changes in phytoplankton species composition, food web dynamics, and/or variability in source nutrients and productivity possibly caused by changes in climate. This study provides insight into nutrient dynamics in the NPSG over the past five millennia, and offers a historical baseline to better analyze the effects of current anthropogenic climate forcing.

  14. 13C/12C and 15N/14N Isotope Analysis to Characterize Natural Degradation of Atrazine: Evidence from Parent and Daughter Compound Values

    NASA Astrophysics Data System (ADS)

    Elsner, Martin; Meyer, Armin

    2013-04-01

    The mobile and still herbicidal metabolites desethylatrazine (DEA) and desisopropylatrazine (DIA) are frequently detected together with its parent compound atrazine (Atz) in the aquatic environment. Interpretation of their transformation state is often difficult with current methods, which are mainly measuring concentrations. Alternatively, compound specific isotope analyses (CSIA) has become a novel tool to detect degradation processes of contaminants in groundwater. The aim of our study was to investigate on the lab scale 13C/12C and 15N/14N isotope trends in parent and daughter compounds associated with different degradation scenarios of atrazine likely to occur in the environment. Thus atrazine was dealkylated with (i) permanganate and (ii) the bacterium Rhodococcus sp. NI86/21. In both transformations, 13C/12C ratios of atrazine increased strongly (epsilon carbon/permanganate = -4.6 ± 0.6 ‰ and epsilon carbon/Rhodoccoccus = -3.8 ± 0.2 ‰) whereas nitrogen isotope fractionation was small. 13C/12C ratios of DEA showed the following trends. (i) When DEA was formed as only product (Atz + permanganate) 13C/12C remained constant, close to the initial value of Atz. (ii) When DEA was formed together with deisopropylatrazine (biodegradation of Atz) 13C/12C increased, but only within 2‰. (iii) When DEA and DIA was further biodegraded, 13C/12C increased for both metabolites up to 9‰. Thus strong enrichment of 13C/12C in the metabolites in comparison to Atz can give strong testimony for further breakdown of the metabolite.

  15. Methylation patterns of aquatic humic substances determined by 13C NMR spectroscopy

    USGS Publications Warehouse

    Thorn, K.A.; Steelink, C.; Wershaw, R. L.

    1987-01-01

    13C NMR spectroscopy is used to examine the hydroxyl group functionality of a series of humic and fulvic acids from different aquatic environments. Samples first are methylated with 13C-labeled diazomethane. The NMR spectra of the diazomethylated samples allow one to distinguish between methyl esters of carboxylic acids, methyl ethers of phenolic hydroxyls, and methyl ethers of phenolic hydroxyls adjacent to two substituents. Samples are then permethylated with 13C-labeled methyl iodide/NaH. 13C NMR spectra of permethylated samples show that a significant fraction of the hydroxyl groups is not methylated with diazomethane alone. In these spectra methyl ethers of carbohydrate and aliphatic hydroxyls overlap with methyl ethers of phenolic hydroxyls. Side reactions of the methyltion procedure including carbon methylation in the CH3I/NaH procedure, are also examined. Humic and fulvic acids from bog, swamp, groundwater, and lake waters showssome differences in their distribution of hydroxyl groups, mainly in the concentrations of phenolic hydroxyls, which may be attributed to their different biogeochemical origins. ?? 1987.

  16. Chemical shift changes and line narrowing in 13C NMR spectra of hydrocarbon clathrate hydrates.

    PubMed

    Kida, Masato; Sakagami, Hirotoshi; Takahashi, Nobuo; Nagao, Jiro

    2013-05-23

    The solid-state (13)C NMR spectra of various guest hydrocarbons (methane, ethane, propane, adamantane) in clathrate hydrates were measured to elucidate the local structural environments around hydrocarbon molecules isolated in guest-host frameworks of clathrate hydrates. The results show that, depending on the cage environment, the trends in the (13)C chemical shift and line width change as a function of temperature. Shielding around the carbons of the guest normal alkanes in looser cage environments tends to decrease with increasing temperature, whereas shielding in tighter cage environments tends to increase continuously with increasing temperature. Furthermore, the (13)C NMR line widths suggest, because of the reorientation of the guest alkanes, that the local structures in structure II are more averaged than those in structure I. The differences between structures I and II tend to be very large in the lower temperature range examined in this study. The (13)C NMR spectra of adamantane guest molecules in structure H hydrate show that the local structures around adamantane guests trapped in structure H hydrate cages are averaged at the same level as in the α phase of solid adamantane. PMID:23607335

  17. (13)C NMR Studies, Molecular Order, and Mesophase Properties of Thiophene Mesogens.

    PubMed

    Veeraprakash, B; Lobo, Nitin P; Narasimhaswamy, T

    2015-12-01

    Three-ring mesogens with a core comprising thiophene linked to one phenyl ring directly and to the other via flexible ester are synthesized with terminal alkoxy chains to probe the mesophase properties and find the molecular order. The phenyl thiophene link in the core offers a comparison of the mesophase features with the molecular shape of the mesogen. The synthesized mesogens display enantiotropic polymesomorphism and accordingly nematic, smectic A, smectic C and smectic B mesophases are perceived depending upon the terminal chain length. For some of the homologues, monotropic higher order smectic phases such as smectic F and crystal E are also witnessed. The existence of polymesomorphism are originally observed by HOPM and DSC and further confirmed by powder X-ray diffraction studies. For the C8 homologue, high resolution solid state (13)C NMR spectroscopy is employed to find the molecular structure in the liquid crystalline phase and using the 2D SLF technique, the (13)C-(1)H dipolar couplings are extracted to calculate the order parameter. By comparing the ratio of local order of thiophene as well as phenyl rings, we establish the bent-core shape of the mesogen. Importantly, for assigning the carbon chemical shifts of the core unit of aligned C8 mesogen, the (13)C NMR measured in mesophase of the synthetic intermediate is employed. Thus, the proposed approach addresses the key step in the spectral assignment of target mesogens with the use of (13)C NMR data of mesomorphic intermediate. PMID:26551439

  18. Identification of natural metabolites in mixture: a pattern recognition strategy based on (13)C NMR.

    PubMed

    Hubert, Jane; Nuzillard, Jean-Marc; Purson, Sylvain; Hamzaoui, Mahmoud; Borie, Nicolas; Reynaud, Romain; Renault, Jean-Hugues

    2014-03-18

    Because of their highly complex metabolite profile, the chemical characterization of bioactive natural extracts usually requires time-consuming multistep purification procedures to achieve the structural elucidation of pure individual metabolites. The aim of the present work was to develop a dereplication strategy for the identification of natural metabolites directly within mixtures. Exploiting the polarity range of metabolites, the principle was to rapidly fractionate a multigram quantity of a crude extract by centrifugal partition extraction (CPE). The obtained fractions of simplified chemical composition were subsequently analyzed by (13)C NMR. After automatic collection and alignment of (13)C signals across spectra, hierarchical clustering analysis (HCA) was performed for pattern recognition. As a result, strong correlations between (13)C signals of a single structure within the mixtures of the fraction series were visualized as chemical shift clusters. Each cluster was finally assigned to a molecular structure with the help of a locally built (13)C NMR chemical shift database. The proof of principle of this strategy was achieved on a simple model mixture of commercially available plant secondary metabolites and then applied to a bark extract of the African tree Anogeissus leiocarpus Guill. & Perr. (Combretaceae). Starting from 5 g of this genuine extract, the fraction series was generated by CPE in only 95 min. (13)C NMR analyses of all fractions followed by pattern recognition of (13)C chemical shifts resulted in the unambiguous identification of seven major compounds, namely, sericoside, trachelosperogenin E, ellagic acid, an epimer mixture of (+)-gallocatechin and (-)-epigallocatechin, 3,3'-di-O-methylellagic acid 4'-O-xylopyranoside, and 3,4,3'-tri-O-methylflavellagic acid 4'-O-glucopyranoside. PMID:24555703

  19. Early life histories of the London poor using δ13C and δ15N stable isotope incremental dentine sampling.

    PubMed

    Henderson, Rowena C; Lee-Thorp, Julia; Loe, Louise

    2014-08-01

    High resolution incremental isotopic analysis of the dentine from early forming teeth, especially first molars (M1s), provides a means to assess the effects of poor childhood nutrition and healthcare on individuals in an assemblage where there are no infants to study. This approach is applied to an 18th and 19th century cemetery population associated with St Saviour's Almshouse burial ground in Southwark, London, to assess whether, or how, early dietary history, including weaning age, influenced health and nutritional status. The results show a general pattern in which non-breast milk foods were introduced before or by 6 months of age, as indicated by elevated δ(15) N during this period. Almost all individuals for which we also have second molar (M2) records, showed lower δ(15) N values from a very young age (>1 year) until approximately 8-10 years, compared to adult values. The overall results show a significant difference in δ(1) (3) C (p = 0 to 4sf, F = 17.327) and a weaker statistical difference in δ(15) N between males and females (p = 0.019, F = 5.581). One possible cause of this is a difference in the diet of males and females early in life, or alternatively, a greater susceptibility of males to nutritional deprivation compared to females. The latter argument is strengthened by a significant difference in the incidence of enamel hypoplasia between the males and females, with 7.7% of male teeth showing defects, compared to 3.9% of females. PMID:24898314

  20. Application of 13C NMR spectroscopy to paratope mapping for larger antigen-Fab complexes.

    PubMed

    Kim, H; Kato, K; Yamato, S; Igarashi, T; Matsunaga, C; Ohtsuka, H; Higuchi, A; Nomura, N; Noguchi, H; Arata, Y

    1994-06-13

    For the purpose of engineering the antibody combining site, mapping residues that are involved in antigen binding provide us with valuable information. By use of 13C NMR spectroscopy with selectively 13C-labeled Fv fragments, we have established a general strategy to identify the residues that are perturbed upon binding of small antigen (hapten) molecules [(1990) Biochemistry 30, 6604-6610]. In the present paper, we demonstrate that this strategy can be extended to molecular structural analyses of the complexes of an Fab fragment and a larger antigen molecule such as Pseudomonas aeruginosa exotoxin A with a molecular mass of 67 kDa. PMID:8013642

  1. Determination of fructose metabolic pathways in normal and fructose-intolerant children: A sup 13 C NMR study using (U- sup 13 C)fructose

    SciTech Connect

    Gopher, A.; Lapidot, A. ); Vaisman, N. ); Mandel, H. )

    1990-07-01

    An inborn deficiency in the ability of aldolase B to split fructose 1-phosphate is found in humans with hereditary fructose intolerance (HFI). A stable isotope procedure to elucidate the mechanism of conversion of fructose to glucose in normal children and in HFI children has been developed. A constant infusion of D-(U-{sup 13}C)fructose was given nasogastrically to control and to HFI children. Hepatic fructose conversion to glucose was estimated by examination of {sup 13}C NMR spectra of plasma glucose. Significantly lower values ({approx}3-fold) for fructose conversion to glucose were obtained for the HFI patients as compared to the controls. A quantitative determination of the metabolic pathways of fructose conversion to glucose was derived from {sup 13}C NMR measurement of plasma ({sup 13}C)glucose isotopomer populations. The finding of isotopomer populations of three adjacent {sup 13}C atoms at glucose C-4 ({sup 13}C{sub 3}-{sup 13}C{sub 4}-{sup 13}C{sub 5}) suggests that there is a direct pathway from fructose, by-passing fructose-1-phosphate aldolase, to fructose 1,6-bisphosphate. The metabolism of fructose by fructose-1-phosphate aldolase activity accounts for only {approx}50% of the total amount of hepatic fructose conversion to glucose. In view of the marked decline by 67% in synthesis of glucose from fructose in HFI subjects found in this study, the extent of ({sup 13}C)glucose formation from a trace amount of (U-{sup 13}C)fructose infused into the patient can be used as a safe and noninvasive diagnostic test for inherent faulty fructose metabolism.

  2. Marine mammals from northeast Atlantic: relationship between their trophic status as determined by delta13C and delta15N measurements and their trace metal concentrations.

    PubMed

    Das, K; Beans, C; Holsbeek, L; Mauger, G; Berrow, S D; Rogan, E; Bouquegneau, J M

    2003-09-01

    The relationship between trophic position through delta13C and delta15N and trace metal concentrations (Zn, Cd, Cu and Hg) was investigated in the tissues of six marine mammal species from the Northeast Atlantic: striped dolphin Stenella coeruleoalba, common dolphin, Delphinus delphis, Atlantic white-sided dolphin Lagenorhynchus acutus, harbour porpoise Phocoena phocoena, white beaked-dolphin Lagenorhynchus albirostris, grey seal Halichoerus grypus stranded on French Channel and Irish coasts. White-beaked dolphins, harbour porpoises, white-sided dolphins, common and striped dolphins display the same relative and decreasing trophic position, as measured by delta15N values, along both the Irish and French channel coasts, reflecting conservative trophic habits between these two places. Hepatic and renal Cd concentrations were significantly correlated to muscle delta13C and delta15N values while Hg, Zn and Cu did not. These results suggest that Cd accumulation is partly linked to the diet while other factors such as age or body condition might explain Hg, Zn or Cu variability in marine mammals. Combined stable isotope and trace metal analyses appear to be useful tools for the study of marine mammal ecology. PMID:12738219

  3. Paleobiological Implications of the Isotopic Signatures ( 13C, 15N) of Fossil Mammal Collagen in Scladina Cave (Sclayn, Belgium)

    NASA Astrophysics Data System (ADS)

    Bocherens, Hervé; Billiou, Daniel; Patou-Mathis, Marylène; Bonjean, Dominique; Otte, Marcel; Mariotti, André

    1997-11-01

    An isotopic investigation of upper Pleistocene mammal bones and teeth from Scladina cave (Sclayn, Belgium) demonstrated the very good quality of collagen preservation. A preliminary screening of the samples used the amount of nitrogen in whole bone and dentine in order to estimate the preserved amount of collagen before starting the extraction process. The isotopic abundances of fossil specimens from still-extant species are consistent with their trophic position. Moreover, the 15N isotopic abundance is higher in dentine than in bone in bears and hyenas, a phenomenon already observed in modern specimens. These results demonstrate that the isotopic compositions of samples from Scladina cave can be interpreted in ecological terms. Mammoths exhibit a high 15N isotopic abundance relative to other herbivores, as was the case in Siberian and Alaskan samples. These results suggest distinctive dietary adaptations in herbivores living in the mammoth steppe. Cave bears are clearly isotopically different from coeval brown bears, suggesting an ecological separation between species, with a pure vegetarian diet for cave bear and an omnivorous diet for brown bear.

  4. Molecular structure of crude beeswax studied by solid-state 13C NMR

    PubMed Central

    Kameda, Tsunenori

    2004-01-01

    13C Solid-state NMR experiments were performed to investigate the structure of beeswax in the native state (crude beeswax) for the first time. From quantitative direct polarization 13C MAS NMR spectrum, it was found that the fraction of internal-chain methylene (int-(CH2)) component compared to other components of crude beeswax was over 95%. The line shape of the int-(CH2) carbon resonance region was comprehensively analyzed in terms of NMR chemical shift. The 13C broad peak component covering from 31 to 35ppm corresponds to int-(CH2) carbons with trans conformation in crystalline domains, whereas the sharp signal at 30.3 ppm corresponds to gauche conformation in the non-crystalline domain. From peak deconvolution of the aliphatic region, it was found that over 85% of the int-(CH2) has a crystal structure and several kinds of molecular packing for int-(CH2), at least three, exist in the crystalline domain. Abbreviation: NMR nuclear magnetic resonance int-(CH2) internal-chain methylene CP cross-polarization MAS magic angle spinning PMID:15861244

  5. Quantification of protein secondary structure by (13)C solid-state NMR.

    PubMed

    Andrade, Fabiana Diuk; Forato, Lucimara Aparecida; Bernardes Filho, Rubens; Colnago, Luiz Alberto

    2016-05-01

    High-resolution (13)C solid-state NMR stands out as one of the most promising techniques to solve the structure of insoluble proteins featuring biological and technological importance. The simplest nuclear magnetic resonance (NMR) spectroscopy method to quantify the secondary structure of proteins uses the areas of carbonyl and alpha carbon peaks. The quantification obtained by fitting procedures depends on the assignment of the peaks to the structure, type of line shape, number of peaks to be used, and other parameters that are set by the operator. In this paper, we demonstrate that the analysis of (13)C NMR spectra by a pattern recognition method-based on the singular value decomposition (SVD) regression, which does not depend on the operator-shows higher correlation coefficients for α-helix and β-sheet (0.96 and 0.91, respectively) than Fourier transform infrared spectroscopy (FTIR) method. Therefore, the use of (13)C solid-state NMR spectra and SVD is a simple and reliable method for quantifying the secondary structures of insoluble proteins in solid-state. PMID:27068694

  6. Direct (13)C-detected NMR experiments for mapping and characterization of hydrogen bonds in RNA.

    PubMed

    Fürtig, Boris; Schnieders, Robbin; Richter, Christian; Zetzsche, Heidi; Keyhani, Sara; Helmling, Christina; Kovacs, Helena; Schwalbe, Harald

    2016-03-01

    In RNA secondary structure determination, it is essential to determine whether a nucleotide is base-paired and not. Base-pairing of nucleotides is mediated by hydrogen bonds. The NMR characterization of hydrogen bonds relies on experiments correlating the NMR resonances of exchangeable protons and can be best performed for structured parts of the RNA, where labile hydrogen atoms are protected from solvent exchange. Functionally important regions in RNA, however, frequently reveal increased dynamic disorder which often leads to NMR signals of exchangeable protons that are broadened beyond (1)H detection. Here, we develop (13)C direct detected experiments to observe all nucleotides in RNA irrespective of whether they are involved in hydrogen bonds or not. Exploiting the self-decoupling of scalar couplings due to the exchange process, the hydrogen bonding behavior of the hydrogen bond donor of each individual nucleotide can be determined. Furthermore, the adaption of HNN-COSY experiments for (13)C direct detection allows correlations of donor-acceptor pairs and the localization of hydrogen-bond acceptor nucleotides. The proposed (13)C direct detected experiments therefore provide information about molecular sites not amenable by conventional proton-detected methods. Such information makes the RNA secondary structure determination by NMR more accurate and helps to validate secondary structure predictions based on bioinformatics. PMID:26852414

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

    PubMed Central

    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. PMID:25923696

  8. 1H to 13C Energy Transfer in Solid State NMR Spectroscopy of Natural Organic Systems

    NASA Astrophysics Data System (ADS)

    Berns, Anne E.; Conte, Pellegrino

    2010-05-01

    Cross polarization (CP) magic angle spinning (MAS) 13C-NMR spectroscopy is a solid state NMR technique widely used to study chemical composition of organic materials with low or no solubility in the common deuterated solvents used to run liquid state NMR experiments. Based on the magnetization transfer from abundant nuclei (with spin of 1 -2) having a high gyromagnetic ratio (γ), such as protons, to the less abundant 13C nuclei with low γ values, 13C-CPMAS NMR spectroscopy is often applied in environmental chemistry to obtain quantitative information on the chemical composition of natural organic matter (NOM) (Conte et al., 2004), although its quantitative assessment is still matter of heavy debates. Many authors (Baldock et al., 1997; Conte et al., 1997, 2002; Dria et al., 2002; Kiem et al., 2000; Kögel-Knabner, 2000; Preston, 2001), reported that the application of appropriate instrument setup as well as the use of special pulse sequences and correct spectra elaboration may provide signal intensities that are directly proportional to the amount of nuclei creating a NMR signal. However, many other papers dealt with the quantitative unsuitability of 13C-CPMAS NMR spectroscopy. Among those, Mao et al. (2000), Smernik and Oades (2000 a,b), and Preston (2001) reported that cross-polarized NMR techniques may fail in a complete excitation of the 13C nuclei. In fact, the amount of observable carbons via 13C-CPMAS NMR spectroscopy appeared, in many cases, lower than that measured by a direct observation of the 13C nuclei. As a consequence, cross-polarized NMR techniques may provide spectra where signal distribution may not be representative of the quantitative distribution of the different natural organic matter components. Cross-polarization is obtained after application of an initial 90° x pulse on protons and a further spin lock pulse (along the y axis) having a fixed length (contact time) for both nuclei (1H and 13C) once the Hartmann-Hahn condition is matched

  9. 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)

    SciTech Connect

    Hodsdon, M.E.; Toner, J.J.; Cistola, D.P.

    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.

  10. Characterization (δ13C, δ15N and TOC/TN) of marine sediments from restored seagrass (Zostera marina) meadows in coastal lagoons of Virginia

    NASA Astrophysics Data System (ADS)

    Egge, N.; Macko, S. A.

    2011-12-01

    Restoration of the Chesapeake Bay and its neighboring coastal areas has been a major focus of scientific research in the last ten years. The decline of the area is often linked with the explosive population growth and subsequent increases in agriculture and urbanization of the region. In the coastal bays of Virginia, large efforts have been made to restore seagrass (Zostera marina) meadows, which were devastated by a combination of events in the 1930s. As a result of the restoration efforts begun in 2001, seagrass has come to dominate portions of South Bay and Hog Island Bay. Sedimentary characterization (δ13C, δ15N, total organic carbon and total nitrogen) is used to assess sources of organic matter in marine environments. Push cores were used to collect samples from sites that have different records of seagrass cover. Sediments dominated by seagrass contain organic matter enriched in 13C relative to autochthonous algal sources. Within the sediment record there are two regions that show heightened 13C content-one near the surface and another at lower depths. Organic matter enriched in 13C near the surface is from current seagrass cover. Similar characteristics of sediment deeper in the core imply that the area was previously dominated comparable vegetation. Results may be used as a proxy to infer the spatial extent of the historic (prior to 1930) presence of seagrass and guide future restoration projects.

  11. Analysis of carbon and nitrogen co-metabolism in yeast by ultrahigh-resolution mass spectrometry applying 13C- and 15N-labeled substrates simultaneously.

    PubMed

    Blank, Lars M; Desphande, Rahul R; Schmid, Andreas; Hayen, Heiko

    2012-06-01

    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 (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., (13)C and (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 (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 (13)C-labeled glucose and (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. PMID:22543713

  12. (1)H, (15)N and (13)C chemical shift assignment of the Gram-positive conjugative transfer protein TraHpIP501.

    PubMed

    Fercher, Christian; Keller, Walter; Zangger, Klaus; Helge Meyer, N

    2016-04-01

    Conjugative transfer of DNA represents the most important transmission pathway in terms of antibiotic resistance and virulence gene dissemination among bacteria. TraH is a putative transfer protein of the type IV secretion system (T4SS) encoded by the Gram-positive (G+) conjugative plasmid pIP501. This molecular machine involves a multi-protein core complex spanning the bacterial envelope thereby serving as a macromolecular secretion channel. Here, we report the near complete (1)H, (13)C and (15)N resonance assignment of a soluble TraH variant comprising the C-terminal domain. PMID:26559076

  13. Sequence-specific (1)H, (13)C and (15)N backbone resonance assignments of the plakin repeat domain of human envoplakin.

    PubMed

    Jeeves, Mark; Fogl, Claudia; Al-Jassar, Caezar; Chidgey, Martyn; Overduin, Michael

    2016-04-01

    The plakin repeat domain is a distinctive hallmark of the plakin superfamily of proteins, which are found within all epithelial tissues. Plakin repeat domains mediate the interactions of these proteins with the cell cytoskeleton and are critical for the maintenance of tissue integrity. Despite their biological importance, no solution state resonance assignments are available for any homologue. Here we report the essentially complete (1)H, (13)C and (15)N backbone chemical shift assignments of the singular 22 kDa plakin repeat domain of human envoplakin, providing the means to investigate its interactions with ligands including intermediate filaments. PMID:26590577

  14. Determination of the δ15N and δ13C of total nitrogen and carbon in solids; RSIL lab code 1832

    USGS Publications Warehouse

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

    2006-01-01

    The purpose of the Reston Stable Isotope Laboratory (RSIL) lab code 1832 is to determine the δ(15N/14N), abbreviated as δ15N, and the δ(13C/12C), abbreviated as δ13C, of total nitrogen and carbon in a solid sample. A Carlo Erba NC 2500 elemental analyzer (EA) is used to convert total nitrogen and carbon in a solid sample into N2 and CO2 gas. The EA is connected to a continuous flow isotope-ratio mass spectrometer (CF-IRMS), which determines the relative difference in stable nitrogen isotope-amount ratio (15N/14N) of the product N2 gas and the relative difference in stable carbon isotope-amount ratio (13C/12C) of the product CO2 gas. The combustion is quantitative; no isotopic fractionation is involved. Samples are placed 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 helium atmosphere containing an excess of oxygen gas. Combustion products are transported by a helium carrier through a reduction furnace to remove excess oxygen and to 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 IRMS through a Finnigan MAT (now Thermo Scientific) ConFlo II interface. The Finnigan MAT ConFlo II interface is used for introducing not only sample into the IRMS but also N2 and CO2 reference gases and helium for sample dilution. The flash combustion is quantitative; no isotopic fractionation is involved. The IRMS is a Thermo Scientific Delta V CF-IRMS. It has a universal triple collector, two wide cups with a narrow cup in the middle; it is capable of measuring mass/charge (m/z) 28, 29, 30 or with a magnet current change 44, 45, 46, simultaneously. The ion beams from these m/z values are as follows: m/z 28 = N2 = 14N/14N; m/z 29 = N2 = 14N/15N primarily; m/z 30 = NO = 14N/16O

  15. (1) H and (13) C NMR data on natural and synthetic capsaicinoids.

    PubMed

    Gómez-Calvario, Víctor; Garduño-Ramírez, María Luisa; León-Rivera, Ismael; Rios, María Yolanda

    2016-04-01

    Capsaicinoids are the compounds responsible for the pungency of chili peppers. These substances have attracted the attention of many research groups in recent decades because of their antinociceptive, analgesic, anti-inflammatory, and anti-obesity properties, among others. There are nearly 160 capsaicinoids reported in the literature. Approximately 25 of them are natural products, while the rest are synthetic or semi-synthetic products. A large amount of NMR data for the capsaicinoids is dispersed throughout literature. Therefore, there is a need to organize all this NMR data in a systematic and orderly way. This review summarizes the (1) H and (13) C NMR data on 159 natural and synthetic capsaicinoids, with a brief discussion of some typical and relevant aspects of these NMR data. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26626418

  16. Solid state 13C NMR characterisation study on fourth generation Ziegler-Natta catalysts.

    PubMed

    Heikkinen, Harri; Liitiä, Tiina; Virkkunen, Ville; Leinonen, Timo; Helaja, Tuulamari; Denifl, Peter

    2012-01-01

    In this study, solid state (13)C NMR spectroscopy was utilised to characterize and identify the metal-ester coordination in active fourth generation (phthalate) Ziegler-Natta catalysts. It is known that different donors affect the active species in ZN catalysts. However, there is still limited data available of detailed molecular information how the donors and the active species are interplaying. One of the main goals of this work was to get better insight into the interactions of donor and active species. Based on the anisotropy tensor values (δ(11), δ(22), δ(33)) from low magic-angle spinning (MAS) (13)C NMR spectra in combination with chemical shift anisotropy (CSA) calculations (δ(aniso) and η), both the coordinative metal (Mg/Ti) and the symmetry of this interaction between metal and the internal donor in the active catalyst (MgCl(2)/TiCl(4)/electron donor) system could be identified. PMID:22425229

  17. Hyperpolarized 13C NMR observation of lactate kinetics in skeletal muscle.

    PubMed

    Park, Jae Mo; Josan, Sonal; Mayer, Dirk; Hurd, Ralph E; Chung, Youngran; Bendahan, David; Spielman, Daniel M; Jue, Thomas

    2015-10-01

    The production of glycolytic end products, such as lactate, usually evokes a cellular shift from aerobic to anaerobic ATP generation and O2 insufficiency. In the classical view, muscle lactate must be exported to the liver for clearance. However, lactate also forms under well-oxygenated conditions, and this has led investigators to postulate lactate shuttling from non-oxidative to oxidative muscle fiber, where it can serve as a precursor. Indeed, the intracellular lactate shuttle and the glycogen shunt hypotheses expand the vision to include a dynamic mobilization and utilization of lactate during a muscle contraction cycle. Testing the tenability of these provocative ideas during a rapid contraction cycle has posed a technical challenge. The present study reports the use of hyperpolarized [1-(13)C]lactate and [2-(13)C]pyruvate in dynamic nuclear polarization (DNP) NMR experiments to measure the rapid pyruvate and lactate kinetics in rat muscle. With a 3 s temporal resolution, (13)C DNP NMR detects both [1-(13)C]lactate and [2-(13)C]pyruvate kinetics in muscle. Infusion of dichloroacetate stimulates pyruvate dehydrogenase activity and shifts the kinetics toward oxidative metabolism. Bicarbonate formation from [1-(13)C]lactate increases sharply and acetyl-l-carnitine, acetoacetate and glutamate levels also rise. Such a quick mobilization of pyruvate and lactate toward oxidative metabolism supports the postulated role of lactate in the glycogen shunt and the intracellular lactate shuttle models. The study thus introduces an innovative DNP approach to measure metabolite transients, which will help delineate the cellular and physiological role of lactate and glycolytic end products. PMID:26347554

  18. A mantle origin for Paleoarchean peridotitic diamonds from the Panda kimberlite, Slave Craton: Evidence from 13C-, 15N- and 33,34S-stable isotope systematics

    NASA Astrophysics Data System (ADS)

    Cartigny, Pierre; Farquhar, James; Thomassot, Emilie; Harris, Jeffrey W.; Wing, Bozwell; Masterson, Andy; McKeegan, Kevin; Stachel, Thomas

    2009-11-01

    In order to address diamond formation and origin in the lithospheric mantle underlying the Central Slave Craton, we report N- and C-stable isotopic compositions and N-contents and aggregation states for 85 diamonds of known paragenesis (73 peridotitic, 8 eclogitic and 4 from lower mantle) from the Panda kimberlite (Ekati Mine, Lac de Gras Area, Canada). For 12 peridotitic and two eclogitic sulfide inclusion-bearing diamonds from this sample set, we also report multiple-sulfur isotope ratios. The 73 peridotitic diamonds have a mean δ13C-value of - 5.2‰ and range from - 6.9 to - 3.0‰, with one extreme value at - 14.1‰. The associated δ15N-values range from - 17.0 to + 8.5‰ with a mean value of - 4.0‰. N-contents range from 0 to 1280 ppm. The 8 eclogitic diamonds have δ13C-values ranging from - 11.2 to - 4.4‰ with one extreme value at - 19.4‰. Their δ15N ranges from - 2.1 to + 7.9‰ and N-contents fall between 0 and 3452 ppm. Four diamonds with an inferred lower mantle origin are all Type II (i.e. nitrogen-free) and have a narrow range of δ13C values, between - 4.5 and - 3.5‰. The δ34S of the 14 analyzed peridotitic and eclogitic sulfide inclusions ranges from - 3.5 to +5.7‰. None of them provide evidence for anomalous δ33S-values; observed variations in δ33S are from +0.19 to - 0.33‰, i.e. within the 2 sigma uncertainties of mantle sulfur ( δ33S = 0‰). At Panda, the N contents and the δ13C of sulfide-bearing peridotitic diamonds show narrower ranges than silicate-bearing peridotitic diamonds. This evidence supports the earlier suggestion established from eclogitic diamonds from the Kaapvaal that sulfide-(±silicate) bearing diamonds sample a more restricted portion of sublithospheric mantle than silicate-(no sulfide) bearing diamonds. Our findings at Panda suggest that sulfide-bearing diamonds should be considered as a specific diamond population on a global-scale. Based on our study of δ34S, Δ 33S, δ15N and δ13C, we find no

  19. Molecular orientational dynamics in C70S48: Investigation by 13C MAS NMR

    NASA Astrophysics Data System (ADS)

    Grell, A.-S.; Talyzin, A.; Pirotte, P.; Masin, F.

    2001-11-01

    At room temperature the MAS 13C NMR spectrum of C70S48 is identical to that of pure C70 above 323 K, except that the 13C line is shifted by 1.7 ppm compared to that of pure C70. From these results, we deduce that our system is mainly of the van der Waals type. A simulation of the low speed MAS spectrum shows that C70 molecules in C70S48 undergo a uniaxial rotation as in pure C70. This new result contradicts what had been previously published. The chemical shift of the 13C line does not vary with temperature, however the rotation of C70 slows down as the temperature is decreased and stops at ca. 150 K. Moreover the 13C spin-lattice relaxation time, T1, can be described by a single correlation time that follows an Arrhenius law with a 900 K activation energy. By NMR no phase transition is observed at 245 K contrary to dielectric relaxation measurements.

  20. δ 13C and δ 15N biogeographic trends in rocky intertidal communities along the coast of South Africa: Evidence of strong environmental signatures

    NASA Astrophysics Data System (ADS)

    Hill, Jaclyn M.; McQuaid, Christopher D.

    2008-11-01

    Ecosystem dynamics driven by top-down controls have been well documented in rocky intertidal communities, while the effects of bottom-up influences are comparatively poorly understood. We hypothesized that large-scale signatures of the physical environment may be identifiable along the South African coastline as it is subject to two very different current systems (Benguela and Agulhas Currents) that profoundly influence primary production and thus both food type and availability. Through stable isotope analysis, we examined biogeographic patterns in multiple trophic levels at four sites along a 1400-km stretch of South African coastline and investigated the dietary role of macroalgal-derived organic carbon in rocky intertidal communities. The general positioning of trophic groups was comparable across all sites, with animals from the same trophic levels grouping together and with a δ 15N fractionation of 1-2‰ between levels. The species found at all sites demonstrated east-west δ 15N enrichment, presumably reflecting a biogeographic shift in nitrogen sources linked to upwelling on the west coast. Filter-feeders gave particularly clear results. Using discriminant analysis, mussels could be categorized into four geographic groups based on carbon and nitrogen signatures: east coast, southeast coast, south-west coast and west coast. Barnacles and polychaetes showed similar geographic groupings to mussels, but with shifts in actual values (1‰ depletion in δ 13C and 3‰ enrichment in δ 15N relative to mussels). This suggests that fractionation varies between species within a trophic level. IsoSource models showed that Ulva sp. made large contributions to the diets of two microalgal grazers ( Siphonaria capensis and Scutellastra granularis) and this dietary dependence increased when moving from west to east coast, along the shoreline. Additionally, IsoSource models determined that relative to phytoplankton, macroalgae accounted for upwards of 60% of suspended

  1. 13C NMR relaxation in neutral and charged tetra- n-alkyl compounds

    NASA Astrophysics Data System (ADS)

    Bordes, B.; Coletta, F.; Ferrarini, A.; Gottardi, F.; Nordio, P. L.

    1998-05-01

    NMR T1 relaxation times have been measured for 13C nuclei in the alkyl chains of symmetric tetra- n-alkylstannanes dissolved in CDCl 3. The results are interpreted in terms of conformational transitions occurring in the aliphatic chains superimposed to rotational diffusion of the whole molecule. A comparison with analogous tetra- n-alkylammonium salts is performed. Differences are ascribed to changes in the overall rotational diffusion deriving from effects of charge upon formation of ion-pairs and larger aggregates.

  2. Using Neural Networks for 13C NMR Chemical Shift Prediction-Comparison with Traditional Methods

    NASA Astrophysics Data System (ADS)

    Meiler, Jens; Maier, Walter; Will, Martin; Meusinger, Reinhard

    2002-08-01

    Interpretation of 13C chemical shifts is essential for structure elucidation of organic molecules by NMR. In this article, we present an improved neural network approach and compare its performance to that of commonly used approaches. Specifically, our recently proposed neural network ( J. Chem. Inf. Comput. Sci. 2000, 40, 1169-1176) is improved by introducing an extended hybrid numerical description of the carbon atom environment, resulting in a standard deviation (std. dev.) of 2.4 ppm for an independent test data set of ˜42,500 carbons. Thus, this neural network allows fast and accurate 13C NMR chemical shift prediction without the necessity of access to molecule or fragment databases. For an unbiased test dataset containing 100 organic structures the accuracy of the improved neural network was compared to that of a prediction method based on the HOSE code ( hierarchically ordered spherical description of environment) using S PECI NFO. The results show the neural network predictions to be of quality (std. dev.=2.7 ppm) comparable to that of the HOSE code prediction (std. dev.=2.6 ppm). Further we compare the neural network predictions to those of a wide variety of other 13C chemical shift prediction tools including incremental methods (C HEMD RAW, S PECT OOL), quantum chemical calculation (G AUSSIAN, C OSMOS), and HOSE code fragment-based prediction (S PECI NFO, ACD/CNMR, P REDICTI T NMR) for the 47 13C-NMR shifts of Taxol, a natural product including many structural features of organic substances. The smallest standard deviations were achieved here with the neural network (1.3 ppm) and S PECI NFO (1.0 ppm).

  3. Quantifying the chemical composition of soil organic carbon with solid-state 13C NMR

    NASA Astrophysics Data System (ADS)

    Baldock, J. A.; Sanderman, J.

    2011-12-01

    The vulnerability of soil organic carbon (SOC) to biological decomposition and mineralisation to CO2 is defined at least partially by its chemical composition. Highly aromatic charcoal-like SOC components are more stable to biological decomposition than other forms of carbon including cellulose. Solid-state 13C NMR has gained wide acceptance as a method capable of defining SOC chemical composition and mathematical fitting processes have been developed to estimate biochemical composition. Obtaining accurate estimates depends on an ability to quantitatively detect all carbon present in a sample. Often little attention has been paid to defining the proportion of organic carbon present in a soil that is observable in solid-state 13C NMR analyses of soil samples. However, if such data is to be used to inform carbon cycling studies, it is critical that quantitative assessments of SOC observability be undertaken. For example, it is now well established that a significant discrimination exists against the detection of the low proton content polyaromatic structures typical of charcoal using cross polarisation 13C NMR analyses. Such discrimination does not exist where direct polarisation analyses are completed. In this study, the chemical composition of SOC as defined by cross polarisation and direct polarisation13C NMR analyses will be compared for Australian soils collected from under a diverse range of agricultural managements and climatic conditions. Results indicate that where significant charcoal C contents exist, it is highly under-represented in the acquired CP spectra. For some soils, a discrimination against alkyl carbon was also evident. The ability to derive correction factors to compensate for such discriminations will be assessed and presented.

  4. Calculation of the 13C NMR shieldings of the C0 2 complexes of aluminosilicates

    NASA Astrophysics Data System (ADS)

    Tossell, J. A.

    1995-04-01

    13C NMR shieldings have been calculated using the random-phase-approximation, localized-orbital local-origins version of ab initio coupled Hartree-Fuck perturbation theory for CO 2 and and for several complexes formed by the reaction of CO 2 with molecular models for aluminosilicate glasses, H 3TOT'H3 3-n, T,T' = Si,Al. Two isomeric forms of the CO 2-aluminosilicate complexes have been considered: (1) "CO 2-like" complexes, in which the CO 2 group is bound through carbon to a bridging oxygen and (2) "CO 3-like" complexes, in which two oxygens of a central CO 3 group form bridging bonds to the two TH 3 groups. The CO 2-like isomer of CO 2-H 3SiOSiH 3 is quite weakly bonded and its 13C isotropic NMR shielding is almost identical to that in free CO 2. As Si is progressively replaced by Al in the - H terminated aluminosilicate model, the CO 2-like isomers show increasing distortion from the free CO 2 geometry and their 13C NMR shieldings decrease uniformly. The calculated 13C shielding value for H 3AlO(CO 2)AlH 3-2 is only about 6 ppm larger than that calculated for point charge stabilized CO 3-2. However, for a geometry of H 3SiO(CO 2) AlH 3-1, in which the bridging oxygen to C bond length has been artificially increased to that found in the - OH terminated cluster (OH) 3SiO(CO 2)Al(OH) 3-1, the calculated 13C shielding is almost identical to that for free CO 2. The CO 3-like isomers of the CO 2-aluminosili-cate complexes show carbonate like geometries and 13C NMR shieldings about 4-9 ppm larger than those of carbonate for all T,T' pairs. For the Si,Si tetrahedral atom pair the CO 2-like isomer is more stable energetically, while for the Si,Al and Al,Al cases the CO 3-like isomer is more stable. Addition of Na + ions to the CO 3-2 or H 3AlO(CO 2)AlH 3-2 complexes reduces the 13C NMR shieldings by about 10 ppm. Complexation with either Na + or CO 2 also reduces the 29Si NMR shieldings of the aluminosilicate models, while the changes in 27Al shielding with Na + or CO 2

  5. (1)H, (15)N and (13)C resonance assignments of translationally-controlled tumor protein from photosynthetic microalga Nannochloropsis oceanica.

    PubMed

    Yao, Xingzhe; Xiao, Yan; Cui, Qiu; Feng, Yingang

    2015-10-01

    Translationally-controlled tumor protein (TCTP) is a eukaryote-conserved protein with crucial roles in cellular growth. It has also been proposed that plant TCTP has functions specific to plant, while no structure of TCTP from photosynthetic organism has been reported. Nannochloropsis is a photosynthetic microalga with high yield of lipid and high-value polyunsaturated fatty acid, which is promising for biodiesel production. Study of growth-related proteins may provide new clue for improving the yield of lipid. TCTP from Nannochloropsis oceanica shares low sequence identity with structure-known TCTPs. Here we reported the NMR resonance assignments of TCTP from N. oceanica for further structural and functional studies. PMID:25680850

  6. Giant quartz vein systems in accretionary orogenic belts: the evidence for a metamorphic fluid origin from δ 15N and δ 13C studies

    NASA Astrophysics Data System (ADS)

    Jia, Yiefei; Kerrich, Robert

    2000-12-01

    Nitrogen isotope compositions of hydrothermal micas in giant structurally hosted quartz vein systems place constraints on the origin of the hydrothermal fluids. The vein systems are from Neoarchean terranes in the Superior Province of Canada, and the Norseman terrane, Western Australia. The four quartz vein systems studied in different terranes formed at metamorphic grades ranging from lower greenschist to lower amphibolite facies, coevally with metamorphism of the host terrane. Nitrogen resides predominantly as NH 4+ in hydrothermal mica. For lower to upper greenschist facies quartz veins, N contents and δ 15N values of micas are between 40 and 200 ppm and 15‰ and 21‰, respectively, whereas in quartz veins formed at the greenschist to amphibolite transition and lower amphibolite facies, micas have N contents of 20-70 ppm and δ 15N of 11-24‰. In contrast, micas and K-feldspars from granitoids in the Neoarchean Abitibi and Red Lake greenstone belts are characterized by systematically lower δ 15N of -5‰ to 5‰, and generally lower N contents of 20-50 ppm, comparable to other granitoids. Carbon and oxygen isotope compositions of hydrothermal ferroan dolomite and calcite show systematic depletions with increasing metamorphic grade. The mean values range from -2.2‰ to -3.7‰ for δ 13C and 13.8‰ to 12.3‰ for δ 18O in veins formed at lower to upper greenschist facies, but from -5.8‰ to -7.1‰ for δ 13C and 11.4‰ to 9.8‰ for δ 18O at higher metamorphic grades. Nitrogen isotope compositions rule out mantle (δ 15N mean=-5‰), magmatic (δ 15N=-5‰ to <10‰), or meteoric fluids (δ 15N mean=4.4±2.0‰) for the quartz veins. Accordingly, the results are consistent with fluids derived from metamorphic dehydration of oceanic crust and sediments within accretionary orogenic belts.

  7. Metabolic pathways for ketone body production. /sup 13/C NMR spectroscopy of rat liver in vivo using /sup 13/C-multilabeled fatty acids

    SciTech Connect

    Pahl-Wostl, C.; Seelig, J.

    1986-11-04

    The hormonal regulation of ketogenesis in the liver of living rat has been studied noninvasively with /sup 13/C nuclear magnetic resonance. The spatial selection for the liver was better than 90%, with extrahepatic adipose tissue contribution only a very small amount of signal. The metabolic activities of the liver were investigated by infusion of /sup 13/C-labeled butyrate in the jugular vein of the anesthetized rat. The rate of butyrate infusion was chosen to be close to the maximum oxidative capacity of the rat liver, and the /sup 13/C signal intensities were enhanced by using doubly labeled (1,3-/sup 13/C)butyrate as a substrate. Different /sup 13/C NMR spectra and hence different metabolites were observed depending on the hormonal state of the animal. The /sup 13/C NMR studies demonstrate that even when rate of acetyl-CoA production are high, the disposal of this compound is not identical in fasted and diabetic animals. This supports previous suggestions that the redox state of the mitochondrion represents the most important factor in regulation. For a given metabolic state of the animal, different signal intensities were obtained depending on whether butyrate was labeled at C-1, C-3, or C-1,3. From the ratios of incorporation of /sup 13/C label into the carbons of 3-hydroxybutyrate, it could be estimated that a large fraction of butyrate evaded ..beta..-oxidation to acetyl-CoA but was converted directly to acetoacetyl-CoA. /sup 13/C-labeled glucose could be detected in vivo in the liver of diabetic rats.

  8. Abrupt recent shift in δ13C and δ15N values in Adélie penguin eggshell in Antarctica

    PubMed Central

    Emslie, Steven D.; Patterson, William P.

    2007-01-01

    Stable isotope values of carbon (δ13C) and nitrogen (δ15N) in blood, feathers, eggshell, and bone have been used in seabird studies since the 1980s, providing a valuable source of information on diet, foraging patterns, and migratory behavior in these birds. These techniques can also be applied to fossil material when preservation of bone and other tissues is sufficient. Excavations of abandoned Adélie penguin (Pygoscelis adeliae) colonies in Antarctica often provide well preserved remains of bone, feathers, and eggshell dating from hundreds to thousands of years B.P. Herein we present an ≈38,000-year time series of δ13C and δ15N values of Adélie penguin eggshell from abandoned colonies located in three major regions of Antarctica. Results indicate an abrupt shift to lower-trophic prey in penguin diets within the past ≈200 years. We posit that penguins only recently began to rely on krill as a major portion of their diet, in conjunction with the removal of baleen whales and krill-eating seals during the historic whaling era. Our results support the “krill surplus” hypothesis that predicts excess krill availability in the Southern Ocean after this period of exploitation. PMID:17620620

  9. Feeding and migration habits of white shark Carcharodon carcharias (Lamniformes: Lamnidae) from Isla Guadalupe inferred by analysis of stable isotopes delta15N and delta13C.

    PubMed

    Jaime-Rivera, Mario; Caraveo-Patiño, Javier; Hoyos-Padilla, Mauricio; Galván-Magaña, Felipe

    2014-06-01

    Stable isotope composition of marine top predator's tissues provides insight information of its trophic ecology and migratory behavior. Previous reports have shown that dermal tissues could record longer patterns of hunting and movement. Based on this, the aim of this study was to describe the feeding and migratory habits of the white shark from Isla Guadalupe, using stable isotopic analysis of dermis. We considered a small subset of many possible prey taxa that the sharks could have eaten throughout their migration: pinnipeds, squid and tuna. We grouped the data in five focal areas: Gulf of California, Coast of California, Isla Guadalupe, SOFA and Hawaii. We performed a Bayesian mixing model to study the trophic ecology of this top predator. Average isotopic values for dermis tissue of white shark were delta13C (-14.5 per thousand) and delta15N (19.1 per thousand). Corrected white shark dermal mean values to resemble muscle were delta13C (-16.6 per thousand) and delta15N (21.2 per thousand). Mixing model data from dermis showed predation in offshore areas such the SOFA and a main importance of pinnipeds as prey of the white shark in Isla Guadalupe. PMID:25102646

  10. Pan-Arctic concentrations of mercury and stable isotope ratios of carbon (δ(13)C) and nitrogen (δ(15)N) in marine zooplankton.

    PubMed

    Pomerleau, Corinne; Stern, Gary A; Pućko, Monika; Foster, Karen L; Macdonald, Robie W; Fortier, Louis

    2016-05-01

    Zooplankton play a central role in marine food webs, dictating the quantity and quality of energy available to upper trophic levels. They act as "keystone" species in transfer of mercury (Hg) up through the marine food chain. Here, we present the first Pan-Arctic overview of total and monomethylmercury concentrations (THg and MMHg) and stable isotope ratios of carbon (δ(13)C) and nitrogen (δ(15)N) in selected zooplankton species by assembling data collected between 1998 and 2012 from six arctic regions (Laptev Sea, Chukchi Sea, southeastern Beaufort Sea, Canadian Arctic Archipelago, Hudson Bay and northern Baffin Bay). MMHg concentrations in Calanus spp., Themisto spp. and Paraeuchaeta spp. were found to increase with higher δ(15)N and lower δ(13)C. The southern Beaufort Sea exhibited both the highest THg and MMHg concentrations. Biomagnification of MMHg between Calanus spp. and two of its known predators, Themisto spp. and Paraeuchaeta spp., was greatest in the southern Beaufort Sea. Our results show large geographical variations in Hg concentrations and isotopic signatures for individual species related to regional ecosystem features, such as varying water masses and freshwater inputs, and highlight the increased exposure to Hg in the marine food chain of the southern Beaufort Sea. PMID:26874765

  11. Effects of Boron Nutrition and Water Stress on Nitrogen Fixation, Seed δ15N and δ13C Dynamics, and Seed Composition in Soybean Cultivars Differing in Maturities

    PubMed Central

    Bellaloui, Nacer; Mengistu, Alemu

    2015-01-01

    Therefore, the objective of the current research was to investigate the effects of foliar B nutrition on seed protein, oil, fatty acids, and sugars under water stress conditions. A repeated greenhouse experiment was conducted using different maturity group (MG) cultivars. Plants were well-watered with no foliar B (W − B), well-watered with foliar B (W + B), water-stressed with no foliar B (WS − B), and water-stressed with foliar B (WS + B). Foliar B was applied at rate of 0.45 kg·ha−1 and was applied twice at flowering and at seed-fill stages. The results showed that seed protein, sucrose, fructose, and glucose were higher in W + B treatment than in W − B, WS + B, and WS − B. The increase in protein in W + B resulted in lower seed oil, and the increase of oleic in WS − B or WS + B resulted in lower linolenic acid. Foliar B resulted in higher nitrogen fixation and water stress resulted in seed δ15N and δ13C alteration. Increased stachyose indicated possible physiological and metabolic changes in carbon and nitrogen pathways and their sources under water stress. This research is beneficial to growers for fertilizer management and seed quality and to breeders to use 15N/14N and 13C/12C ratios and stachyose to select for drought tolerance soybean. PMID:25667936

  12. Total-body protein turnover in parenterally fed neonates: effects of energy source studied by using [15N]glycine and [1-13C]leucine.

    PubMed

    Pencharz, P; Beesley, J; Sauer, P; Van Aerde, J; Canagarayar, U; Renner, J; McVey, M; Wesson, D; Swyer, P

    1989-12-01

    The effects of nonprotein energy source (ie, glucose only vs glucose and lipid) on nitrogen retention and total-body protein turnover were studied in 20 parenterally fed newborn infants. All infants received approximately 3 g amino acids and 80-90 kcal.kg body wt.d. Total-body protein synthesis was estimated by using three constant-infusion, end-product methods: enrichment of urinary urea and ammonia in response to a [15N]glycine label and exhaled carbon dioxide enrichment in response to a [1-13C]leucine label. No differences were seen in nitrogen retention between the two energy sources. The estimate of total-body protein turnover obtained from the 13C label was similar to that obtained with the [15N]urea label. No differences in turnover rates were observed between the two diet groups. Use of the glucose-plus-lipid fuel system enhanced energy storage and the reutilization of amino acid for protein synthesis. PMID:2512806

  13. Submillimeter Observations of Titan: Global Measures of Stratospheric Temperature, CO, HCN, HC3N, and the Isotopic Ratios 12C/13C and 14N/15N

    NASA Astrophysics Data System (ADS)

    Gurwell, Mark A.

    2004-11-01

    Interferometric observations of the atmosphere of Titan were performed with the Submillimeter Array on two nights in 2004 February to investigate the global average vertical distributions of several molecular species above the tropopause. Rotational transitions of CO, isomers of HCN, and HC3N were simultaneously recorded. The abundance of CO is determined to be 51+/-4 parts per million (ppm), constant with altitude. The vertical profile of HCN is dependent on the assumed temperature but generally increases from 30 parts per billion at the condensation altitude (~83 km) to 5 ppm at ~300 km. Furthermore, the central core of the HCN emission is strong and can be reproduced only if the upper stratospheric temperature increases with altitude. The isotopic ratios are determined to be 12C/13C=132+/-25 and 14N/15N=94+/-13 assuming the Coustenis & Bézard temperature profile. If the Lellouch temperature profile is assumed, the ratios decrease to 12C/13C=108+/-20 and 14N/15N=72+/-9. The vertical profile of HC3N is consistent with that derived by Marten et al.

  14. (1)H, (13)C and (15)N resonance assignment of the cytosolic dithiol glutaredoxin 1 from the pathogen Trypanosoma brucei.

    PubMed

    Stefani, Monica; Sturlese, Mattia; Manta, Bruno; Löhr, Frank; Mammi, Stefano; Comini, Marcelo; Bellanda, Massimo

    2016-04-01

    Trypanosomatids are parasites responsible for several tropical and subtropical diseases, such as Chaga's disease, sleeping sickness and Leishmaniasis. In contrast to the mammalian host, the thiol-redox metabolism of these pathogens depends on trypanothione [bis-glutathionylspermidine, T(SH)2] instead of glutathione (GSH) providing a set of lineage-specific proteins as drug target candidates. Glutaredoxins (Grx) are ubiquitous small thiol-disulfide oxidoreductases that belong to the thioredoxin-fold family. They play a central role in redox homeostasis and iron sulfur-cluster biogenesis. Each species, including trypanosomes, possesses its own set of isoforms distributed in different subcellular compartments. The genome of trypanosomatids encodes for two class I (dithiolic) Grxs named 2-C-Grx1 and 2-C-Grx2. Both proteins were shown to efficiently reduce different disulfides at the expenses of T(SH)2 using a mechanism that involves the two cysteines in the active site. Moreover, the cytosolic Trypanosoma brucei 2-C-Grx1 but not the mitochondrial 2-C-Grx2 was able to coordinate an iron-sulfur cluster with T(SH)2 or GSH as ligand. As a first step to unravel the structural basis for the specificity observed in the trypanosomal glutaredoxins, we present here the NMR resonance assignment of 2-C-Grx1 from the parasite T. brucei brucei. PMID:26386962

  15. Late Holocene monsoon climate of northeastern Taiwan inferred from elemental (C, N) and isotopic (δ13C, δ15N) data in lake sediments

    NASA Astrophysics Data System (ADS)

    Selvaraj, Kandasamy; Wei, Kuo-Yen; Liu, Kon-Kee; Kao, Shuh-Ji

    2012-03-01

    Little information exists about centennial-scale climate variability on oceanic islands in the western Pacific where the East Asian monsoon (EAM) strongly influences the climate, mountain ecosystem and the society. In this study, we investigate a 168 cm long sediment core recovered from Emerald Peak Lake in subalpine NE Taiwan for the contents of grain size, total organic carbon (TOC), C/N ratio, and stable isotopes (δ13C and δ15N) to reconstruct the monsoon climate and vegetation density during the late Holocene. Six radiocarbon (14C) ages obtained on plant remains used for the chronology indicate that the sediment core has been accumulated since ˜3770 cal BP with a mean sedimentation rate of 44.6 cm/ka. The sub-centennial resolution of our proxy records reveals strong fluctuations of the EAM and vegetation density for the past ˜3770 cal BP. The greater contents of coarse and medium sediments with overall decreasing trends from 3770 to 2000 cal BP suggest an increasing fine sediment influx from the catchment likely due to an increasing lake water level. Although low TOC content, C/N ratio, and enriched δ13C values in bulk and fine sediments during this interval suggest a sparsely vegetated catchment, increasing trends of TOC content and C/N ratio together with decreasing trends of δ13C and δ15N values indicate a strengthening pattern of summer monsoon. This is in contrast to a decreasing monsoon strength inferred from Dongge Cave δ18O record at that time, supporting the idea of anti-phasing of summer EAM and Indian summer monsoon. Since 2000 cal BP, higher content of fine sediments with high TOC content and C/N ratio but relatively depleted δ13C and low δ15N values suggest a high but stable lake water level and dense C3 plants, consistent with a stronger summer monsoon in a wet climate. Within this general trend, we interpret a prominent change of proxy parameters in sediments from ˜560 to 150 cal BP, as subtropical evidence for the Little Ice Age in NE

  16. Microsolvation of methylmercury: structures, energies, bonding and NMR constants ((199)Hg, (13)C and (17)O).

    PubMed

    Flórez, Edison; Maldonado, Alejandro F; Aucar, Gustavo A; David, Jorge; Restrepo, Albeiro

    2016-01-21

    Hartree-Fock (HF) and second order perturbation theory (MP2) calculations within the scalar and full relativistic frames were carried out in order to determine the equilibrium geometries and interaction energies between cationic methylmercury (CH3Hg(+)) and up to three water molecules. A total of nine structures were obtained. Bonding properties were analyzed using the Quantum Theory of Atoms In Molecules (QTAIM). The analyses of the topology of electron densities reveal that all structures exhibit a partially covalent HgO interaction between methylmercury and one water molecule. Consideration of additional water molecules suggests that they solvate the (CH3HgOH2)(+) unit. Nuclear magnetic shielding constants σ((199)Hg), σ((13)C) and σ((17)O), as well as indirect spin-spin coupling constants J((199)Hg-(13)C), J((199)Hg-(17)O) and J((13)C-(17)O), were calculated for each one of the geometries. Thermodynamic stability and the values of NMR constants correlate with the ability of the system to directly coordinate oxygen atoms of water molecules to the mercury atom in methylmercury and with the formation of hydrogen bonds among solvating water molecules. Relativistic effects account for 11% on σ((13)C) and 14% on σ((17)O), which is due to the presence of Hg (heavy atom on light atom, HALA effect), while the relativistic effects on σ((199)Hg) are close to 50% (heavy atom on heavy atom itself, HAHA effect). J-coupling constants are highly influenced by relativity when mercury is involved as in J((199)Hg-(13)C) and J((199)Hg-(17)O). On the other hand, our results show that the values of NMR constants for carbon and oxygen, atoms which are connected through mercury (C-HgO), are highly correlated and are greatly influenced by the presence of water molecules. Water molecules introduce additional electronic effects to the relativistic effects due to the mercury atom. PMID:26670708

  17. Soil organic matter stability in agricultural land: New insights using δ15N, δ13C and C:N ratio

    NASA Astrophysics Data System (ADS)

    Mao, Yanling; Heiling, Maria; De Clercq, Tim; Resch, Christian; Aigner, Martina; Mayr, Leo; Vanlauwe, Bernard; Thuita, Moses; Steier, Peter; Leifeld, Jens; Merckx, Roel; Spiegel, Heide; Cepuder, Peter; Nguyen, Minh-Long; Zaman, Mohammad; Dercon, Gerd

    2014-05-01

    Soil organic matter (SOM) contains three times more carbon than in the atmosphere or terrestrial vegetation. This major pool of organic carbon is sensitive to climate change, but the mechanisms for carbon stabilization in soils are still not well understood and the ultimate potential for carbon stabilization is unknown. For predicting SOM dynamics, it is necessary to gain information on the turnover rates or stability of different soil organic carbon pools. The common method to determine stability and age of SOM is the 14C radio carbon technique, which is very expensive and therefore limited in use. Conen et al. (2008) developed a model to estimate the SOM stability based on the isotopic discrimination of 15N natural abundance by soil micro-organisms, and the decreasing C:N ratio during organic matter decomposition. This model has been developed for permanent grasslands in the Swiss Alps under steady-state conditions. The objective of our study was to validate whether this model could be used or adapted, in combination with 13C isotope signatures of SOM, to predict the relative age and stability of SOM fractions in more disturbed agricultural ecosystems. The present study was carried out on soils collected from six long-term experimental trials (from 12 to 50 years) under different agricultural management practices (e.g. no tillage vs conventional tillage, and mulch, fertilizer, green or animal manure application), located in Austria, Belgium, Kenya and China. Top and subsoil were sampled until 80-100 cm depth. Particulate organic matter (POM) fraction was obtained by wet sieving (> 63μm) after sonification and density separation (<1.8 g cm-3). Carbon and nitrogen contents and their stable isotopic ratios (i.e. 15N and 13C) were measured in POM and bulk soils. The mineral associated matter fraction (mOM), as the protected carbon, was calculated by difference to the bulk soil organic carbon. The relative age of the SOM was calculated using the Conen model and

  18. Quantitative analysis of deuterium using the isotopic effect on quaternary (13)C NMR chemical shifts.

    PubMed

    Darwish, Tamim A; Yepuri, Nageshwar Rao; Holden, Peter J; James, Michael

    2016-07-13

    Quantitative analysis of specifically deuterated compounds can be achieved by a number of conventional methods, such as mass spectroscopy, or by quantifying the residual (1)H NMR signals compared to signals from internal standards. However, site specific quantification using these methods becomes challenging when dealing with non-specifically or randomly deuterated compounds that are produced by metal catalyzed hydrothermal reactions in D2O, one of the most convenient deuteration methods. In this study, deuterium-induced NMR isotope shifts of quaternary (13)C resonances neighboring deuterated sites have been utilized to quantify the degree of isotope labeling of molecular sites in non-specifically deuterated molecules. By probing (13)C NMR signals while decoupling both proton and deuterium nuclei, it is possible to resolve (13)C resonances of the different isotopologues based on the isotopic shifts and the degree of deuteration of the carbon atoms. We demonstrate that in different isotopologues, the same quaternary carbon, neighboring partially deuterated carbon atoms, are affected to an equal extent by relaxation. Decoupling both nuclei ((1)H, (2)H) resolves closely separated quaternary (13)C signals of the different isotopologues, and allows their accurate integration and quantification under short relaxation delays (D1 = 1 s) and hence fast accumulative spectral acquisition. We have performed a number of approaches to quantify the deuterium content at different specific sites to demonstrate a convenient and generic analysis method for use in randomly deuterated molecules, or in cases of specifically deuterated molecules where back-exchange processes may take place during work up. PMID:27237841

  19. Variations of soil δ13C and δ15N across a precipitation gradient in a savanna ecosystem: Implications of climate change on the carbon cycle

    NASA Astrophysics Data System (ADS)

    Dintwe, K.; Gilhooly, W., III; Wang, L.; O'Donnell, F. C.; Bhattachan, A.; D'Odorico, P.; Okin, G. S.

    2015-12-01

    Savannas are the third largest terrestrial carbon pool after only tropical and borealforests. They are highly productive ecosystems and contribute about 30% of the globalterrestrial net primary productivity and potentially contain 20% of the world's soilorganic carbon. Global circulation models have predicted that many savannas willbecome warmer and drier during the twenty-first century. The impacts of the projectedclimatic trend on the productivity and biogeochemical cycles of savannas are not fullyunderstood. Here, we assessed the abundance of stable carbon (δ13C) and nitrogen (δ15N)isotopes in soil profiles at four sites along a 1000 km transect with a strong south-northprecipitation gradient in southern Africa. The south receives about 180 mm of rainfall peryear and dominated by grass species (C4) whereas the north receives 540 mm·yr-1 anddominated by woody plants (C3). Soil surface δ13C showed that woody vegetation contributedmore than 75% of soil carbon input in the wet sites whereas grasses contributed about65% of soil carbon input in the dry sites. The soil profile δ13C indicated that intermediatesites have shifted from grass dominated to woody-shrub-dominated statesduring recent past. The dry sites had relatively higher δ15N (~10‰) compared to the wetsites (~5‰) indicating significantly greater N2 fixation in the wetter sites or high rates ofNH3 volatilization in the drier sites. Our results suggest that as savannas become warmerand drier due to climate change, woody shrubs are likely to be the dominant form ofvegetation structure, a process that could alter biogeochemical processes and results insavannas becoming net carbon sink or source.

  20. Variation in δ13C and δ15N diet–vibrissae trophic discrimination factors in a wild population of California sea otters

    USGS Publications Warehouse

    Newsome, Seth D.; Bentall, Gena B.; Tinker, M. Tim; Oftedal, Olav T.; Ralls, Katherine; Estes, James A.; Fogel, Marilyn L.

    2010-01-01

    The ability to quantify dietary inputs using stable isotope data depends on accurate estimates of isotopic differences between a consumer (c) and its diet (d), commonly referred to as trophic discrimination factors (TDFs) and denoted by Δc-d. At present, TDFs are available for only a few mammals and are usually derived in captive settings. The magnitude of TDFs and the degree to which they vary in wild populations is unknown. We determined δ13C and δ15N TDFs for vibrissae (i.e., whiskers), a tissue that is rapidly becoming an informative isotopic substrate for ecologists, of a wild population of sea otters for which individual diet has been quantified through extensive observational study. This is one of the very few studies that report TDFs for free-living wild animals feeding on natural diets. Trophic discrimination factors of 2.2‰ ± 0.7‰ for δ13C and 3.5‰ ± 0.6‰ for δ15N (mean ± SD) were similar to those reported for captive carnivores, and variation in individual δ13C TDFs was negatively but significantly related to sea urchin consumption. This pattern may relate to the lipid-rich diet consumed by most sea otters in this population and suggests that it may not be appropriate to lipid-extract prey samples when using the isotopic composition of keratinaceous tissues to examine diet in consumers that frequently consume lipid-rich foods, such as many marine mammals and seabirds. We suggest that inherent variation in TDFs should be included in isotopically based estimates of trophic level, food chain length, and mixing models used to quantify dietary inputs in wild populations; this practice will further define the capabilities and limitations of isotopic approaches in ecological studies.

  1. Variation in delta13C and delta15N diet-vibrissae trophic discrimination factors in a wild population of California sea otters.

    PubMed

    Newsome, Seth D; Bentall, Gena B; Tinker, M Tim; Oftedal, Olav T; Ralls, Katherine; Estes, James A; Fogel, Marilyn L

    2010-09-01

    The ability to quantify dietary inputs using stable isotope data depends on accurate estimates of isotopic differences between a consumer (c) and its diet (d), commonly referred to as trophic discrimination factors (TDFs) and denoted by delta(c-d). At present, TDFs are available for only a few mammals and are usually derived in captive settings. The magnitude of TDFs and the degree to which they vary in wild populations is unknown. We determined delta13C and delta15N TDFs for vibrissae (i.e., whiskers), a tissue that is rapidly becoming an informative isotopic substrate for ecologists, of a wild population of sea otters for which individual diet has been quantified through extensive observational study. This is one of the very few studies that report TDFs for free-living wild animals feeding on natural diets. Trophic discrimination factors of 2.2 per thousand +/- 0.7 per thousand for delta13C and 3.5 per thousand +/- 0.6 per thousand for delta15N (mean +/- SD) were similar to those reported for captive carnivores, and variation in individual delta13C TDFs was negatively but significantly related to sea urchin consumption. This pattern may relate to the lipid-rich diet consumed by most sea otters in this population and suggests that it may not be appropriate to lipid-extract prey samples when using the isotopic composition of keratinaceous tissues to examine diet in consumers that frequently consume lipid-rich foods, such as many marine mammals and seabirds. We suggest that inherent variation in TDFs should be included in isotopically based estimates of trophic level, food chain length, and mixing models used to quantify dietary inputs in wild populations; this practice will further define the capabilities and limitations of isotopic approaches in ecological studies. PMID:20945772

  2. Balancing the (carbon) budget: Using linear inverse models to estimate carbon flows and mass-balance 13C:15N labelling experiments in low oxygen sediments.

    NASA Astrophysics Data System (ADS)

    Hunter, William Ross; Van Oevelen, Dick; Witte, Ursula

    2013-04-01

    Over 1 million km2 of seafloor experience permanent low-oxygen conditions within oxygen minimum zones (OMZs). OMZs are predicted to grow as a consequence of climate change, potentially affecting oceanic biogeochemical cycles. The Arabian Sea OMZ impinges upon the western Indian continental margin at bathyal depths (150 - 1500m) producing a strong depth dependent oxygen gradient at the sea floor. The influence of the OMZ upon the short term processing of organic matter by sediment ecosystems was investigated using in situ stable isotope pulse chase experiments. These deployed doses of 13C:15N labeled organic matter onto the sediment surface at four stations from across the OMZ (water depth 540 - 1100 m; [O2] = 0.35 - 15 μM). In order to prevent experimentally anoxia, the mesocosms were not sealed. 13C and 15N labels were traced into sediment, bacteria, fauna and 13C into sediment porewater DIC and DOC. However, the DIC and DOC flux to the water column could not be measured, limiting our capacity to obtain mass-balance for C in each experimental mesocosm. Linear Inverse Modeling (LIM) provides a method to obtain a mass-balanced model of carbon flow that integrates stable-isotope tracer data with community biomass and biogeochemical flux data from a range of sources. Here we present an adaptation of the LIM methodology used to investigate how ecosystem structure influenced carbon flow across the Indian margin OMZ. We demonstrate how oxygen conditions affect food-web complexity, affecting the linkages between the bacteria, foraminifera and metazoan fauna, and their contributions to benthic respiration. The food-web models demonstrate how changes in ecosystem complexity are associated with oxygen availability across the OMZ and allow us to obtain a complete carbon budget for the stationa where stable-isotope labelling experiments were conducted.

  3. Improved Carbohydrate Structure Generalization Scheme for (1)H and (13)C NMR Simulations.

    PubMed

    Kapaev, Roman R; Toukach, Philip V

    2015-07-21

    The improved Carbohydrate Structure Generalization Scheme has been developed for the simulation of (13)C and (1)H NMR spectra of oligo- and polysaccharides and their derivatives, including those containing noncarbohydrate constituents found in natural glycans. Besides adding the (1)H NMR calculations, we improved the accuracy and performance of prediction and optimized the mathematical model of the precision estimation. This new approach outperformed other methods of chemical shift simulation, including database-driven, neural net-based, and purely empirical methods and quantum-mechanical calculations at high theory levels. It can process structures with rarely occurring and noncarbohydrate constituents unsupported by the other methods. The algorithm is transparent to users and allows tracking used reference NMR data to original publications. It was implemented in the Glycan-Optimized Dual Empirical Spectrum Simulation (GODESS) web service, which is freely available at the platform of the Carbohydrate Structure Database (CSDB) project ( http://csdb.glycoscience.ru). PMID:26087011

  4. The response of the foliar antioxidant system and stable isotopes (δ(13)C and δ(15)N) of white willow to low-level air pollution.

    PubMed

    Wuytack, Tatiana; AbdElgawad, Hamada; Staelens, Jeroen; Asard, Han; Boeckx, Pascal; Verheyen, Kris; Samson, Roeland

    2013-06-01

    In this study we aimed to determine and elucidate the effect of ambient air pollution on the foliar antioxidant system and stable carbon and nitrogen isotopes of white willow (Salix alba L.). We grew white willow in uniform potting soil in the near vicinity of sixteen air quality monitoring stations in Belgium where nitrogen dioxide (NO2), ozone, sulfur dioxide and particulate matter concentrations were continuously measured. The trees were exposed to ambient air during six months (April-September 2011), and, thereafter, the degree of lipid peroxidation and foliar content of antioxidant molecules (ascorbate, glutathione, polyphenols, flavonoids), antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, peroxidase) and foliar stable carbon (δ(13)C) and nitrogen (δ(15)N) isotopes were measured. We found that lipid peroxidation was caused by air pollution stress, arising from high ambient NO2 concentrations, as shown by an increased amount of malondialdehyde. The antioxidant system was activated by increasing the amount of polyphenols at monitoring stations with a high atmospheric NO2 and low O3 concentration, while no increase of key enzymes (e.g., ascorbate, glutathione) was observed. The δ(13)C also decreased with increasing NO2 concentrations and decreasing O3 concentrations, probably reflecting a decreased net photosynthesis and/or a concomitant decrease of (13)CO2 in the atmosphere. Shade also influenced foliar δ(13)C and the content of leaf ascorbate and glutathione. PMID:23562799

  5. Cross-basin heterogeneity in lanternfish (family Myctophidae) assemblages and isotopic niches (δ13C and δ15N) in the southern Tasman Sea abyssal basin

    NASA Astrophysics Data System (ADS)

    Flynn, A. J.; Kloser, R. J.

    2012-11-01

    A cross-basin (longitudinal) study of lanternfishes in the southern Tasman Sea abyssal basin during the austral winter of 2008 and 2009 found that mean biomass in the Western sector was higher than that in the Eastern sector, corresponding with cross-basin patterns in oceanographic heterogeneity and productivity. Dominant species over the abyssal basin differed from those previously recorded over the neighbouring continental slope. Vertical biomass profiles indicated diffuse night-time distributions in the Central sector and extensive diel vertical migrations in the Eastern sector. In the Western sector, macrocrustacean δ13C values were significantly higher, and δ15N significantly lower, than those in the Eastern sector. The results indicate a cross-basin difference in the primary productivity environment and 15N enrichment at the base of the foodweb. The cross-basin pattern in lanternfish δ15N values mirrored that for macrocrustaceans and was not correlated with standard length. Lanternfish δ13C values did not differ between sectors, but there were depth-wise differences, with values in the shallowest stratum (0-200 m) significantly higher than those in the deepest stratum (800-1000 m). Calculated trophic levels (TLs) of lanternfishes spanned the third trophic level and marked niche segregation was evident in the Eastern (mean TL 3.0-3.9) and Central (mean TL 2.5-3.6) sectors. Together, the results suggest that the Eastern and Western sectors are distinct sub-basin scale pelagic habitats, with implications for ecosystem modelling and future monitoring.

  6. Neuroprotective effects of caffeine in MPTP model of Parkinson's disease: A (13)C NMR study.

    PubMed

    Bagga, Puneet; Chugani, Anup N; Patel, Anant B

    2016-01-01

    Parkinson's disease (PD) is a neurodegenerative disorder characterized by degeneration of nigrostriatal dopaminergic neurons with an accompanying neuroinflammation leading to loss of dopamine in the basal ganglia. Caffeine, a well-known A2A receptor antagonist is reported to slow down the neuroinflammation caused by activated microglia and reduce the extracellular glutamate in the brain. In this study, we have evaluated the neuroprotective effect of caffeine in the MPTP model of PD by monitoring the region specific cerebral energy metabolism. Adult C57BL6 mice were treated with caffeine (30 mg/kg, i.p.) 30 min prior to MPTP (25 mg/kg, i.p.) administration for 8 days. The paw grip strength of mice was assessed in order to evaluate the motor function after various treatments. For metabolic studies, mice were infused with [1,6-(13)C2]glucose, and (13)C labeling of amino acids was monitored using ex vivo(1)H-[(13)C]-NMR spectroscopy. The paw grip strength was found to be reduced following the MPTP treatment. The caffeine pretreatment showed significant protection against the reduction of paw grip strength in MPTP treated mice. The levels of GABA and myo-inositol were found to be elevated in the striatum of MPTP treated mice. The (13)C labeling of GluC4, GABAC2 and GlnC4 from [1,6-(13)C2]glucose was decreased in the cerebral cortex, striatum, olfactory bulb, thalamus and cerebellum suggesting impaired glutamatergic and GABAergic neuronal activity and neurotransmission of the MPTP treated mice. Most interestingly, the pretreatment of caffeine maintained the (13)C labeling of amino acids to the control values in cortical, olfactory bulb and cerebellum regions while it partially retained in striatal and thalamic regions in MPTP treated mice. The pretreatment of caffeine provides a partial neuro-protection against severe striatal degeneration in the MPTP model of PD. PMID:26626997

  7. Isotopic variation ( δ15N, δ13C, and δ34S) with body size in post-larval estuarine consumers

    NASA Astrophysics Data System (ADS)

    Marie Wilson, Rachel; Chanton, Jeffrey; Lewis, Graham; Nowacek, Douglas

    2009-07-01

    Stable isotope analyses were conducted on fish and crabs in Apalachicola Bay, Florida (USA) to determine whether δ15N values are correlated with length within these species. Our objective was to define the smallest trophic unit of the system as a prelude to further isotopic evaluations of consumer trophic level in Apalachicola Bay. In some cases, the smallest trophic unit is the individual species; however, previous gut content analysis on organisms in this system indicated that subdivision by size class within species might be required. However, over the size ranges observed, there was no systematic change in the δ15N values of individual consumers. Analysis of the δ13C and δ34S values of these species reveal no obvious shifts in the utilization of organic matter sources with length that could be obscuring a trend in the δ15N signal. We find no isotopic evidence for a systematic trophic level increase within the species examined over the size ranges observed in this estuary. These results indicate that species rather than size class is the best predictor of trophic habit among these Apalachicola Bay consumers.

  8. Dynamic 13C NMR analysis of oxidative metabolism in the in vivo canine myocardium.

    PubMed

    Robitaille, P M; Rath, D P; Abduljalil, A M; O'Donnell, J M; Jiang, Z; Zhang, H; Hamlin, R L

    1993-12-15

    Oxidative metabolism in the in vivo canine myocardium was studied noninvasively using 13C-enriched acetate and non-steady state 13C NMR techniques. Under low workload conditions, the myocardium oxidized the infused [2-13C]acetate and incorporated the labeled carbon into the glutamate pool as expected. This conclusion stems from the rapid enrichment of the C-2, C-3, and C-4 carbons of glutamic acid both under in vivo conditions and in extracts. Surprisingly, [2-13C]acetate uptake was not observed at high workloads as reflected by an absence of glutamate pool enrichment at these rate pressure products. Rather, the myocardium selected its substrate from an endogenous pool. Since free acetate can directly cross the inner mitochondrial membrane and be converted to acetyl-CoA through acetyl-CoA synthetase, these results support workload-dependent regulation of substrate access to the mitochondrial CoASH pool. As such, we advance the hypothesis that the selection of substrate for condensation with CoASH and subsequent oxidation in the tricarboxylic acid cycle is regulated kinetically through the Km values of the appropriate condensation enzymes and through the absolute levels of free CoASH in the mitochondria. PMID:8253751

  9. The complete set of spin observables for the (13)C(polarized proton, polarized neutron)(13)N and (15)N(polarized proton, polarized neutron)(15)O reactions

    NASA Astrophysics Data System (ADS)

    Du, Qun Qun

    1998-12-01

    The 13C(p,n)13N and 15N(p,n)15O reactions have been a puzzle for more than ten years. The ground state transitions are Jπ=1/2- to Jπ=1/2-. These are 'mixed' transitions because they can involve quantum number changes either (/Delta T=1,/ /Delta J=0,/ /Delta/pi=0,/ /Delta S=0), or (/Delta T=1,/ /Delta J=1,/ /Delta/pi=0,/ /Delta S=1); these quantum number changes are refered to as 'Fermi' and 'Gamow-Teller' respectively. Because the quantum number changes are the same as for Fermi and Gamow-Teller beta decay. From the systematics of (p,n) and (n,p) reactions on pure Fermi transitions (e.g. 0 + to 0+) and pure Gamow-Teller transitions (e.g. 0+ to 1+), calibrations have been established of cross section per unit B(F) or unit B(GT), where 'B' refers to doubly reduced matrix elements extracted from beta decay. However, cross sections for the 13C(p,n)13N(g.s.) and 15N(p,n)15O(g.s.) reactions are substantially larger than one would then predict from the known B(F)s and B(GT)s for these transitions. To explore this anomaly, spin observables were used to extract separately the Fermi and Gamow-Teller cross sections for these reactions. To acquire the complete sets of polarization- transfer observables, a new neutron polarimeter was designed, built, commissioned and calibrated. This polarimeter, call the '2π polarimeter' because of its complete azimuthal coverage for scattered neutrons, has very good position and timing resolution (354 ps). The complete sets of spin-transfer coefficients Dij for 13C(p,n)13N (at 0o , 5.5o , and 11o ) and 15N(p,n)15O (at 0o ) at 135 MeV were measured. Following the formalism of Ichimura and Kawahigashi, we extracted the spin-longitudinal, and spin-transverse and spin-independent responses D0,/ Dq,/ Dn and Dp from the measured Dijs. The F and GT fractions of the (p,n) cross sections are then extracted as f F=D0 and fGT=Dn+Dp+Dq=1- d0. Values of Dk for both the 13C(p,n)13N(g.s) and 15N(p,n)15O(g.s.) were extracted. From these responses, we

  10. Determination of Spin-Lattice Relaxation of Time Using (Super 13)C NMR: An Undergraduate Physical Chemistry Laboratory Experiment

    ERIC Educational Resources Information Center

    Gasyna, Zbigniew L.; Jurkiewicz, Antoni

    2004-01-01

    An experiment designed for the physical chemistry laboratory where (super 13)C NMR is applied to determine the spin-lattice relaxation time for carbon atoms in n-hexanol is proposed. It is concluded that students learn the principles and concepts of NMR spectroscopy as well as dynamic NMR experiments.

  11. Analysis of Hydroperoxides in Solid Polyethylene by MAS (13)C NMR and EPR

    SciTech Connect

    ASSINK,ROGER A.; CELINA,MATHIAS C.; DUNBAR,TIMOTHY D.; ALAM,TODD M.; CLOUGH,ROGER LEE; GILLEN,KENNETH T.

    1999-11-19

    {sup 13}C-enriched polyethylene was subjected to {gamma}-irradiation in the presence of air at 25 and 80 C for total doses ranging from 71 to 355 kGy. Significant quantities of hydroperoxides were detected in the 25 C irradiated sample by {sup 13}C magic angle spinning NMR spectroscopy. This method of detection was performed on the solid polymer and required no chemical derivatization or addition of solvent. The chemical stability and subsequent products of the hydroperoxide species were studied by annealing the irradiated samples in air at temperatures ranging from 22 to 110 C. A time-temperature superposition analysis provided an activation energy of 108 kJ/mol for the hydroperoxide decomposition process. The primary products of hydroperoxide decomposition were ketones and secondary alcohols with lesser amounts of acids and esters. EPR measurements suggest that the reactive hydroperoxide species reside in the amorphous phase of polyethylene, consistent with degradation occurring in the amorphous phase.

  12. 13C NMR spectroscopy of the insoluble carbon of carbonaceous chondrites

    NASA Technical Reports Server (NTRS)

    Cronin, J. R.; Pizzarello, S.; Frye, J. S.

    1987-01-01

    13C NMR spectra have been obtained of the insoluble carbon residues resulting from HF-digestion of three carbonaceous chondrites, Orgueil (C1), Murchison (CM2), and Allende (CV3). Spectra obtained using the cross polarization magic-angle spinning technique show two major features attributable respectively to carbon in aliphatic/olefinic structures. The spectrum obtained from the Allende sample was weak, presumably as a consequence of its low hydrogen content. Single pulse excitation spectra, which do not depend on 1H-13C polarization transfer for signal enhancement were also obtained. These spectra, which may be more representative of the total carbon in the meteorite samples, indicate a greater content of carbon in aromatic/olefinic structures. These results suggest that extensive polycyclic aromatic sheets are important structural features of the insoluble carbon of all three meteorites. The Orgueil and Murchison materials contain additional hydrogenated aromatic/olefinic and aliphatic groups.

  13. NMR analyses of the cold cataract. III. /sup 13/C acrylamide studies

    SciTech Connect

    Lerman, S.; Megaw, J.M.; Moran, M.N.

    1985-10-01

    /sup 13/C-enriched acrylamide was employed to further delineate the action of this compound in preventing the cold cataract phenomenon when it is incorporated (in vitro) into young human and rabbit lenses. The extent of acrylamide incorporation, in the dark and with concurrent UV exposure, was monitored by /sup 13/C NMR spectroscopy. These studies provide further evidence that UV exposure causes permanent acrylamide photobinding within the lens. In such lenses, the gamma crystallin fraction of the soluble lens proteins is affected to the greatest extent. It appears to become aggregated and/or combined with the alpha and beta fractions resulting in an apparent loss of most of the gamma monomers. There is also an age-related effect with respect to the amount of acrylamide that can be incorporated into the lens. The decrease in acrylamide incorporation with age directly parallels the age-related decline in gamma crystallin levels.

  14. Characterisation of coke from FCC catalysts by solid state {sup 13}C NMR and mass spectrometry

    SciTech Connect

    Andresen, J.M.; McGhee, B.; Snape, C.E.

    1995-12-31

    Coke has been concentrated by demineralisation from deactivated FCC catalysts from both refinery operations with actual feeds and MAT tests using n-hexadecane to facilitate detailed characterisation by solid state {sup 13}C NMR and mass spectrometry. All the catalysts investigated contained about 1% w/w carbon. As for solid fuels, the use of a low-field spectrometer for solid state {sup 13}C NMR in conjunction with the single pulse excitation (SPE or Bloch decay) technique has enabled quantitative carbon skeletal parameters to be obtained for the cokes. Internal standard measurements demonstrated that most of the carbon was observed by SPE and, therefore, NMR-invisible graphitic layers are not thought to be major structural features of the cokes. Differences in feedstock composition were reflected in the structure of the refinery cokes with the aromatic nuclei from a residue feed (5% Conradson carbon) corresponding to 15-20 peri-condensed aromatic rings and being more highly condensed than those from a hydrotreated vacuum gas oil. Mass spectrometry (EI, CI and FIMS) has confirmed that the refinery cokes are highly condensed, but those obtained from n-hexadecane in the MAT tests displayed significant aliphatic character.

  15. Hyperpolarized 13C NMR lifetimes in the liquid-state: relating structures and T1 relaxation times

    NASA Astrophysics Data System (ADS)

    Parish, Christopher; Niedbalski, Peter; Hashami, Zohreh; Fidelino, Leila; Kovacs, Zoltan; Lumata, Lloyd

    Among the various attempts to solve the insensitivity problem in nuclear magnetic resonance (NMR), the physics-based technique dissolution dynamic nuclear polarization (DNP) is probably the most successful method of hyperpolarization or amplifying NMR signals. Using this technique, liquid-state NMR signal enhancements of several thousand-fold are expected for low-gamma nuclei such as carbon-13. The lifetimes of these hyperpolarized 13C NMR signals are directly related to their 13C spin-lattice relaxation times T1. Depending upon the 13C isotopic location, the lifetimes of hyperpolarized 13C compounds can range from a few seconds to minutes. In this study, we have investigated the hyperpolarized 13C NMR lifetimes of several 13C compounds with various chemical structures from glucose, acetate, citric acid, naphthalene to tetramethylallene and their deuterated analogs at 9.4 T and 25 deg C. Our results show that the 13C T1s of these compounds can range from a few seconds to more than 60 s at this field. Correlations between the chemical structures and T1 relaxation times will be discussed and corresponding implications of these results on 13C DNP experiments will be revealed. US Dept of Defense Award No. W81XWH-14-1-0048 and Robert A. Welch Foundation Grant No. AT-1877.

  16. Biogeochemical Indicators in High- and Low-Arctic Marine and Terrestrial Avian Community Changes: Comparative Isotopic (13C, 15N, and 34S) Studies in Alaska and Greenland

    NASA Astrophysics Data System (ADS)

    Causey, D.; Bargmann, N. A.; Burnham, K. K.; Burnham, J. L.; Padula, V. M.; Johnson, J. A.; Welker, J. M.

    2011-12-01

    Understanding the complex dynamics of environmental change in northern latitudes is of paramount importance today, given documented rapid shifts in sea ice, plant phenology, temperatures, deglaciation, and habitat fidelity. This knowledge is particularly critical for Arctic avian communities, which are integral components by which biological teleconnections are maintained between the mid and northern latitudes. Furthermore, Arctic birds are fundamental to Native subsistence lifestyles and a focus for conservation activities. Avian communities of marine and terrestrial Arctic environments represent a broad spectrum of trophic levels, from herbivores (eg., geese Chen spp.), planktivores (eg., auklets Aethia spp.), and insectivores (eg., passerines: Wheatears Oenanthe spp., Longspurs Calcarius spp.), to predators of marine invertebrates (eg., eiders Somateria spp.), nearshore and offshore fish (eg., cormorants Phalacrocorax spp, puffins Fratercula spp.), even other bird species (eg., gulls Larus spp., falcons Peregrinus spp.). This diversity of trophic interconnections is an integral factor in the dynamics of Arctic ecosystem ecology, and they are key indicators for the strength and trajectories of change. We are especially interested in their feeding ecology, using stable isotope-diet relations to examine historical diets and to predict future feeding ecology by this range of species. Since 2009, we have been studying the foodweb ecology using stable isotopes (δ13C, δ15N, δ34S) of contemporaneous coastal and marine bird communities in High Arctic (Northwest Greenland) and Low Arctic (western Aleutian Islands, AK). We are quantifying the isotopic values of blood, organ tissues, and feathers, and have carried out comparisons between native and lipid-extracted samples. Although geographically distant, these communities comprise similar taxonomic and ecological congeners, including several species common to both (eg., Common Eider, Black-legged Kittiwake, Northern

  17. Application of δ13C and δ15N isotopic signatures of organic matter fractions sequentially separated from adjacent arable and forest soils to identify carbon stabilization mechanisms

    NASA Astrophysics Data System (ADS)

    Kayler, Z. E.; Kaiser, M.; Gessler, A.; Ellerbrock, R. H.; Sommer, M.

    2011-03-01

    Identifying the chemical mechanisms behind soil carbon bound in organo-mineral complexes is necessary to determine the degree to which soil organic carbon is stabilized belowground. We used the δ13C and δ15N isotopic signatures from two organic matter (OM) fractions from soil to identify the likely binding mechanisms involved. We used OM fractions hypothesized to contain carbon stabilized through organo-mineral complexes: (1) OM separated chemically with sodium pyrophosphate (OM(PY)) and (2) OM stabilized in microstructures found in the chemical extraction residue (OM(ER)). Furthermore, because the OM fractions were separated from five different soils with paired forest and arable land use histories, we could address the impact of land use change on carbon binding and processing mechanisms within these soils. We used partial least squares regression to analyze patterns in the isotopic signature of OM with established proxies of different binding mechanisms. Parsing soil OM into different fractions is a systematic method of dissection, however, we are primarily interested in how OM is bound in soil as a whole, requiring a means of re-assembly. Thus, we implemented the recent zonal framework described by Kleber et al. (2007) to relate our findings to undisturbed soil. The δ15N signature of OM fractions served as a reliable indicator for microbial processed carbon in both arable and forest land use types. The δ13C signature of OM fractions in arable sites did not correlate well with proxies of soil mineral properties while a consistent pattern of enrichment was seen in the δ13C of OM fractions in the forest sites. We found a significant difference in δ13C of pooled OM fractions between the forest and arable land use type although it was relatively small (<1‰). We found different binding mechanisms predominate in each land use type. The isotopic signatures of OM fractions from arable soils were highly related to the clay and silt size particles amount while

  18. The Influence of Plant Litter on Soil Water Repellency: Insight from 13C NMR Spectroscopy.

    PubMed

    Cesarano, Gaspare; Incerti, Guido; Bonanomi, Giuliano

    2016-01-01

    Soil water repellency (SWR, i.e. reduced affinity for water owing to the presence of organic hydrophobic coatings on soil particles) has relevant hydrological implications because low rates of infiltration enhance water runoff, and untargeted diffusion of fertilizers and pesticides. Previous studies investigated the occurrence of SWR in ecosystems with different vegetation cover but did not clarify its relationships with litter biochemical quality. Here, we investigated the capability of different plant litter types to induce SWR by using fresh and decomposed leaf materials from 12 species, to amend a model sandy soil over a year-long microcosm experiment. Water repellency, measured by the Molarity of an Ethanol Droplet (MED) test, was tested for the effects of litter species and age, and compared with litter quality assessed by 13C-CPMAS NMR in solid state and elemental chemical parameters. All litter types were highly water repellent, with MED values of 18% or higher. In contrast, when litter was incorporated into the soil, only undecomposed materials induced SWR, but with a large variability of onset and peak dynamics among litter types. Surprisingly, SWR induced by litter addition was unrelated to the aliphatic fraction of litter. In contrast, lignin-poor but labile C-rich litter, as defined by O-alkyl C and N-alkyl and methoxyl C of 13C-CPMAS NMR spectral regions, respectively, induced a stronger SWR. This study suggests that biochemical quality of plant litter is a major controlling factor of SWR and, by defining litter quality with 13C-CPMAS NMR, our results provide a significant novel contribution towards a full understanding of the relationships between plant litter biochemistry and SWR. PMID:27022916

  19. The Influence of Plant Litter on Soil Water Repellency: Insight from 13C NMR Spectroscopy

    PubMed Central

    Cesarano, Gaspare; Incerti, Guido; Bonanomi, Giuliano

    2016-01-01

    Soil water repellency (SWR, i.e. reduced affinity for water owing to the presence of organic hydrophobic coatings on soil particles) has relevant hydrological implications because low rates of infiltration enhance water runoff, and untargeted diffusion of fertilizers and pesticides. Previous studies investigated the occurrence of SWR in ecosystems with different vegetation cover but did not clarify its relationships with litter biochemical quality. Here, we investigated the capability of different plant litter types to induce SWR by using fresh and decomposed leaf materials from 12 species, to amend a model sandy soil over a year-long microcosm experiment. Water repellency, measured by the Molarity of an Ethanol Droplet (MED) test, was tested for the effects of litter species and age, and compared with litter quality assessed by 13C-CPMAS NMR in solid state and elemental chemical parameters. All litter types were highly water repellent, with MED values of 18% or higher. In contrast, when litter was incorporated into the soil, only undecomposed materials induced SWR, but with a large variability of onset and peak dynamics among litter types. Surprisingly, SWR induced by litter addition was unrelated to the aliphatic fraction of litter. In contrast, lignin-poor but labile C-rich litter, as defined by O-alkyl C and N-alkyl and methoxyl C of 13C-CPMAS NMR spectral regions, respectively, induced a stronger SWR. This study suggests that biochemical quality of plant litter is a major controlling factor of SWR and, by defining litter quality with 13C-CPMAS NMR, our results provide a significant novel contribution towards a full understanding of the relationships between plant litter biochemistry and SWR. PMID:27022916

  20. (13)C-(1)H NMR relaxation and fluorescence anisotropy decay study of tyrosine dynamics in motilin.

    PubMed Central

    Damberg, Peter; Jarvet, Jüri; Allard, Peter; Mets, Ulo; Rigler, Rudolf; Gräslund, Astrid

    2002-01-01

    Tyrosine ring dynamics of the gastrointestinal hormone motilin was studied using two independent physical methods: fluorescence polarization anisotropy decay and NMR relaxation. Motilin, a 22-residue peptide, was selectively (13)C labeled in the ring epsilon-carbons of the single tyrosine residue. To eliminate effects of differences in peptide concentration, the same motilin sample was used in both experiments. NMR relaxation rates of the tyrosine ring C(epsilon)-H(epsilon) vectors, measured at four magnetic field strengths (9.4, 11.7, 14.1, and 18.8 Tesla) were used to map the spectral density function. When the data were analyzed using dynamic models with the same number of components, the dynamic parameters from NMR and fluorescence are in excellent agreement. However, the estimated rotational correlation times depend on the choice of dynamic model. The correlation times estimated from the two-component model-free approach and the three-component models were significantly different (1.7 ns and 2.2 ns, respectively). Various earlier studies of protein dynamics by NMR and fluorescence were compared. The rotational correlation times estimated by NMR for samples with high protein concentration were on average 18% longer for folded monomeric proteins than the corresponding times estimated by fluorescence polarization anisotropy decay, after correction for differences in viscosity due to temperature and D(2)O/H(2)O ratio. PMID:12414713

  1. sup 13 C and sup 31 P NMR studies of myocardial metabolism

    SciTech Connect

    Laughlin, M.R.

    1988-01-01

    The fluxes through two enzyme systems have been measured in perfused or in in vivo heart using NMR: phosphocreatine kinase, and glycogen synthase and phosphorylase. The rates of synthesis and degradation of glycogen were monitored in vivo in fed, fasted, and diabetic rat heart during infusions of {sup 13}C-1-glucose and insulin using proton-decoupled {sup 13}C-NMR at 1.9 and 4.7 tesla. The enzyme activities of glycogen synthase and glycogen phosphorylase were also measured in this tissue which had been freeze clamped at the end of the experiment, for comparison with the synthetic rates. For normal fed, fasted, and diabetic animals, synthesis rates were 0.28, 0.16, and 0.15 {mu}mol/min.gww respectively. Glycogen synthase i activity was 0.23, 0.14, and 0.14 {mu}mol/min.gww in these hearts at the end of the experiment, when measured at appropriate substrate and activator concentrations, and follow activation time courses that are consistent with being the main rate determinant for net synthesis in all cases. Turnover of glycogen was studied by observing the preformed {sup 13}C-1-glycogen signal during infusion of {sup 12}C-glucose and insulin, and was found to be close to zero. Extracted phosphorylase a activity was approximately ten times that of synthase i under these circumstances. In order to fully interpret the turnover studies, glycogenolysis of preformed {sup 13}C-glycogen was observed after a bolus of glucagon. The glycogen had either been synthesized from {sup 13}C-1-glucose for a single hour, or during an hour of {sup 13}C-glucose and a subsequent hour of {sup 12}C-glucose infusion. The author observed that breakdown follows an exponential time course related to the phosphorylase a activation state and that the last synthesized glycogen breaks down at the rate of 2.5 {mu}mol/min.gww, five times faster than that synthesized an hour earlier.

  2. Cucurbitacins from Cayaponia racemosa: isolation and total assignment of 1H and 13C NMR spectra.

    PubMed

    Chaves, Davina C; Assunção, João Carlos C; Braz-Filho, Raimundo; Lemos, Telma L G; Monte, Francisco J Q

    2007-05-01

    Two new cucurbitane-type triterpenoids, 2beta,3beta,16alpha,20(R),25-pentahydroxy-9-methyl-19-norlanost-5-en-7,22-dione and 2beta,3beta,16alpha,20(R),25-pentahydroxy-9-methyl-19-norlanost-5-en-7,11,22-trione, were isolated from fruits of Cayaponia racemosa. The total (1)H and (13)C chemical shift assignment of these two closely related compounds is described, making use of one- and two-dimensional NMR techniques. PMID:17372957

  3. The binding of metal ions and angiotensin converting enzyme (ACE) inhibitor by 13C NMR

    NASA Astrophysics Data System (ADS)

    Sakamoto, Yohko; Sakamoto, Yuko; Ishii, Tomoko; Ohmoto, Taichi

    1991-06-01

    Enalaprilat (MK-422, 1- [ N- [1 (S)-carboxy-3-phenylpropyl]- L-alanyl]- L-proline (1)) and Lisinopril (MK521, N- N- [ (s)-l-carboxy-3- phenylpropyl]- L-lysyl- L-proline, (2)) exhibit the capacity to act as a chelate, unidentate or bridge towards metal ions in aqueous solution, as determined by 13C NMR. By adding metal ions, in the series of Zn 2+, Ni 2+, Pb 2+, Pd 2+ and Cd 2+, the active site of the ACE inhibitor was well defined. MK-521 was more influenced by nuclei that were distant from the active site than MK-422.

  4. 13C NMR spectral characterization of epimeric rotenone and some related tetrahydrobenzopyranofurobenzopyranones

    USGS Publications Warehouse

    Abidi, S.L.; Abidi, M.S.

    1983-01-01

    The 13C nuclear magnetic resonance (nmr) spectra of epimers of rotenone and four 12a-hydroxy-analogues were examined to determine the stereochemical effect of the B/C ring fusion involving the 6a- and 12a-carbon centers. Chemical shift differences between the epimeric carbon resonances of cis- and trans-6a,12a-compounds were notably larger than those of diastereoisomers derived from the same B/C ring junction stereochemistry. Results of the spectral analysis have been useful for the quantification of mixtures of epimers and for the measurement of rates of epimerization and oxygenation.

  5. Compartmentalized Cerebral Metabolism of [1,6-13C]Glucose Determined by in vivo 13C NMR Spectroscopy at 14.1 T

    PubMed Central

    Duarte, João M. N.; Lanz, Bernard; Gruetter, Rolf

    2011-01-01

    Cerebral metabolism is compartmentalized between neurons and glia. Although glial glycolysis is thought to largely sustain the energetic requirements of neurotransmission while oxidative metabolism takes place mainly in neurons, this hypothesis is matter of debate. The compartmentalization of cerebral metabolic fluxes can be determined by 13C nuclear magnetic resonance (NMR) spectroscopy upon infusion of 13C-enriched compounds, especially glucose. Rats under light α-chloralose anesthesia were infused with [1,6-13C]glucose and 13C enrichment in the brain metabolites was measured by 13C NMR spectroscopy with high sensitivity and spectral resolution at 14.1 T. This allowed determining 13C enrichment curves of amino acid carbons with high reproducibility and to reliably estimate cerebral metabolic fluxes (mean error of 8%). We further found that TCA cycle intermediates are not required for flux determination in mathematical models of brain metabolism. Neuronal tricarboxylic acid cycle rate (VTCA) and neurotransmission rate (VNT) were 0.45 ± 0.01 and 0.11 ± 0.01 μmol/g/min, respectively. Glial VTCA was found to be 38 ± 3% of total cerebral oxidative metabolism, accounting for more than half of neuronal oxidative metabolism. Furthermore, glial anaplerotic pyruvate carboxylation rate (VPC) was 0.069 ± 0.004 μmol/g/min, i.e., 25 ± 1% of the glial TCA cycle rate. These results support a role of glial cells as active partners of neurons during synaptic transmission beyond glycolytic metabolism. PMID:21713114

  6. (13) C-TmDOTA as versatile thermometer compound for solid-state NMR of hydrated lipid bilayer membranes.

    PubMed

    Umegawa, Yuichi; Tanaka, Yuya; Nobuaki, Matsumori; Murata, Michio

    2016-03-01

    Recent advances in solid-state nuclear magnetic resonance (NMR) techniques, such as magic angle spinning and high-power decoupling, have dramatically increased the sensitivity and resolution of NMR. However, these NMR techniques generate extra heat, causing a temperature difference between the sample in the rotor and the variable temperature gas. This extra heating is a particularly crucial problem for hydrated lipid membrane samples. Thus, to develop an NMR thermometer that is suitable for hydrated lipid samples, thulium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTA) was synthesized and labeled with (13) C (i.e., (13) C-TmDOTA) to increase the NMR sensitivity. The complex was mixed with a hydrated lipid membrane, and the system was subjected to solid-state NMR and differential scanning calorimetric analyses. The physical properties of the lipid bilayer and the quality of the NMR spectra of the membrane were negligibly affected by the presence of (13) C-TmDOTA, and the (13) C chemical shift of the complex exhibited a large-temperature dependence. The results demonstrated that (13) C-TmDOTA could be successfully used as a thermometer to accurately monitor temperature changes induced by (1) H decoupling pulses and/or by magic angle spinning and the temperature distribution of the sample inside the rotor. Thus, (13) C-TmDOTA was shown to be a versatile thermometer for hydrated lipid assemblies. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26460094

  7. Quantification of soy protein using the isotope method (δ(13)C and δ(15)N) for commercial brands of beef hamburger.

    PubMed

    Ducatti, Rhani; de Almeida Nogueira Pinto, José Paes; Sartori, Maria Márcia Pereira; Ducatti, Carlos

    2016-12-01

    Hamburgers (beef patties) may be adulterated through the overuse of protein extenders. Among vegetables, soy protein is the best substitute for animal protein. These ingredients help to reduce the cost of producing a final product, and they maximize profits for fraudulent industries. Moreover, the ingestion of soy or other non-meat proteins by allergic individuals may present a health risk. In addition, monitoring by supervisory bodies is hampered by a lack of appropriate analytical methodologies. Within this context, the aim of this study was to determine and quantify the levels of added soy protein by determination of (15)N and (13)C stable isotopes. A total of 100 beef hamburger samples from 10 commercial brands were analyzed. Only three samples of the G brand were within the standards set the Brazilian legislation. The remaining 97 samples from 10 commercial brands contained >4% soy protein; therefore, they are adulterated and not in compliance with the current legislation. PMID:27501234

  8. Trace elements (Cu, Zn, and Hg) and δ(13)C/δ(15)N in seabird subfossils from three islands of the South China Sea and its implications.

    PubMed

    Xu, Liqiang; Liu, Xiaodong; Nie, Yaguang

    2016-05-01

    Seabird subfossils were collected on three islands of the Xisha Archipelago, South China Sea. Via elemental analysis, we identified that bird guano was a significant source for heavy metals Cu, Zn, and Hg. Cu and Zn levels in these guano samples are comparable to their levels in wildbird feces, but guano Hg was lower than previously reported. Trophic positions significantly impacted transfer efficiency of heavy metals by seabirds. Despite of a common source, trace elements, as well as stable isotopes (i.e., guano δ(13)C and collagen δ(15)N), showed island-specific characteristics. Bird subfossils on larger island had relatively greater metal concentrations and revealed higher trophic positions. Partition of element and isotope levels among the islands suggested that transfer efficacy of seabirds on different islands was different, and bird species were probably unevenly distributed among the islets. Island area is possibly a driving factor for distributions of seabird species. PMID:27056479

  9. Assessment of trace elements, POPs, (210)Po and stable isotopes ((15)N and (13)C) in a rare filter-feeding shark: The megamouth.

    PubMed

    Moura, Jailson Fulgencio de; Merico, Agostino; Montone, Rosalinda Carmela; Silva, Josilene; Seixas, Tércia Guedes; Godoy, José Marcus de Oliveira; Saint'Pierre, Tatiana Dillenburg; Hauser-Davis, Rachel Ann; Di Beneditto, Ana Paula Madeira; Reis, Estéfane Cardinot; Tavares, Davi Castro; Lemos, Leila Soledade; Siciliano, Salvatore

    2015-06-15

    With less than 60 records being reported worldwide, the megamouth (Megachasma pelagios) is today one of the least known shark species inhabiting our oceans. Therefore, information concerning the biology and ecology of this enigmatic organism is very scarce and limited to feeding behaviour and preferred habitat. The present work reports new data on the concentrations of trace elements, organic mercury, POPs and (210)Po in hepatic and muscular tissues of a specimen found stranded in the southeastern coast of Brazil. Additionally, we provide new evidence based on stable isotope analysis (δ(15)N and δ(13)C) confirming the preference for the pelagic habitat and the zooplanktivorous feeding behaviour of the megamouth. These results are consistent with the low concentrations of organic pollutant compounds and other elements measured in our samples. PMID:25858662

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

    PubMed

    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. PMID:26878852

  11. Determination of [{sup 13}C]pyrene sequestration in sediment microcosms using flash pyrolysis--GC--MS and {sup 13}C NMR

    SciTech Connect

    Guthrie, E.A.; Bortiatynski, J.M.; Hardy, K.S.; Kovach, E.M.; Van Heemst, J.D.H.; Hatcher, P.G.; Richman, J.E.

    1999-01-01

    In this study, the use of a {sup 13}C-labeled pollutant probe, [{sup 13}C]pyrene, and the application of flash pyrolysis--GC--MS and CPMAS {sup 13}C NMR provided analytical capabilities to study pyrene interactions with soluble and insoluble compartments of sedimentary organic matter (S{sub D}OM) during whole sediments incubations in aerated microcosms. Surface sediments were collected from a site of previous hydrocarbon contamination in New Orleans, LA. Over a period of 60 days, humic acid and humin fractions of S{sub D}OM accumulated increasing amounts of pyrene that were resistant to exhaustive extraction with organic solvents. The sequestered pyrene was evident in CPMAS {sup 13}C NMR spectra of humin fractions. The amount of sequestered pyrene in humic materials was quantified by flash pyrolysis--GC--MS, a technique that destroys the three-dimensional structure of macromolecular S{sub D}OM. Noncovalent binding of pyrene to humic materials in S{sub D}OM was greater in sediments incubated with biological activity than biocide-treated sediments. The combined analytical approaches demonstrate that the sequestered pyrene, or bound residue, is noncovalently associated with S{sub D}OM and has not undergone structural alteration. Implications of these data are discussed in reference to S{sub D}OM diagenesis and long-term availability of bound pollutant residues in sediments.

  12. Stabilization dynamics of root versus needle-derived 13C and 15N during 10 years in a temperate forest soil.

    NASA Astrophysics Data System (ADS)

    Bird, J. A.; Hatton, P. J.; Castanha, C.; Torn, M. S.

    2012-12-01

    Belowground plant carbon (C) allocation as fine roots can result in greater retention of C in soils compared with aboveground litter in temperate forest ecosystems. However, much of our understanding of the fate of fine root C and nitrogen (N) in soils comes from short-term studies, often lasting only a few months to a few years. In 2011, we concluded a 10-year field study that compared the fate of dual-labeled (13C/15N) Ponderosa pine fine roots (<2 mm diameter) with needles in a temperate forest soil of the Sierra Nevada, CA, USA. The 13C- and 15N-labeled fine roots or needles were added to mesocosms at two soil depths (top of O or A horizon) to compare C and N stabilization in mineral versus organic soil horizons. We will present data on retention of litter C and N in soil after 0.5, 1.5, 5 and 10 years in situ. For soil samples recovered after 5 years, litter-derived C and N recovered in the mineral soil was partitioned into several operationally-defined physical and chemical soil organic matter (SOM) fractions, which were also characterized by natural abundance 14C. In addition, we compared two fractionation methods (i.e., with and without occluded light fraction isolation) on the partitioning of litter-derived C and N in mineral soil. After 5 years in situ the retention of fine root C in soil (59.9±3.8%) was significantly greater than that of added needle C (38.4±2.0%); however the depth of litter placement in the soil did not affect total litter C or N recovery. Our results provide a direct, decade-scale measure of stabilization of above- and belowground plant inputs to soil, including a portrait of the dominant stabilization mechanisms.

  13. Effect of age and ration on diet-tissue isotopic (Δ13C, Δ15N) discrimination in striped skunks (Mephitis mephitis).

    PubMed

    Hobson, Keith A; Quirk, Travis W

    2014-01-01

    An important prerequisite for the effective use of stable isotopes in animal ecology is the accurate assessment of isotopic discrimination factors linking animals to their diets for a multitude of tissue types. Surprisingly, these values are poorly known in general and especially for mammalian carnivores and omnivores in particular. Also largely unknown are the factors that influence diet-tissue isotopic discrimination such as nutritional quality and age. We raised adult and juvenile striped skunks (Mephitis mephitis) in captivity on a constant omnivore diet (Mazuri Omnivore A 5635). Adults (n=6) and juveniles (n=3) were kept for 7 months and young (n=7) to the age of 50 days. We then examined individuals for stable carbon (δ(13)C) and nitrogen (δ(15)N) isotope values of hair, nails, lipid, liver, muscle, bone collagen and the plasma, and cellular fractions of blood. Discrimination values differed among age groups and were significantly higher for young compared with their mothers, likely due to the effects of weaning. Δ(15)N isotopic discrimination factors ranged from 3.14 (nails) to 5.6‰ (plasma) in adults and 4.3 (nails) to 5.8‰ (liver) for young. For Δ(13)C, values ranged from-3.3 (fat) to 3.0‰ (collagen) in adults and from-3.3 (fat) to 2.0‰ (collagen) in young. Our data provide an important tool for predicting diets and source of feeding for medium-sized mammalian omnivorous adults integrated over short (e.g. liver, plasma) through long (e.g. collagen) periods and underline the potential effects of age on isotopic values in omnivore diets. PMID:24506487

  14. Quantitative solid-state 13C NMR with signal enhancement by multiple cross polarization

    NASA Astrophysics Data System (ADS)

    Johnson, Robert L.; Schmidt-Rohr, Klaus

    2014-02-01

    A simple new method is presented that yields quantitative solid-state magic-angle spinning (MAS) 13C NMR spectra of organic materials with good signal-to-noise ratios. It achieves long (>10 ms) cross polarization (CP) from 1H without significant magnetization losses due to relaxation and with a moderate duty cycle of the radio-frequency irradiation, by multiple 1-ms CP periods alternating with 1H spin-lattice relaxation periods that repolarize the protons. The new method incorporates previous techniques that yield less distorted CP/MAS spectra, such as a linear variation (“ramp”) of the radio-frequency field strength, and it overcomes their main limitation, which is T1ρ relaxation of the spin-locked 1H magnetization. The ramp of the radio-frequency field strength and the asymptotic limit of cross polarization makes the spectral intensity quite insensitive to the exact field strengths used. The new multiCP pulse sequence is a “drop-in” replacement for previous CP methods and produces no additional data-processing burden. Compared to the only reliable quantitative 13C NMR method for unlabeled solids previously available, namely direct-polarization NMR, the measuring time is reduced by more than a factor of 50, enabling higher-throughput quantitative NMR studies. The new multiCP technique is validated with 14-kHz MAS on amino-acid derivatives, plant matter, a highly aromatic humic acid, and carbon materials made by low-temperature pyrolysis.

  15. δ(13)C and δ(15)N in deep-living fishes and shrimps after the Deepwater Horizon oil spill, Gulf of Mexico.

    PubMed

    Quintana-Rizzo, Ester; Torres, Joseph J; Ross, Steve W; Romero, Isabel; Watson, Kathleen; Goddard, Ethan; Hollander, David

    2015-05-15

    The blowout of the Deepwater Horizon (DWH) drill-rig produced a surface oil layer, dispersed micro-droplets throughout the water column, and sub-surface plumes. We measured stable carbon and nitrogen isotopes in mesopelagic fishes and shrimps in the vicinity of DWH collected prior to, six weeks after, and one year after the oil spill (2007, 2010 and 2011). In 2010, the year of the oil spill, a small but significant depletion of δ(13)C was found in two mesopelagic fishes (Gonostoma elongatum and Chauliodus sloani) and one shrimp (Systellaspis debilis); a significant δ(15)N enrichment was identified in the same shrimp and in three fish species (G. elongatum, Ceratoscopelus warmingii, and Lepidophanes guentheri). The δ(15)N change did not suggest a change of trophic level, but did indicate a change in diet. The data suggest that carbon from the Deepwater Horizon oil spill was incorporated into the mesopelagic food web of the Gulf of Mexico. PMID:25778549

  16. delta 15N and non-carbonate delta 13C values for two petroleum source rock reference materials and a marine sediment reference material

    USGS Publications Warehouse

    Dennen, Kristin O.; Johnson, Craig A.; Otter, Marshall L.; Silva, Steven R.; Wandless, Gregory A.

    2006-01-01

    Samples of United States Geological Survey (USGS) Certified Reference Materials USGS Devonian Ohio Shale (SDO-1), and USGS Eocene Green River Shale (SGR-1), and National Research Council Canada (NRCC) Certified Marine Sediment Reference Material (PACS-2), were sent for analysis to four separate analytical laboratories as blind controls for organic rich sedimentary rock samples being analyzed from the Red Dog mine area in Alaska. The samples were analyzed for stable isotopes of carbon (delta13Cncc) and nitrogen (delta15N), percent non-carbonate carbon (Wt % Cncc) and percent nitrogen (Wt % N). SDO-1, collected from the Huron Member of the Ohio Shale, near Morehead, Kentucky, and SGR-1, collected from the Mahogany zone of the Green River Formation are petroleum source rocks used as reference materials for chemical analyses of sedimentary rocks. PACS-2 is modern marine sediment collected from the Esquimalt, British Columbia harbor. The results presented in this study are, with the exceptions noted below, the first published for these reference materials. There are published information values for the elemental concentrations of 'organic' carbon (Wt % Corg measured range is 8.98 - 10.4) and nitrogen (Wt % Ntot 0.347 with SD 0.043) only for SDO-1. The suggested values presented here should be considered 'information values' as defined by the NRCC Institute for National Measurement Reference Materials and should be useful for the analysis of 13C, 15N, C and N in organic material in sedimentary rocks.

  17. Trophic ecology of the supralittoral rocky shore (Roscoff, France): A dual stable isotope (δ 13C, δ 15N) and experimental approach

    NASA Astrophysics Data System (ADS)

    Laurand, Sandrine; Riera, Pascal

    2006-07-01

    The present study investigates the trophic transfers on the upper littoral rocky shore (i.e. the supralittoral zone together with the upper midlittoral and adlittoral) of northern Brittany. The population mainly consists of four invertebrate species: the littorinids Littorina saxatilis and Melarhaphe neritoides, the isopod Ligia oceanica and the insect Petrobius maritimus. The utilisation of food sources available to these grazers was examined in a laboratory microcosm feeding experiment and a field study using stable isotopes (δ 13C, δ 15N). The results indicated that although Ligia oceanica preferentially occurs in the supralittoral zone, its trophic subsidies originate mostly from the adlittoral and lower intertidal zones. The stable isotope data also suggested that adlittoral terrestrial organic material may be the major food source of Petrobius maritimus. δ 15N of Littorina saxatilis indicated a highly variable diet consisting of supralittoral lichens, midlittoral macroalgae and other food sources (e.g. microalgae). Both feeding experiments and stable isotope data show that only Melarhaphe neritoides has a clearly identifiable diet based on a mixture of lichens, mostly Verrucaria maura and Caloplaca marina, as estimated by an isotopic mixing model. Hence, the food web of this intertidal zone appears largely based on trophic subsidies from other habitats (i.e. upper and lower intertidal zones).

  18. Sedimentary records of δ(13)C, δ(15)N and organic matter accumulation in lakes receiving nutrient-rich mine waters.

    PubMed

    Widerlund, Anders; Chlot, Sara; Öhlander, Björn

    2014-07-01

    Organic C and total N concentrations, C/N ratios, δ(15)N and δ(13)C values in (210)Pb-dated sediment cores were used to reconstruct historical changes in organic matter (OM) accumulation in three Swedish lakes receiving nutrient-rich mine waters. Ammonium-nitrate-based explosives and sodium cyanide (NaCN) used in gold extraction were the major N sources, while lesser amounts of P originated from apatite and flotation chemicals. The software IsoSource was used to model the relative contribution of soil, terrestrial and littoral vegetation, and phytoplankton detritus in the lake sediments. In one lake the IsoSource modelling failed, suggesting the presence of additional, unknown OM sources. In two of the lakes sedimentary detritus of littoral vegetation and phytoplankton had increased by 15-20% and 20-35%, respectively, since ~1950, when N- and P-rich mine waters began to reach the lakes. Today, phytoplankton is the dominating OM component in these lake sediments, which appears to be a eutrophication effect related to mining operations. Changes in the N isotopic composition of biota, lake water, and sediments related to the use of ammonium-nitrate-based explosives and NaCN were evident in the two studied systems. However, N isotope signals in the receiving waters (δ(15)N~+9‰ to +19‰) were clearly shifted from the primary signal in explosives (δ(15)N-NO3=+3.4±0.3‰; δ(15)N-NH4=-8.0±0.3‰) and NaCN (δ(15)N=+1.1±0.5‰), and direct tracing of the primary N isotope signals in mining chemicals was not possible in the receiving waters. Systems where mine waters with a well known discharge history are a major point source of N with well-defined isotopic composition should, however, be suitable for further studies of processes controlling N isotope signatures and their transformation in aquatic systems receiving mine waters. PMID:24727038

  19. 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

    SciTech Connect

    Hansen, A.P.; Petros, A.M.; Meadows, R.P.; Mazar, A.P.; Nettesheim, D.G.; Pederson, T.M.; Fesik, S.W.

    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.

  20. Establishing spatial trends in water chemistry and stable isotopes (δ15N and δ13C) in the Elwha River prior to dam removal and salmon recolonization

    USGS Publications Warehouse

    Duda, J.J.; Coe, H.J.; Morley, S.A.; Kloehn, K.K.

    2011-01-01

    Two high-head dams on the Elwha River in Washington State (USA) have changed the migratory patterns of resident and anadromous fish, limiting Pacific salmon to the lower 7.9 km of a river that historically supported large Pacific salmon runs. To document the effects of the dams prior to their removal, we measured carbon and nitrogen stable isotope ratios of primary producers, benthic macroinvertebrates, and fish, and water chemistry above, between and below the dams. We found that δ15N was significantly higher in fish, stoneflies, black flies, periphyton and macroalgae where salmon still have access. Fish and chloroperlid stoneflies were enriched in δ13C, but the values were more variable than in δ15N. For some taxa, there were also differences between the two river sections that lack salmon, suggesting that factors other than marine-derived nutrients are structuring longitudinal isotopic profiles. Consistent with trophic theory, macroalgae had the lowest δ15N, followed by periphyton, macroinvertebrates and fish, with a range of 6.9, 6.2 and 7.7‰ below, between, and above the dams, respectively. Water chemistry analyses confirmed earlier reports that the river is oligotrophic. Phosphorous levels in the Elwha were lower than those found in other regional rivers, with significant differences among regulated, unregulated and reference sections. The removal of these dams, among the largest of such projects ever attempted, is expected to facilitate the return of salmon and their marine-derived nutrients (MDN) throughout the watershed, possibly altering the food web structure, nutrient levels and stable isotope values that we documented.

  1. Purity analysis of hydrogen cyanide, cyanogen chloride and phosgene by quantitative (13)C NMR spectroscopy.

    PubMed

    Henderson, Terry J; Cullinan, David B

    2007-11-01

    Hydrogen cyanide, cyanogen chloride and phosgene are produced in tremendously large quantities today by the chemical industry. The compounds are also particularly attractive to foreign states and terrorists seeking an inexpensive mass-destruction capability. Along with contemporary warfare agents, therefore, the US Army evaluates protective equipment used by warfighters and domestic emergency responders against the compounds, and requires their certification at > or = 95 carbon atom % before use. We have investigated the (13)C spin-lattice relaxation behavior of the compounds to develop a quantitative NMR method for characterizing chemical lots supplied to the Army. Behavior was assessed at 75 and 126 MHz for temperatures between 5 and 15 degrees C to hold the compounds in their liquid states, dramatically improving detection sensitivity. T(1) values for cyanogen chloride and phosgene were somewhat comparable, ranging between 20 and 31 s. Hydrogen cyanide values were significantly shorter at 10-18 s, most likely because of a (1)H--(13)C dipolar contribution to relaxation not possible for the other compounds. The T(1) measurements were used to derive relaxation delays for collecting the quantitative (13)C data sets. At 126 MHz, only a single data acquisition with a cryogenic probehead gave a signal-to-noise ratio exceeding that necessary for certifying the compounds at > or = 95 carbon atom % and 99% confidence. Data acquired at 75 MHz with a conventional probehead, however, required > or = 5 acquisitions to reach this certifying signal-to-noise ratio for phosgene, and >/= 12 acquisitions were required for the other compounds under these same conditions. In terms of accuracy and execution time, the NMR method rivals typical chromatographic methods. PMID:17924355

  2. Hydrogen Bonds in Crystalline Imidazoles Studied by 15N NMR and ab initio MO Calculations

    NASA Astrophysics Data System (ADS)

    Ueda, Takahiro; Nagatomo, Shigenori; Masui, Hirotsugu; Nakamura, Nobuo; Hayashi, Shigenobu

    1999-07-01

    Intermolecular hydrogen bonds of the type N-H...N in crystals of imidazole and its 4-substituted and 4,5-disubstituted derivatives were studied by 15N CP/MAS NMR and an ab initio molecular orbital (MO) calculation. In the 15N CP/MAS NMR spectrum of each of the imidazole derivatives, two peaks due to the two different functional groups, >NH and =N-, were observed. The value of the 15N isotropic chemical shift for each nitrogen atom depends on both the length of the intermolecular hydrogen bond and the kind of the substituent or substituents. It was found that the difference between the experimen-tal chemical shifts of >NH and =N-varies predominantly with the hydrogen bond length but does not show any systematic dependence on the kind of substituent. The ab initio MO calculations suggest that the hydrogen bond formation influences the 15N isotropic chemical shift predominantly, and that the difference between the 15N isotropic chemical shift of >NH and =N-varies linearly with the hydrogen bond length.

  3. Spectral density mapping at multiple magnetic fields suitable for 13C NMR relaxation studies

    NASA Astrophysics Data System (ADS)

    Kadeřávek, Pavel; Zapletal, Vojtěch; Fiala, Radovan; Srb, Pavel; Padrta, Petr; Přecechtělová, Jana Pavlíková; Šoltésová, Mária; Kowalewski, Jozef; Widmalm, Göran; Chmelík, Josef; Sklenář, Vladimír; Žídek, Lukáš

    2016-05-01

    Standard spectral density mapping protocols, well suited for the analysis of 15N relaxation rates, introduce significant systematic errors when applied to 13C relaxation data, especially if the dynamics is dominated by motions with short correlation times (small molecules, dynamic residues of macromolecules). A possibility to improve the accuracy by employing cross-correlated relaxation rates and on measurements taken at several magnetic fields has been examined. A suite of protocols for analyzing such data has been developed and their performance tested. Applicability of the proposed protocols is documented in two case studies, spectral density mapping of a uniformly labeled RNA hairpin and of a selectively labeled disaccharide exhibiting highly anisotropic tumbling. Combination of auto- and cross-correlated relaxation data acquired at three magnetic fields was applied in the former case in order to separate effects of fast motions and conformational or chemical exchange. An approach using auto-correlated relaxation rates acquired at five magnetic fields, applicable to anisotropically moving molecules, was used in the latter case. The results were compared with a more advanced analysis of data obtained by interpolation of auto-correlated relaxation rates measured at seven magnetic fields, and with the spectral density mapping of cross-correlated relaxation rates. The results showed that sufficiently accurate values of auto- and cross-correlated spectral density functions at zero and 13C frequencies can be obtained from data acquired at three magnetic fields for uniformly 13C -labeled molecules with a moderate anisotropy of the rotational diffusion tensor. Analysis of auto-correlated relaxation rates at five magnetic fields represents an alternative for molecules undergoing highly anisotropic motions.

  4. Spectral density mapping at multiple magnetic fields suitable for (13)C NMR relaxation studies.

    PubMed

    Kadeřávek, Pavel; Zapletal, Vojtěch; Fiala, Radovan; Srb, Pavel; Padrta, Petr; Přecechtělová, Jana Pavlíková; Šoltésová, Mária; Kowalewski, Jozef; Widmalm, Göran; Chmelík, Josef; Sklenář, Vladimír; Žídek, Lukáš

    2016-05-01

    Standard spectral density mapping protocols, well suited for the analysis of (15)N relaxation rates, introduce significant systematic errors when applied to (13)C relaxation data, especially if the dynamics is dominated by motions with short correlation times (small molecules, dynamic residues of macromolecules). A possibility to improve the accuracy by employing cross-correlated relaxation rates and on measurements taken at several magnetic fields has been examined. A suite of protocols for analyzing such data has been developed and their performance tested. Applicability of the proposed protocols is documented in two case studies, spectral density mapping of a uniformly labeled RNA hairpin and of a selectively labeled disaccharide exhibiting highly anisotropic tumbling. Combination of auto- and cross-correlated relaxation data acquired at three magnetic fields was applied in the former case in order to separate effects of fast motions and conformational or chemical exchange. An approach using auto-correlated relaxation rates acquired at five magnetic fields, applicable to anisotropically moving molecules, was used in the latter case. The results were compared with a more advanced analysis of data obtained by interpolation of auto-correlated relaxation rates measured at seven magnetic fields, and with the spectral density mapping of cross-correlated relaxation rates. The results showed that sufficiently accurate values of auto- and cross-correlated spectral density functions at zero and (13)C frequencies can be obtained from data acquired at three magnetic fields for uniformly (13)C-labeled molecules with a moderate anisotropy of the rotational diffusion tensor. Analysis of auto-correlated relaxation rates at five magnetic fields represents an alternative for molecules undergoing highly anisotropic motions. PMID:27003380

  5. Spectroscopic separation of (13) C NMR spectra of complex isomeric mixtures by the CSSF-TOCSY-INEPT experiment.

    PubMed

    Yang, Lu; Moreno, Aitor; Fieber, Wolfgang; Brauchli, Robert; Sommer, Horst

    2015-04-01

    Isomeric mixtures from synthetic or natural origins can pose fundamental challenges for their chromatographic separation and spectroscopic identification. A novel 1D selective NMR experiment, chemical shift selective filter (CSSF)-TOCSY-INEPT, is presented that allows the extraction of (13) C NMR subspectra of discrete isomers in complex mixtures without physical separation. This is achieved via CSS excitation of proton signals in the (1) H NMR mixture spectrum, propagation of the selectivity by polarization transfer within coupled (1) H spins, and subsequent relaying of the magnetization from (1) H to (13) C by direct INEPT transfer to generate (13) C NMR subspectra. Simple consolidation of the subspectra yields (13) C NMR spectra for individual isomers. Alternatively, CSSF-INEPT with heteronuclear long-range transfer can correlate the isolated networks of coupled spins and therefore facilitate the reconstruction of the (13) C NMR spectra for isomers containing multiple spin systems. A proof-of-principle validation of the CSSF-TOCSY-INEPT experiment is demonstrated on three mixtures with different spectral and structural complexities. The results show that CSSF-TOCSY-INEPT is a versatile, powerful tool for deconvoluting isomeric mixtures within the NMR tube with unprecedented resolution and offers unique, unambiguous spectral information for structure elucidation. PMID:25616134

  6. (13)C NMR-based metabolomics for the classification of green coffee beans according to variety and origin.

    PubMed

    Wei, Feifei; Furihata, Kazuo; Koda, Masanori; Hu, Fangyu; Kato, Rieko; Miyakawa, Takuya; Tanokura, Masaru

    2012-10-10

    (13)C NMR-based metabolomics was demonstrated as a useful tool for distinguishing the species and origins of green coffee bean samples of arabica and robusta from six different geographic regions. By the application of information on (13)C signal assignment, significantly different levels of 14 metabolites of green coffee beans were identified in the classifications, including sucrose, caffeine, chlorogenic acids, choline, amino acids, organic acids, and trigonelline, as captured by multivariate analytical models. These studies demonstrate that the species and geographical origin can be quickly discriminated by evaluating the major metabolites of green coffee beans quantitatively using (13)C NMR-based metabolite profiling. PMID:22989016

  7. 1H and 13C resonance designation of antimycin A1 by two-dimensional NMR spectroscopy

    USGS Publications Warehouse

    Abidi, S.L.; Adams, B.R.

    1987-01-01

    Complete 1H and 13C resonance assignments of antimycin A1 were accomplished by two-dimensional NMR techniques, viz. 1H homonuclear COSY correlation, heteronuclear 13C-1H chemical shift correlation and long-range heteronuclear 13C-1H COLOC correlation. Antimycin A1 was found to consist of two isomeric components in a 2:1 ratio based on NMR spectroscopic evidence. The structure of the major component was newly assigned as the 8-isopentanoic acid ester. The spectra of the minor component were consistent with the known structure of antimycin A1.

  8. Spectroscopic characterization of the 1-substituted 3,3-diphenyl-4-(2'-hydroxyphenyl)azetidin-2-ones: Application of 13C NMR, 1H- 13C COSY NMR and mass spectroscopy

    NASA Astrophysics Data System (ADS)

    Singh, Girija S.; Pheko, Tshepo

    2008-08-01

    The article deals with spectroscopic characterization of azetidin-2-ones. The presence of substituents like hydroxyl, fluoro, methoxy and benzhydryl, etc., on the azetidin-2-one ring significantly affects the IR absorption and 13C NMR frequencies of the carbonyl group present in these compounds. The presence of an ester carbonyl group or too many methine protons in the molecule has been observed to limit the scope of IR and 1H NMR spectroscopy in unambiguous assignment of the structure. The application of 13C NMR, 2D NMR ( 1H- 13C COSY) and mass spectroscopy in characterization of complex azetidin-2-ones is discussed. An application of the latter two techniques is described in deciding unequivocally between an azetidin-2-one ring and chroman-2-one ring structure for the product obtained by treatment of the 1-substituted 3,3-diphenyl-4-[2'-( O-diphenylacyl)hydroxyphenyl]-2-azetidinones with ethanolic sodium hydroxide at room temperature.

  9. 1H and 13C NMR investigation of 20-hydroxyecdysone dioxolane derivatives, a novel group of MDR modulator agents.

    PubMed

    Balázs, Attila; Hunyadi, Attila; Csábi, József; Jedlinszki, Nikoletta; Martins, Ana; Simon, András; Tóth, Gábor

    2013-12-01

    The synthesis, structure elucidation and the complete (1)H and (13)C signal assignment of a series of dioxolane derivatives of 20-hydroxyecdysone, synthesized as novel modulators of multidrug resistance, are presented. The structures and NMR signal assignment were established by comprehensive one-dimensional and two-dimensional NMR spectroscopy supported by mass spectrometry. PMID:24114927

  10. Topotecan dynamics, tautomerism and reactivity--1H/13C NMR and ESI MS study.

    PubMed

    Hyz, Karolina; Kawecki, Robert; Bednarek, Elzbieta; Bocian, Wojciech; Sitkowski, Jerzy; Kozerski, Lech

    2010-08-01

    Topotecan (TPT) is in clinical use as an antitumor agent, hycamtin. Because of this, it requires both biologically and chemically useful information to be available. TPT acts by binding to the covalent complex formed by nicked DNA and topoisomerase I. This has a poisonous effect since inserted into the single-strand nick and TPT inhibits its religation. We used NMR to trace TPT dynamics, tautomerism and solvolysis products in various solvents and conditions. Chemical stability was assessed in methanol and DMSO as compared to water, and the regioselectivity of the N- and O-methylation was studied using various alkylating agents. The reaction products of quaternization of the nitrogen atom and methylation of the oxygen atom were characterized by means of ESI MS, (1)H/(13)C-HMBC and -HSQCAD NMR. We have focused on the NMR characterization of TPT with an anticipation that its aggregation, tumbling properties and the intramolecular dipolar interactions will be a common feature for other compounds described in this article. These features can also be useful in tracing the interactions of this class of topoisomerase I (TopoI) poisons with DNA. Moreover, the results explained shed light on the recently disclosed problem of lack of stability of TPT in the heart tissue homogenate samples using the analytical assays developed for this class of compounds carried out in the presence of methanol. PMID:20623719

  11. Determination of glucan phosphorylation using heteronuclear 1H, 13C double and 1H, 13C, 31P triple-resonance NMR spectra.

    PubMed

    Schmieder, Peter; Nitschke, Felix; Steup, Martin; Mallow, Keven; Specker, Edgar

    2013-10-01

    Phosphorylation and dephosphorylation of starch and glycogen are important for their physicochemical properties and also their physiological functions. It is therefore desirable to reliably determine the phosphorylation sites. Heteronuclear multidimensional NMR-spectroscopy is in principle a straightforward analytical approach even for complex carbohydrate molecules. With heterogeneous samples from natural sources, however, the task becomes more difficult because a full assignment of the resonances of the carbohydrates is impossible to obtain. Here, we show that the combination of heteronuclear (1) H,(13) C and (1) H,(13) C,(31) P techniques and information derived from spectra of a set of reference compounds can lead to an unambiguous determination of the phosphorylation sites even in heterogeneous samples. PMID:23913630

  12. Chlorophyll a specific Δ14C, δ13C and δ15N values in stream periphyton: implications for aquatic food web studies

    NASA Astrophysics Data System (ADS)

    Ishikawa, N. F.; Yamane, M.; Suga, H.; Ogawa, N. O.; Yokoyama, Y.; Ohkouchi, N.

    2015-07-01

    We determined the isotopic composition of chlorophyll a in periphytic algae attached to a streambed substrate (periphyton). The samples were collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, pure aquatic primary producer (Cladophora sp.) and terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Mayfly larva, Epeorus latifolium), suggesting that periphyton δ13C values do not faithfully trace carbon transfer between primary producers and primary consumers. Periphyton Δ14Cchl values (-258 ‰ in April and -190 ‰ in October) were slightly lower than Δ14Cbulk values (-228 ‰ in April and -179 ‰ in October), but were close to the Δ14C value for dissolved inorganic carbon (DIC) (-217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = -1000 ‰) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (-215 ‰ in April and -199 ‰ in October) and Cladophora sp. (-210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

  13. Nicotine, acetanilide and urea multi-level2H-,13C- and15N-abundance reference materials for continuous-flow isotope ratio mass spectrometry

    USGS Publications Warehouse

    Schimmelmann, A.; Albertino, A.; Sauer, P.E.; Qi, H.; Molinie, R.; Mesnard, F.

    2009-01-01

    Accurate determinations of stable isotope ratios require a calibration using at least two reference materials with different isotopic compositions to anchor the isotopic scale and compensate for differences in machine slope. Ideally, the S values of these reference materials should bracket the isotopic range of samples with unknown S values. While the practice of analyzing two isotopically distinct reference materials is common for water (VSMOW-SLAP) and carbonates (NBS 19 and L-SVEC), the lack of widely available organic reference materials with distinct isotopic composition has hindered the practice when analyzing organic materials by elemental analysis/isotope ratio mass spectrometry (EA-IRMS). At present only L-glutamic acids USGS40 and USGS41 satisfy these requirements for ??13C and ??13N, with the limitation that L-glutamic acid is not suitable for analysis by gas chromatography (GC). We describe the development and quality testing of (i) four nicotine laboratory reference materials for on-line (i.e. continuous flow) hydrogen reductive gas chromatography-isotope ratio mass-spectrometry (GC-IRMS), (ii) five nicotines for oxidative C, N gas chromatography-combustion-isotope ratio mass-spectrometry (GC-C-IRMS, or GC-IRMS), and (iii) also three acetanilide and three urea reference materials for on-line oxidative EA-IRMS for C and N. Isotopic off-line calibration against international stable isotope measurement standards at Indiana University adhered to the 'principle of identical treatment'. The new reference materials cover the following isotopic ranges: ??2Hnicotine -162 to -45%o, ??13Cnicotine -30.05 to +7.72%, ?? 15Nnicotine -6.03 to +33.62%; ??15N acetanilide +1-18 to +40.57%; ??13Curea -34.13 to +11.71%, ??15Nurea +0.26 to +40.61% (recommended ?? values refer to calibration with NBS 19, L-SVEC, IAEA-N-1, and IAEA-N-2). Nicotines fill a gap as the first organic nitrogen stable isotope reference materials for GC-IRMS that are available with different ??13N

  14. Effects of preservation methods of muscle tissue from upper-trophic level reef fishes on stable isotope values (δ13C and δ15N)

    PubMed Central

    Nelson, James A.; Rozar, Katherine L.; Adams, Charles S.; Wall, Kara R.; Switzer, Theodore S.; Winner, Brent L.; Hollander, David J.

    2015-01-01

    Research that uses stable isotope analysis often involves a delay between sample collection in the field and laboratory processing, therefore requiring preservation to prevent or reduce tissue degradation and associated isotopic compositions. Although there is a growing literature describing the effects of various preservation techniques, the results are often contextual, unpredictable and vary among taxa, suggesting the need to treat each species individually. We conducted a controlled experiment to test the effects of four preservation methods of muscle tissue from four species of upper trophic-level reef fish collected from the eastern Gulf of Mexico (Red Grouper Epinephelus morio, Gag Mycteroperca microlepis, Scamp Mycteroperca phenax, and Red Snapper Lutjanus campechanus). We used a paired design to measure the effects on isotopic values for carbon and nitrogen after storage using ice, 95% ethanol, and sodium chloride (table salt), against that in a liquid nitrogen control. Mean offsets for both δ13C and δ15N values from controls were lowest for samples preserved on ice, intermediate for those preserved with salt, and highest with ethanol. Within species, both salt and ethanol significantly enriched the δ15N values in nearly all comparisons. Ethanol also had strong effects on the δ13C values in all three groupers. Conversely, for samples preserved on ice, we did not detect a significant offset in either isotopic ratio for any of the focal species. Previous studies have addressed preservation-induced offsets in isotope values using a mass balance correction that accounts for changes in the isotope value to that in the C/N ratio. We tested the application of standard mass balance corrections for isotope values that were significantly affected by the preservation methods and found generally poor agreement between corrected and control values. The poor performance by the correction may have been due to preferential loss of lighter isotopes and corresponding

  15. Effects of preservation methods of muscle tissue from upper-trophic level reef fishes on stable isotope values (δ (13)C and δ (15)N).

    PubMed

    Stallings, Christopher D; Nelson, James A; Rozar, Katherine L; Adams, Charles S; Wall, Kara R; Switzer, Theodore S; Winner, Brent L; Hollander, David J

    2015-01-01

    Research that uses stable isotope analysis often involves a delay between sample collection in the field and laboratory processing, therefore requiring preservation to prevent or reduce tissue degradation and associated isotopic compositions. Although there is a growing literature describing the effects of various preservation techniques, the results are often contextual, unpredictable and vary among taxa, suggesting the need to treat each species individually. We conducted a controlled experiment to test the effects of four preservation methods of muscle tissue from four species of upper trophic-level reef fish collected from the eastern Gulf of Mexico (Red Grouper Epinephelus morio, Gag Mycteroperca microlepis, Scamp Mycteroperca phenax, and Red Snapper Lutjanus campechanus). We used a paired design to measure the effects on isotopic values for carbon and nitrogen after storage using ice, 95% ethanol, and sodium chloride (table salt), against that in a liquid nitrogen control. Mean offsets for both δ (13)C and δ (15)N values from controls were lowest for samples preserved on ice, intermediate for those preserved with salt, and highest with ethanol. Within species, both salt and ethanol significantly enriched the δ (15)N values in nearly all comparisons. Ethanol also had strong effects on the δ (13)C values in all three groupers. Conversely, for samples preserved on ice, we did not detect a significant offset in either isotopic ratio for any of the focal species. Previous studies have addressed preservation-induced offsets in isotope values using a mass balance correction that accounts for changes in the isotope value to that in the C/N ratio. We tested the application of standard mass balance corrections for isotope values that were significantly affected by the preservation methods and found generally poor agreement between corrected and control values. The poor performance by the correction may have been due to preferential loss of lighter isotopes and

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

    SciTech Connect

    Jones, R.A.

    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.

  17. Stable Isotope (δ13C, δ15N, δ34S) Analysis and Satellite Telemetry Depict the Complexity of Gray Wolf (Canis lupus) Diets in Southwest Alaska

    NASA Astrophysics Data System (ADS)

    Stanek, A.; Watts, D. E.; Cohn, B. R.; Spencer, P.; Mangipane, B.; Welker, J. M.

    2010-12-01

    Throughout Alaska, gray wolves (Canis lupus) are a top predator of large ungulates. While they primarily rely on ungulates such as moose (Alces alces) and caribou (Rangifer tarandus) as food, they are opportunistic and use alternative resources. The variation and supplemental protein sources in wolf diet has not been studied extensively on live animals currently using the landscape. With large seasonal influxes of Pacific salmon (Oncorhynchus sp.) into Alaska, terrestrial carnivore use of marine species is of particular interest. Using stable isotope (δ13C, δ15N, δ34S) analysis of wolf guard hair and blood, this study aims to determine the proportion of marine derived nutrients (MDN) in the diet of wolf packs within and surrounding Lake Clark National Park and Preserve and Alaska Peninsula and Becharof National Wildlife Refuges in Southwest Alaska. Satellite telemetry from the animals sampled facilitates quantification of landscape use patterns in correspondence with isotopic traits. Wolf pack territories within and surrounding the Lake Clark region appear to vary in spatial extent and in availability of MDN, such as salmon. Initial analysis shows that two packs with smaller home ranges, centrally located around areas with greater salmon availability, have enriched δ15N values compared to packs that have larger home ranges not centralized around salmon spawning waters. This pattern of isotopic enrichment is found in red blood cells, blood serum and hair, representing diets over different time scales. The enrichment in both blood and hair indicates a sustained use of MDN over the previous six to nine months. In the Lake Clark region, simple mixing model estimates suggest that up to 30% of wolf pack diets may be from marine sources. In contrast, packs with larger home ranges and less access to salmon have stable isotope values representative of a terrestrial diet.

  18. Inter- and intrahabitat dietary variability of chacma baboons (Papio ursinus) in South African savannas based on fecal delta13C, delta15N, and %N.

    PubMed

    Codron, Daryl; Lee-Thorp, Julia A; Sponheimer, Matt; de Ruiter, Darryl; Codron, Jacqueline

    2006-02-01

    Baboons are dietary generalists, consuming a wide range of food items in varying proportions. It is thus difficult to quantify and explain the dietary behavior of these primates. We present stable carbon (delta(13)C) and nitrogen (delta(15)N) isotopic data, and percentage nitrogen (%N), of feces from chacma baboons (Papio ursinus) living in two savanna environments of South Africa: the mountainous Waterberg region and the low-lying Kruger National Park. Baboons living in the more homogeneous landscapes of the Waterberg consume a more isotopically heterogeneous diet than their counterparts living in Kruger Park. Grasses and other C(4)-based foods comprise between approximately 10-20% (on average) of the bulk diet of Kruger Park baboons. Carbon isotopic data from the Waterberg suggest diets of approximately 30-50% grass, which is higher than generally reported for baboons across the African savanna. Based on observations of succulent-feeding, we propose that baboons in the Waterberg consume a mix of C(4) grasses and CAM-photosynthesizing succulents in combined proportions varying between approximately 5-75% (average, approximately 35%). Fecal delta(15)N of baboons is lower than that of sympatric ungulates, which may be due to a combination of low levels of faunivory, foraging on subterranean plant parts, or the use of human foods in the case of Kruger Park populations. Fecal N levels in baboons are consistently higher than those of sympatric ungulate herbivores, indicating that baboons consume a greater proportion of protein-rich foods than do other savanna mammals. These data suggest that chacma baboons adapt their dietary behavior so as to maximize protein intake, regardless of their environment. PMID:16247809

  19. ^13C Solid NMR Study of Devulcanization and Revulcanization of SBR Ne

    NASA Astrophysics Data System (ADS)

    Massey, J.; Levin, V.; Isayev, A.; von Meerwall, E.

    1996-03-01

    As part of a larger effort in support of recycling of rubber-based composites, we have used ^13C CP-MAS NMR spectroscopy and relaxation to study molecular and segmental mobilities in styrene-butadiene random copolymers before and after sulfur crosslinking, after subsequent devulcanization using a thermal ultrasound technique, and following revulcanization. Tracking the cis-trans ratio indicates that overall network crosslink density increases during each of these steps, including devulcanization, which produces mesoscale network aggregates and substantial amounts of sol. This observation is confirmed by the transverse (T_2) relaxation times, which show that molecular/segmental mobilities monotonically decrease in the same sequence. Analysis of these effects requires the invocation of alterations in sulfur crosslinking, i.e. density, distribution, and functionality, including extensive cyclization. Measurements of the glass transition temperatures in melt, network, sol , and revulcanizate are in accord with this picture.

  20. 13C-NMR study of acetate assimilation in Thermoproteus neutrophilus.

    PubMed

    Schäfer, S; Paalme, T; Vilu, R; Fuchs, G

    1989-12-22

    Acetate assimilation into amino acids and the functioning of central biosynthetic pathways in the extremely thermophilic anaerobic archaebacterium Thermoproteus neutrophilus was investigated using 13C NMR as the method for determination of the labelling patterns. Acetate was assimilated via reductive carboxylation of acetyl-CoA to pyruvate and pyruvate conversion to phosphoenolpyruvate which was further carboxylated to oxaloacetate. 2-Oxoglutarate was mainly formed via citrate. However, the labelling patterns of glutamic acid and alanine were in agreement with the concurrent synthesis of about 15% 2-oxoglutarate and 5% pyruvate through the reductive citric acid cycle. A scrambling phenomenon occurring in aspartate and all amino acids derived through oxaloacetate was observed. The labelling patterns of amino acids were in agreement with their standard biosynthetic pathways, with two remarkable exceptions: isoleucine was synthesized via the citramalate pathway and lysine was synthesized via the 2-aminoadipate pathway which has previously been reported only in eukaryotic microorganisms. PMID:2514097

  1. The effect of sample hydration on 13C CPMAS NMR spectra of fulvic acids

    USGS Publications Warehouse

    Hatcher, P.G.; Wilson, M.A.

    1991-01-01

    Three fulvic acids, two of which have been well studied by a number of other groups (Armadale and Suwannee river fulvic acids) have been examined by high resolution solid-state 13C-NMR techniques to delineate the effect of absorbed water. Two main effects of absorbed water were observed: (1) changes in spin lattice relaxation times in the rotating frame and cross polarization times and (2) total loss of signal so that some fulvic acid is effectively in solution. These results suggest that discrepancies in the literature concerning observed relative signal intensities from different structural groups are due to absorbed water and emphasize the necessity for proper precautionary drying before spectroscopic analysis. ?? 1991.

  2. 13C solid-state NMR analysis of heterogeneous structure of beeswax in native state

    NASA Astrophysics Data System (ADS)

    Kameda, Tsunenori

    2005-12-01

    I investigated the molecular structure of natural wax from Japanese bees (Apis cerana japonica) in its native state (neither purified nor recrystallized) by 13C and 1H solid-state NMR. Two strong 13C peaks at 32.9 and 34.0 ppm were attributed to signals from internal-chain methylene carbons [int-(CH2)] in two types of crystal form. The peak at 32.9 ppm was assigned to an orthorhombic crystal form, and that at 34.0 ppm was assigned to a triclinic or monoclinic form. In both crystalline regions, bi-exponential decay of 13C spin-lattice relaxation [T1(C)] for the crystalline peaks due to chain diffusion was observed. 1H spin-lattice relaxation [T1(H)] values for protons of the CH3 group and for int-(CH2) in the crystalline and amorphous regions were identical; this was interpreted as being due to averaging of the T1(H) relaxation rates via spin diffusion. In contrast, although the T_{{1}_{\\rho}}(H) decay curves for protons of the CH3 group and for int-(CH2) in the amorphous and orthorhombic forms were almost identical, those of the triclinic or monoclinic forms were different. This unhomogeneous character of T_{{1}_{\\rho}}(H) was interpreted as resulting from differences in the molecular composition of each crystal form. Moreover, two components with long and short 1H spin-spin relaxation [T2(H)] values, arising from the mobile and rigid phases, respectively, were observed at above about -30 °C.

  3. Partial 13C isotopic enrichment of nucleoside monophosphates: useful reporters for NMR structural studies

    PubMed Central

    Kishore, Anita I.; Mayer, Michael R.; Prestegard, James H.

    2005-01-01

    Analysis of the 13C isotopic labeling patterns of nucleoside monophosphates (NMPs) extracted from Escherichia coli grown in a mixture of C-1 and C-2 glucose is presented. By comparing our results to previous observations on amino acids grown in similar media, we have been able to rationalize the labeling pattern based on the well-known biochemistry of nucleotide biosynthesis. Except for a few notable absences of label (C4 in purines and C3′ in ribose) and one highly enriched site (C1′ in ribose), most carbons are randomly enriched at a low level (an average of 13%). These sparsely labeled NMPs give less complex NMR spectra than their fully isotopically labeled analogs due to the elimination of most 13C–13C scalar couplings. The spectral simplicity is particularly advantageous when working in ordered systems, as illustrated with guanosine diphosphate (GDP) bound to ADP ribosylation factor 1 (ARF1) aligned in a liquid crystalline medium. In this system, the absence of scalar couplings and additional long-range dipolar couplings significantly enhances signal to noise and resolution. PMID:16254075

  4. NMR characterization of 13C-benzene sorbed to natural and prepared charcoals.

    PubMed

    Smernik, Ronald J; Kookana, Rai S; Skjemstad, Jan O

    2006-03-15

    We investigated how the NMR properties of uniformly 13C-labeled benzene molecules are influenced by sorption to charcoals produced in the laboratory and collected from the field following wildfires. Uniformly 13C-labeled benzene was sorbed to two charcoals produced in the laboratory at 450 and 850 degrees C. The chemical shift of benzene sorbed to the higher-temperature charcoal was 5-6 ppm lower than that of benzene sorbed to the lower-temperature charcoal. This difference was attributed to stronger diamagnetic ring currents (which cause a shift to lower ppm values) in the more condensed or "graphitic" high-temperature charcoal. The chemical shift of benzene sorbed to two charcoals collected from the field following wildfires indicated a degree of charcoal graphitization intermediate between that of the two laboratory-prepared charcoals. Variable contact time and dipolar dephasing experiments showed that the molecular mobility of sorbed benzene molecules increased with increasing charcoal graphitization, and also increased with increasing benzene concentration. We propose that the chemical shift displacement of molecules sorbed to charcoal could be used to identify molecules sorbed to black carbon in heterogeneous matrixes such as soils and sediments, and to establish how condensed or "graphitic" the black carbon is. PMID:16570595

  5. Cigarette Butt Decomposition and Associated Chemical Changes Assessed by 13C CPMAS NMR

    PubMed Central

    Bonanomi, Giuliano; Incerti, Guido; Cesarano, Gaspare; Gaglione, Salvatore A.; Lanzotti, Virginia

    2015-01-01

    Cigarette butts (CBs) are the most common type of litter on earth, with an estimated 4.5 trillion discarded annually. Apart from being unsightly, CBs pose a serious threat to living organisms and ecosystem health when discarded in the environment because they are toxic to microbes, insects, fish and mammals. In spite of the CB toxic hazard, no studies have addressed the effects of environmental conditions on CB decomposition rate. In this study we investigate the interactive effects of substrate fertility and N transfer dynamics on CB decomposition rate and carbon quality changes. We carried out an experiment using smoked CBs and wood sticks, used as a slow decomposing standard organic substrate, incubated in both laboratory and field conditions for two years. CB carbon quality changes during decomposition was assessed by 13C CPMAS NMR. Our experiment confirmed the low degradation rate of CBs which, on average, lost only 37.8% of their initial mass after two years of decomposition. Although a net N transfer occurred from soil to CBs, contrary to our hypothesis, mass loss in the medium-term (two years) was unaffected by N availability in the surrounding substrate. The opposite held for wood sticks, in agreement with the model that N-rich substrates promote the decomposition of other N-poor natural organic materials with a high C/N ratio. As regards CB chemical quality, after two years of decomposition 13C NMR spectroscopy highlighted very small changes in C quality that are likely to reflect a limited microbial attack. PMID:25625643

  6. Cigarette butt decomposition and associated chemical changes assessed by 13C CPMAS NMR.

    PubMed

    Bonanomi, Giuliano; Incerti, Guido; Cesarano, Gaspare; Gaglione, Salvatore A; Lanzotti, Virginia

    2015-01-01

    Cigarette butts (CBs) are the most common type of litter on earth, with an estimated 4.5 trillion discarded annually. Apart from being unsightly, CBs pose a serious threat to living organisms and ecosystem health when discarded in the environment because they are toxic to microbes, insects, fish and mammals. In spite of the CB toxic hazard, no studies have addressed the effects of environmental conditions on CB decomposition rate. In this study we investigate the interactive effects of substrate fertility and N transfer dynamics on CB decomposition rate and carbon quality changes. We carried out an experiment using smoked CBs and wood sticks, used as a slow decomposing standard organic substrate, incubated in both laboratory and field conditions for two years. CB carbon quality changes during decomposition was assessed by 13C CPMAS NMR. Our experiment confirmed the low degradation rate of CBs which, on average, lost only 37.8% of their initial mass after two years of decomposition. Although a net N transfer occurred from soil to CBs, contrary to our hypothesis, mass loss in the medium-term (two years) was unaffected by N availability in the surrounding substrate. The opposite held for wood sticks, in agreement with the model that N-rich substrates promote the decomposition of other N-poor natural organic materials with a high C/N ratio. As regards CB chemical quality, after two years of decomposition 13C NMR spectroscopy highlighted very small changes in C quality that are likely to reflect a limited microbial attack. PMID:25625643

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

    PubMed

    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. PMID:26590548

  8. Identifying the African Wintering Grounds of Hybrid Flycatchers Using a Multi–Isotope (δ2H, δ13C, δ15N) Assignment Approach

    PubMed Central

    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. PMID:24847717

  9. Chemical weathering and the role of sulfuric and nitric acids in carbonate weathering: Isotopes (13C, 15N, 34S, and 18O) and chemical constraints

    NASA Astrophysics Data System (ADS)

    Li, Cai; Ji, Hongbing

    2016-05-01

    Multiple isotopes (13C-DIC, 34S and 18O-SO42-, 15N and 18O-NO3-) and water chemistry were used to evaluate weathering rates and associated CO2 consumption by carbonic acid and strong acids (H2SO4 and HNO3) in a typical karst watershed (Wujiang River, Southwest China). The dual sulfate isotopes indicate that sulfate is mainly derived from sulfide oxidation in coal stratum and sulfide-containing minerals, and dual nitrate isotopes indicate that nitrate is mainly derived from soil N and nitrification. The correlation between isotopic compositions and water chemistry suggests that sulfuric and nitric acids, in addition to carbonic acid, are involved in carbonate weathering. The silicate and carbonate weathering rates are 7.2 t km-2 yr-1 and 76 t km-2 yr-1, respectively. In comparison with carbonate weathering rates (43 t km-2 yr-1) by carbonic acid alone, the subsequent increase in rates indicates significant enhancement of weathering when combined with sulfuric and nitric acids. Therefore, the role of sulfuric and nitric acids in the rock weathering should be considered in the global carbon cycle.

  10. Polyamine flux analysis by determination of heavy isotope incorporation from 13C, 15N-enriched amino acids into polyamines by LC-MS/MS.

    PubMed

    Cerrada-Gimenez, Marc; Häkkinen, Merja R; Vepsäläinen, Jouko; Auriola, Seppo; Alhonen, Leena; Keinänen, Tuomo A

    2012-02-01

    The study of polyamine flux, i.e. the circulating flow of polyamines through the interconnected biosynthetic and catabolic pathways, is of considerable interest because of the established links between the polyamine metabolism and many diseases, such as cancer and diabetes. To study polyamine flux in detail, a novel method based on following the label incorporation from the (13)C, (15)N-labeled polyamine precursors, arginine, methionine and ornithine, into polyamines by LC-MS/MS was implemented. This methodology was tested on three distinct cell lines with different spermidine/spermine-N (1)-acetyltransferase (SSAT) expression levels, i.e. non-transgenic, transgenic and knockout. These trials allowed the identification of the critical conditions for the successful polyamine flux measurement, such as the functional time frame of label incorporation, until plateau phase with the selected precursor is reached. The novel LC-MS/MS-based method for polyamine flux overcame the limitations of previous existing methodologies, with baseline separation of the different polyamine species and the exact quantification of the incorporated label. Moreover, the obtained results clearly show that the increased SSAT expression is associated with accelerated polyamine flux. PMID:21818565

  11. Quantitative 13C NMR of whole and fractionated Iowa Mollisols for assessment of organic matter composition

    NASA Astrophysics Data System (ADS)

    Fang, Xiaowen; Chua, Teresita; Schmidt-Rohr, Klaus; Thompson, Michael L.

    2010-01-01

    Both the concentrations and the stocks of soil organic carbon vary across the landscape. Do the amounts of recalcitrant components of soil organic matter (SOM) vary with landscape position? To address this question, we studied four Mollisols in central Iowa, two developed in till and two developed in loess. Two of the soils were well drained and two were poorly drained. We collected surface-horizon samples and studied organic matter in the particulate organic matter (POM) fraction, the clay fractions, and the whole, unfractionated samples. We treated the soil samples with 5 M HF at ambient temperature or at 60 °C for 30 min to concentrate the SOM. To assess the composition of the SOM, we used solid-state nuclear magnetic resonance (NMR) spectroscopy, in particular, quantitative 13C DP/MAS (direct-polarization/magic-angle spinning), with and without recoupled dipolar dephasing. Spin counting by correlation of the integral NMR intensity with the C concentration by elemental analysis showed that NMR was ⩾85% quantitative for the majority of the samples studied. For untreated whole-soil samples with <2.5 wt.% C, which is considerably less than in most previous quantitative NMR analyses of SOM, useful spectra that reflected ⩾65% of all C were obtained. The NMR analyses allowed us to conclude (1) that the HF treatment (with or without heat) had low impact on the organic C composition in the samples, except for protonating carboxylate anions to carboxylic acids, (2) that most organic C was observable by NMR even in untreated soil materials, (3) that esters were likely to compose only a minor fraction of SOM in these Mollisols, and (4) that the aromatic components of SOM were enriched to ˜53% in the poorly drained soils, compared with ˜48% in the well drained soils; in plant tissue and particulate organic matter (POM) the aromaticities were ˜18% and ˜32%, respectively. Nonpolar, nonprotonated aromatic C, interpreted as a proxy for charcoal C, dominated the

  12. Dynamic nuclear polarization-enhanced 1H-13C double resonance NMR in static samples below 20 K

    NASA Astrophysics Data System (ADS)

    Potapov, Alexey; Thurber, Kent R.; Yau, Wai-Ming; Tycko, Robert

    2012-08-01

    We demonstrate the feasibility of one-dimensional and two-dimensional 1H-13C double resonance NMR experiments with dynamic nuclear polarization (DNP) at 9.4 T and temperatures below 20 K, including both 1H-13C cross-polarization and 1H decoupling, and discuss the effects of polarizing agent type, polarizing agent concentration, temperature, and solvent deuteration. We describe a two-channel low-temperature DNP/NMR probe, capable of carrying the radio-frequency power load required for 1H-13C cross-polarization and high-power proton decoupling. Experiments at 8 K and 16 K reveal a significant T2 relaxation of 13C, induced by electron spin flips. Carr-Purcell experiments and numerical simulations of Carr-Purcell dephasing curves allow us to determine the effective correlation time of electron flips under our experimental conditions. The dependence of the DNP signal enhancement on electron spin concentration shows a maximum near 80 mM. Although no significant difference in the absolute DNP enhancements for triradical (DOTOPA-TEMPO) and biradical (TOTAPOL) dopants was found, the triradical produced greater DNP build-up rates, which are advantageous for DNP experiments. Additionally the feasibility of structural measurements on 13C-labeled biomolecules was demonstrated with a two-dimensional 13C-13C exchange spectrum of selectively 13C-labeled β-amyloid fibrils.

  13. A stable isotope ( δ13C, δ15N) model for the North Water food web: implications for evaluating trophodynamics and the flow of energy and contaminants

    NASA Astrophysics Data System (ADS)

    Hobson, Keith A.; Fisk, Aaron; Karnovsky, Nina; Holst, Meike; Gagnon, Jean-Marc; Fortier, Martin

    fundamentally in transferring energy and carbon flux to higher trophic-level seabirds and marine mammals. We measured PCB 153 among selected organisms to investigate the behavior of bioaccumulating contaminants within the food web. Our isotopic model confirmed the trophic magnification of PCB 153 in this high-Arctic food web due to a strong correlation between contaminant concentration and organism δ15N values, demonstrating the utility of combining isotopic and contaminant approaches to food-web studies. Stable-carbon isotope analysis confirmed an enrichment in 13C between POM and ice algae (-22.3 vs. -17.7‰). Benthic organisms were generally enriched in 13C compared to pelagic species. We discuss individual species isotopic data and the general utility of our stable isotope model for defining carbon flux and contaminant flow through the North Water food web.

  14. 13C CPMAS NMR studies and DFT calculations of triterpene xylosides isolated from Actaea racemosa

    NASA Astrophysics Data System (ADS)

    Jamróz, Marta K.; Paradowska, Katarzyna; Gliński, Jan A.; Wawer, Iwona

    2011-05-01

    13C CPMAS NMR spectra of four triterpene glycosides: cimigenol xyloside ( 1), 26-deoxyactein ( 2), cimicifugoside H-1 ( 3) and 24-acethylhydroshengmanol xyloside ( 4) were recorded and analyzed to characterize their solid-state structure. Experimental data were supported by theoretical calculations of NMR shielding constants with the GIAO/6-31G**-su1 approach. A number of methods for the conformational search and a number of functionals for the DFT calculations were applied to ( 1). The best method was proven to be MMFF or MMFFAQ for the conformational search and the PBE1PBE functional for the DFT calculations. Extra calculations simulating C16 dbnd O⋯HOH hydrogen bond yield the isotropic shielding closer to the experimental solid-state value. For all the compounds CP kinetics parameters were calculated using either the I-S or the I-I*-S model. The analysis of CP kinetics data for methyl groups revealed differences in the T2 time constant for two methyl groups (C29 and C30) linked at C4.

  15. Covalent binding of reduced metabolites of [{sup 15}N{sub 3}]TNT to soil organic matter during a bioremediation process analyzed by {sup 15}N NMR spectroscopy

    SciTech Connect

    Achtnich, C.; Fernandes, E.; Bollag, J.M.; Knackmuss, H.J.; Lenke, H.

    1999-12-15

    Evidence is presented for the covalent binding of biologically reduced metabolites of 2,4,6-{sup 15}N{sub 3}-trinitrotoluene (TNT) to different soil fractions, using liquid {sup 15}N NMR spectroscopy. A silylation procedure was used to release soil organic matter from humin and whole soil for spectroscopic measurements. TNT-contaminated soil was spiked with 2,4,6-{sup 15}N{sub 3}-trinitrotoluene and {sup 14}C-ring labeled TNT, before treatment in a soil slurry reactor. During the anaerobic/aerobic incubation the amount of radioactivity detected in the fulvic and humic acid fractions did not change significantly whereas the radioactivity bound to humin increased to 71%. The {sup 15}N NMR spectra of the fulvic acid samples were dominated by a large peak that corresponded to aliphatic amines or ammonia. In the early stages of incubation, {sup 15}N NMR analysis of the humic acids indicated bound azoxy compounds. The signals arising from nitro and azoxy groups disappeared with further anaerobic treatment. At the end of incubation, the NMR shifts showed that nitrogen was covalently bound to humic acid as substituted amines and amides. The NMR spectra of the silylated humin suggest formation of azoxy compounds and imine linkages. Bound metabolites possessing nitro groups were also detected. Primary amines formed during the anaerobic incubation disappeared during the aerobic treatment. Simultaneously, the amount of amides and tertiary amines increased. Nitro and azoxy groups of bound molecules were still present in humin at the end of the incubation period. Formation of azoxy compounds from partially reduced TNT followed by binding and further reduction appears to be an important mechanism for the immobilization of metabolites of TNT to soil.

  16. Identification of histidine tautomers in proteins by 2D 1H/13C(delta2) one-bond correlated NMR.

    PubMed

    Sudmeier, James L; Bradshaw, Elizabeth M; Haddad, Kristin E Coffman; Day, Regina M; Thalhauser, Craig J; Bullock, Peter A; Bachovchin, William W

    2003-07-16

    If the 13Cdelta2 chemical shift of neutral ("high pH") histidine is >122 ppm, primarily Ndelta1-H tautomer (2) is indicated; if it is <122 ppm, primarily Nepsilon2-H tautomer (1) is indicated. His resonances from the catalytic triad of active serine proteases, for example, are readily distinguished from those of denatured enzyme. The 13Cdelta2 chemical shifts increased by 6.2 ppm for the catalytic histidines in both alpha-lytic protease and subtilisin BPN' in raising the pH from that of imidazolium cation to that of tautomer 2. This tautomer identification method is easy to implement, requiring only bioincorporation of [U-13C] (or the more readily available [U-13C,15N])-histidine. Standard 1H/13C correlation HMQC or HSQC NMR pulse programs then yield the 13Cdelta2 chemical shifts with the benefit of high 1H sensitivity. Because of large one-bond spin-couplings (1JCH approximately 200 Hz), the method should extend to proteins having large 1H and 13C line widths, including very high molecular weights. PMID:12848537

  17. 13C and 15N—Chemical Shift Anisotropy of Ampicillin and Penicillin-V Studied by 2D-PASS and CP/MAS NMR

    NASA Astrophysics Data System (ADS)

    Antzutkin, Oleg N.; Lee, Young K.; Levitt, Malcolm H.

    1998-11-01

    The principal values of the chemical shift tensors of all13C and15N sites in two antibiotics, ampicillin and penicillin-V, were determined by 2-dimensionalphaseadjustedspinningsideband (2D-PASS) and conventional CP/MAS experiments. The13C and15N chemical shift anisotropies (CSA), and their confidence limits, were evaluated using a Mathematica program. The CSA values suggest a revised assignment of the 2-methyl13C sites in the case of ampicillin. We speculate on a relationship between the chemical shift principal values of many of the13C and15N sites and the β-lactam ring conformation.

  18. Characterization of a Mixture of CO2 Adsorption Products in Hyperbranched Aminosilica Adsorbents by (13)C Solid-State NMR.

    PubMed

    Moore, Jeremy K; Sakwa-Novak, Miles A; Chaikittisilp, Watcharop; Mehta, Anil K; Conradi, Mark S; Jones, Christopher W; Hayes, Sophia E

    2015-11-17

    Hyperbranched amine polymers (HAS) grown from the mesoporous silica SBA-15 (hereafter "SBA-15-HAS") exhibit large capacities for CO2 adsorption. We have used static in situ and magic-angle spinning (MAS) ex situ (13)C nuclear magnetic resonance (NMR) to examine the adsorption of CO2 by SBA-15-HAS. (13)C NMR distinguishes the signal of gas-phase (13)CO2 from that of the chemisorbed species. HAS polymers possess primary, secondary, and tertiary amines, leading to multiple chemisorption reaction outcomes, including carbamate (RnNCOO(-)), carbamic acid (RnNCOOH), and bicarbonate (HCO3(-)) moieties. Carbamates and bicarbonate fall within a small (13)C chemical shift range (162-166 ppm), and a mixture was observed including carbamic acid and carbamate, the former disappearing upon evacuation of the sample. By examining the (13)C-(14)N dipolar coupling through low-field (B0 = 3 T) (13)C{(1)H} cross-polarization MAS NMR, carbamate is confirmed through splitting of the (13)C resonance. A third species that is either bicarbonate or a second carbamate is evident from bimodal T2 decay times of the ∼163 ppm peak, indicating the presence of two species comprising that single resonance. The mixture of products suggests that (1) the presence of amines and water leads to bicarbonate being present and/or (2) the multiple types of amine sites in HAS permit formation of chemically distinct carbamates. PMID:26477882

  19. 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

    NASA Astrophysics Data System (ADS)

    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 < 0.05). By contrast, δ 15N signatures of C. nasus in ZS, CM, and JJ groups were significantly lower than those in PYL group ( P < 0.05). Basing on δ 13C and δ 15N signatures, we could distinguish anadromous (ZS, CM, and JJ) and non-anadromous (PYL) groups. The trophic level (TL) of anadromous C. nasus ranged from 2.90 to 3.04, whereas that of non-anadromous C. nasus was 4.38. C. nasus occupied the middle and top 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.

  20. Benthic macroinvertebrates and the use of stable isotopes (δ13C and δ15N) in the impact assessment of peatland use on boreal stream ecosystems

    NASA Astrophysics Data System (ADS)

    Nieminen, Mika L.; Daza Secco, Emmanuela; Nykänen, Hannu; Meissner, Kristian

    2013-04-01

    Stable isotope analysis (SIA) can provide insights into carbon flow dynamics and trophic positions of consumers in food webs. SIA is used in this study, where we assess the possible changes in the basal resources of Finnish boreal stream ecosystems and differences in the impact of two forms of peatland use, forestry and peat mining. About 30% of the total land area of Finland is classified as peatland, of which about 55% has been drained for forestry and about 0.6% is in peat production. Unlike forestry, peat production is regionally less scattered and can thus have measurable local impacts although the total area of peat production is small. Three watersheds were used as study areas. Within each watershed, one stream drains a subcatchment affected only by peat mining, whereas the other stream flows through a subcatchment affected by forestry. The two subcatchment streams merge to form a single stream flowing into a lake. Studied watersheds were subject to no other forms of land use. In addition to the impacted sites, we used two pristine natural mire and two natural forest catchments as controls. We analysed the stable isotopes of carbon (δ13C) and nitrogen (δ15N) from benthic macroinvertebrates, stream bank soil, stream sediment, and dissolved organic carbon (DOC) in stream water. Samples for stable isotope analyses were collected in the summer of 2011 and samples for invertebrate community analyses in the autumn of 2011. Upon sampling we measured several physical parameters at each sampling site. In addition, stream water samples collected in summer and autumn 2012 were analysed for CH4 and CO2 gas concentrations and autumn gas samples also for their δ13C values. Our initial SIA results of invertebrates suggest some degree of discrimination between different sources of OM and possible effects on feeding habits, presumably due to the quality of the basal resources. We will explore this result further by examining not only taxonomical structure, but also the

  1. Whole-body protein turnover in preterm appropriate for gestational age and small for gestational age infants: comparison of [15N]glycine and [1-(13)C]leucine administered simultaneously.

    PubMed

    Van Goudoever, J B; Sulkers, E J; Halliday, D; Degenhart, H J; Carnielli, V P; Wattimena, J L; Sauer, P J

    1995-04-01

    Measurements of whole-body protein turnover in preterm infants have been made using different stable isotope methods. Large variation in results has been found, which could be due to different clinical conditions and/or the use of different tracers. We studied 14 appropriate for gestational age and nine small for gestational age orally fed preterm infants using [15N]glycine and [1-(13)C]leucine simultaneously, which allowed us to make a comparison of commonly used methods to calculate whole-body protein turnover. Whole-body protein turnover was calculated from 15N enrichment in urinary ammonia and urea after [15N]-glycine administration and from the 13C enrichment in expired CO2 after administration of [1-(13)C]leucine. Enrichment of alpha-ketoisocaproic acid after [1-(13)C]leucine constant infusion was measured as a direct parameter of whole-body protein turnover. Group means for whole-body protein turnover using [15N]glycine or [1-(13)C]leucine ranged from 10 to 14 g.kg-1.d-1, except when using the end product method that assumes a correlation between leucine oxidation and total nitrogen excretion. We found very low 15N enrichment of urinary urea in the majority of small for gestational age infants. These infants also had a lower nitrogen excretion in urine and oxidized less leucine. Nitrogen balance was higher in small for gestational age infants (416 +/- 25 mg.kg-1.d-1) compared with appropriate for gestational age infants (374 +/- 41 mg.kg-1.d-1, p = 0.003). [15N]Glycine does not seem to exchange its label with the body nitrogen pool to a significant degree and is therefore not always suitable as a carrier for 15N in protein turnover studies in premature infants. PMID:7596675

  2. Dynamic nuclear polarization-enhanced 1H–13C double resonance NMR in static samples below 20 K

    PubMed Central

    Potapov, Alexey; Thurber, Kent R.; Yau, Wai-Ming; Tycko, Robert

    2012-01-01

    We demonstrate the feasibility of one-dimensional and two-dimensional 1H–13C double resonance NMR experiments with dynamic nuclear polarization (DNP) at 9.4 T and temperatures below 20 K, including both 1H–13C cross-polarization and 1H decoupling, and discuss the effects of polarizing agent type, polarizing agent concentration, temperature, and solvent deuteration. We describe a two-channel low-temperature DNP/NMR probe, capable of carrying the radio-frequency power load required for 1H–13C cross-polarization and high-power proton decoupling. Experiments at 8 K and 16 K reveal a significant T2 relaxation of 13C, induced by electron spin flips. Carr–Purcell experiments and numerical simulations of Carr–Purcell dephasing curves allow us to determine the effective correlation time of electron flips under our experimental conditions. The dependence of the DNP signal enhancement on electron spin concentration shows a maximum near 80 mM. Although no significant difference in the absolute DNP enhancements for triradical (DOTOPA-TEMPO) and biradical (TOTAPOL) dopants was found, the triradical produced greater DNP build-up rates, which are advantageous for DNP experiments. Additionally the feasibility of structural measurements on 13C-labeled biomolecules was demonstrated with a two-dimensional 13C–13C exchange spectrum of selectively 13C-labeled β-amyloid fibrils. PMID:22743540

  3. Spatio-temporal isotopic signatures (δ13 C and δ15 N) reveal that two sympatric West African mullet species do not feed on the same basal production sources.

    PubMed

    Le Loc'h, F; Durand, J-D; Diop, K; Panfili, J

    2015-04-01

    Potential trophic competition between two sympatric mullet species, Mugil cephalus and Mugil curema, was explored in the hypersaline estuary of the Saloum Delta (Senegal) using δ(13) C and δ(15) N composition of muscle tissues. Between species, δ(15) N compositions were similar, suggesting a similar trophic level, while the difference in δ(13) C compositions indicated that these species did not feed from exactly the same basal production sources or at least not in the same proportions. This result provides the first evidence of isotopic niche segregation between two limno-benthophageous species belonging to the geographically widespread, and often locally abundant, Mugilidae family. PMID:25846862

  4. Experimentally Derived δ13C and δ15N Discrimination Factors for Gray Wolves and the Impact of Prior Information in Bayesian Mixing Models

    PubMed Central

    Bucci, Melanie E.; Callahan, Peggy; Koprowski, John L.; Polfus, Jean L.; Krausman, Paul R.

    2015-01-01

    Stable isotope analysis of diet has become a common tool in conservation research. However, the multiple sources of uncertainty inherent in this analysis framework involve consequences that have not been thoroughly addressed. Uncertainty arises from the choice of trophic discrimination factors, and for Bayesian stable isotope mixing models (SIMMs), the specification of prior information; the combined effect of these aspects has not been explicitly tested. We used a captive feeding study of gray wolves (Canis lupus) to determine the first experimentally-derived trophic discrimination factors of C and N for this large carnivore of broad conservation interest. Using the estimated diet in our controlled system and data from a published study on wild wolves and their prey in Montana, USA, we then investigated the simultaneous effect of discrimination factors and prior information on diet reconstruction with Bayesian SIMMs. Discrimination factors for gray wolves and their prey were 1.97‰ for δ13C and 3.04‰ for δ15N. Specifying wolf discrimination factors, as opposed to the commonly used red fox (Vulpes vulpes) factors, made little practical difference to estimates of wolf diet, but prior information had a strong effect on bias, precision, and accuracy of posterior estimates. Without specifying prior information in our Bayesian SIMM, it was not possible to produce SIMM posteriors statistically similar to the estimated diet in our controlled study or the diet of wild wolves. Our study demonstrates the critical effect of prior information on estimates of animal diets using Bayesian SIMMs, and suggests species-specific trophic discrimination factors are of secondary importance. When using stable isotope analysis to inform conservation decisions researchers should understand the limits of their data. It may be difficult to obtain useful information from SIMMs if informative priors are omitted and species-specific discrimination factors are unavailable. PMID:25803664

  5. Chlorophyll a-specific Δ14C, δ13C and δ15N values in stream periphyton: implications for aquatic food web studies

    NASA Astrophysics Data System (ADS)

    Ishikawa, N. F.; Yamane, M.; Suga, H.; Ogawa, N. O.; Yokoyama, Y.; Ohkouchi, N.

    2015-11-01

    Periphytic algae attached to a streambed substrate (periphyton) are an important primary producer in stream ecosystems. We determined the isotopic composition of chlorophyll a in periphyton collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, a pure aquatic primary producer (Cladophora sp.) and a terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Epeorus latifolium). Periphyton Δ14Cchl values (-258 ‰ in April and -190 ‰ in October) were slightly lower than Δ14Cbulk values (-228 ‰ in April and -179 ‰ in October) but were close to the Δ14C value for dissolved inorganic carbon (DIC; -217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = -1000 ‰), CO2 derived from aquatic and terrestrial organic matters (variable Δ14C) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (-215 ‰ in April and -199 ‰ in October) and Cladophora sp. (-210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

  6. Major Sources of Organic Matter in a Complex Coral Reef Lagoon: Identification from Isotopic Signatures (δ13C and δ15N).

    PubMed

    Briand, Marine J; Bonnet, Xavier; Goiran, Claire; Guillou, Gaël; Letourneur, Yves

    2015-01-01

    A wide investigation was conducted into the main organic matter (OM) sources supporting coral reef trophic networks in the lagoon of New Caledonia. Sampling included different reef locations (fringing, intermediate and barrier reef), different associated ecosystems (mangroves and seagrass beds) and rivers. In total, 30 taxa of macrophytes, plus pools of particulate and sedimentary OM (POM and SOM) were sampled. Isotopic signatures (C and N) of each OM sources was characterized and the composition of OM pools assessed. In addition, spatial and seasonal variations of reef OM sources were examined. Mangroves isotopic signatures were the most C-depleted (-30.17 ± 0.41 ‰) and seagrass signatures were the most C-enriched (-4.36 ± 0.72 ‰). Trichodesmium spp. had the most N-depleted signatures (-0.14 ± 0.03 ‰) whereas mangroves had the most N-enriched signatures (6.47 ± 0.41 ‰). The composition of POM and SOM varied along a coast-to-barrier reef gradient. River POM and marine POM contributed equally to coastal POM, whereas marine POM represented 90% of the POM on barrier reefs, compared to 10% river POM. The relative importance of river POM, marine POM and mangroves to the SOM pool decreased from fringing to barrier reefs. Conversely, the relative importance of seagrass, Trichodesmium spp. and macroalgae increased along this gradient. Overall, spatial fluctuations in POM and SOM were much greater than in primary producers. Seasonal fluctuations were low for all OM sources. Our results demonstrated that a large variety of OM sources sustain coral reefs, varying in their origin, composition and role and suggest that δ13C was a more useful fingerprint than δ15N in this endeavour. This study also suggested substantial OM exchanges and trophic connections between coral reefs and surrounding ecosystems. Finally, the importance of accounting for environmental characteristics at small temporal and spatial scales before drawing general patterns is highlighted. PMID

  7. Investigations of (Delta)14C, (delta)13C, and (delta)15N in vertebrae of white shark (Carcharodon carcharias) from the eastern North Pacific Ocean

    SciTech Connect

    Kerr, L A; Andrews, A H; Cailliet, G M; Brown, T A; Coale, K H

    2006-06-08

    The white shark (Carcharodon carcharias) has a complex life history that is characterized by large scale movements and a highly variable diet. Estimates of age and growth for the white shark from the eastern North Pacific Ocean indicate they have a slow growth rate and a relatively high longevity. Age, growth, and longevity estimates useful for stock assessment and fishery models, however, require some form of validation. By counting vertebral growth band pairs, ages can be estimated, but because not all sharks deposit annual growth bands and many are not easily discernable, it is necessary to validate growth band periodicity with an independent method. Radiocarbon ({sup 14}C) age validation uses the discrete {sup 14}C signal produced from thermonuclear testing in the 1950s and 1960s that is retained in skeletal structures as a time-specific marker. Growth band pairs in vertebrae, estimated as annual and spanning the 1930s to 1990s, were analyzed for {Delta}{sup 14}C and stable carbon and nitrogen isotopes ({delta}{sup 13}C and {delta}{sup 15}N). The aim of this study was to evaluate the utility of {sup 14}C age validation for a wide-ranging species with a complex life history and to use stable isotope measurements in vertebrae as a means of resolving complexity introduced into the {sup 14}C chronology by ontogenetic shifts in diet and habitat. Stable isotopes provided useful trophic position information; however, validation of age estimates was confounded by what may have been some combination of the dietary source of carbon to the vertebrae, large-scale movement patterns, and steep {sup 14}C gradients with depth in the eastern North Pacific Ocean.

  8. 1H, 13C and 15N resonance assignments of a highly-soluble murine interleukin-3 analogue with wild-type bioactivity.

    PubMed

    Yao, Shenggen; Murphy, James M; Low, Andrew; Norton, Raymond S

    2010-04-01

    Interleukin-3 (IL-3) is a cytokine that acts as a critical mediator of inflammation and immune responses to infections. IL-3, like interleukin-5 (IL-5) and granulocyte-macrophage colony stimulating factor (GM-CSF), exerts its effects on target cells via receptors composed of cytokine-specific alpha-subunits and a common beta-subunit (betac-subunit, shared with IL-5 and GM-CSF). In contrast to humans, mice also possess an additional beta-receptor, beta(IL-3), that can specifically bind IL-3. Except for a study carried out on an analogue of human IL-3 that contains 14 mutations, structure-related studies of IL-3 have been very limited, largely because of its poor solution behaviour. Here we report (1)H, (13)C, and (15)N chemical shift assignments of murine IL-3 comprising residues 33-156 (SWISS-PROT accession number: P01586), in which the only mutation is an alanine substitution of Cys105. The mIL-3 construct used in the present study was engineered by eliminating residues 27-32 of the N-terminus (the first 26 residues of the primary sequence of mIL-3 are cleaved in vivo during secretion), the C-terminal 10 residues (157-166), and a disulfide bond between Cys105 and Cys166 that is poorly conserved in orthologue sequences. The new construct vastly improves the solubility of murine IL-3 while maintaining its wild-type biological activity. PMID:20174897

  9. Major Sources of Organic Matter in a Complex Coral Reef Lagoon: Identification from Isotopic Signatures (δ13C and δ15N)

    PubMed Central

    Briand, Marine J.; Bonnet, Xavier; Goiran, Claire; Guillou, Gaël; Letourneur, Yves

    2015-01-01

    A wide investigation was conducted into the main organic matter (OM) sources supporting coral reef trophic networks in the lagoon of New Caledonia. Sampling included different reef locations (fringing, intermediate and barrier reef), different associated ecosystems (mangroves and seagrass beds) and rivers. In total, 30 taxa of macrophytes, plus pools of particulate and sedimentary OM (POM and SOM) were sampled. Isotopic signatures (C and N) of each OM sources was characterized and the composition of OM pools assessed. In addition, spatial and seasonal variations of reef OM sources were examined. Mangroves isotopic signatures were the most C-depleted (-30.17 ± 0.41 ‰) and seagrass signatures were the most C-enriched (-4.36 ± 0.72 ‰). Trichodesmium spp. had the most N-depleted signatures (-0.14 ± 0.03 ‰) whereas mangroves had the most N-enriched signatures (6.47 ± 0.41 ‰). The composition of POM and SOM varied along a coast-to-barrier reef gradient. River POM and marine POM contributed equally to coastal POM, whereas marine POM represented 90% of the POM on barrier reefs, compared to 10% river POM. The relative importance of river POM, marine POM and mangroves to the SOM pool decreased from fringing to barrier reefs. Conversely, the relative importance of seagrass, Trichodesmium spp. and macroalgae increased along this gradient. Overall, spatial fluctuations in POM and SOM were much greater than in primary producers. Seasonal fluctuations were low for all OM sources. Our results demonstrated that a large variety of OM sources sustain coral reefs, varying in their origin, composition and role and suggest that δ13C was a more useful fingerprint than δ15N in this endeavour. This study also suggested substantial OM exchanges and trophic connections between coral reefs and surrounding ecosystems. Finally, the importance of accounting for environmental characteristics at small temporal and spatial scales before drawing general patterns is highlighted. PMID

  10. Discovering [superscript 13]C NMR, [superscript 1]H NMR, and IR Spectroscopy in the General Chemistry Laboratory through a Sequence of Guided-Inquiry Exercises

    ERIC Educational Resources Information Center

    Iler, H. Darrell; Justice, David; Brauer, Shari; Landis, Amanda

    2012-01-01

    This sequence of three guided-inquiry labs is designed for a second-semester general chemistry course and challenges students to discover basic theoretical principles associated with [superscript 13]C NMR, [superscript 1]H NMR, and IR spectroscopy. Students learn to identify and explain basic concepts of magnetic resonance and vibrational…

  11. Natural abundance 14N and 15N solid-state NMR of pharmaceuticals and their polymorphs

    DOE PAGESBeta

    Veinberg, Stanislav L.; Johnston, Karen E.; Jaroszewicz, Michael J.; Kispal, Brianna M.; Mireault, Christopher R.; Kobayashi, Takeshi; Pruski, Marek; Schurko, Robert W.

    2016-06-08

    14N ultra-wideline (UW), 1H{15N} indirectly-detected HETCOR (idHETCOR) and 15N dynamic nuclear polarization (DNP) solid-state NMR (SSNMR) experiments, in combination with plane-wave density functional theory (DFT) calculations of 14N EFG tensors, were utilized to characterize a series of nitrogen-containing active pharmaceutical ingredients (APIs), including HCl salts of scopolamine, alprenolol, isoprenaline, acebutolol, dibucaine, nicardipine, and ranitidine. Here, a case study applying these methods for the differentiation of polymorphs of bupivacaine HCl is also presented. All experiments were conducted upon samples with naturally-abundant nitrogen isotopes. For most of the APIs, it was possible to acquire frequency-stepped UW 14N SSNMR spectra of stationarymore » samples, which display powder patterns corresponding to pseudo-tetrahedral (i.e., RR'R"NH+ and RR'NH2+) or other (i.e., RNH2 and RNO2) nitrogen environments.« less

  12. Backbone 1H, 13C, and 15N assignments for the tandem ubiquitin binding domains of signal transducing adapter molecule 1.

    PubMed

    Lim, Jongsoo; Hong, Yoon-Hun; Lee, Bong-Jin; Ahn, Hee-Chul

    2011-04-01

    Signal transducing adapter molecule (STAM) forms the endosomal sorting complex required for transport-0 (ESCRT-0) complex with hepatocyte growth factor-regulated substrate (Hrs) to sort the ubiquitinated cargo proteins from the early endosomes to the ESCRT-1 complex. ESCRT-0 complex, STAM and Hrs, contains multiple ubiquitin binding domains, in which STAM has two ubiquitin binding domains, Vps27/Hrs/Stam (VHS) and ubiquitin interacting motif (UIM) at its N-terminus. By the cooperation of the multiple ubiquitin binding domains, the ESCRT-0 complex recognizes poly-ubiquitin, especially Lys63-linked ubiquitin. Here, we report the backbone resonance assignments and the secondary structure of the N-terminal 191 amino acids of the human STAM1 which includes the VHS domain and UIM. The {(1)H}-(15)N heteronuclear NOE experiments revealed that an unstructured and flexible loop region connects the VHS domain and UIM. Our work provides the basic information for the further NMR investigation of the interaction between STAM1 and poly-ubiquitin. PMID:20927613

  13. 500-fold enhancement of in situ 13C liquid state NMR using gyrotron-driven temperature-jump DNP

    NASA Astrophysics Data System (ADS)

    Yoon, Dongyoung; Soundararajan, Murari; Caspers, Christian; Braunmueller, Falk; Genoud, Jérémy; Alberti, Stefano; Ansermet, Jean-Philippe

    2016-09-01

    A 550-fold increase in the liquid state 13C NMR signal of a 50 μL sample was obtained by first hyperpolarizing the sample at 20 K using a gyrotron (260 GHz), then, switching its frequency in order to apply 100 W for 1.5 s so as to melt the sample, finally, turning off the gyrotron to acquire the 13C NMR signal. The sample stays in its NMR resonator, so the sequence can be repeated with rapid cooling as the entire cryostat stays cold. DNP and thawing of the sample are performed only by the switchable and tunable gyrotron without external devices. Rapid transition from DNP to thawing in one second time scale was necessary especially in order to enhance liquid 1H NMR signal.

  14. 500-fold enhancement of in situ (13)C liquid state NMR using gyrotron-driven temperature-jump DNP.

    PubMed

    Yoon, Dongyoung; Soundararajan, Murari; Caspers, Christian; Braunmueller, Falk; Genoud, Jérémy; Alberti, Stefano; Ansermet, Jean-Philippe

    2016-09-01

    A 550-fold increase in the liquid state (13)C NMR signal of a 50μL sample was obtained by first hyperpolarizing the sample at 20K using a gyrotron (260GHz), then, switching its frequency in order to apply 100W for 1.5s so as to melt the sample, finally, turning off the gyrotron to acquire the (13)C NMR signal. The sample stays in its NMR resonator, so the sequence can be repeated with rapid cooling as the entire cryostat stays cold. DNP and thawing of the sample are performed only by the switchable and tunable gyrotron without external devices. Rapid transition from DNP to thawing in one second time scale was necessary especially in order to enhance liquid (1)H NMR signal. PMID:27490302

  15. Spectrally edited 2D 13Csbnd 13C NMR spectra without diagonal ridge for characterizing 13C-enriched low-temperature carbon materials

    NASA Astrophysics Data System (ADS)

    Johnson, Robert L.; Anderson, Jason M.; Shanks, Brent H.; Fang, Xiaowen; Hong, Mei; Schmidt-Rohr, Klaus

    2013-09-01

    Two robust combinations of spectral editing techniques with 2D 13Csbnd 13C NMR have been developed for characterizing the aromatic components of 13C-enriched low-temperature carbon materials. One method (exchange with protonated and nonprotonated spectral editing, EXPANSE) selects cross peaks of protonated and nearby nonprotonated carbons, while the other technique, dipolar-dephased double-quantum/single-quantum (DQ/SQ) NMR, selects signals of bonded nonprotonated carbons. Both spectra are free of a diagonal ridge, which has many advantages: Cross peaks on the diagonal or of small intensity can be detected, and residual spinning sidebands or truncation artifacts associated with the diagonal ridge are avoided. In the DQ/SQ experiment, dipolar dephasing of the double-quantum coherence removes protonated-carbon signals; this approach also eliminates the need for high-power proton decoupling. The initial magnetization is generated with minimal fluctuation by combining direct polarization, cross polarization, and equilibration by 13C spin diffusion. The dipolar dephased DQ/SQ spectrum shows signals from all linkages between aromatic rings, including a distinctive peak from polycondensed aromatics. In EXPANSE NMR, signals of protonated carbons are selected in the first spectral dimension by short cross polarization combined with dipolar dephasing difference. This removes ambiguities of peak assignment to overlapping signals of nonprotonated and protonated aromatic carbons, e.g. near 125 ppm. Spin diffusion is enhanced by dipolar-assisted rotational resonance. Before detection, Csbnd H dipolar dephasing by gated decoupling is applied, which selects signals of nonprotonated carbons. Thus, only cross peaks due to magnetization originating from protonated C and ending on nearby nonprotonated C are retained. Combined with the chemical shifts deduced from the cross-peak position, this double spectral editing defines the bonding environment of aromatic, COO, and Cdbnd O carbons

  16. Stable Carbon and Nitrogen isoscapes of the California Coast: integrated δ15N and δ13C of suspended particulate organic matter inferred from tissues of the California Mussel (mytilus californianus)

    NASA Astrophysics Data System (ADS)

    Vokhshoori, N. L.; McCarthy, M. D.

    2011-12-01

    Spatial maps of isotopic variability in a single species, or isoscapes, can characterize the natural variability in carbon (C) and nitrogen (N) isotope ratios across ecosystems on broad spatial scales, trace the signature of a source across a given area, as well as constrain animal migration patterns (Graham et al. 2002). In this study, isoscapes of stable carbon (13C) and nitrogen (15N) isotopes were constructed using intertidal mussels for northeast Pacific coastal waters of California. In this region biogeochemical cycling is primarily controlled by upwelling intensity and large-scale transport of the California Current System (CCS). We hypothesize that sampling specific tissues of filter-feeding organisms can provide an integrated measure of variation in 15N and 13C of the suspended particulate organic matter (POM) pool vs. latitude within the CCS, as well indicate main sources of both organic C and N to littoral food webs. California mussels (mytilus californianus) were collected from 28 sites between Coos Bay, OR and La Jolla, CA in the winter of 2009-2010 and summer of 2011, and adductor tissue was analyzed for δ13C and δ15N. Mussel size classes were chosen to provide ~ 1 yr integrated signal. Spatial trends in δ15N from the winter sampling show a strong linear trend in increasing δ15N values with latitude north to south (δ15N values range from 7 % to 12%) consistent with slowly attenuating northward transport of 15N-depleted nitrate via California Undercurrent (Altabet et al. 1999). The δ13C values have no strong north to south correlation, but exhibit strong location-specific variability. The δ13C values range between -13 % and -18%. We propose the site-specific signature of δ13C indicates relative source of primary productin to POM at a given region (i.e. kelp, phytoplankton, zooplankton). Overall, these results suggest that isoscapes for filter-feeding organisms may offer a more accurate integrated picture of 15N and 13C values of POM than is

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

    PubMed

    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. PMID:26154586

  18. Characterization of Stratum Corneum Molecular Dynamics by Natural-Abundance 13C Solid-State NMR

    PubMed Central

    Bouwstra, Joke A.; Sparr, Emma; Topgaard, Daniel

    2013-01-01

    Despite the enormous potential for pharmaceutical applications, there is still a lack of understanding of the molecular details that can contribute to increased permeability of the stratum corneum (SC). To investigate the influence of hydration and heating on the SC, we record the natural-abundance 13C signal of SC using polarization transfer solid-state NMR methods. Resonance lines from all major SC components are assigned. Comparison of the signal intensities obtained with the INEPT and CP pulse sequences gives information on the molecular dynamics of SC components. The majority of the lipids are rigid at 32°C, and those lipids co-exist with a small pool of mobile lipids. The ratio between mobile and rigid lipids increases with hydration. An abrupt change of keratin filament dynamics occurs at RH = 80–85%, from completely rigid to a structure with rigid backbone and mobile protruding terminals. Heating has a strong effect on the lipid mobility, but only a weak influence on the keratin filaments. The results provide novel molecular insight into how the SC constituents are affected by hydration and heating, and improve the understanding of enhanced SC permeability, which is associated with elevated temperatures and SC hydration. PMID:23626744

  19. (13)C-NMR-Based Metabolomic Profiling of Typical Asian Soy Sauces.

    PubMed

    Kamal, Ghulam Mustafa; Yuan, Bin; Hussain, Abdullah Ijaz; Wang, Jie; Jiang, Bin; Zhang, Xu; Liu, Maili

    2016-01-01

    It has been a strong consumer interest to choose high quality food products with clear information about their origin and composition. In the present study, a total of 22 Asian soy sauce samples have been analyzed in terms of (13)C-NMR spectroscopy. Spectral data were analyzed by multivariate statistical methods in order to find out the important metabolites causing the discrimination among typical soy sauces from different Asian regions. It was found that significantly higher concentrations of glutamate in Chinese red cooking (CR) soy sauce may be the result of the manual addition of monosodium glutamate (MSG) in the final soy sauce product. Whereas lower concentrations of amino acids, like leucine, isoleucine and valine, observed in CR indicate the different fermentation period used in production of CR soy sauce, on the other hand, the concentration of some fermentation cycle metabolites, such as acetate and sucrose, can be divided into two groups. The concentrations of these fermentation cycle metabolites were lower in CR and Singapore Kikkoman (SK), whereas much higher in Japanese shoyu (JS) and Taiwan (China) light (TL), which depict the influence of climatic conditions. Therefore, the results of our study directly indicate the influences of traditional ways of fermentation, climatic conditions and the selection of raw materials and can be helpful for consumers to choose their desired soy sauce products, as well as for researchers in further authentication studies about soy sauce. PMID:27598115

  20. 1H, 13C and 29Si NMR of tetramethylsilane in liquid crystals

    NASA Astrophysics Data System (ADS)

    Hiltunen, Y.; Jokisaari, J.

    1990-12-01

    The 1H, 13C and 29Si NMR spectra of tetramethylsilane (TMS) dissolved in two nematic liquid crystals (LC) and in their three mixtures were recorded. The proton—proton, proton—carbon and proton—silicon dipolar couplings, which arise from molecular deformation in the LC environment, were determined. The results for the 2DHH, 4DHH, 3DCH and 2DSiH couplings show only a small variation as a function of the composition of the LC mixture. On the contrary, the one-bond CH dipolar coupling is markedly solvent dependent: it varies from -6.22 Hz (in ZLI 1167) to +3.63 Hz (in phase IV). The 1DCH coupling of TMS vanishes in a certain mixture of the two liquid crystals; this mixture, however, is not the same as that in which the corresponding coupling of methane was earlier observed to vanish. This different behaviour of TMS and methane may be due to the additional torques which act on the SiC bonds of TMS.

  1. Biosynthesis of the antibiotic maduramicin. Origin of the carbon and oxygen atoms as well as the 13C NMR assignments.

    PubMed

    Tsou, H; Rajan, S; Fiala, R; Mowery, P C; Bullock, M W; Borders, D B; James, J C; Martin, J H; Morton, G O

    1984-12-01

    The biosynthesis of maduramicin alpha and beta in a culture of Actinomadura yumaensis has been studied using 13C, 14C and 18O labeled precursors. The alpha component of this recently discovered polyether antibiotic, containing forty-seven carbon atoms in a seven-ring system, is derived from eight acetate, seven propionate and four methionine molecules. The beta component which is missing one methoxy group incorporates three methionine methyl groups. The carbohydrate moiety was enriched by methionine, but not significantly by acetate or propionate. Studies of the incorporation of 13C labeled precursors permit the 13C NMR assignment of maduramicin. The origin of oxygen atoms of maduramicin has been examined by feeding [1-13C, 18O2]acetate and [1-13C, 18O2]propionate separately in the fermentation culture and the resulting doubly labeled maduramicin samples were analyzed by the isotopic shifts in the 13C NMR spectra. These results are consistent with the initial formation of a triene, which is converted to maduramicin by cyclization of the triepoxide. PMID:6526733

  2. NMR studies of bent DNA using {sup 13}C-enriched samples

    SciTech Connect

    Zimmer, D.P.; Crothers, D.M.

    1994-12-01

    Bending of the DNA double helix can be brought about by introducing runs of adenines (A-tracts) in phase with the helical repeat of the DNA. The requirements for bending of DNA by A-tracts are that the length of the A-tract be greater than 3 base pairs and that the A-tracts must be in phase with the helical repeat (every 10 or 11 bp). Other factors, such as the number of adenines in the run, flanking sequences, and whether the A-tracts are phased with respect to the 5{prime}A or the 3{prime}A, have effects upon the degree of bending as assayed by electrophoretic mobility on native polyacrylamide gels. There are a number of models for bending A-tract DNA. The junction-bending model postulates that the structure of A-tracts is similar to the fiber diffraction structure of poly A, in which there is a significant degree of base pair tilt with respect to the helix axis. In this model, bending occurs at the junction between the A-tract and the B-form helix to allow favorable stacking interactions to occur. The bend of the helix could arise as a result of some other perturbation of B-form DNA by A-tracts, such as propeller twist; bending also could be due to a combination of factors. Our goal is to find the structural features of A-tracts responsible for bending of the helix by performing NMR on oligonucleotides containing A-tracts to obtain higher resolution structural data. One of the problems encountered in NMR structure determination of nucleic acids and other macromolecules is the assignment of resonances to nuclei. This procedure can be greatly facilitated through the use of {sup 13}C-enriched nucleic acid samples. We are developing a technique for the enzymatic synthesis of labeled DNA for NMR. The technique we are developing is similar to RNA labeling techniques already in use. The technique involves growth of methylotrophic bacteria on {sup 13}CH{sub 3}OH.

  3. Spinning-frequency-dependent linewidths in 1H-decoupled 13C magic-angle spinning NMR spectra

    NASA Astrophysics Data System (ADS)

    Nakai, Toshihito; McDowell, Charles A.

    1994-09-01

    The broadenings observed in 13C MAS NMR spectra, which depend on the sample-spinning speed, were studied, using polycrystalline adamantane. Not only was a monotonic increase of the linewidths with the increase of the spinning frequency observed, but also a novel resonant feature was found. The phenomena were interpreted as originating from rotary-resonance 13C 1H recoupling.

  4. Complete assignment of (1)H and (13)C NMR spectra of standard neo-iota-carrabiose oligosaccharides.

    PubMed

    Jouanneau, Diane; Boulenguer, Patrick; Mazoyer, Jacques; Helbert, William

    2010-02-26

    Standard Eucheuma denticulatum iota-carrageenan was degraded with the Alteromonas fortis iota-carrageenase. The most abundant products, the neo-iota-carratetraose and neo-iota-carrahexaose were purified by permeation gel chromatography, and their corresponding (1)H and (13)C NMR spectra were fully assigned. PMID:20038459

  5. Conformational changes of alamethicin induced by solvent and temperature. A 13C-NMR and circular-dichroism study.

    PubMed

    Jung, G; Dubischar, N

    1975-06-01

    13C nuclear magnetic resonance (NMR) and circular dichroism (CD) have been used for studies on the conformation of alamethicin. The 13C NMR spectrum is assigned with the aid of signals of synthetic partial sequences and selective proton decoupling. The solvent and temperature-dependence of the 13C NMR spectra, T1 measurements and the use of lanthanide-shift reagents allow the differentiation between the amino acids belonging to a rigid alpha-helical portion of the alamethicin sequence and those belonging to a more flexible part. The 13C NMR results are in agreement with results obtained from extended solvent and temperature-dependent CD studies which indicate a highly stabilized nonpolar and intrachenar alpha-helical part. The concentration-dependence of the CD spectrum of alamethicin in a nematic phase revealed aggregation phenomena which might simulate those observed in natural and synthetic membranes. After dissolving alamethicin in aqueous alcohol there is a time-dependence of the ellipticity of the Cotton effects showing a sort of memory effect on the mode of dissolution. Four different conformations can be characterized by CD spectra depending on the solvent and concentration. A model illustrating the dynamic conformations and aggregation phenomena within a membrane is proposed. PMID:1175592

  6. Enzymatic 13C Labeling and Multidimensional NMR Analysis of Miltiradiene Synthesized by Bifunctional Diterpene Cyclase in Selaginella moellendorffii*

    PubMed Central

    Sugai, Yoshinori; Ueno, Yohei; Hayashi, Ken-ichiro; Oogami, Shingo; Toyomasu, Tomonobu; Matsumoto, Sadamu; Natsume, Masahiro; Nozaki, Hiroshi; Kawaide, Hiroshi

    2011-01-01

    Diterpenes show diverse chemical structures and various physiological roles. The diversity of diterpene is primarily established by diterpene cyclases that catalyze a cyclization reaction to form the carbon skeleton of cyclic diterpene. Diterpene cyclases are divided into two types, monofunctional and bifunctional cyclases. Bifunctional diterpene cyclases (BDTCs) are involved in hormone and defense compound biosyntheses in bryophytes and gymnosperms, respectively. The BDTCs catalyze the successive two-step type-B (protonation-initiated cyclization) and type-A (ionization-initiated cyclization) reactions of geranylgeranyl diphosphate (GGDP). We found that the genome of a lycophyte, Selaginella moellendorffii, contains six BDTC genes with the majority being uncharacterized. The cDNA from S. moellendorffii encoding a BDTC-like enzyme, miltiradiene synthase (SmMDS), was cloned. The recombinant SmMDS converted GGDP to a diterpene hydrocarbon product with a molecular mass of 272 Da. Mutation in the type-B active motif of SmMDS abolished the cyclase activity, whereas (+)-copalyl diphosphate, the reaction intermediate from the conversion of GGDP to the hydrocarbon product, rescued the cyclase activity of the mutant to form a diterpene hydrocarbon. Another mutant lacking type-A activity accumulated copalyl diphosphate as the reaction intermediate. When the diterpene hydrocarbon was enzymatically synthesized from [U-13C6]mevalonate, all carbons were labeled with 13C stable isotope (>99%). The fully 13C-labeled product was subjected to 13C-13C COSY NMR spectroscopic analyses. The direct carbon-carbon connectivities observed in the multidimensional NMR spectra demonstrated that the hydrocarbon product by SmMDS is miltiradiene, a putative biosynthetic precursor of tanshinone identified from the Chinese medicinal herb Salvia miltiorrhiza. Hence, SmMDS functions as a bifunctional miltiradiene synthase in S. moellendorffii. In this study, we demonstrate that one-dimensional and

  7. Secondary structure and side-chain sup 1 H and sup 13 C resonance assignments of calmodulin in solution by heteronuclear multidimensional NMR spectroscopy

    SciTech Connect

    Ikura, Mitsuhiko; Spera, S.; Barbato, G.; Kay, L.E.; Bax, A. ); Krinks, M. )

    1991-09-24

    Heteronuclear 2D and 3D NMR experiments were carried out on recombinant Drosophila calmodulin (CaM), a protein of 148 residues and with molecular mass of 16.7 kDa, that is uniformly labeled with {sup 15}N and {sup 13}C to a level of > 95%. Nearly complete {sup 1}H and {sup 13}C side-chain assignments for all amino acid residues are obtained by using the 3D HCCH-COSY and HCCH-TOCSY experiments that rely on large heteronuclear one-bond scalar couplings to transfer magnetization and establish through-bond connectivities. The secondary structure of this protein in solution has been elucidated by a qualitative interpretation of nuclear Overhauser effects, hydrogen exchange data, and {sup 3}J{sub HNH{alpha}} coupling constants. A clear correlation between the {sup 13}C{alpha} chemical shift and secondary structure is found. The secondary structure in the two globular domains of Drosophila CaM in solution is essentially identical with that of the X-ray crystal structure of mammalian CaM which consists of two pairs of a helix-loop-helix motif in each globular domain. The existence of a short antiparallel {beta}-sheet between the two loops in each domain has been confirmed. The eight {alpha}-helix segments identified from the NMR data are located at Glu-6 to Phe-19, thr-29 to Ser-38, Glu-45 to Glu-54, Phe-65 to Lys-77, Glu-82 to Asp-93, Ala-102 to Asn-111, Asp-118 to Glu-127, and Tyr-138 to Thr-146. Although the crystal structure has a long central helix from Phe-65 to Phe-92 that connects the two globular domains, NMR data indicate that residues Asp-78 to Ser-81 of this central helix adopt a nonhelical conformation with considerable flexibility.

  8. Characterization of cerebral glutamine uptake from blood in the mouse brain: implications for metabolic modeling of 13C NMR data

    PubMed Central

    Bagga, Puneet; Behar, Kevin L; Mason, Graeme F; De Feyter, Henk M; Rothman, Douglas L; Patel, Anant B

    2014-01-01

    13C Nuclear Magnetic Resonance (NMR) studies of rodent and human brain using [1-13C]/[1,6-13C2]glucose as labeled substrate have consistently found a lower enrichment (∼25% to 30%) of glutamine-C4 compared with glutamate-C4 at isotopic steady state. The source of this isotope dilution has not been established experimentally but may potentially arise either from blood/brain exchange of glutamine or from metabolism of unlabeled substrates in astrocytes, where glutamine synthesis occurs. In this study, the contribution of the former was evaluated ex vivo using 1H-[13C]-NMR spectroscopy together with intravenous infusion of [U-13C5]glutamine for 3, 15, 30, and 60 minutes in mice. 13C labeling of brain glutamine was found to be saturated at plasma glutamine levels >1.0 mmol/L. Fitting a blood–astrocyte–neuron metabolic model to the 13C enrichment time courses of glutamate and glutamine yielded the value of glutamine influx, VGln(in), 0.036±0.002 μmol/g per minute for plasma glutamine of 1.8 mmol/L. For physiologic plasma glutamine level (∼0.6 mmol/L), VGln(in) would be ∼0.010 μmol/g per minute, which corresponds to ∼6% of the glutamine synthesis rate and rises to ∼11% for saturating blood glutamine concentrations. Thus, glutamine influx from blood contributes at most ∼20% to the dilution of astroglial glutamine-C4 consistently seen in metabolic studies using [1-13C]glucose. PMID:25074745

  9. Using magnetic coupling to implement 1H, 19F, 13C experiments in routine high resolution NMR probes

    NASA Astrophysics Data System (ADS)

    Bowyer, Paul; Finnigan, Jim; Marsden, Brian; Taber, Bob; Zens, Albert

    2015-12-01

    We report in this paper the design of 1H, 19F, 13C circuitry using magnetic coupling which can do on demand experiments where one of the three nuclei is observed and the other two are decoupled. The implementation of this circuitry in routine NMR probes is compared with capacitive coupling methods where it was found that by using magnetic coupling the performance of the routine NMR probe was not impacted by the addition of this circuitry. It is surmised that using this type of circuitry would be highly desirable for those chemists doing routine 19F NMR.

  10. Effects of foliar boron application on seed composition, cell wall boron, and seed δ15N and δ13C isotopes in water-stressed soybean plants

    PubMed Central

    Bellaloui, Nacer; Hu, Yanbo; Mengistu, Alemu; Kassem, My A.; Abel, Craig A.

    2013-01-01

    Limited information is available on the effects of foliar boron (B) application on soybean seed composition. The objective of this research was to investigate the effects of foliar B on seed composition (protein, oil, fatty acids, and sugars). Our hypothesis was that since B is involved in nitrogen and carbon metabolism, it may impact seed composition. A repeated greenhouse experiment was conducted where half of the soybean plants was exposed to water stress (WS) and the other half was well-watered. Foliar boron (FB) in the form of boric acid was applied twice at a rate of 1.1 kg ha−1. The first application was during flowering stage, and the second application was during seed-fill stage. Treatments were water stressed plants with no FB (WS–B); water stressed plants with FB (WS+B); watered plants without FB (W–B), and watered plants with FB (W+B). The treatment W–B was used as a control. Comparing with WS–B plants, B concentration was the highest in leaves and seed of W+B plants (84% increase in leaves and 73% in seed). Seeds of W+B plants had higher protein (11% increase), oleic acid (27% increase), sucrose (up to 40% increase), glucose, and fructose comparing with W–B. However, seed stachyose concentrations increased by 43% in WS–B plants seed compared with W–B plants. Cell wall (structural) B concentration in leaves was higher in all plants under water stress, especially in WS–B plants where the percentage of cell wall B reached up to 90%. Water stress changed seed δ15N and δ13C values in both B applied and non-B applied plants, indicating possible effects on nitrogen and carbon metabolism. This research demonstrated that FB increased B accumulation in leaves and seed, and altered seed composition of well-watered and water stressed plants, indicating a possible involvement of B in seed protein, and oleic and linolenic fatty acids. Further research is needed to explain mechanisms of B involvement in seed protein and fatty acids. PMID:23888163

  11. Effects of foliar boron application on seed composition, cell wall boron, and seed δ(15)N and δ(13)C isotopes in water-stressed soybean plants.

    PubMed

    Bellaloui, Nacer; Hu, Yanbo; Mengistu, Alemu; Kassem, My A; Abel, Craig A

    2013-01-01

    Limited information is available on the effects of foliar boron (B) application on soybean seed composition. The objective of this research was to investigate the effects of foliar B on seed composition (protein, oil, fatty acids, and sugars). Our hypothesis was that since B is involved in nitrogen and carbon metabolism, it may impact seed composition. A repeated greenhouse experiment was conducted where half of the soybean plants was exposed to water stress (WS) and the other half was well-watered. Foliar boron (FB) in the form of boric acid was applied twice at a rate of 1.1 kg ha(-1). The first application was during flowering stage, and the second application was during seed-fill stage. Treatments were water stressed plants with no FB (WS-B); water stressed plants with FB (WS+B); watered plants without FB (W-B), and watered plants with FB (W+B). The treatment W-B was used as a control. Comparing with WS-B plants, B concentration was the highest in leaves and seed of W+B plants (84% increase in leaves and 73% in seed). Seeds of W+B plants had higher protein (11% increase), oleic acid (27% increase), sucrose (up to 40% increase), glucose, and fructose comparing with W-B. However, seed stachyose concentrations increased by 43% in WS-B plants seed compared with W-B plants. Cell wall (structural) B concentration in leaves was higher in all plants under water stress, especially in WS-B plants where the percentage of cell wall B reached up to 90%. Water stress changed seed δ(15)N and δ(13)C values in both B applied and non-B applied plants, indicating possible effects on nitrogen and carbon metabolism. This research demonstrated that FB increased B accumulation in leaves and seed, and altered seed composition of well-watered and water stressed plants, indicating a possible involvement of B in seed protein, and oleic and linolenic fatty acids. Further research is needed to explain mechanisms of B involvement in seed protein and fatty acids. PMID:23888163

  12. Shifts in Ross Sea food web structure as indicated by δ15N and δ13C values of fossil Antarctic seals

    NASA Astrophysics Data System (ADS)

    Leopold, A.; Brault, E.; McMahon, K.

    2013-12-01

    As climate change continues to mount, there is a growing need for understanding its effects on biological-physical interactions of marine ecosystems. Assessing the effects of anthropogenic activities on the coastal marine ecosystem involves understanding the underlying mechanisms driving these changes as well as establishing baselines of the natural system. Preliminary findings have indicated shifts in bulk carbon (C) and nitrogen (N) isotopic values of southern elephant seal (Mirounga leonina) samples, collected in the Dry Valleys of Antarctica in the Ross Sea region, over approximately the last 7,000 years. These shifts could result from 1) seals changing their foraging location and/or diet over this time, 2) climate change-induced shifts in the biogeochemistry at the base of the food web, or 3) some combination of both processes. We explored the patterns of long-term change in Ross Sea food web structure by examining the stable isotope values of three top predators in this system, Weddell seals (Leptonychotes weddellii), leopard seals (Hydrurga leptonyx), and crabeater seals (Lobodon carcinophagus). Fossil seal samples were collected in the Dry Valleys during the austral summer of 2012/13 and then analyzed for bulk C and N isotopes via an elemental analyzer/isotope-ratio mass spectrometer (EA/IRMS). Our initial findings indicate that C isotopic values of fossil seal samples from Weddell, leopard, and crabeater seals were more enriched than isotopic values of modern seals of the same species (e.g., δ13C = -22.79 × 0.92 ‰ and -26.71 × 0.50 ‰ for fossil and modern crabeater seals, respectively). Given the relatively consistent diet of crabeater seals, these findings suggest a shift in baseline food web structure occurred over the last 10,000 years, either through changes in foraging location or local shifts in biogeochemistry. For all species, N isotopic values are widely variable (e.g., 7.28 to 16.0 δ15N ‰ for the Weddell seal), which may be a result of

  13. Relaxation-Compensated Difference Spin Diffusion NMR for Detecting 13C-13C Long-Range Correlations in Proteins and Polysaccharides

    PubMed Central

    Wang, Tuo; Williams, Jonathan K.; Schmidt-Rohr, Klaus; Hong, Mei

    2015-01-01

    The measurement of long-range distances remains a challenge in solid-state NMR structure determination of biological macromolecules. In 2D and 3D correlation spectra of uniformly 13C-labeled biomolecules, inter-residue, inter-segmental, and intermolecular 13C-13C cross peaks that provide important long-range distance constraints for three-dimensional structures often overlap with short-range cross peaks that only reflect the covalent structure of the molecule. It is therefore desirable to develop new approaches to obtain spectra containing only long-range cross peaks. Here we show that a relaxation-compensated modification of the commonly used 2D 1H-driven spin diffusion (PDSD) experiment allows the clean detection of such long-range cross peaks. By adding a z-filter to keep the total z-period of the experiment constant, we compensate for 13C T1 relaxation. As a result, the difference spectrum between a long- and a scaled short-mixing time spectrum show only long-range correlation signals. We show that one- and two-bond cross peaks equalize within a few tens of milliseconds. Within ~200 ms, the intensity equilibrates within an amino acid residue and a monosaccharide to a value that reflects the number of spins in the local network. With T1 relaxation compensation, at longer mixing times, inter-residue and inter-segmental cross peaks increase in intensity whereas intra-segmental cross-peak intensities remain unchanged relative to each other and can all be subtracted out. Without relaxation compensation, the difference 2D spectra exhibit both negative and positive intensities due to heterogeneous T1 relaxation in most biomolecules, which can cause peak cancellation. We demonstrate this relaxation-compensated difference PDSD approach on amino acids, monosaccharides, a crystalline model peptide, a membrane-bound peptide and a plant cell wall sample. The resulting difference spectra yield clean multi-bond, inter-residue and intermolecular correlation peaks, which are

  14. Natural abundance (14)N and (15)N solid-state NMR of pharmaceuticals and their polymorphs.

    PubMed

    Veinberg, Stanislav L; Johnston, Karen E; Jaroszewicz, Michael J; Kispal, Brianna M; Mireault, Christopher R; Kobayashi, Takeshi; Pruski, Marek; Schurko, Robert W

    2016-06-29

    (14)N ultra-wideline (UW), (1)H{(15)N} indirectly-detected HETCOR (idHETCOR) and (15)N dynamic nuclear polarization (DNP) solid-state NMR (SSNMR) experiments, in combination with plane-wave density functional theory (DFT) calculations of (14)N EFG tensors, were utilized to characterize a series of nitrogen-containing active pharmaceutical ingredients (APIs), including HCl salts of scopolamine, alprenolol, isoprenaline, acebutolol, dibucaine, nicardipine, and ranitidine. A case study applying these methods for the differentiation of polymorphs of bupivacaine HCl is also presented. All experiments were conducted upon samples with naturally-abundant nitrogen isotopes. For most of the APIs, it was possible to acquire frequency-stepped UW (14)N SSNMR spectra of stationary samples, which display powder patterns corresponding to pseudo-tetrahedral (i.e., RR'R''NH(+) and RR'NH2(+)) or other (i.e., RNH2 and RNO2) nitrogen environments. Directly-excited (14)N NMR spectra were acquired using the WURST-CPMG pulse sequence, which incorporates WURST (wideband, uniform rate, and smooth truncation) pulses and a CPMG (Carr-Purcell Meiboom-Gill) refocusing protocol. In certain cases, spectra were acquired using (1)H → (14)N broadband cross-polarization, via the BRAIN-CP (broadband adiabatic inversion - cross polarization) pulse sequence. These spectra provide (14)N electric field gradient (EFG) tensor parameters and orientations that are particularly sensitive to variations in local structure and intermolecular hydrogen-bonding interactions. The (1)H{(15)N} idHETCOR spectra, acquired under conditions of fast magic-angle spinning (MAS), used CP transfers to provide (1)H-(15)N chemical shift correlations for all nitrogen environments, except for two sites in acebutolol and nicardipine. One of these two sites (RR'NH2(+) in acebutolol) was successfully detected using the DNP-enhanced (15)N{(1)H} CP/MAS measurement, and one (RNO2 in nicardipine) remained elusive due to the absence of

  15. Changes in dynamics of SRE-RNA on binding to the VTS1p-SAM domain studied by 13C NMR relaxation.

    PubMed

    Oberstrass, Florian C; Allain, Frédéric H-T; Ravindranathan, Sapna

    2008-09-10

    RNA recognition by proteins is often accompanied by significant changes in RNA dynamics in addition to conformational changes. However, there are very few studies which characterize the changes in molecular motions in RNA on protein binding. We present a quantitative (13)C NMR relaxation study of the changes in RNA dynamics in the pico-nanosecond time scale and micro-millisecond time scale resulting from interaction of the stem-loop SRE-RNA with the VTS1p-SAM domain. (13)C relaxation rates of the protonated carbons of the nucleotide base and anomeric carbons have been analyzed by employing the model-free formalism, for a fully (13)C/(15)N-labeled sample of the SRE-RNA in the free and protein-bound forms. In the free RNA, the nature of molecular motions are found to be distinctly different in the stem and the loop region. On binding to the protein, the nature of motions becomes more homogeneous throughout the RNA, with many residues showing increased flexibility at the aromatic carbon sites, while the anomeric carbon sites become more rigid. Surprisingly, we also observe indications of a slow collective motion of the RNA in the binding pocket of the protein. The observation of increased motions on binding is interesting in the context of growing evidence that binding does not always lead to motional restrictions and the resulting entropy gain could favor the free energy of association. PMID:18698768

  16. (15)N CSA tensors and (15)N-(1)H dipolar couplings of protein hydrophobic core residues investigated by static solid-state NMR.

    PubMed

    Vugmeyster, Liliya; Ostrovsky, Dmitry; Fu, Riqiang

    2015-10-01

    In this work, we assess the usefulness of static (15)N NMR techniques for the determination of the (15)N chemical shift anisotropy (CSA) tensor parameters and (15)N-(1)H dipolar splittings in powder protein samples. By using five single labeled samples of the villin headpiece subdomain protein in a hydrated lyophilized powder state, we determine the backbone (15)N CSA tensors at two temperatures, 22 and -35 °C, in order to get a snapshot of the variability across the residues and as a function of temperature. All sites probed belonged to the hydrophobic core and most of them were part of α-helical regions. The values of the anisotropy (which include the effect of the dynamics) varied between 130 and 156 ppm at 22 °C, while the values of the asymmetry were in the 0.32-0.082 range. The Leu-75 and Leu-61 backbone sites exhibited high mobility based on the values of their temperature-dependent anisotropy parameters. Under the assumption that most differences stem from dynamics, we obtained the values of the motional order parameters for the (15)N backbone sites. While a simple one-dimensional line shape experiment was used for the determination of the (15)N CSA parameters, a more advanced approach based on the "magic sandwich" SAMMY pulse sequence (Nevzorov and Opella, 2003) was employed for the determination of the (15)N-(1)H dipolar patterns, which yielded estimates of the dipolar couplings. Accordingly, the motional order parameters for the dipolar interaction were obtained. It was found that the order parameters from the CSA and dipolar measurements are highly correlated, validating that the variability between the residues is governed by the differences in dynamics. The values of the parameters obtained in this work can serve as reference values for developing more advanced magic-angle spinning recoupling techniques for multiple labeled samples. PMID:26367322

  17. Method for determining molar concentrations of metabolites in complex solutions from two-dimensional 1H-13C NMR spectra.

    PubMed

    Lewis, Ian A; Schommer, Seth C; Hodis, Brendan; Robb, Kate A; Tonelli, Marco; Westler, William M; Sussman, Michael R; Markley, John L

    2007-12-15

    One-dimensional (1D) (1)H nuclear magnetic resonance (NMR) spectroscopy is used extensively for high-throughput analysis of metabolites in biological fluids and tissue extracts. Typically, such spectra are treated as multivariate statistical objects rather than as collections of quantifiable metabolites. We report here a two-dimensional (2D) (1)H-(13)C NMR strategy (fast metabolite quantification, FMQ, by NMR) for identifying and quantifying the approximately 40 most abundant metabolites in biological samples. To validate this technique, we prepared mixtures of synthetic compounds and extracts from Arabidopsis thaliana, Saccharomyces cerevisiae, and Medicago sativa. We show that accurate (technical error 2.7%) molar concentrations can be determined in 12 min using our quantitative 2D (1)H-(13)C NMR strategy. In contrast, traditional 1D (1)H NMR analysis resulted in 16.2% technical error under nearly ideal conditions. We propose FMQ by NMR as a practical alternative to 1D (1)H NMR for metabolomics studies in which 50-mg (extract dry weight) samples can be obtained. PMID:17985927

  18. /sup 31/P NMR saturation-transfer and /sup 13/C NMR kinetic studies of glycolytic regulation during anaerobic and aerobic glycolysis

    SciTech Connect

    Campbell-Burk, S.L.; den Hollander, J.A.; Alger, J.R.; Shulman, R.G.

    1987-11-17

    /sup 31/P NMR saturation-transfer techniques have been employed in glucose-gown derepressed yeast to determine unidirectional fluxes in the upper part of the Embden-Meyerhof-Parnas pathway. The experiments were performed during anaerobic and aerobic glycolysis by saturating the ATP/sub ..gamma../ resonances and monitoring changes in the phosphomonoester signals from glucose 6-phosphate and fructose 1,6-bisphosphate. These experiments were supplemented with /sup 13/C NMR measurements of glucose utilization rates and /sup 13/C NMR label distribution studies. Combined with data obtained previously from radioisotope measurement, these /sup 31/P and /sup 13/C NMR kinetic studies allowed estimation of the net glycolytic flow in addition to relative flows through phosphofructokinase (PFK) and Fru-1,6-P/sub 2/ase during anaerobic and aerobic glycolysis. The /sup 31/P NMR saturation-transfer results are consistent with previous results obtained from measurements of metabolite levels, radioisotope data, and /sup 13/C NMR studies, providing additional support for in vivo measurement of the flows during glycolysis.

  19. Comparison of vertebral δ13C and δ15N records with organism-based isoscapes to identify fish migration, site fidelity and food-web preferences of fishes in the eastern Gulf of Mexico, USA

    NASA Astrophysics Data System (ADS)

    Radabaugh, K. R.; Wallace, A. A.; Huelster, S. A.; Hollander, D. J.; Peebles, E. B.

    2011-12-01

    Geographic variation in stable isotopic composition of dissolved and particulate nitrogen and carbon enables the use of stable isotopes as endogenous markers to track the origin and migration of motile marine species. Variation in river discharge, the light environment, and biological activity result in δ13C and δ15N spatial heterogeneity within coastal marine food webs. SEAMAP (Southeast Area Monitoring and Assessment Program) groundfish surveys at more than 130 locations were used to acquire samples from >1,600 samples of fish muscle and primary producers for bulk δ13C and δ15N analysis, allowing the creation of isoscapes for the West Florida Shelf (eastern Gulf of Mexico, USA). The δ15N isoscapes exhibited strong latitudinal and longitudinal isotopic gradients for all fish species examined, with high interannual and seasonal stability. δ15N was 3-4 % higher in the northwest region of the study compared to the southeast for both Syacium papillosum (dusky flounder) and Synodus foetens (inshore lizardfish). Low individual variability in δ15N values revealed strong site fidelities for these trawl-caught fishes. δ13C isoscapes exhibited depth gradients with greater seasonal and interannual variability. This study assesses the utility of combining isotopic analyses of fish vertebrae with organism-based isoscapes to investigate the migration routes of individual fishes. Unlike muscle isotopes that provide an integrated perspective of recent spatial and temporal environmental variation (at a scale dependent on muscle turnover rates), analysis of δ13C and δ15N along the growth radii of fish vertebrae enables chronological reconstruction of individual histories. When corrected for trophic level and metabolic fractionation, comparison of isotopic life history to the established isoscapes may enable reconstruction of migration routes and changing food-web positions of commercial and recreational fisheries species.

  20. {sup 13}C chemical shift anisotropies for carbonate ions in cement minerals and the use of {sup 13}C, {sup 27}Al and {sup 29}Si MAS NMR in studies of Portland cement including limestone additions

    SciTech Connect

    Sevelsted, Tine F.; Herfort, Duncan

    2013-10-15

    {sup 13}C isotropic chemical shifts and chemical shift anisotropy parameters have been determined for a number of inorganic carbonates relevant in cement chemistry from slow-speed {sup 13}C MAS or {sup 13}C({sup 1}H) CP/MAS NMR spectra (9.4 T or 14.1 T) for {sup 13}C in natural abundance. The variation in the {sup 13}C chemical shift parameters is relatively small, raising some doubts that different carbonate species in Portland cement-based materials may not be sufficiently resolved in {sup 13}C MAS NMR spectra. However, it is shown that by combining {sup 13}C MAS and {sup 13}C({sup 1}H) CP/MAS NMR carbonate anions in anhydrous and hydrated phases can be distinguished, thereby providing valuable information about the reactivity of limestone in cement blends. This is illustrated for three cement pastes prepared from an ordinary Portland cement, including 0, 16, and 25 wt.% limestone, and following the hydration for up to one year. For these blends {sup 29}Si MAS NMR reveals that the limestone filler accelerates the hydration for alite and also results in a smaller fraction of tetrahedrally coordinated Al incorporated in the C-S-H phase. The latter result is more clearly observed in {sup 27}Al MAS NMR spectra of the cement–limestone blends and suggests that dissolved aluminate species in the cement–limestone blends readily react with carbonate ions from the limestone filler, forming calcium monocarboaluminate hydrate. -- Highlights: •{sup 13}C chemical shift anisotropies for inorganic carbonates from {sup 13}C MAS NMR. •Narrow {sup 13}C NMR chemical shift range (163–171 ppm) for inorganic carbonates. •Anhydrous and hydrated carbonate species by {sup 13}C MAS and {sup 13}C({sup 1}H) CP/MAS NMR. •Limestone accelerates the hydration for alite in Portland – limestone cements. •Limestone reduces the amount of aluminium incorporated in the C-S-H phase.

  1. DFT study of molecular structures and 13C NMR parameters of two fluorinated biphenyls and their η6-tricarbonylchromium complexes

    NASA Astrophysics Data System (ADS)

    Gryff-Keller, Adam; Szczeciński, Przemysław

    2015-07-01

    The molecular structures of 2,2‧-difluoro-6,6‧-dimethylbiphenyl, 4,5-difluoro-9,10-dihydrophenanthrene and of their η6-tricarbonylchromium complexes have been discussed in the light of the results of molecular energy calculations. Also the isotropic magnetic shielding constants and carbon-fluorine spin-spin coupling constants for these objects have been calculated and compared with the experimental values of 13C NMR chemical shifts and J constants. The calculational methods used were: DFT/BHandH/6-311++G(2d,p) and/or DFT/B3LYP/6-311++G(2d,p). It has been confirmed that experimental 13C NMR chemical shifts for η6-arene tricarbonylchromium complexes can be satisfactorily predicted using both methods, although the method exploiting BHandH functional is not able to reproduce the 13C NMR chemical shifts of Cr(CO)3 carbon atoms. On the other hand, this method provides the J(13C, 19F) values which are close to the experimental ones.

  2. 13C NMR of methane in an AlPO4-11 molecular sieve: Exchange effects and shielding anisotropy

    NASA Astrophysics Data System (ADS)

    Koskela, Tuomas; Ylihautala, Mika; Jokisaari, Jukka; Vaara, Juha

    1998-12-01

    13C NMR spectra of 13CH4 in an AlPO4-11 molecular sieve reveal exchange effects between adsorbed and nonadsorbed methane gas. An application of pulsed field gradients is introduced to decrease nonadsorbed and exchanging gas signals in order to extract the chemical shift anisotropy line shape of the adsorbed gas. The resulting 13C shielding anisotropy of methane is compared to existing value for methane in related SAPO-11 material. Less anisotropic shielding is observed in AlPO4-11, most likely due to the lack of charge-compensating cations.

  3. A facile method for expression and purification of (15)N isotope-labeled human Alzheimer's β-amyloid peptides from E. coli for NMR-based structural analysis.

    PubMed

    Sharma, Sudhir C; Armand, Tara; Ball, K Aurelia; Chen, Anna; Pelton, Jeffrey G; Wemmer, David E; Head-Gordon, Teresa

    2015-12-01

    Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting millions of people worldwide. AD is characterized by the presence of extracellular plaques composed of aggregated/oligomerized β-amyloid peptides with Aβ42 peptide representing a major isoform in the senile plaques. Given the pathological significance of Aβ42 in the progression of AD, there is considerable interest in understanding the structural ensembles for soluble monomer and oligomeric forms of Aβ42. This report describes an efficient method to express and purify high quality (15)N isotope-labeled Aβ42 for structural studies by NMR. The protocol involves utilization of an auto induction system with (15)N isotope labeled medium, for high-level expression of Aβ42 as a fusion with IFABP. After the over-expression of the (15)N isotope-labeled IFABP-Aβ42 fusion protein in the inclusion bodies, pure (15)N isotope-labeled Aβ42 peptide is obtained following a purification method that is streamlined and improved from the method originally developed for the isolation of unlabeled Aβ42 peptide (Garai et al., 2009). We obtain a final yield of ∼ 6 mg/L culture for (15)N isotope-labeled Aβ42 peptide. Mass spectrometry and (1)H-(15)N HSQC spectra of monomeric Aβ42 peptide validate the uniform incorporation of the isotopic label. The method described here is equally applicable for the uniform isotope labeling with (15)N and (13)C in Aβ42 peptide as well as its other variants including any Aβ42 peptide mutants. PMID:26231074

  4. Low-temperature solid-state /sup 13/C NMR studies of the retinal chromophore in rhodopsin

    SciTech Connect

    Smith, S.O.; Palings, I.; Copie, V.; Raleigh, D.P.; Courtin, J.; Pardoen, J.A.; Lugtenburg, J.; Mathies, R.A.; Griffin, R.G.

    1987-03-24

    Magic angle sample spinning (MASS) /sup 13/C NMR spectra have been obtained of bovine rhodopsin regenerated with retinal prosthetic groups isotopically enriched with /sup 13/C at C-5 and C-14. In order to observe the /sup 13/C retinal chromophore resonances, it was necessary to employ low temperatures (-15 ..-->.. -35/sup 0/C) to restrict rotational diffusion of the protein. The isotropic chemical shift and principal values of the chemical shift tensor of the /sup 13/C-5 label indicate that the retinal chromophore is in the twisted 6-s-cis conformation in rhodopsin, in contrast to the planar 6-s-trans confirmation found in bacteriorhodopsin. The /sup 13/C-14 isotropic shift and shift tensor principal values show that the Schiff base C=N bond is anti. Furthermore, the /sup 13/C-14 chemical shift (121.2 ppm) is within the range of values (120-123 ppm) exhibited by protonated (C=N anti) Schiff base model compounds, indicating that the C=N linkage is protonated. The results are discussed with regard to the mechanism of wavelength regulation in rhodopsin.

  5. Complexation of oxygen ligands with dimeric rhodium(II) tetrakistrifluoroacetate in chloroform: 1H, 13C NMR and DFT studies

    NASA Astrophysics Data System (ADS)

    Głaszczka, Rafał; Jaźwiński, Jarosław

    2013-03-01

    The complexation of dimeric rhodium(II) tetrakistrifluoroacetylate with 25 ligands containing oxygen atoms: alcohols, ethers, ketones, aldehydes, carboxylic acids and esters in chloroform solution have been investigated by 1H and 13C NMR spectroscopy and Density Functional Theory (DFT) methods. Investigated ligands form 1:1 adducts in our experimental conditions, with stability constants in the order of several hundred mol-1. The exchange of ligands in solution is fast on the NMR spectroscopic timescale. The decrease of longitudinal relaxation times T1 in ligands in the presence of rhodium salt has been tested as the means of determination of the complexation site in ligands. The influence of complexation on chemical shifts in ligands was evaluated by a parameter complexation shift Δδ (Δδ = δadd - δlig). These parameters were positive (>0 ppm) and did not exceed 1 ppm for 1H NMR; and varied from ca. -5 to +15 ppm in the case of 13C NMR. The calculation by DFT methods using the B3LYP functional (structure optimization, electronic energy) and B3PW91 functional (shielding), and combinations of the (6-31G(2d), 6-311G++(2d,p), and LANL2DZ basis sets, followed by scaling procedures reproduced satisfactorily 1H and 13C chemical shifts and, with some limitations, allowed to estimate Δδ parameters.

  6. Use of 13C and 15N mass spectrometry to study the decomposition of Calamagrostis epigeios in soil column experiments with and without ash additions.

    PubMed

    Ludwig, B; Heil, B; Flessa, H; Beese, F

    2000-01-01

    The dynamics of C and N in terrestrial ecosystems are not completely understood and the use of stable isotopes may be useful to gain further insight in the pathways of CO2 emissions and leaching of dissolved organic carbon (DOC) and nitrogen (DON) during decomposition of litter. Objectives were (i) to study the decomposition dynamics of Calamagrostis epigeios, a common grass species in forests, using 13C-depleted and 15N-enriched plants and (ii) to quantify the effect wood ash addition on the decomposition and leaching of DOC and DON. Decomposition was studied for 128 days under aerobic conditions at 8 degrees C and moisture close to field capacity in a spodic dystric Cambisol with mor-moder layer. Variants included control plots and additions of (i) Calamagrostis litter and (ii) Calamagrostis litter plus 4 kg ash m-2. (i) Decomposition of Calamagrostis resulted in a CO2 production of 76.2 g CO2-C m-2 (10% of added C) after 128 days and cumulative DOC production was 14.0 g C m-2 out of which 0.9 g C m-2 was Calamagrostis-derived (0.1% of added C). The specific CO2 formation and specific DOC production from Calamagrostis were 6 times higher (CO2) and 4 times smaller (DOC) than those from the organic layer. The amount of Calamagrostis-derived total N (NH4+, NO3-, DON) leached was 0.7 g N m-2 (4.8% of added N). Cumulative DON production was 0.8 g N m-2 which was slightly higher than for the control. During soil passage, much of the DOC and DON was removed due to sorption or decomposition. DOC and DON releases from the mineral soil (17 cm depth) were 6.3 g C m-2 and 0.5 g N m-2. (ii) Addition of ash resulted in a complete fixing of CO2 for 40 days due to carbonatisation. Afterwards, the CO2 production rates were similar to the variant without ash addition. Production of DOC (98.6 g C m-2) and DON (2.5 g N m-2) was marked, mainly owing to humus decay. However, Calamagrostis-derived DOC and Calamagrostis-derived total N were only 3.9 g C m-2 (0.5% of added C) and 0.5 g N

  7. Properties of bridgehead-substituted polycycloalkanes. Synthesis and NMR analysis of /sup 15/N-labeled 1-aminobicycloalkanes and their hydrochlorides

    SciTech Connect

    Della, E.W.; Kasum, B.; Kirkbride, K.P.

    1987-04-29

    NMR analysis of adamantane and four bicycloalkanes substituted at the bridgehead with /sup 15/N-labeled amino and ammonio groups is described. It is found that where most of the one-bond carbon-nitrogen coupling constants are relatively large, those in 1-aminobicyclo (1.1.1)pentane and its hydrochloride are significantly reduced; in fact, in the latter compound one-bond /sup 13/C-/sup 15/N coupling could not even be detected. Values of experimentally determined vicinal couplings were in accord with those expected on the basis of the number of three-bond pathways available for transmission of spin-spin information; INDO calculations, however, suggest that in the more highly strained systems there is a substantial contribution to /sup 3/J(CN) arising from through-space interactions and that these oppose through-bond effects. Large four-bond /sup 15/N-/sup 1/H couplings were found to occur in 1-aminobicyclo(1.1.1)pentane and its hydrochloride; MO calculations indicate that through-space interactions constitute the predominant mechanism contributing to /sup 4/J(/sup 15/N-/sup 1/H), although in this case through-bond and through-space effects reinforce each other. The nitrogen-15 chemical shifts of the amine hydrochlorides were determined, and they appear to occur in random fashion.

  8. Covalent binding of aniline to humic substances. 2. 15N NMR studies of nucleophilic addition reactions

    USGS Publications Warehouse

    Thorn, K.A.; Pettigrew, P.J.; Goldenberg, W.S.; Weber, E.J.

    1996-01-01

    Aromatic amines are known to undergo covalent binding with humic substances in the environment. Although previous studies have examined reaction conditions and proposed mechanisms, there has been no direct spectroscopic evidence for the covalent binding of the amines to the functional groups in humic substances. In order to further elucidate the reaction mechanisms, the Suwannee River and IHSS soil fulvic and humic acids were reacted with 15N-labeled aniline at pH 6 and analyzed using 15N NMR spectrometry. Aniline underwent nucleophilic addition reactions with the quinone and other carbonyl groups in the samples and became incorporated in the form of anilinohydroquinone, anilinoquinone, anilide, imine, and heterocyclic nitrogen, the latter comprising 50% or more of the bound amine. The anilide and anilinohydroquinone nitrogens were determined to be susceptible to chemical exchange by ammonia. In the case of Suwannee River fulvic acid, reaction under anoxic conditions and pretreatment with sodium borohydride or hydroxylamine prior to reaction under oxic conditions resulted in a decrease in the proportion of anilinohydroquinone nitrogen incorporated. The relative decrease in the incorporation of anilinohydroquinone nitrogen with respect to anilinoquinone nitrogen under anoxic conditions suggested that inter- or intramolecular redox reactions accompanied the nucleophilic addition reactions.

  9. Thermal maturity of type II kerogen from the New Albany Shale assessed by13C CP/MAS NMR

    USGS Publications Warehouse

    Werner-Zwanziger, U.; Lis, G.; Mastalerz, Maria; Schimmelmann, A.

    2005-01-01

    Thermal maturity of oil and gas source rocks is typically quantified in terms of vitrinite reflectance, which is based on optical properties of terrestrial woody remains. This study evaluates 13C CP/MAS NMR parameters in kerogen (i.e., the insoluble fraction of organic matter in sediments and sedimentary rocks) as proxies for thermal maturity in marine-derived source rocks where terrestrially derived vitrinite is often absent or sparse. In a suite of samples from the New Albany Shale (Middle Devonian to the Early Mississippian, Illinois Basin) the abundance of aromatic carbon in kerogen determined by 13C CP/MAS NMR correlates linearly well with vitrinite reflectance. ?? 2004 Elsevier Inc. All rights reserved.

  10. Large structure rearrangement of colicin ia channel domain after membrane binding from 2D 13C spin diffusion NMR.

    PubMed

    Luo, Wenbin; Yao, Xiaolan; Hong, Mei

    2005-05-01

    One of the main mechanisms of membrane protein folding is by spontaneous insertion into the lipid bilayer from the aqueous environment. The bacterial toxin, colicin Ia, is one such protein. To shed light on the conformational changes involved in this dramatic transfer from the polar to the hydrophobic milieu, we carried out 2D magic-angle spinning (13)C NMR experiments on the water-soluble and membrane-bound states of the channel-forming domain of colicin Ia. Proton-driven (13)C spin diffusion spectra of selectively (13)C-labeled protein show unequivocal attenuation of cross-peaks after membrane binding. This attenuation can be assigned to distance increases but not reduction of the diffusion coefficient. Analysis of the statistics of the interhelical and intrahelical (13)C-(13)C distances in the soluble protein structure indicates that the observed cross-peak reduction is well correlated with a high percentage of short interhelical contacts in the soluble protein. This suggests that colicin Ia channel domain becomes open and extended upon membrane binding, thus lengthening interhelical distances. In comparison, cross-peaks with similar intensities between the two states are dominated by intrahelical contacts in the soluble state. This suggests that the membrane-bound structure of colicin Ia channel domain may be described as a "molten globule", in which the helical secondary structure is retained while the tertiary structure is unfolded. This study demonstrates that (13)C spin diffusion NMR is a valuable tool for obtaining qualitative long-range distance constraints on membrane protein folding. PMID:15853348

  11. Optimized Spectral Editing of 13C MAS NMR Spectra of Rigid Solids Using Cross-Polarization Methods

    NASA Astrophysics Data System (ADS)

    Sangill, R.; Rastrupandersen, N.; Bildsoe, H.; Jakobsen, H. J.; Nielsen, N. C.

    Combinations of 13C magic-angle spinning (MAS) NMR experiments employing cross polarization (CP), cross polarization-depolarization (CPD), and cross polarization-depolarization-repolarization are analyzed quantitatively to derive simple and general procedures for optimized spectral editing of 13C CP/MAS NMR spectra of rigid solids by separation of the 13C resonances into CH n subspectra ( n = 0, 1, 2, and 3). Special attention is devoted to a differentiation by CPD/MAS of CH and CH 2 resonances since these groups behave quite similarly during spin lock under Hartmann-Hahn match and are therefore generally difficult to distinguish unambiguously. A general procedure for the design of subexperiments and linear combinations of their spectra to provide optimized signal-to-noise ratios for the edited subspectra is described. The technique is illustrated by a series of edited 13C CP/MAS spectra for a number of rigid solids ranging from simple organic compounds (sucrose and l-menthol) to complex pharmaceutical products (calcipotriol monohydrate and vitamin D 3) and polymers (polypropylene, polyvinyl alcohol, polyvinyl chloride, and polystyrene).

  12. High-field 13C NMR spectroscopy of tissue in Vivo. A double-resonance surface-coil probe

    NASA Astrophysics Data System (ADS)

    Reo, Nicholas V.; Ewy, Coleen S.; Siegfried, Barry A.; Ackerman, Joseph J. H.

    A double-resonance surface-coil NMR probe is described for performance of high-field (8.5 T) proton decoupled carbon-13 experiments with tissue in vivo. The probe may be accommodated in standard, 89 mm i.d. clear bore, commercial spectrometers and is suitable for studies utilizing small laboratory animals such as mice, hamsters, and rats. A coaxial coil design is employed (10 mm diameter 13C coil, 20 mm diameter 1H coil) which provides ca. 40 dB attenuation between the 13C observe and 1H decouple channels. The inherent efficiency of the surface-coil configuration provides a sensitivity comparable to a commercial probe of the same nominal dimension (10 mm Helmholtz coil) and assures adequate decoupling in conductive samples with ca. 3-5 W power. In the absence of 13C isotopic enrichment, NMR spectra of rat leg, liver, and brain in vivo provide signalto-noise sufficient for 10 min time resolution. Administration of 100 mg of 90% 13C-labeled glucose into a peripheral vein of a ca. 300 g rat resulted in a liver glucose resonance which could be monitored with good signal-to-noise and 3 min time resolution.

  13. Structure and dynamics of homoleptic beryllocenes: a solid-state 9Be and 13C NMR study.

    PubMed

    Hung, Ivan; Macdonald, Charles L B; Schurko, Robert W

    2004-11-19

    The correlation between anisotropic 9Be NMR (quadrupolar and chemical shielding) interactions and the structure and dynamics in [Cp2Be], [Cp2*Be], and [(C5Me4H)2Be] is examined by solid-state 9Be NMR spectroscopy, as well as by ab initio and hybrid density functional theory calculations. The 9Be quadrupole coupling constants in the three compounds correspond well to the relative degrees of spherical ground-state electronic symmetry of the environment about beryllium. Theoretical computations of NMR interaction tensors are in excellent agreement with experimental values and aid in understanding the origins of NMR interaction tensors and their correlation to molecular symmetry. Variable-temperature (VT) 9Be and 13C NMR experiments reveal a highly fluxional structure in the condensed phase of [Cp2Be]. In particular, the pathway by which the Cp rings of [Cp2Be] 'invert' coordination modes is examined in detail using hybrid density functional theory in order to inspect variations of the 9Be NMR interaction tensors. The activation energy for the 'inversion' process is found to be 36.9 kJ mol(-1) from chemical exchange analysis of 13C VT CP/MAS NMR spectra. The low-temperature (ca. -100 degrees C) X-ray crystal structures of all three compounds have been collected and refined, and are in agreement with previously reported structures. In addition, the structure of the same Cp2Be crystal was determined at 20 degrees C and displays features consistent with increased intramolecular motion, supporting observations by 9Be VT NMR spectroscopy. PMID:15484199

  14. 1H, 13C MAS NMR and GIAO-CPHF calculations of chloramphenicol, thiamphenicol and their pyrrole analogues

    NASA Astrophysics Data System (ADS)

    Żołek, Teresa; Paradowska, Katarzyna; Krajewska, Dorota; Różański, Andrzej; Wawer, Iwona

    2003-02-01

    The 13C CP MAS and 1H MAS NMR and ab initio (GIAO-CPHF) calculations were used to obtain structural information on two known antibiotics: chloramphenicol, and thiamphenicol, and two new analogues: DL- threo-1-(1-methyl-4-nitro-pyrrole-2-yl)-2-dichloroacetamidopropane-1,3-diol and DL- threo-1-(1-methylsulfonylpyrrole-3-yl)-2-dichloroacetamidopropane-1,3-diol.

  15. Molecular structure of actein: 13C CPMAS NMR, IR, X-ray diffraction studies and theoretical DFT-GIAO calculations

    NASA Astrophysics Data System (ADS)

    Jamróz, Marta K.; Bąk, Joanna; Gliński, Jan A.; Koczorowska, Agnieszka; Wawer, Iwona

    2009-09-01

    Actein is a prominent triterpene glycoside occurring in Actaea racemosa. The triterpene glycosides are believed to be responsible for the estrogenic activity of an extract prepared from this herb. We determined in the crystal structure of actein by X-ray crystallography to be monoclinic P2(1) chiral space group. Refining the disorder, we determined 70% and 30% of contributions of ( S)- and ( R)-actein, respectively. The IR and Raman spectra suggest that actein forms at least four different types of hydrogen bonds. The 13C NMR spectra of actein were recorded both in solution and solid state. The 13C CPMAS spectrum of actein displays multiplet signals, in agreement with the crystallographic data. The NMR shielding constants were calculated for actein using GIAO approach and a variety of basis sets: 6-31G**, 6-311G**, 6-31+G**, cc-pVDZ, cc-pVDZ-su1 and 6-31G**-su1, as well as IGLO approach combined with the IGLO II basis set. The best results (RMSD of 1.6 ppm and maximum error of 3.4 ppm) were obtained with the 6-31G**-su1 basis set. The calculations of the shielding constants are helpful in the interpretation of the 13C CPMAS NMR spectra of actein and actein's analogues.

  16. Unified and isomer-specific NMR metabolomics database for the accurate analysis of (13)C-(1)H HSQC spectra.

    PubMed

    Bingol, Kerem; Li, Da-Wei; Bruschweiler-Li, Lei; Cabrera, Oscar A; Megraw, Timothy; Zhang, Fengli; Brüschweiler, Rafael

    2015-02-20

    A new metabolomics database and query algorithm for the analysis of (13)C-(1)H HSQC spectra is introduced, which unifies NMR spectroscopic information on 555 metabolites from both the Biological Magnetic Resonance Data Bank (BMRB) and Human Metabolome Database (HMDB). The new database, termed Complex Mixture Analysis by NMR (COLMAR) (13)C-(1)H HSQC database, can be queried via an interactive, easy to use web interface at http://spin.ccic.ohio-state.edu/index.php/hsqc/index . Our new HSQC database separately treats slowly exchanging isomers that belong to the same metabolite, which permits improved query in cases where lowly populated isomers are below the HSQC detection limit. The performance of our new database and query web server compares favorably with the one of existing web servers, especially for spectra of samples of high complexity, including metabolite mixtures from the model organisms Drosophila melanogaster and Escherichia coli. For such samples, our web server has on average a 37% higher accuracy (true positive rate) and a 82% lower false positive rate, which makes it a useful tool for the rapid and accurate identification of metabolites from (13)C-(1)H HSQC spectra at natural abundance. This information can be combined and validated with NMR data from 2D TOCSY-type spectra that provide connectivity information not present in HSQC spectra. PMID:25333826

  17. Diet-tissue discrimination factors (Δ(13) C and Δ(15) N) and turnover rate in somatic tissues of a neotropical detritivorous fish on C3 and C4 diets.

    PubMed

    Sacramento, P A; Manetta, G I; Benedito, E

    2016-07-01

    In this study, diet-tissue discrimination factors and turnover rates were determined from the somatic tissues of a detritivorous fish Prochilodus lineatus. The carbon (Δ(13) C) and nitrogen (Δ(15) N) diet-tissue discrimination factors varied for all feed rations with a range of Δ(13) C values between -1·9 and 3·6‰ and Δ(15) N between 3·3 and 5·7‰. Carbon turnover rate in the blood was 23·1 days for the C3 ration and 34·7 days for the C4 ration, in the liver was 9·9 days under the C3 ration and nitrogen turnover rate was the same (23·1 days) in the liver for both C4 and C3 -C4 rations, and 13·9 days in the muscle for C3 -C4 ration. PMID:26728338

  18. A 13C{31P} REDOR NMR Investigation of the Role of Glutamic Acid Residues in Statherin-Hydroxyapatite Recognition

    PubMed Central

    Ndao, Moise; Ash, Jason T.; Breen, Nicholas F.; Goobes, Gil; Stayton, Patrick S.; Drobny, Gary P.

    2011-01-01

    The side chain carboxyl groups of acidic proteins found in the extra-cellular matrix (ECM) of mineralized tissues play a key role in promoting or inhibiting the growth of minerals such as hydroxyapatite (HAP), the principal mineral component of bone and teeth. Among the acidic proteins found in the saliva is statherin, a 43-residue tyrosine-rich peptide that is a potent lubricant in the salivary pellicle and an inhibitor of both HAP crystal nucleation and growth. Three acidic amino acids – D1, E4, and E5 – are located in the N-terminal 15 amino acid segment, with a fourth amino acid, E26, located outside the N-terminus. We have utilized 13C{31P} REDOR NMR to analyze the role played by acidic amino acids in the binding mechanism of statherin to the HAP surface by measuring the distance between the δ-carboxyl 13C spins of the three glutamic acid side chains of statherin (residues E4, E5, E26) and 31P spins of the phosphate groups at the HAP surface. 13C{31P} REDOR studies of glutamic-5-13C acid incorporated at positions E4 and E26 indicate a 13C–31P distance of more than 6.5 Å between the side chain carboxyl 13C spin of E4 and the closest 31P in the HAP surface. In contrast, the carboxyl 13C spin at E5 has a much shorter 13C–31P internuclear distance of 4.25±0.09 Å, indicating that the carboxyl group of this side chain interacts directly with the surface. 13C T1ρ and slow-spinning MAS studies indicate that the motions of the side chains of E4 and E5 are more restricted than that of E26. Together, these results provide further insight into the molecular interactions of statherin with HAP surfaces. PMID:19678690

  19. (1)H, (13)C and (15)N backbone and side-chain resonance assignment of the LAM-RRM1 N-terminal module of La protein from Dictyostelium discoideum.

    PubMed

    Chasapis, Christos T; Argyriou, Aikaterini I; Apostolidi, Maria; Konstantinidou, Parthena; Bentrop, Detlef; Stathopoulos, Constantinos; Spyroulias, Georgios A

    2015-10-01

    The N-terminal half of La protein consists of two concatenated motifs: La motif (LAM) and the N-terminal RNA recognition motif (RRM1) both of which are responsible for poly(U) RNA binding. Here, we present the backbone and side-chain assignments of the (1)H, (13)C and (15)N resonances of the 191-residue LAM-RRM1 region of the La protein from the lower eukaryote Dictyostelium discoideum and its secondary structure prediction. PMID:25687647

  20. Changing gull diet in a changing world: a 150-year stable isotope (δ13C, δ15N) record from feathers collected in the Pacific Northwest of North America.

    PubMed

    Blight, Louise K; Hobson, Keith A; Kyser, T Kurt; Arcese, Peter

    2015-04-01

    The world's oceans have undergone significant ecological changes following European colonial expansion and associated industrialization. Seabirds are useful indicators of marine food web structure and can be used to track multidecadal environmental change, potentially reflecting long-term human impacts. We used stable isotope (δ(13)C, δ(15)N) analysis of feathers from glaucous-winged gulls (Larus glaucescens) in a heavily disturbed region of the northeast Pacific to ask whether diets of this generalist forager changed in response to shifts in food availability over 150 years, and whether any detected change might explain long-term trends in gull abundance. Sampled feathers came from birds collected between 1860 and 2009 at nesting colonies in the Salish Sea, a transboundary marine system adjacent to Washington, USA and British Columbia, Canada. To determine whether temporal trends in stable isotope ratios might simply reflect changes to baseline environmental values, we also analysed muscle tissue from forage fishes collected in the same region over a multidecadal timeframe. Values of δ(13)C and δ(15)N declined since 1860 in both subadult and adult gulls (δ(13)C, ~ 2-6‰; δ(15)N, ~4-5‰), indicating that their diet has become less marine over time, and that birds now feed at a lower trophic level than previously. Conversely, forage fish δ(13)C and δ(15)N values showed no trends, supporting our conclusion that gull feather values were indicative of declines in marine food availability rather than of baseline environmental change. Gradual declines in feather isotope values are consistent with trends predicted had gulls consumed less fish over time, but were equivocal with respect to whether gulls had switched to a more garbage-based diet, or one comprising marine invertebrates. Nevertheless, our results suggest a long-term decrease in diet quality linked to declining fish abundance or other anthropogenic influences, and may help to explain regional

  1. Compositional differences among Chinese soy sauce types studied by (13)C NMR spectroscopy coupled with multivariate statistical analysis.

    PubMed

    Kamal, Ghulam Mustafa; Wang, Xiaohua; Bin Yuan; Wang, Jie; Sun, Peng; Zhang, Xu; Liu, Maili

    2016-09-01

    Soy sauce a well known seasoning all over the world, especially in Asia, is available in global market in a wide range of types based on its purpose and the processing methods. Its composition varies with respect to the fermentation processes and addition of additives, preservatives and flavor enhancers. A comprehensive (1)H NMR based study regarding the metabonomic variations of soy sauce to differentiate among different types of soy sauce available on the global market has been limited due to the complexity of the mixture. In present study, (13)C NMR spectroscopy coupled with multivariate statistical data analysis like principle component analysis (PCA), and orthogonal partial least square-discriminant analysis (OPLS-DA) was applied to investigate metabonomic variations among different types of soy sauce, namely super light, super dark, red cooking and mushroom soy sauce. The main additives in soy sauce like glutamate, sucrose and glucose were easily distinguished and quantified using (13)C NMR spectroscopy which were otherwise difficult to be assigned and quantified due to serious signal overlaps in (1)H NMR spectra. The significantly higher concentration of sucrose in dark, red cooking and mushroom flavored soy sauce can directly be linked to the addition of caramel in soy sauce. Similarly, significantly higher level of glutamate in super light as compared to super dark and mushroom flavored soy sauce may come from the addition of monosodium glutamate. The study highlights the potentiality of (13)C NMR based metabonomics coupled with multivariate statistical data analysis in differentiating between the types of soy sauce on the basis of level of additives, raw materials and fermentation procedures. PMID:27343582

  2. 13C NMR study of the generation of C2- and C3-deuterated lactic acid by tumoral pancreatic islet cells exposed to D-[1-13C]-, D-[2-13C]- and D-[6-13C]-glucose in 2H2O.

    PubMed

    Willem, R; Biesemans, M; Kayser, F; Malaisse, W J

    1994-03-01

    Tumoral pancreatic islet cells of the RIN5mF line were incubated for 120 min in media prepared in 2H2O and containing D-[1-13C]glucose, D-[2-13C]glucose, and D-[6-13C]glucose. The generation of C2- and C3-deuterated lactic acid was assessed by 13C NMR. The interpretation of experimental results suggests that a) the efficiency of deuteration on the C1 of D-fructose 6-phosphate does not exceed about 47% and 4% in the phosphoglucoisomerase and phosphomannoisomerase reactions, respectively; b) approximately 38% of the molecules of D-glyceraldehyde 3-phosphate generated from D-glucose escape deuteration in the sequence of reactions catalyzed by triose phosphate isomerase and aldolase; and c) about 41% of the molecules of pyruvate generated by glycolysis are immediately converted to lactate, the remaining 59% of pyruvate molecules undergoing first a single or double back-and-forth interconversion with L-alanine. It is proposed that this methodological approach, based on high resolution 13C NMR spectroscopy, may provide novel information on the regulation of back-and-forth interconversion of glycolytic intermediates in intact cells as modulated, for instance, by enzyme-to-enzyme tunneling. PMID:8057796

  3. Combination of the (87)Sr/(86)Sr ratio and light stable isotopic values (δ(13)C, δ(15)N and δD) for identifying the geographical origin of winter wheat in China.

    PubMed

    Liu, Hongyan; Wei, Yimin; Lu, Hai; Wei, Shuai; Jiang, Tao; Zhang, Yingquan; Guo, Boli

    2016-12-01

    This study aims to investigate whether isotopic signatures can be used to develop reliable fingerprints for discriminating the geographical origin of Chinese winter wheat, and to evaluate the discrimination effects of δ(13)C, δ(15)N and δD, alone or with (87)Sr/(86)Sr. In this study, the values of δ(13)C, δ(15)N and δD, and the (87)Sr/(86)Sr ratios of wheat and provenance soils from three regions were determined. Significant differences were found in all parameters of wheat and (87)Sr/(86)Sr in soil extract (reflecting the bioavailable fraction of soil) among different regions. A significantly positive correlation was observed between the (87)Sr/(86)Sr ratios of wheat and soil extracts. An overall correct classification rate of 77.8% was obtained for discriminating wheat from three regions based on light stable isotopes (δ(13)C, δ(15)N, and δD). The correct classification rate of 98.1% could be obtained with the combination of the (87)Sr/(86)Sr ratio and the light stable isotopic values. PMID:27374544

  4. Detailed 1H and 13C NMR structural assignment and relative stereochemistry determination for three new and one known semi-synthetic sesquiterpene lactones

    NASA Astrophysics Data System (ADS)

    Sass, Daiane Cristina; Heleno, Vladimir Constantino Gomes; Soares, Ana Carolina Ferreira; Lopes, João Luis Callegari; Constantino, Mauricio Gomes

    2012-01-01

    In this work is described a complete 1H and 13C NMR analysis for a group of four sesquiterpene lactones, three previously unknown. The unequivocal assignments were achieved by 1H NMR, 13C{ 1H} NMR, J-resolved, gCOSY, gHMQC, gHMBC and NOESY experiments and no ambiguities were left behind. All hydrogen coupling constants were measured, clarifying all hydrogen signals multiplicities.

  5. Solid-state 13C NMR analysis of Lower Cretaceous Baganuur (Mongolia) lignite

    NASA Astrophysics Data System (ADS)

    Erdenetsogt, B.; Lee, I.; Lee, S.; Ko, Y.

    2009-12-01

    The transformation of plant matter into peat and coal has two steps, called the biochemical and geochemical stages of coalification. Biochemical coalification begins with the accumulation of dead vegetable matter and ends at the rank of subbituminous coal. The rank of Baganuur lignite ranges from lignite to subbituminous coal. It is transition between biochemical and physico-chemical coalification stages. The changes of chemical structure of coal during the transition between above mentioned two stages were studied by solid state CP/MAS 13C NMR. The most predominant alteration is the disappearance of the resonances from oxygenated aliphatic carbons (63 ppm), protonated aromatic carbons (114 ppm), oxygen-substituted aromatic carbons (144 ppm) and carbonyl carbons (195 ppm). In addition, the intensity of resonances from methoxyl carbons (56 ppm) and oxygenated aliphatic carbons (72 ppm) decreased. While the intensities of resonance from aliphatic (30 ppm), protonated aromatic (125 ppm) and carboxyl carbon (174 ppm) increased or remained almost constant. The relative percent of O-substituted aromatic carbons decreased by ~25% mainly due to the intensity loss of the peak at 144 ppm, indicating removal of O-containing functional groups substituted to aromatic carbons. It is consistent with the decreased relative percent (~75%) of the peak at 114 ppm from protonated aromatic carbons nearby oxygen-substituted aromatic carbons. In addition, the resonance from 125 ppm was shifted to 128 ppm and its relative area increased by ~20%, indicating replacement of O-substituent of aromatic rings by hydrogen or carbon. Protonated aromatic carbons at least two bond away from an oxygen-substituted aromatic carbons give a resonance at 125 ppm and carbon-substituted aromatic carbons give a resonance at 130-132 ppm. With the increase relative percent of C-substituted aromatic carbons, their resonance were overlapped with protonated aromatic carbons and shifted to higher ppm. A decreasing

  6. Photochemically Induced Dynamic Nuclear Polarization Observed by Solid-State NMR in a Uniformly (13)C-Isotope-Labeled Photosynthetic Reaction Center.

    PubMed

    Paul, Shubhajit; Bode, Bela E; Matysik, Jörg; Alia, A

    2015-10-29

    A sample of solubilized and quinone-depleted reaction centers from the purple bacterium Rhodobacter (R.) sphaeroides wild type has been prepared entirely (13)C and (15)N isotope labeled at all positions of the protein as well as of the cofactors. In this sample, the occurrence of the solid-state photo-CIDNP (photochemically induced dynamic nuclear polarization) effect has been probed by (13)C solid-state magic-angle spinning NMR under illumination. Under continuous illumination, signal intensities are modified by the three-spin mixing (TSM) mechanism. Time-resolved illumination experiments reveal the occurrence of light-induced nuclear polarization on the time scale of hundreds of microseconds, initially dominated by the transient polarization of the singlet branch of the radical-pair mechanism. A first kinetic analysis shows that the lifetime of the polarization from the singlet branch, indicated by the enhanced absorptive intensities of the signals from aliphatic carbons, is significantly extended. Upon arrival of the polarization from the triplet decay branch, emissive polarization caused by the TSM mechanism is observed. Also, this arrival is significantly delayed. The decay of TSM polarization occurs in two steps, assigned to intra- and intermolecular spin diffusion. PMID:26110356

  7. 13C/15N‐Enriched l‐Dopa as a Triple‐Resonance NMR Probe to Monitor Neurotransmitter Dopamine in the Brain and Liver Extracts of Mice

    PubMed Central

    Kameda, Tetsuro; Kimura, Yu; Imai, Hirohiko; Matsuda, Tetsuya; Sando, Shinsuke; Toshimitsu, Akio

    2015-01-01

    Abstract In an attempt to monitor μm‐level trace constituents, we applied here 1H‐{13C‐15N} triple‐resonance nuclear magnetic resonance (NMR) to 13C/15N‐enriched l‐Dopa as the inevitable precursor of the neurotransmitter dopamine in the brain. The perfect selectivity (to render endogenous components silent) and μm‐level sensitivity (700 MHz spectrometer equipped with a cryogenic probe) of triple‐resonance allowed the unambiguous and quantitative metabolic and pharmacokinetic analyses of administered l‐Dopa/dopamine in the brain and liver of mice. The level of dopamine generated in the brain (within the range 7–76 μm, which covers the typical stimulated level of ∼30 μm) could be clearly monitored ex vivo, but was slightly short of the detection limit of a 7 T MR machine for small animals. This work suggests that μm‐level trace constituents are potential targets of ex vivo monitoring as long as they contain N atom(s) and their appropriate 13C/15N‐enrichment is synthetically accessible. PMID:27308224

  8. Area per Lipid and Cholesterol Interactions in Membranes from Separated Local-Field 13C NMR Spectroscopy

    PubMed Central

    Leftin, Avigdor; Molugu, Trivikram R.; Job, Constantin; Beyer, Klaus; Brown, Michael F.

    2014-01-01

    Investigations of lipid membranes using NMR spectroscopy generally require isotopic labeling, often precluding structural studies of complex lipid systems. Solid-state 13C magic-angle spinning NMR spectroscopy at natural isotopic abundance gives site-specific structural information that can aid in the characterization of complex biomembranes. Using the separated local-field experiment DROSS, we resolved 13C-1H residual dipolar couplings that were interpreted with a statistical mean-torque model. Liquid-disordered and liquid-ordered phases were characterized according to membrane thickness and average cross-sectional area per lipid. Knowledge of such structural parameters is vital for molecular dynamics simulations, and provides information about the balance of forces in membrane lipid bilayers. Experiments were conducted with both phosphatidylcholine (dimyristoylphosphatidylcholine (DMPC) and palmitoyloleoylphosphatidylcholine (POPC)) and egg-yolk sphingomyelin (EYSM) lipids, and allowed us to extract segmental order parameters from the 13C-1H residual dipolar couplings. Order parameters were used to calculate membrane structural quantities, including the area per lipid and bilayer thickness. Relative to POPC, EYSM is more ordered in the ld phase and experiences less structural perturbation upon adding 50% cholesterol to form the lo phase. The loss of configurational entropy is smaller for EYSM than for POPC, thus favoring its interaction with cholesterol in raftlike lipid systems. Our studies show that solid-state 13C NMR spectroscopy is applicable to investigations of complex lipids and makes it possible to obtain structural parameters for biomembrane systems where isotope labeling may be prohibitive. PMID:25418296

  9. A 13C NMR study of the adsorbed states of CO on Rh dispersed on Al2O3

    NASA Astrophysics Data System (ADS)

    Duncan, T. M.; Yates, J. T.; Vaughan, R. W.

    1980-07-01

    The results of nuclear magnetic resonance (NMR) spectroscopy have been analyzed with respect to previous infrared studies of CO adsorbed on Rh dispersed on Al2O3 to quantify the site distribution and to describe the adsorbed state. The 13C NMR spectra account for all the 13CO adsorbed on a 2.2% Rh on Al2O3 substrate. Although the spectra from the different adsorbed states of CO overlap, the line shapes may be separated into two components based on differences in the 13C spin-lattice relaxation times. These two components have been assigned to the 13CO dicarbonyl formed on single Rh atoms and to 13CO adsorbed on Rh rafts. The component attributed to the CO adsorbed on the raft sites is further separated into linear and bridged CO state contributions based on chemical shift information, yielding a quantitative distribution of the three adsorbed states of CO on Rh. The 13CO distribution is used to estimate the molar integrated intensities of the infrared spectrum of 13CO on Rh at high coverage and to determine the degree of dispersion of Rh on the Al2O3. The 13C NMR line shapes of CO adsorbed on Rh are different from the powder pattern of Rh2Cl2(CO)4. It is suggested that the line shape of the dicarbonyl surface species is narrowed to a Lorentzian curve by reorientation at the site and the line shape of CO on the Rh rafts is modulated by exchange between sites on a single raft. The 13C relaxation time distribution provides further evidence for the existence of isolated Rh atoms on the Al2O3 surface.

  10. Influence of different organic fertilizers on quality parameters and the delta(15)N, delta(13)C, delta(2)H, delta(34)S, and delta(18)O values of orange fruit (Citrus sinensis L. Osbeck).

    PubMed

    Rapisarda, Paolo; Camin, Federica; Fabroni, Simona; Perini, Matteo; Torrisi, Biagio; Intrigliolo, Francesco

    2010-03-24

    To investigate the influence of different types of fertilizers on quality parameters, N-containing compounds, and the delta(15)N, delta(13)C, delta(2)H, delta (34)S, and delta(18)O values of citrus fruit, a study was performed on the orange fruit cv. 'Valencia late' (Citrus sinensis L. Osbeck), which was harvested in four plots (three organic and one conventional) located on the same farm. The results demonstrated that different types of organic fertilizers containing the same amount of nitrogen did not effect important changes in orange fruit quality parameters. The levels of total N and N-containing compounds such as synephrine in fruit juice were not statistically different among the different treatments. The delta(15)N values of orange fruit grown under fertilizer derived from animal origin as well as from vegetable compost were statistically higher than those grown with mineral fertilizer. Therefore, delta(15)N values can be used as an indicator of citrus fertilization management (organic or conventional), because even when applied organic fertilizers are of different origins, the natural abundance of (15)N in organic citrus fruit remains higher than in conventional ones. These treatments also did not effect differences in the delta(13)C, delta(2)H, delta(34)S, and delta(18)O values of fruit. PMID:20184327

  11. Chemoselective detection and discrimination of carbonyl-containing compounds in metabolite mixtures by 1H-detected 15N NMR

    PubMed Central

    Lane, Andrew N.; Arumugam, Sengodagounder; Lorkiewicz, Pawel K.; Higashi, Richard M.; Laulhé, Sébastien; Nantz, Michael H.; Moseley, Hunter N.B.; Fan, Teresa W.-M.

    2015-01-01

    NMR spectra of mixtures of metabolites extracted from cells or tissues are extremely complex, reflecting the large number of compounds that are present over a wide range of concentrations. Although multidimensional NMR can greatly improve resolution as well as improve reliability of compound assignments, lower abundance metabolites often remain hidden. We have developed a carbonyl selective aminooxy probe that specifically reacts with free keto and aldehyde functions, but not carboxylates. By incorporating 15N in the aminooxy functional group, 15N-edited NMR was used to select exclusively those metabolites that contain a free carbonyl function while all other metabolites are rejected. Here we demonstrate that the chemical shifts of the aminooxy adducts of ketones and aldehydes are very different, which can be used to discriminate between aldoses and ketoses for example. Utilizing the 2 or 3 bond 15N-1H couplings, the 15N-edited NMR analysis was optimized first with authentic standards and then applied to an extract of the lung adenocarcinoma cell line A549. More than 30 carbonyl containing compounds at NMR detectable levels, 6 of which we have assigned by reference to our database. As the aminooxy probe contains a permanently charged quaternary ammonium group, the adducts are also optimized for detection by mass spectrometry. Thus, this sample preparation technique provides a better link between the two structural determination tools, thereby paving the way to faster and more reliable identification of both known and unknown metabolites directly in crude biological extracts. PMID:25616249

  12. Local structure and molecular motions in imidazolium hydrogen malonate crystal as studied by 2H and 13C NMR

    NASA Astrophysics Data System (ADS)

    Mizuno, M.; Chizuwa, M.; Umiyama, T.; Kumagai, Y.; Miyatou, T.; Ohashi, R.; Ida, T.; Tansho, M.; Shimizu, T.

    2015-04-01

    The local structure and molecular motion of the imidazolium hydrogen malonate crystal were investigated using solid-state 2H and 13C NMR. The imidazolium ion undergoes isotropic rotation, which is correlated with a defect in the crystal, as observed by 2H NMR broadline spectra above 263 K. A 180∘ flip of the imidazolium ion in the regular site was observed from 2H NMR quadrupole Carr-Purcell-Meiboom-Gill (QCPMG) spectra. The Grotthuss mechanism was accompanied by a 180∘ flip of the imidazolium ion in regular sites. Moreover, the proton transfer associated with the imidazolium ion of the defective crystal is important for proton conductivity of the imidazolium hydrogen malonate crystal.

  13. Backbone dynamics of the oligomerization domain of p53 determined from 15N NMR relaxation measurements.

    PubMed

    Clubb, R T; Omichinski, J G; Sakaguchi, K; Appella, E; Gronenborn, A M; Clore, G M

    1995-05-01

    The backbone dynamics of the tetrameric p53 oligomerization domain (residues 319-360) have been investigated by two-dimensional inverse detected heteronuclear 1H-15N NMR spectroscopy at 500 and 600 MHz. 15N T1, T2, and heteronuclear NOEs were measured for 39 of 40 non-proline backbone NH vectors at both field strengths. The overall correlation time for the tetramer, calculated from the T1/T2 ratios, was found to be 14.8 ns at 35 degrees C. The correlation times and amplitudes of the internal motions were extracted from the relaxation data using the model-free formalism (Lipari G, Szabo A, 1982, J Am Chem Soc 104:4546-4559). The internal dynamics of the structural core of the p53 oligomerization domain are uniform and fairly rigid, with residues 327-354 exhibiting an average generalized order parameter (S2) of 0.88 +/- 0.08. The N- and C-termini exhibit substantial mobility and are unstructured in the solution structure of p53. Residues located at the N- and C-termini, in the beta-sheet, in the turn between the alpha-helix and beta-sheet, and at the C-terminal end of the alpha-helix display two distinct internal motions that are faster than the overall correlation time. Fast internal motions (< or = 20 ps) are within the extreme narrowing limit and are of uniform amplitude. The slower motions (0.6-2.2 ns) are outside the extreme narrowing limit and vary in amplitude.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7663341

  14. Dual element ((15)N/(14)N, (13)C/(12)C) isotope analysis of glyphosate and AMPA by derivatization-gas chromatography isotope ratio mass spectrometry (GC/IRMS) combined with LC/IRMS.

    PubMed

    Mogusu, Emmanuel O; Wolbert, J Benjamin; Kujawinski, Dorothea M; Jochmann, Maik A; Elsner, Martin

    2015-07-01

    To assess sources and degradation of the herbicide glyphosate [N-(phosphonomethyl) glycine] and its metabolite AMPA (aminomethylphosphonic acid), concentration measurements are often inconclusive and even (13)C/(12)C analysis alone may give limited information. To advance isotope ratio analysis of an additional element, we present compound-specific (15)N/(14)N analysis of glyphosate and AMPA by a two step derivatization in combination with gas chromatography/isotope ratio mass spectrometry (GC/IRMS). The N-H group was derivatized with isopropyl chloroformate (iso-PCF), and remaining acidic groups were subsequently methylated with trimethylsilyldiazomethane (TMSD). Iso-PCF treatment at pH <10 gave too low (15)N/(14)N ratios indicating an incomplete derivatization; in contrast, too high (15)N/(14)N ratios at pH >10 indicated decomposition of the derivative. At pH 10, and with an excess of iso-PCF by 10-24, greatest yields and accurate (15)N/(14)N ratios were obtained (deviation from elemental analyzer-IRMS: -0.2 ± 0.9% for glyphosate; -0.4 ± 0.7% for AMPA). Limits for accurate δ(15)N analysis of glyphosate and AMPA were 150 and 250 ng injected, respectively. A combination of δ(15)N and δ(13)C analysis by liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) (1) enabled an improved distinction of commercial glyphosate products and (2) showed that glyphosate isotope values during degradation by MnO2 clearly fell outside the commercial product range. This highlights the potential of combined carbon and nitrogen isotopes analysis to trace sources and degradation of glyphosate. PMID:25967147

  15. Metabolic engineering applications of in vivo sup 31 P and sup 13 C NMR studies of Saccharomyces cerevisiae

    SciTech Connect

    Shanks, J.V.

    1989-01-01

    With intent to quantify NMR measurements as much as possible, analysis techniques of the in vivo {sup 31}P NMR spectrum are developed. A systematic procedure is formulated for estimating the relative intracellular concentrations of the sugar phosphates in S. cerevisiae from the {sup 31}P NMR spectrum. In addition, in vivo correlation of inorganic phosphate chemical shift with the chemical shifts of 3-phosphoglycerate, {beta}-fructose 1,6-diphosphate, fructose 6-phosphate, and glucose 6-phosphate are determined. Also, a method was developed for elucidation of the cytoplasmic and vacuolar components of inorganic phosphate in the {sup 31}P NMR spectrum of S. cerevisiae. An in vivo correlation relating the inorganic phosphate chemical shift of the vacuole with the chemical shift of the resonance for pyrophosphate and the terminal phosphate of polyphosphate (PP{sub 1}) is established. Transient measurements provided by {sup 31}P NMR are applied to reg1 mutant and standard strains. {sup 31}P and {sup 13}C NMR measurements are used to analyze the performance of recombinant strains in which the glucose phosphorylation step had been altered.

  16. Uranyl nitrate inhibits lactate gluconeogenesis in isolated human and mouse renal proximal tubules: A {sup 13}C-NMR study

    SciTech Connect

    Renault, Sophie; Faiz, Hassan; Gadet, Rudy; Ferrier, Bernard; Martin, Guy; Baverel, Gabriel; Conjard-Duplany, Agnes

    2010-01-01

    As part of a study on uranium nephrotoxicity, we investigated the effect of uranyl nitrate in isolated human and mouse kidney cortex tubules metabolizing the physiological substrate lactate. In the millimolar range, uranyl nitrate reduced lactate removal and gluconeogenesis and the cellular ATP level in a dose-dependent fashion. After incubation in phosphate-free Krebs-Henseleit medium with 5 mM L-[1-{sup 13}C]-, or L-[2-{sup 13}C]-, or L-[3-{sup 13}C]lactate, substrate utilization and product formation were measured by enzymatic and NMR spectroscopic methods. In the presence of 3 mM uranyl nitrate, glucose production and the intracellular ATP content were significantly reduced in both human and mouse tubules. Combination of enzymatic and NMR measurements with a mathematical model of lactate metabolism revealed an inhibition of fluxes through lactate dehydrogenase and the gluconeogenic enzymes in the presence of 3 mM uranyl nitrate; in human and mouse tubules, fluxes were lowered by 20% and 14% (lactate dehydrogenase), 27% and 32% (pyruvate carboxylase), 35% and 36% (phosphoenolpyruvate carboxykinase), and 39% and 45% (glucose-6-phosphatase), respectively. These results indicate that natural uranium is an inhibitor of renal lactate gluconeogenesis in both humans and mice.

  17. Toward dynamic isotopomer analysis in the rat brain in vivo: automatic quantitation of 13C NMR spectra using LCModel.

    PubMed

    Henry, Pierre-Gilles; Oz, Gülin; Provencher, Stephen; Gruetter, Rolf

    2003-01-01

    The LCModel method was adapted to analyze localized in vivo (13)C NMR spectra obtained from the rat brain in vivo at 9.4 T. Prior knowledge of chemical-shifts, J-coupling constants and J-evolution was included in the analysis. Up to 50 different isotopomer signals corresponding to 10 metabolites were quantified simultaneously in 400 microl volumes in the rat brain in vivo during infusion of [1,6-(13)C(2)]glucose. The analysis remained accurate even at low signal-to-noise ratio of the order of 3:1. The relative distribution of isotopomers in glutamate, glutamine and aspartate determined in vivo in 22 min was in excellent agreement with that measured in brain extracts. Quantitation of time series of (13)C spectra yielded time courses of total (13)C label incorporation into up to 16 carbon positions, as well as time courses of individual isotopomer signals, with a temporal resolution as low as 5 min (dynamic isotopomer analysis). The possibility of measuring in vivo a wealth of information that was hitherto accessible only in extracts is likely to expand the scope of metabolic studies in the intact brain. PMID:14679502

  18. Solid-State Selective 13C Excitation and Spin Diffusion NMR to Resolve Spatial Dimensions in Plant Cell Walls

    SciTech Connect

    Foston, M.; Katahira, R.; Gjersing, E.; Davis, M. F.; Ragauskas, A. J.

    2012-02-15

    The average spatial dimensions between major biopolymers within the plant cell wall can be resolved using a solid-state NMR technique referred to as a {sup 13}C cross-polarization (CP) SELDOM (selectively by destruction of magnetization) with a mixing time delay for spin diffusion. Selective excitation of specific aromatic lignin carbons indicates that lignin is in close proximity to hemicellulose followed by amorphous and finally crystalline cellulose. {sup 13}C spin diffusion time constants (T{sub SD}) were extracted using a two-site spin diffusion theory developed for {sup 13}C nuclei under magic angle spinning (MAS) conditions. These time constants were then used to calculate an average lower-limit spin diffusion length between chemical groups within the plant cell wall. The results on untreated {sup 13}C enriched corn stover stem reveal that the lignin carbons are, on average, located at distances {approx}0.7-2.0 nm from the carbons in hemicellulose and cellulose, whereas the pretreated material had larger separations.

  19. A closer look at the nitrogen next door: 1H-15N NMR methods for glycosaminoglycan structural characterization

    NASA Astrophysics Data System (ADS)

    Langeslay, Derek J.; Beni, Szabolcs; Larive, Cynthia K.

    2012-03-01

    Recently, experimental conditions were presented for the detection of the N-sulfoglucosamine (GlcNS) NHSO3- or sulfamate 1H and 15N NMR resonances of the pharmaceutically and biologically important glycosaminoglycan (GAG) heparin in aqueous solution. In the present work, we explore further the applicability of nitrogen-bound proton detection to provide structural information for GAGs. Compared to the detection of 15N chemical shifts of aminosugars through long-range couplings using the IMPACT-HNMBC pulse sequence, the more sensitive two-dimensional 1H-15N HSQC-TOCSY experiments provided additional structural data. The IMPACT-HNMBC experiment remains a powerful tool as demonstrated by the spectrum measured for the unsubstituted amine of 3-O-sulfoglucosamine (GlcN(3S)), which cannot be observed with the 1H-15N HSQC-TOCSY experiment due to the fast exchange of the amino group protons with solvent. The 1H-15N HSQC-TOCSY NMR spectrum reported for the mixture of model compounds GlcNS and N-acetylglucosamine (GlcNAc) demonstrate the broad utility of this approach. Measurements for the synthetic pentasaccharide drug Arixtra® (Fondaparinux sodium) in aqueous solution illustrate the power of this NMR pulse sequence for structural characterization of highly similar N-sulfoglucosamine residues in GAG-derived oligosaccharides.

  20. A (13)C NMR analysis of the effects of electron radiation on graphite/polyetherimide composites

    NASA Technical Reports Server (NTRS)

    Ferguson, Milton W.

    1989-01-01

    Initial investigations have been made into the use of high resolution nuclear magnetic resonance (NMR) for the characterization of radiation effects in graphite and Kevlar fibers, polymers, and the fiber/matrix interface in graphite/polyetherimide composites. Sample preparation techniques were refined. Essential equipment has been procured. A new NMR probe was constructed to increase the proton signal-to-noise ratio. Problem areas have been identified and plans developed to resolve them.

  1. Testing the use of bulk organic δ13C, δ15N, and Corg:Ntot ratios to estimate subsidence during the 1964 great Alaska earthquake

    USGS Publications Warehouse

    Bender, Adrian M; Witter, Robert C.; Rogers, Matthew

    2015-01-01

    During the Mw 9.2 1964 great Alaska earthquake, Turnagain Arm near Girdwood, Alaska subsided 1.7 ± 0.1 m based on pre- and postearthquake leveling. The coseismic subsidence in 1964 caused equivalent sudden relative sea-level (RSL) rise that is stratigraphically preserved as mud-over-peat contacts where intertidal silt buried peaty marsh surfaces. Changes in intertidal microfossil assemblages across these contacts have been used to estimate subsidence in 1964 by applying quantitative microfossil transfer functions to reconstruct corresponding RSL rise. Here, we review the use of organic stable C and N isotope values and Corg:Ntot ratios as alternative proxies for reconstructing coseismic RSL changes, and report independent estimates of subsidence in 1964 by using δ13C values from intertidal sediment to assess RSL change caused by the earthquake. We observe that surface sediment δ13C values systematically decrease by ∼4‰ over the ∼2.5 m increase in elevation along three 60- to 100-m-long transects extending from intertidal mud flat to upland environments. We use a straightforward linear regression to quantify the relationship between modern sediment δ13C values and elevation (n = 84, R2 = 0.56). The linear regression provides a slope–intercept equation used to reconstruct the paleoelevation of the site before and after the earthquake based on δ13C values in sandy silt above and herbaceous peat below the 1964 contact. The regression standard error (average = ±0.59‰) reflects the modern isotopic variability at sites of similar surface elevation, and is equivalent to an uncertainty of ±0.4 m elevation with respect to Mean Higher High Water. To reduce potential errors in paleoelevation and subsidence estimates, we analyzed multiple sediment δ13C values in nine cores on a shore-perpendicular transect at Bird Point. Our method estimates 1.3 ± 0.4 m of coseismic RSL rise across the 1964 contact by taking the arithmetic mean of the

  2. 1H and 13C NMR study of substituted 3-OH pyridines

    NASA Astrophysics Data System (ADS)

    De Kowalewski, D. G.; De Los Santos, C.

    1990-04-01

    nJHH, nJCH and δ 13C values have been measured for a series of X substituted 3-hydroxypyridines (X = 2-NH 2, 2-NO 2, 5-Cl, 6-CH 3, 2-Cl, 2-Br, 2-I). The results show that the additivity of δ 13C provides a valuable criterion to differentiate the phenolic from the zwitterion structure. This conclusion is based on the fact that in the first case, for 2-NH 2-, 2-NO 2-, 5-Cl- and 6-CH 3-3-hydroxypyridines, there is agreement between the experimental and the additivity δ 13C values, while in the three halogen derivatives (2-Cl-, 2-Br- and 2-I-3-hydroxypyridines) the δ exp—addC3 values of -4.95, -7.25 and 9.05 are probably due to the negative charge present on the three position of the zwitterion. Since the additivity of 1JCH values holds in all substances examined (unlike the case of the 2-pyridone derivatives) it is not possible to use that criterion to differentiate between the phenolic and dipolar structures. The above conclusions are in agreement with IR, p K, NQR, RX, kinetics experiments and quantum chemical calculations of other authors.

  3. Backbone and side-chain (1)H, (15)N, (13)C assignment and secondary structure of BPSL1445 from Burkholderia pseudomallei.

    PubMed

    Quilici, Giacomo; Berardi, Andrea; Gaudesi, Davide; Gourlay, Louise J; Bolognesi, Martino; Musco, Giovanna

    2015-10-01

    BPSL1445 is a lipoprotein produced by the Gram-negative bacterium Burkholderia pseudomallei (B. pseudomallei), the etiological agent of melioidosis. Immunodetection assays against sera patients using protein microarray suggest BPSL1445 involvement in melioidosis. Herein we report backbone, side chain NMR assignment and secondary structure for the recombinant protein. PMID:25893672

  4. 13C, 2h NMR studies of structural and dynamical modifications of glucose-exposed porcine aortic elastin.

    PubMed

    Silverstein, Moshe C; Bilici, Kübra; Morgan, Steven W; Wang, Yunjie; Zhang, Yanhang; Boutis, Gregory S

    2015-04-01

    Elastin, the principal component of the elastic fiber of the extracellular matrix, imparts to vertebrate tissues remarkable resilience and longevity. This work focuses on elucidating dynamical and structural modifications of porcine aortic elastin exposed to glucose by solid-state NMR spectroscopic and relaxation methodologies. Results from macroscopic stress-strain tests are also presented and indicate that glucose-treated elastin is mechanically stiffer than the same tissue without glucose treatment. These measurements show a large hysteresis in the stress-strain behavior of glucose-treated elastin-a well-known signature of viscoelasticity. Two-dimensional relaxation NMR methods were used to investigate the correlation time, distribution, and population of water in these samples. Differences are observed between the relative populations of water, whereas the measured correlation times of tumbling motion of water across the samples were similar. (13)C magic-angle-spinning NMR methods were applied to investigate structural and dynamical modifications after glucose treatment. Although some overall structure is preserved, the process of glucose exposure results in more heterogeneous structures and slower mobility. The correlation times of tumbling motion of the (13)C-(1)H internuclear vectors in the glucose-treated sample are larger than in untreated samples, pointing to their more rigid structure. The (13)C cross-polarization spectra reveal a notably increased α-helical character in the alanine motifs after glucose exposure. Results from molecular dynamics simulations are provided that add further insight into dynamical and structural changes of a short repeat, [VPGVG]5, an alanine pentamer, desmosine, and isodesmosine sites with and without glucose. The simulations point to changes in the entropic and energetic contributions in the retractive forces of VPGVG and AAAAA motifs. The most notable change is the increase of the energetic contribution in the retractive

  5. Hydrothermal carbon from biomass: structural differences between hydrothermal and pyrolyzed carbons via 13C solid state NMR.

    PubMed

    Falco, Camillo; Perez Caballero, Fernando; Babonneau, Florence; Gervais, Christel; Laurent, Guillaume; Titirici, Maria-Magdalena; Baccile, Niki

    2011-12-01

    The objective of this paper is to better describe the structure of the hydrothermal carbon (HTC) process and put it in relationship with the more classical pyrolytic carbons. Indeed, despite the low energetic impact and the number of applications described so far for HTC, very little is known about the structure, reaction mechanism, and the way these materials relate to coals. Are HTC and calcination processes equivalent? Are the structures of the processed materials related to each other in any way? Which is the extent of polyaromatic hydrocarbons (PAH) inside HTC? In this work, the effect of hydrothermal treatment and pyrolysis are compared on glucose, a good model carbohydrate; a detailed single-quantum double-quantum (SQ-DQ) solid state (13)C NMR study of the HTC and calcined HTC is used to interpret the spectral region corresponding to the signal of furanic and arene groups. These data are compared to the spectroscopic signatures of calcined glucose, starch, and xylose. A semiquantitative analysis of the (13)C NMR spectra provides an estimation of the furanic-to-arene ratio which varies from 1:1 to 4:1 according to the processing conditions and carbohydrate employed. In addition, we formulate some hypothesis, validated by DFT (density functional theory) modeling associated with (13)C NMR chemical shifts calculations, about the possible furan-rich structural intermediates that occur in the coalification process leading to condensed polyaromatic structures. In combination with a broad parallel study on the HTC processing conditions effect on glucose, cellulose, and raw biomass (Falco, C.; Baccile, N.; Titirici, M.-M. Green Chem., 2011, DOI: 10.1039/C1GC15742F), we propose a broad reaction scheme and in which we show that, through HTC, it is possible to tune the furan-to-arene ratio composing the aromatic core of the produced HTC carbons, which is not possible if calcination is used alone, in the temperature range below 350 °C. PMID:22050004

  6. Phase behavior and 13C NMR spectroscopic analysis of the mixed methane + ethane + propane hydrates in mesoporous silica gels.

    PubMed

    Lee, Seungmin; Cha, Inuk; Seo, Yongwon

    2010-11-25

    In this study, the phase behavior and quantitative determination of hydrate composition and cage occupancy for the mixed CH(4) + C(2)H(6) + C(3)H(8) hydrates were closely investigated through the experimental measurement of three-phase hydrate (H)-water-rich liquid (L(W))-vapor (V) equilibria and (13)C NMR spectra. To examine the effect of pore size and salinity, we measured hydrate phase equilibria for the quaternary CH(4) (90%) + C(2)H(6) (7%) + C(3)H(8) (3%) + water mixtures in silica gel pores of nominal diameters of 6.0, 15.0, and 30.0 nm and for the quinary CH(4) (90%) + C(2)H(6) (7%) + C(3)H(8) (3%) + NaCl + water mixtures of two different NaCl concentrations (3 and 10 wt %) in silica gel pores of a nominal 30.0 nm diameter. The value of hydrate-water interfacial tension for the CH(4) (90%) + C(2)H(6) (7%) + C(3)H(8) (3%) hydrate was found to be 47 ± 4 mJ/m(2) from the relation of the dissociation temperature depression with the pore size of silica gels at a given pressure. At a specified temperature, three-phase H-L(W)-V equilibrium curves of pore hydrates were shifted to higher pressure regions depending on pore sizes and NaCl concentrations. From the cage-dependent (13)C NMR chemical shifts of enclathrated guest molecules, the mixed CH(4) (90%) + C(2)H(6) (7%) + C(3)H(8) (3%) gas hydrate was confirmed to be structure II. The cage occupancies of each guest molecule and the hydration number of the mixed gas hydrates were also estimated from the (13)C NMR spectra. PMID:20964277

  7. 13C, 2H NMR Studies of Structural and Dynamical Modifications of Glucose-Exposed Porcine Aortic Elastin

    PubMed Central

    Silverstein, Moshe C.; Bilici, Kübra; Morgan, Steven W.; Wang, Yunjie; Zhang, Yanhang; Boutis, Gregory S.

    2015-01-01

    Elastin, the principal component of the elastic fiber of the extracellular matrix, imparts to vertebrate tissues remarkable resilience and longevity. This work focuses on elucidating dynamical and structural modifications of porcine aortic elastin exposed to glucose by solid-state NMR spectroscopic and relaxation methodologies. Results from macroscopic stress-strain tests are also presented and indicate that glucose-treated elastin is mechanically stiffer than the same tissue without glucose treatment. These measurements show a large hysteresis in the stress-strain behavior of glucose-treated elastin—a well-known signature of viscoelasticity. Two-dimensional relaxation NMR methods were used to investigate the correlation time, distribution, and population of water in these samples. Differences are observed between the relative populations of water, whereas the measured correlation times of tumbling motion of water across the samples were similar. 13C magic-angle-spinning NMR methods were applied to investigate structural and dynamical modifications after glucose treatment. Although some overall structure is preserved, the process of glucose exposure results in more heterogeneous structures and slower mobility. The correlation times of tumbling motion of the 13C-1H internuclear vectors in the glucose-treated sample are larger than in untreated samples, pointing to their more rigid structure. The 13C cross-polarization spectra reveal a notably increased α-helical character in the alanine motifs after glucose exposure. Results from molecular dynamics simulations are provided that add further insight into dynamical and structural changes of a short repeat, [VPGVG]5, an alanine pentamer, desmosine, and isodesmosine sites with and without glucose. The simulations point to changes in the entropic and energetic contributions in the retractive forces of VPGVG and AAAAA motifs. The most notable change is the increase of the energetic contribution in the retractive force

  8. Carbon-13, sup 15 N, and sup 31 P NMR studies on 6-hydroxy-L-nicotine oxidase from Arthrobacter oxidans

    SciTech Connect

    Pust, S.; Vervoort, J.; Decker, K.; Bacher, A.; Mueller, F. )

    1989-01-24

    The interaction between the apoprotein of 6-hydroxy-L-nicotine oxidase from Arthrobacter oxidans and the prosthetic group FAD has been investigated by {sup 13}C, {sup 15}N and {sup 31}P NMR techniques. The FAD prosthetic group was selectively enriched in {sup 13}C and {sup 15}N isotopes by adding isotopically labeled riboflavin derivatives to the growth medium of riboflavin-requiring mutant cells. In the oxidized state the chemical shift of the C(7) and C(8) atoms indicates that the xylene moiety of the isoalloxazine ring is embedded in a hydrophobic environment. The binding of the competitive inhibitor, 6-hydroxy-D-nicotine, influences the resonances of the C(4a) and the N(5) atom strongly. It is suggested that these shifts are due to a strong hydrogen-bonding interaction between the N(5) atom and the inhibitor. On reduction all resonances, except those of the C(10a) and the N(1) atoms, shift upfield, indicating the increased electron density in the ring system. It can unambiguously be concluded from the chemical shift of the N(1) atom that the reduced flavin is anionic. The doublet character of the N(3) and N(5) resonances suggests that bulk water has no access to the active center. The strong downfield shift of the N(1) position indicates that this atom is embedded in a polar environment, but it does not indicate the presence of a positively charged residue. The {sup 31}P NMR spectra show that the resonances of the pyrophosphate group of the bound FAD differ slightly from those of free FAD. Besides the {sup 31}P resonances from FAD, four peaks around 0 ppm are observed that belongs to bound phosphorus residues. The residues are not located close to the isoalloxazine ring.

  9. Use of solid-state 13C NMR in structural studies of humic acids and humin from Holocene sediments

    USGS Publications Warehouse

    Hatcher, P.G.; VanderHart, D.L.; Earl, W.L.

    1980-01-01

    13C NMR spectra of solid humic substances in Holocene sediments have been obtained using cross polarization with magic-angle sample spinning techniques. The results demonstrate that this technique holds great promise for structural characterizations of complex macromolecular substances such as humin and humic acids. Quantifiable distinctions can be made between structural features of aquatic and terrestrial humic substances. The aliphatic carbons of the humic substances are dominant components suggestive of input from lipid-like materials. An interesting resemblance is also noted between terrestrial humic acid and humin spectra. ?? 1980.

  10. Characterization of aging in organic materials on atomic-, meso- and macro-length scales by {sup 13}C NMR spectroscopy

    SciTech Connect

    Assink, R.A.; Jamison, G.M.; Alam, T.M.; Gillen, K.T.

    1997-10-01

    A fundamental understanding of aging in an organic material requires that one understand how aging affects the chemical structure of a material, and how these chemical changes are related to the material`s macroscopic properties. This level of understanding is usually achieved by examining the material on a variety of length scales ranging from atomic to meso-scale to macroscopic. The authors are developing and applying several {sup 13}C nuclear magnetic resonance (NMR) spectroscopy experiments to characterize the aging process of organic materials over a broad range of length scales. Examples of studies which range from atomic to macroscopic will be presented.

  11. Biosynthesis of pyrroloquinoline quinone. 1. Identification of biosynthetic precursors using /sup 13/C labeling and NMR spectroscopy

    SciTech Connect

    Houck, D.R.; Hanners, J.L.; Unkefer, C.J.

    1988-09-28

    The biosynthesis of pyrroloquinoline quinone (PQQ) in the methylotropic bacterium methylobacterium AM1 has been investigated using /sup 13/C-labelling of the products and NMR spectroscopy. The data indicated that the quinoline portion of PQQ is formed by a novel condensation of N-1, C-2, -3, and -4 of glutamate with a symmetrical six-carbon ring derived from the shikimate pathway. It is postulated that tyrosine is the shikimate-derived percursor, since pyrrole could be formed by the internal cyclization of the amino acid backbone. 18 references, 2 figures, 2 tables.

  12. A stable isotope (δ13C and δ15N) perspective on human diet on Rapa Nui (Easter Island) ca. AD 1400-1900.

    PubMed

    Commendador, Amy S; Dudgeon, John V; Finney, Bruce P; Fuller, Benjamin T; Esh, Kelley S

    2013-10-01

    Ecological and environmental evidence suggests that Rapa Nui was among the most marginally habitable islands in Eastern Polynesia, with only a fraction of the biotic diversity found on archipelagos to the west, and capable of sustaining many fewer cultigens traditionally transported by Polynesian colonizers. However, archaeological evidence for human dietary adaptations under such restrictions is limited. Little is known about the particulars of the subsistence base and dietary changes on Rapa Nui that may be associated with a hypothesized late prehistoric decline in the quality and diversity of food sources. To better understand prehistoric Rapa Nui diet we examined stable carbon and nitrogen isotope compositions of human teeth along with archaeological faunal material thought to comprise the Rapa Nui food web. Our results indicate that contrary to previous zooarchaeological studies diet was predominantly terrestrial throughout the entire sequence of occupation, with reliance on rats, chickens and C3 plants. While a few individuals may have had access to higher trophic level marine resources, this is evident only later in time (generally post-AD 1600). A decline in (15)N through time was observed, and may be attributed to declines in available terrestrial proteins; however, presently we cannot rule out the effect of changing soil and plant baseline δ(15)N. Our results also suggest differential access to higher trophic level marine resources among contemporaneous populations, but more research is required to clarify this observation. PMID:23996514

  13. Vibrational, XRD and (13)C NMR studies of DL-phenylglycinium methanesulfonate crystal.

    PubMed

    Wołoszyn, Łukasz; Ilczyszyn, Marek; Ilczyszyn, Maria M; Haupa, Karolina

    2016-11-01

    A new crystal formed by DL-phenylglycine and methanesulfonic acid (PGLYMS) was obtained and studied by X-ray diffraction, IR and Raman spectroscopy, solid state NMR and DSC methods. Additionally, theoretical computations for the phenylglycine cation were carried out (DFT/B3LYP/aug-cc-pVDZ). Our results show that PGLYMS does not exhibit any phase transitions and crystallizes in the P21/c space group of monoclinic system (Z=4). Detailed analysis of its structure and its IR, Raman and NMR spectra is presented. PMID:27289350

  14. 13C NMR Metabolomic Evaluation of Immediate and Delayed Mild Hypothermia in Cerebrocortical Slices After Oxygen-Glucose Deprivation

    PubMed Central

    Liu, Jia; Segal, Mark; Kelly, Mark J.S.; Pelton, Jeffrey G.; Kim, Myungwon; James, Thomas L.; Litt, Lawrence

    2013-01-01

    Background Mild brain hypothermia (32°C–34°C) after human neonatal asphyxia improves neurodevelopmental outcomes. Astrocytes but not neurons have pyruvate carboxylase (PC) and an acetate uptake transporter. 13C NMR spectroscopy of rodent brain extracts after administering [1-13C]glucose and [1,2-13C]acetate can distinguish metabolic differences between glia and neurons, and tricarboxylic acid cycle (TCA cycle) entry via pyruvate dehydrogenase (PDH) and PC. Methods Neonatal rat cerebrocortical slices receiving a 13C-acetate/glucose mixture underwent a 45-min asphyxia simulation via oxygen-glucose-deprivation (OGD) followed by 6 h of recovery. Protocols in three groups of N = 3 experiments were identical except for temperature management. The three temperature groups were: normothermia (37°C), hypothermia (32°C for 3.75 h beginning at OGD start), and delayed hypothermia (32°C for 3.75 h, beginning 15 min after OGD start). Multivariate analysis of nuclear magnetic resonance metabolite quantifications included principal component analyses and the L1-Penalized Regularized Regression algorithm known as the Least Absolute Shrinkage and Selection Operator (LASSO). Results The most significant metabolite difference (p < 0.0056) was [2-13C]glutamine’s higher final/control ratio for the Hypothermia group (1.75 ± 0.12) compared to ratios for the Delayed (1.12 ± 0.12) and Normothermia group (0.94 ± 0.06), implying a higher PC/PDH ratio for glutamine formation. LASSO found the most important metabolites associated with adenosine triphosphate preservation: [3,4-13C]glutamate—produced via PDH entry, [2-13C]taurine--an important osmolyte, and phosphocreatine. Final principal component analyses scores plots suggested separate cluster formation for the hypothermia group, but with insufficient data for statistical significance. Conclusions Starting mild hypothermia simultaneously with OGD, compared with delayed starting or no hypothermia, has higher PC throughput

  15. Use of dipolar 1H-15N and 1H-13C couplings in the structure determination of magnetically oriented macromolecules in solution.

    PubMed

    Tjandra, N; Omichinski, J G; Gronenborn, A M; Clore, G M; Bax, A

    1997-09-01

    Anisotropy of the molecular magnetic susceptibility gives rise to a small degree of alignment. The resulting residual dipolar couplings, which can now be measured with the advent of higher magnetic fields in NMR, contain information on the orientation of the internuclear vectors relative to the molecular magnetic susceptibility tensor, thereby providing information on long range order that is not accessible by any of the solution NMR parameters currently used in structure determination. Thus, the dipolar couplings constitute unique and powerful restraints in determining the structures of magnetically oriented macromolecules in solution. The method is demonstrated on a complex of the DNA-binding domain of the transcription factor GATA-1 with a 16 base pair oligodeoxyribonucleotide. PMID:9303001