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Sample records for 13c tracer experiments

  1. Analysis of 13C labeling enrichment in microbial culture applying metabolic tracer experiments using gas chromatography-combustion-isotope ratio mass spectrometry.

    PubMed

    Heinzle, Elmar; Yuan, Yongbo; Kumar, Sathish; Wittmann, Christoph; Gehre, Matthias; Richnow, Hans-Herrmann; Wehrung, Patrick; Adam, Pierre; Albrecht, Pierre

    2008-09-15

    The applicability of gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) for the quantification of 13C enrichment of proteinogenic amino acids in metabolic tracer experiments was evaluated. Measurement of the 13C enrichment of proteinogenic amino acids from cell hydrolyzates of Corynebacterium glutamicum growing on different mixtures containing between 0.5 and 10% [1-13C]glucose shows the significance of kinetic isotope effects in metabolic flux studies at low degree of labeling. We developed a method to calculate the 13C enrichment. The approach to correct for these effects in metabolic flux studies using delta13C measurement by GC-C-IRMS uses two parallel experiments applying substrate with natural abundance and 13C-enriched tracer substrate, respectively. The fractional enrichment obtained in natural substrate is subtracted from that of the enriched one. Tracer studies with C. glutamicum resulted in a statistically identical relative fractional enrichment of 13C in proteinogenic amino acids over the whole range of applied concentrations of [1-13C]glucose. The current findings indicate a great potential of GC-C-IRMS for labeling quantification in 13C metabolic flux analysis with low labeling degree of tracer substrate directly in larger scale bioreactors.

  2. Evaluation of 13C isotopic tracers for metabolic flux analysis in mammalian cells

    PubMed Central

    Metallo, Christian M.; Walther, Jason L.; Stephanopoulos, Gregory

    2009-01-01

    13C metabolic flux analysis (MFA) is the most comprehensive means of characterizing cellular metabolic states. Uniquely labeled isotopic tracers enable more focused analyses to probe specific reactions within the network. As a result, the choice of tracer largely determines the precision with which one can estimate metabolic fluxes, especially in complex mammalian systems that require multiple substrates. Here we have experimentally determined metabolic fluxes in a tumor cell line, successfully recapitulating the hallmarks of cancer cell metabolism. Using these data, we computationally evaluated specifically labeled 13C glucose and glutamine tracers for their ability to precisely and accurately estimate fluxes in central carbon metabolism. These methods enabled us to to identify the optimal tracer for analyzing individual fluxes, specific pathways, and central carbon metabolism as a whole. [1,2-13C2]glucose provided the most precise estimates for glycolysis, the pentose phosphate pathway, and the overall network. Tracers such as [2-13C]glucose and [3-13C]glucose also outperformed the more commonly used [1-13C]glucose. [U-13C5]glutamine emerged as the preferred isotopic tracer for analysis of the tricarboxylic acid (TCA) cycle. These results provide valuable, quantitative information on the performance of 13C-labeled substrates and can aid in the design of more informative MFA experiments in mammalian cell culture. PMID:19622376

  3. Uptake of algal carbon and the synthesis of an "essential" fatty acid by Uvigerina ex. gr. semiornata (Foraminifera) within the Pakistan margin oxygen minimum zone: evidence from fatty acid biomarker and 13C tracer experiments

    NASA Astrophysics Data System (ADS)

    Larkin, K. E.; Gooday, A. J.; Woulds, C.; Jeffreys, R.; Schwartz, M.; Cowie, G.; Whitcraft, C.; Levin, L.; Dick, J. R.; Pond, D. W.

    2014-01-01

    Foraminifera are an important component of benthic communities in oxygen depleted settings, where they potentially play a~significant role in the processing of organic matter. We tracked the uptake of a 13C-labeled algal food source into individual fatty acids in the benthic foraminiferal species, Uvigerina ex. gr. semiornata, from the Arabian Sea oxygen minimum zone (OMZ). The tracer experiments were conducted on the Pakistan Margin during the late/post monsoon period (August-October 2003). A monoculture of the diatom Thalassiosira weisflogii was 13C-labeled and used to simulate a pulse of phytoplankton in two complementary experiments. A lander system was used for in situ incubations at 140 m and for 2.5 days duration, whilst a laboratory incubation used an oxystat system to maintain ambient dissolved oxygen concentrations. These shipboard experiments were terminated after 5 days. Uptake of diatoms was rapid, with high incorporation of diatom fatty acids into foraminifera after ~2 days in both experiments. Ingestion of the diatom food source was indicated by the increase over time in the quantity of diatom biomarker fatty acids in the foraminifera and by the high percentage of 13C in many of the fatty acids present at the endpoint of both in~situ and laboratory-based experiments. These results indicate that U. ex. gr. semiornata rapidly ingested the diatom food source and that this foraminifera will play an important role in the short-term cycling of organic matter within this OMZ environment. The experiments also suggested that U. ex. gr. semiornata consumed non-labeled bacterial food items, particularly bacteria, and synthesised the polyunsaturated fatty acid 20:4(n-6) de novo. 20:4(n-6) is often abundant in benthic fauna yet its origins and function have remained unclear. This study demonstrates that U. ex. gr. semiornata is capable of de novo synthesis of this "essential fatty acid" and is potentially a major source of this dietary nutrient in benthic food

  4. Uptake of algal carbon and the likely synthesis of an "essential" fatty acid by Uvigerina ex. gr. semiornata (Foraminifera) within the Pakistan margin oxygen minimum zone: evidence from fatty acid biomarker and 13C tracer experiments

    NASA Astrophysics Data System (ADS)

    Larkin, K. E.; Gooday, A. J.; Woulds, C.; Jeffreys, R. M.; Schwartz, M.; Cowie, G.; Whitcraft, C.; Levin, L.; Dick, J. R.; Pond, D. W.

    2014-07-01

    Foraminifera are an important component of benthic communities in oxygen-depleted settings, where they potentially play a significant role in the processing of organic matter. We tracked the uptake of a 13C-labelled algal food source into individual fatty acids in the benthic foraminiferal species Uvigerina ex. gr. semiornata from the Arabian Sea oxygen minimum zone (OMZ). The tracer experiments were conducted on the Pakistan margin during the late/post monsoon period (August-October 2003). A monoculture of the diatom Thalassiosira weisflogii was 13C-labelled and used to simulate a pulse of phytoplankton in two complementary experiments. A lander system was used for in situ incubations at 140 m water depth and for 2.5 days in duration. Shipboard laboratory incubations of cores collected at 140 m incorporated an oxystat system to maintain ambient dissolved oxygen concentrations and were terminated after 5 days. Uptake of diatoms was rapid, with a high incorporation of diatom fatty acids into foraminifera after ~ 2 days in both experiments. Ingestion of the diatom food source was indicated by the increase over time in the quantity of diatom biomarker fatty acids in the foraminifera and by the high percentage of 13C in many of the fatty acids present at the endpoint of both in situ and laboratory-based experiments. These results indicate that experiments, possibly because it was synthesised de novo. This "essential fatty acid" is often abundant in benthic fauna, yet

  5. EDGE2D Simulations of JET{sup 13}C Migration Experiments

    SciTech Connect

    J.D. Strachan; J.P. Coad; G. Corrigan; G.F. Matthews; J. Spence

    2004-06-16

    Material migration has received renewed interest due to tritium retention associated with carbon transport to remote vessel locations. Those results influence the desirability of carbon usage on ITER. Subsequently, additional experiments have been performed, including tracer experiments attempting to identify material migration from specific locations. In this paper, EDGE2D models a well-diagnosed JET{sup 13}C tracer migration experiment. The role of SOL flows upon the migration patterns is identified.

  6. Measuring and modeling C flux rates through the central metabolic pathways in microbial communities using position-specific 13C-labeled tracers

    NASA Astrophysics Data System (ADS)

    Dijkstra, P.; van Groenigen, K.; Hagerty, S.; Salpas, E.; Fairbanks, D. E.; Hungate, B. A.; KOCH, G. W.; Schwartz, E.

    2012-12-01

    The production of energy and metabolic precursors occurs in well-known processes such as glycolysis and Krebs cycle. We use position-specific 13C-labeled metabolic tracers, combined with models of microbial metabolic organization, to analyze the response of microbial community energy production, biosynthesis, and C use efficiency (CUE) in soils, decomposing litter, and aquatic communities. The method consists of adding position-specific 13C -labeled metabolic tracers to parallel soil incubations, in this case 1-13C and 2,3-13C pyruvate and 1-13C and U-13C glucose. The measurement of CO2 released from the labeled tracers is used to calculate the C flux rates through the various metabolic pathways. A simplified metabolic model consisting of 23 reactions is solved using results of the metabolic tracer experiments and assumptions of microbial precursor demand. This new method enables direct estimation of fundamental aspects of microbial energy production, CUE, and soil organic matter formation in relatively undisturbed microbial communities. We will present results showing the range of metabolic patterns observed in these communities and discuss results from testing metabolic models.

  7. A Polymer-Based Magnetic Resonance Tracer for Visualization of Solid Tumors by 13C Spectroscopic Imaging

    PubMed Central

    Suzuki, Yoshikazu; Iida, Mitsuru; Miura, Iwao; Inubushi, Toshiro; Morikawa, Shigehiro

    2014-01-01

    Morphological imaging precedes lesion-specific visualization in magnetic resonance imaging (MRI) because of the superior ability of this technique to depict tissue morphology with excellent spatial and temporal resolutions. To achieve lesion-specific visualization of tumors by MRI, we investigated the availability of a novel polymer-based tracer. Although the 13C nucleus is a candidate for a detection nucleus because of its low background signal in the body, the low magnetic resonance sensitivity of the nucleus needs to be resolved before developing a 13C-based tracer. In order to overcome this problem, we enriched polyethylene glycol (PEG), a biocompatible polymer, with 13C atoms. 13C-PEG40,000 (13C-PEG with an average molecular weight of 40 kDa) emitted a single 13C signal with a high signal-to-noise ratio due to its ability to maintain signal sharpness, as was confirmed by in vivo investigation, and displayed a chemical shift sufficiently distinct from that of endogenous fat. 13C-PEG40,000 intravenously injected into mice showed long retention in circulation, leading to its effective accumulation in tumors reflecting the well-known phenomenon that macromolecules accumulate in tumors because of leaky tumor capillaries. These properties of 13C-PEG40,000 allowed visualization of tumors in mice by 13C spectroscopic imaging. These findings suggest that a technique based on 13C-PEG is a promising strategy for tumor detection. PMID:25007334

  8. Experimental design principles for isotopically instationary 13C labeling experiments.

    PubMed

    Nöh, Katharina; Wiechert, Wolfgang

    2006-06-01

    13C metabolic flux analysis (MFA) is a well-established tool in Metabolic Engineering that found numerous applications in recent years. However, one strong limitation of the current method is the requirement of an-at least approximate-isotopic stationary state at sampling time. This requirement leads to a principle lower limit for the duration of a 13C labeling experiment. A new methodological development is based on repeated sampling during the instationary transient of the 13C labeling dynamics. The statistical and computational treatment of such instationary experiments is a completely new terrain. The computational effort is very high because large differential equations have to be solved and, moreover, the intracellular pool sizes play a significant role. For this reason, the present contribution works out principles and strategies for the experimental design of instationary experiments based on a simple example network. Hereby, the potential of isotopically instationary experiments is investigated in detail. Various statistical results on instationary flux identifiability are presented and possible pitfalls of experimental design are discussed. Finally, a framework for almost optimal experimental design of isotopically instationary experiments is proposed which provides a practical guideline for the analysis of large-scale networks.

  9. 13C Tracers for Glucose Degrading Pathway Discrimination in Gluconobacter oxydans 621H

    PubMed Central

    Ostermann, Steffen; Richhardt, Janine; Bringer, Stephanie; Bott, Michael; Wiechert, Wolfgang; Oldiges, Marco

    2015-01-01

    Gluconobacter oxydans 621H is used as an industrial production organism due to its exceptional ability to incompletely oxidize a great variety of carbohydrates in the periplasm. With glucose as the carbon source, up to 90% of the initial concentration is oxidized periplasmatically to gluconate and ketogluconates. Growth on glucose is biphasic and intracellular sugar catabolism proceeds via the Entner–Doudoroff pathway (EDP) and the pentose phosphate pathway (PPP). Here we studied the in vivo contributions of the two pathways to glucose catabolism on a microtiter scale. In our approach we applied specifically 13C labeled glucose, whereby a labeling pattern in alanine was generated intracellularly. This method revealed a dynamic growth phase-dependent pathway activity with increased activity of EDP in the first and PPP in the second growth phase, respectively. Evidence for a growth phase-independent decarboxylation-carboxylation cycle around the pyruvate node was obtained from 13C fragmentation patterns of alanine. For the first time, down-scaled microtiter plate cultivation together with 13C-labeled substrate was applied for G. oxydans to elucidate pathway operation, exhibiting reasonable labeling costs and allowing for sufficient replicate experiments. PMID:26404385

  10. Anaerobic Methane Oxidation in Soils - revealed using 13C-labelled methane tracers

    NASA Astrophysics Data System (ADS)

    Riekie, G. J.; Baggs, E. M.; Killham, K. S.; Smith, J. U.

    2008-12-01

    In marine sediments, anaerobic methane oxidation is a significant biogeochemical process limiting methane flux from ocean to atmosphere. To date, evidence for anaerobic methane oxidation in terrestrial environments has proved elusive, and its significance is uncertain. In this study, an isotope dilution method specifically designed to detect the process of anaerobic methane oxidation in methanogenic wetland soils is applied. Methane emissions of soils from three contrasting permanently waterlogged sites in Scotland are investigated in strictly anoxic microcosms to which 13C- labelled methane is added, and changes in the concentration and 12C/13C isotope ratios of methane and carbon dioxide are subsequently measured and used to calculate separate the separate components of the methane flux. The method used takes into account the 13C-methane associated with methanogenesis, and the amount of methane dissolved in the soil. The calculations make no prior assumptions about the kinetics of methane production or oxidation. The results indicate that methane oxidation can take place in anoxic soil environments. The clearest evidence for anaerobic methane oxidation is provided by soils from a minerotrophic fen site (pH 6.0) in Bin Forest underlain by ultra-basic and serpentine till. In the fresh soil anoxic microcosms, net consumption methane was observed, and the amount of headspace 13C-CO2 increased at a greater rate than the 12+13C-CO2, further proof of methane oxidation. A net increase in methane was measured in microcosms of soil from Murder Moss, an alkaline site, pH 6.5, with a strong calcareous influence. However, the 13C-CH4 data provided evidence of methane oxidation, both in the disappearance of C- CH4 and appearance of smaller quantities of 13C-CO2. The least alkaline (pH 5.5) microcosms, of Gateside Farm soil - a granitic till - exhibited net methanogenesis and the changes in 13C-CH4 and 13C-CO2 here followed the pattern expected if no methane is consumed

  11. Effects of trichloroacetic acid on the nitrogen metabolism of Pinus sylvestris--a 13C/15N tracer study.

    PubMed

    Hafner, Christoph; Jung, Klaus; Schüürmann, Gerrit

    2002-01-01

    Trichloroacetic acid (TCA) can be found in various environmental compartments like air, rain and plants all over the world. It is assumed that TCA is an atmospheric degradation product of volatile chloroorganic hydrocarbons. The herbicide effect of TCA in higher concentrations is well known, but not much is known about the phytotoxic effects in environmentally relevant concentrations. It can be shown in this study by using the 13C/15N stable isotope tracer technique that [13C]TCA is taken up by roots of two-year-old seedlings of Pinus sylvestris L. and transported into the needles. At the same time the effect of the substance on nitrogen metabolism can be analyzed by measuring the incorporation of 15NO3- into different nitrogen fractions of the plant. The more [13C]TCA incorporation, the higher the synthesis of 15N labelled amino acids and proteins is. These effects on the nitrogen metabolism are probably based on the activation of stress- and detoxification metabolism. It has to be assumed that there is an influence on N metabolism of Pinus sylvestris caused by the deposition of environmentally relevant TCA concentrations.

  12. Forward Modeling of Fluctuating Dietary 13C Signals to Validate 13C Turnover Models of Milk and Milk Components from a Diet-Switch Experiment

    PubMed Central

    Braun, Alexander; Schneider, Stephan; Auerswald, Karl; Bellof, Gerhard; Schnyder, Hans

    2013-01-01

    Isotopic variation of food stuffs propagates through trophic systems. But, this variation is dampened in each trophic step, due to buffering effects of metabolic and storage pools. Thus, understanding of isotopic variation in trophic systems requires knowledge of isotopic turnover. In animals, turnover is usually quantified in diet-switch experiments in controlled conditions. Such experiments usually involve changes in diet chemical composition, which may affect turnover. Furthermore, it is uncertain if diet-switch based turnover models are applicable under conditions with randomly fluctuating dietary input signals. Here, we investigate if turnover information derived from diet-switch experiments with dairy cows can predict the isotopic composition of metabolic products (milk, milk components and feces) under natural fluctuations of dietary isotope and chemical composition. First, a diet-switch from a C3-grass/maize diet to a pure C3-grass diet was used to quantify carbon turnover in whole milk, lactose, casein, milk fat and feces. Data were analyzed with a compartmental mixed effects model, which allowed for multiple pools and intra-population variability, and included a delay between feed ingestion and first tracer appearance in outputs. The delay for milk components and whole milk was ∼12 h, and that of feces ∼20 h. The half-life (t½) for carbon in the feces was 9 h, while lactose, casein and milk fat had a t½ of 10, 18 and 19 h. The 13C kinetics of whole milk revealed two pools, a fast pool with a t½ of 10 h (likely representing lactose), and a slower pool with a t½ of 21 h (likely including casein and milk fat). The diet-switch based turnover information provided a precise prediction (RMSE ∼0.2 ‰) of the natural 13C fluctuations in outputs during a 30 days-long period when cows ingested a pure C3 grass with naturally fluctuating isotope composition. PMID:24392000

  13. 13C-Tracer and Gas Chromatography-Mass Spectrometry Analyses Reveal Metabolic Flux Distribution in the Oleaginous Microalga Chlorella protothecoides1[C][W][OA

    PubMed Central

    Xiong, Wei; Liu, Lixia; Wu, Chao; Yang, Chen; Wu, Qingyu

    2010-01-01

    The green alga Chlorella protothecoides has received considerable attention because it accumulates neutral triacylglycerols, commonly regarded as an ideal feedstock for biodiesel production. In order to gain a better understanding of its metabolism, tracer experiments with [U-13C]/[1-13C]glucose were performed with heterotrophic growth of C. protothecoides for identifying the metabolic network topology and estimating intracellular fluxes. Gas chromatography-mass spectrometry analysis tracked the labeling patterns of protein-bound amino acids, revealing a metabolic network consisting of the glycolysis, the pentose phosphate pathway, and the tricarboxylic acid cycle with inactive glyoxylate shunt. Evidence of phosphoenolpyruvate carboxylase, phosphoenolpyruvate carboxykinase, and malic enzyme activity was also obtained. It was demonstrated that the relative activity of the pentose phosphate pathway to glycolysis under nitrogen-limited environment increased, reflecting excess NADPH requirements for lipid biosynthesis. Although the growth rate and cellular oil content were significantly altered in response to nitrogen limitation, global flux distribution of C. protothecoides remained stable, exhibiting the rigidity of central carbon metabolism. In conclusion, quantitative knowledge on the metabolic flux distribution of oleaginous alga obtained in this study may be of value in designing strategies for metabolic engineering of desirable bioproducts. PMID:20720172

  14. Double cross polarization /sup 13/C-NMR experiment in solid fossil fuel structure analysis

    SciTech Connect

    Hagaman, E.W.; Woody, M.C.

    1988-01-01

    The Double Cross Polarization /sup 13/C-MAS/NMR experiment has been used to derive a new operational classification of solid fossil fuels based on chemical reactivity. The method requires labeling reactive sites in the organic matrix with a magnetically active isotope not present in the precursor material, and using the local, isolated dipole-dipole interaction between this nucleus and nearby /sup 13/C nuclei to detect via cross polarization the carbon centers in the vicinity of the label. The technique is a marriage of chemistry and spectroscopy and the information content of the DCP spectra is defined by both partners. /sup 1/H-/sup 13/C-/sup 31/P DCP/MAS /sup 13/C-NMR spectroscopy has been used to statistically describe phenolic ortho-substitution patterns of coals via their aryl phosphinate or phosphate derivatives. In these applications of DCP NMR the new, detailed structure and/or reactivity information is realized by detection of carbon resonances one or more bonds removed from the reaction center, but in a volume element of intramolecular dimensions. To the extent that intermolecular contributions to the spectrum are detected, and not recognized as such, the structure/reactivity correlation is weakened. Direct substitution of phosphorus on the aromatic rings in the organic matrix of the coal is not readily accomplished. This environment potentially can be labeled with fluorine in a selective fashion using newly developed reagents. The possibility of determining the changes in average ring substitution patterns as a function of chemical treatment or coal diagenesis emerges. Recent developments in the field of DCP /sup 13/C NMR are presented.

  15. Triple resonance experiments for aligned sample solid-state NMR of 13C and 15N labeled proteins

    PubMed Central

    Sinha, Neeraj; Grant, Christopher V.; Park, Sang Ho; Brown, Jonathan Miles; Opella, Stanley J.

    2013-01-01

    Initial steps in the development of a suite of triple-resonance 1H/13C/15N solid-state NMR experiments applicable to aligned samples of 13C and 15N labeled proteins are described. The experiments take advantage of the opportunities for 13C detection without the need for homonuclear 13C/13C decoupling presented by samples with two different patterns of isotopic labeling. In one type of sample, the proteins are ~20% randomly labeled with 13C in all backbone and side chain carbon sites and ~100% uniformly 15N labeled in all nitrogen sites; in the second type of sample, the peptides and proteins are 13C labeled at only the α-carbon and 15N labeled at the amide nitrogen of a few residues. The requirement for homonuclear 13C/13C decoupling while detecting 13C signals is avoided in the first case because of the low probability of any two 13C nuclei being bonded to each other; in the second case, the labeled 13Cα sites are separated by at least three bonds in the polypeptide chain. The experiments enable the measurement of the 13C chemical shift and 1H–13C and 15N–13C heteronuclear dipolar coupling frequencies associated with the 13Cα and 13C′ backbone sites, which provide orientation constraints complementary to those derived from the 15N labeled amide backbone sites. 13C/13C spin-exchange experiments identify proximate carbon sites. The ability to measure 13C–15N dipolar coupling frequencies and correlate 13C and 15N resonances provides a mechanism for making backbone resonance assignments. Three-dimensional combinations of these experiments ensure that the resolution, assignment, and measurement of orientationally dependent frequencies can be extended to larger proteins. Moreover, measurements of the 13C chemical shift and 1H–13C heteronuclear dipolar coupling frequencies for nearly all side chain sites enable the complete three-dimensional structures of proteins to be determined with this approach. PMID:17293139

  16. Measurement of 13C chemical shift tensor principal values with a magic-angle turning experiment.

    PubMed

    Hu, J Z; Orendt, A M; Alderman, D W; Pugmire, R J; Ye, C; Grant, D M

    1994-08-01

    The magic-angle turning (MAT) experiment introduced by Gan is developed into a powerful and routine method for measuring the principal values of 13C chemical shift tensors in powdered solids. A large-volume MAT probe with stable rotation frequencies down to 22 Hz is described. A triple-echo MAT pulse sequence is introduced to improve the quality of the two-dimensional baseplane. It is shown that measurements of the principal values of chemical shift tensors in complex compounds can be enhanced by using either short contact times or dipolar dephasing pulse sequences to isolate the powder patterns from protonated or non-protonated carbons, respectively. A model compound, 1,2,3-trimethoxybenzene, is used to demonstrate these techniques, and the 13C principal values in 2,3-dimethylnaphthalene and Pocahontas coal are reported as typical examples.

  17. Pathway analysis using (13) C-glycerol and other carbon tracers reveals a bipartite metabolism of Legionella pneumophila.

    PubMed

    Häuslein, Ina; Manske, Christian; Goebel, Werner; Eisenreich, Wolfgang; Hilbi, Hubert

    2016-04-01

    Amino acids represent the prime carbon and energy source for Legionella pneumophila, a facultative intracellular pathogen, which can cause a life-threatening pneumonia termed Legionnaires' disease. Genome, transcriptome and proteome studies indicate that L. pneumophila also utilizes carbon substrates other than amino acids. We show here that glycerol promotes intracellular replication of L. pneumophila in amoeba or macrophages (but not extracellular growth) dependent on glycerol-3-phosphate dehydrogenase, GlpD. An L. pneumophila mutant strain lacking glpD was outcompeted by wild-type bacteria upon co-infection of amoeba, indicating an important role of glycerol during infection. Isotopologue profiling studies using (13) C-labelled substrates were performed in a novel minimal defined medium, MDM, comprising essential amino acids, proline and phenylalanine. In MDM, L. pneumophila utilized (13) C-labelled glycerol or glucose predominantly for gluconeogenesis and the pentose phosphate pathway, while the amino acid serine was used for energy generation via the citrate cycle. Similar results were obtained for L. pneumophila growing intracellularly in amoeba fed with (13) C-labelled glycerol, glucose or serine. Collectively, these results reveal a bipartite metabolism of L. pneumophila, where glycerol and carbohydrates like glucose are mainly fed into anabolic processes, while serine serves as major energy supply. PMID:26691313

  18. BEBEtr and BUBI: J-compensated concurrent shaped pulses for 1H-13C experiments

    NASA Astrophysics Data System (ADS)

    Ehni, Sebastian; Luy, Burkhard

    2013-07-01

    Shaped pulses designed for broadband excitation, inversion and refocusing are important tools in modern NMR spectroscopy to achieve robust pulse sequences especially in heteronuclear correlation experiments. A large variety of mostly computer-optimized pulse shapes exist for different desired bandwidths, available rf-field strengths, and tolerance to B1-inhomogeneity. They are usually derived for a single spin 1/2, neglecting evolution due to J-couplings. While pulses with constant resulting phase are selfcompensated for heteronuclear coupling evolution as long as they are applied exclusively on a single nucleus, the situation changes for concurrently applied pulse shapes. Using the example of a 1H,13C two spin system, two J-compensated pulse pairs for the application in INEPT-type transfer elements were optimized: a point-to-point pulse sandwich called BEBEtr, consisting of a broadband excitation and time-reversed excitation pulse, and a combined universal rotation and point-to-point pulse pair called BUBI, which acts as a refocusing pulse on 1H and a corresponding inversion pulse on 13C. After a derivation of quality factors and optimization protocols, a theoretical and experimental comparison with conventionally derived BEBOP, BIBOP, and BURBOP-180° pulses is given. While the overall transfer efficiency of a single pulse pair is only reduced by approximately 0.1%, resulting transfer to undesired coherences is reduced by several percent. In experiments this can lead to undesired phase distortions for pairs of uncompensated pulse shapes and even differences in signal intensities of 5-10% in HSQC and up to 68% in more complex COB-HSQC experiments.

  19. Effect of light and brine shrimp on skeletal δ 13C in the Hawaiian coral Porites compressa: a tank experiment

    NASA Astrophysics Data System (ADS)

    Grottoli, Andréa G.

    2002-06-01

    Previous experimental fieldwork showed that coral skeletal δ 13C values decreased when solar intensity was reduced, and increased in the absence of zooplankton. However, actual seasonal changes in solar irradiance levels are typically less pronounced than those used in the previous experiment and the effect of increases in the consumption of zooplankton in the coral diet on skeletal δ 13C remains relatively unknown. In the present study, the effects of four different light and heterotrophy regimes on coral skeletal δ 13C values were measured. Porites compressa corals were grown in outdoor flow-through tanks under 112%, 100%, 75%, and 50% light conditions at the Hawaii Institute of Marine Biology, Hawaii. In addition, corals were fed either zero, low, medium, or high concentrations of brine shrimp. Decreases in light from 100% resulted in significant decreases in δ 13C that is most likely due to a corresponding decrease in photosynthesis. Increases in light to 112% also resulted in a decrease in δ 13C values. This latter response may be a consequence of photoinhibition. The overall curved response in δ 13C values was described by a significant quadratic function. Increases in brine shrimp concentrations resulted in increased skeletal δ 13C levels. This unexpected outcome appears to be attributable to enhanced nitrogen supply associated with the brine shrimp diet which led to increased zooxanthellae concentrations, increased photosynthesis rates, and thus increased δ 13C values. This result highlights the potential influence of nutrients from heterotrophically acquired carbon in maintaining the zooxanthellae-host symbiosis in balance. In addition, evidence is presented that suggests that coral skeletal growth and δ 13C are decoupled. These results increase our knowledge of how light and heterotrophy affects the δ 13C of coral skeletons.

  20. IsoDesign: a software for optimizing the design of 13C-metabolic flux analysis experiments.

    PubMed

    Millard, Pierre; Sokol, Serguei; Letisse, Fabien; Portais, Jean-Charles

    2014-01-01

    The growing demand for (13) C-metabolic flux analysis ((13) C-MFA) in the field of metabolic engineering and systems biology is driving the need to rationalize expensive and time-consuming (13) C-labeling experiments. Experimental design is a key step in improving both the number of fluxes that can be calculated from a set of isotopic data and the precision of flux values. We present IsoDesign, a software that enables these parameters to be maximized by optimizing the isotopic composition of the label input. It can be applied to (13) C-MFA investigations using a broad panel of analytical tools (MS, MS/MS, (1) H NMR, (13) C NMR, etc.) individually or in combination. It includes a visualization module to intuitively select the optimal label input depending on the biological question to be addressed. Applications of IsoDesign are described, with an example of the entire (13) C-MFA workflow from the experimental design to the flux map including important practical considerations. IsoDesign makes the experimental design of (13) C-MFA experiments more accessible to a wider biological community. IsoDesign is distributed under an open source license at http://metasys.insa-toulouse.fr/software/isodes/

  1. Absolute bioavailability of evacetrapib in healthy subjects determined by simultaneous administration of oral evacetrapib and intravenous [13C8]‐evacetrapib as a tracer

    PubMed Central

    Aburub, Aktham; Ward, Chris; Hinds, Chris; Czeskis, Boris; Ruterbories, Kenneth; Suico, Jeffrey G.; Royalty, Jane; Ortega, Demetrio; Pack, Brian W.; Begum, Syeda L.; Annes, William F.; Lin, Qun; Small, David S.

    2015-01-01

    This open‐label, single‐period study in healthy subjects estimated evacetrapib absolute bioavailability following simultaneous administration of a 130‐mg evacetrapib oral dose and 4‐h intravenous (IV) infusion of 175 µg [13C8]‐evacetrapib as a tracer. Plasma samples collected through 168 h were analyzed for evacetrapib and [13C8]‐evacetrapib using high‐performance liquid chromatography/tandem mass spectrometry. Pharmacokinetic parameter estimates following oral and IV doses, including area under the concentration‐time curve (AUC) from zero to infinity (AUC[0‐∞]) and to the last measureable concentration (AUC[0‐tlast]), were calculated. Bioavailability was calculated as the ratio of least‐squares geometric mean of dose‐normalized AUC (oral : IV) and corresponding 90% confidence interval (CI). Bioavailability of evacetrapib was 44.8% (90% CI: 42.2–47.6%) for AUC(0‐∞) and 44.3% (90% CI: 41.8–46.9%) for AUC(0‐tlast). Evacetrapib was well tolerated with no reports of clinically significant safety assessment findings. This is among the first studies to estimate absolute bioavailability using simultaneous administration of an unlabeled oral dose with a 13C‐labeled IV microdose tracer at about 1/1000th the oral dose, with measurement in the pg/mL range. This approach is beneficial for poorly soluble drugs, does not require additional toxicology studies, does not change oral dose pharmacokinetics, and ultimately gives researchers another tool to evaluate absolute bioavailability. PMID:26639670

  2. Detection and quantification of chlordecone in contaminated soils from the French West Indies by GC-MS using the 13C10-chlordecone stable isotope as a tracer.

    PubMed

    Martin-Laurent, Fabrice; Sahnoun, Mehdi M; Merlin, Chloé; Vollmer, Guy; Lübke, Markus

    2014-04-01

    Chlordecone is an organochlorine insecticide that has been widely used to control banana weevil in the French West Indies. As a result of this intense use, up to 20,000 ha are contaminated by this insecticide in the French West Indies, and this causes environmental damage and health problems. A scenario of exposure was drawn by French authorities, based on land usage records. Many efforts have been made to monitor the occurrence of chlordecone and its main metabolites using different analytical methods, including GC, GC/MS, LC/MS, and NIRS. Although these different methods allow for the detection and quantification of chlordecone from soils, none of them estimate the bottleneck caused by extraction of this organochlorine from soils with high adsorption ability. In this study, we used (13)C10-chlordecone as a tracer to estimate chlordecone extraction yield and to quantify chlordecone in soil extracts based on the (13)C/(12)C isotope dilution. We report the optimization of (13)C10-chlordecone extraction from an Andosol. The method was found to be linear from 0.118 to 43 mg kg(-1) in the Andosol, with an instrumental detection limit estimated at 8.84 μg kg(-1). This method showed that chlordecone ranged from 35.4 down to 0.18 mg kg(-1) in Andosol, Nitisol, Ferralsol, and Fluvisol soil types. Traces of the metabolite β-monohydrochlordecone were detected in the Andosol, Nitisol, and Ferralsol soil samples. This last result indicates that this method could be useful to monitor the fate of chlordecone in soils of the French West Indies. PMID:23733305

  3. 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

  4. Following Glycolysis Using 13C NMR: An Experiment Adaptable to Different Undergraduate Levels

    NASA Astrophysics Data System (ADS)

    Mega, T. L.; Carlson, C. B.; Cleary, D. A.

    1997-12-01

    This paper describes a laboratory exercise where the glycolysis of [1-13C] glucose under anaerobic conditions was followed using 13C NMR spectroscopy. The exercise is described in terms of its suitability for a variety of different undergraduate levels, although the emphasis in this paper is on its use in a n advanced chemistry laboratory course. The kinetics of the loss of glucose and the production of ethanol were investigated and found not to fit simple first or second order kinetics. In addition, the relative reaction rates of the two anomeric forms of glucose were analyzed, and it was determined that the a anomeric form reacted faster than the β anomeric form. Using proton-coupled 13C NMR, some of the metabolites were identified including ethanol (major) and glycerol (minor). Reaction and spectroscopic details are included.

  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. 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.

  7. Estimation of continuous anthropogenic CO2: model-based evaluation of CO2, CO, δ13C(CO2) and Δ14C(CO2) tracer methods

    NASA Astrophysics Data System (ADS)

    Vardag, S. N.; Gerbig, C.; Janssens-Maenhout, G.; Levin, I.

    2015-11-01

    We investigate different methods for estimating anthropogenic CO2 using modeled continuous atmospheric concentrations of CO2 alone, as well as CO2 in combination with the surrogate tracers CO, δ13C(CO2) and Δ14C(CO2). These methods are applied at three hypothetical stations representing rural, urban and polluted conditions. We find that, independent of the tracer used, an observation-based estimate of continuous anthropogenic CO2 is not yet feasible at rural measurement sites due to the low signal-to-noise ratio of anthropogenic CO2 estimates at such settings. The tracers δ13C(CO2) and CO provide an accurate possibility to determine anthropogenic CO2 continuously, only if all CO2 sources in the catchment area are well characterized or calibrated with respect to their isotopic signature and CO to anthropogenic CO2 ratio. We test different calibration strategies for the mean isotopic signature and CO to CO2 ratio using precise Δ14C(CO2) measurements on monthly integrated as well as on grab samples. For δ13C(CO2), a calibration with annually averaged 14C(CO2) grab samples is most promising, since integrated sampling introduces large biases into anthropogenic CO2 estimates. For CO, these biases are smaller. The precision of continuous anthropogenic CO2 determination using δ13C(CO2) depends on measurement precision of δ13C(CO2) and CO2, while the CO method is mainly limited by the variation in natural CO sources and sinks. At present, continuous anthropogenic CO2 could be determined using the tracers δ13C(CO2) and/or CO with a precision of about 30 %, a mean bias of about 10 % and without significant diurnal discrepancies. Hypothetical future measurements of continuous Δ14C(CO2) with a precision of 5 ‰ are promising for anthropogenic CO2 determination (precision ca. 10-20 %) but are not yet available. The investigated tracer-based approaches open the door to improving, validating and reducing biases of highly resolved emission inventories using atmospheric

  8. SOIL RESPIRED D13C SIGNATURES REFLECT ROOT EXUDATE OR ROOT TURNOVER SIGNATURES IN AN ELEVATED CO2 AND OZONE MESOCOSM EXPERIMENT

    EPA Science Inventory

    Bulk tissue and root and soil respired d13C signatures were measured throughout the soil profile in a Ponderosa Pine mesocosm experiment exposed to ambient and elevated CO2 concentrations. For the ambient treatment, root (0-1mm, 1-2mm, and >2mm) and soil d13C signatures were ?24...

  9. 13C direct detected COCO-TOCSY: A tool for sequence specific assignment and structure determination in protonless NMR experiments

    NASA Astrophysics Data System (ADS)

    Balayssac, Stéphane; Jiménez, Beatriz; Piccioli, Mario

    2006-10-01

    A novel experiment is proposed to provide inter-residue sequential correlations among carbonyl spins in 13C detected, protonless NMR experiments. The COCO-TOCSY experiment connects, in proteins, two carbonyls separated from each other by three, four or even five bonds. The quantitative analysis provides structural information on backbone dihedral angles ϕ as well as on the side chain dihedral angles of Asx and Glx residues. This is the first dihedral angle constraint that can be obtained via a protonless approach. About 75% of backbone carbonyls in Calbindin D 9K, a 75 aminoacid dicalcium protein, could be sequentially connected via a COCO-TOCSY spectrum. 49 3J values were measured and related to backbone ϕ angles. Structural information can be extended to the side chain orientation of aminoacids containing carbonyl groups. Additionally, long range homonuclear coupling constants, 4JCC and 5JCC, could be measured. This constitutes an unprecedented case for proteins of medium and small size.

  10. 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.

  11. BEBE(tr) and BUBI: J-compensated concurrent shaped pulses for 1H-13C experiments.

    PubMed

    Ehni, Sebastian; Luy, Burkhard

    2013-07-01

    Shaped pulses designed for broadband excitation, inversion and refocusing are important tools in modern NMR spectroscopy to achieve robust pulse sequences especially in heteronuclear correlation experiments. A large variety of mostly computer-optimized pulse shapes exist for different desired bandwidths, available rf-field strengths, and tolerance to B1-inhomogeneity. They are usually derived for a single spin 1/2, neglecting evolution due to J-couplings. While pulses with constant resulting phase are selfcompensated for heteronuclear coupling evolution as long as they are applied exclusively on a single nucleus, the situation changes for concurrently applied pulse shapes. Using the example of a (1)H,(13)C two spin system, two J-compensated pulse pairs for the application in INEPT-type transfer elements were optimized: a point-to-point pulse sandwich called BEBE(tr), consisting of a broadband excitation and time-reversed excitation pulse, and a combined universal rotation and point-to-point pulse pair called BUBI, which acts as a refocusing pulse on (1)H and a corresponding inversion pulse on (13)C. After a derivation of quality factors and optimization protocols, a theoretical and experimental comparison with conventionally derived BEBOP, BIBOP, and BURBOP-180° pulses is given. While the overall transfer efficiency of a single pulse pair is only reduced by approximately 0.1%, resulting transfer to undesired coherences is reduced by several percent. In experiments this can lead to undesired phase distortions for pairs of uncompensated pulse shapes and even differences in signal intensities of 5-10% in HSQC and up to 68% in more complex COB-HSQC experiments.

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

    PubMed Central

    Takeuchi, Koh; Gal, Maayan; Takahashi, Hideo; Shimada, Ichio

    2011-01-01

    Described here is a set of three-dimensional (3D) NMR experiments that rely on CACA-TOCSY magnetization transfer via the weak 3JCαCα coupling. These pulse sequences, which resemble recently described 13C detected CACA-TOCSY (Takeuchi et al. 2010) experiments, are recorded in 1H2O, and use 1H excitation and detection. These experiments require alternate 13C-12C labeling together with perdeuteration, which allows utilizing the small 3JCαCα scalar coupling that is otherwise masked by the stronger 1JCC couplings in uniformly 13C labeled samples. These new experiments provide a unique assignment ladder-mark that yields bidirectional supra-sequential information and can readily straddle proline residues. Unlike the conventional HNCA experiment, which contains only sequential information to the 13Cα of the preceding residue, the 3D hnCA-TOCSY-caNH experiment can yield sequential correlations to alpha carbons in positions i−1, i + 1 and i−2. Furthermore, the 3D hNca-TOCSY-caNH and Hnca-TOC-SY-caNH experiments, which share the same magnetization pathway but use a different chemical shift encoding, directly couple the 15N-1H spin pair of residue i to adjacent amide protons and nitrogens at positions i−2, i−1, i + 1 and i + 2, respectively. These new experimental features make protein backbone assignments more robust by reducing the degeneracy problem associated with the conventional 3D NMR experiments. PMID:21110064

  13. Atmospheric tracer experiments for regional dispersion studies

    SciTech Connect

    Heffter, J.L.; Ferber, G.J.

    1980-01-01

    Tracer experiments are being conducted to verify atmospheric transport and dispersion calculations at distances from tens to hundreds of km from pollutant sources. In one study, a 2 1/2 year sampling program has been carried out at 13 sites located 30 to 140 km from a source of /sup 85/Kr at the Savannah River Plant in South Carolina. Average weekly concentrations as well as twice-daily concentrations were obtained. Sampling data and meteorological data, including surface, tower, and rawinsonde observations are available on magnetic tape for model verification studies. Some verification results for the Air Resources Laboratories Atmospheric Transort and Dispersion Model (ARL-ATAD) are shown for averaging periods from one week to two years.

  14. (13)C-detected NMR experiments for automatic resonance assignment of IDPs and multiple-fixing SMFT processing.

    PubMed

    Dziekański, Paweł; Grudziąż, Katarzyna; Jarvoll, Patrik; Koźmiński, Wiktor; Zawadzka-Kazimierczuk, Anna

    2015-06-01

    Intrinsically disordered proteins (IDPs) have recently attracted much interest, due to their role in many biological processes, including signaling and regulation mechanisms. High-dimensional (13)C direct-detected NMR experiments have proven exceptionally useful in case of IDPs, providing spectra with superior peak dispersion. Here, two such novel experiments recorded with non-uniform sampling are introduced, these are 5D HabCabCO(CA)NCO and 5D HNCO(CA)NCO. Together with the 4D (HACA)CON(CA)NCO, an extension of the previously published 3D experiments (Pantoja-Uceda and Santoro in J Biomol NMR 59:43-50, 2014. doi: 10.1007/s10858-014-9827-1), they form a set allowing for complete and reliable resonance assignment of difficult IDPs. The processing is performed with sparse multidimensional Fourier transform based on the concept of restricting (fixing) some of spectral dimensions to a priori known resonance frequencies. In our study, a multiple-fixing method was developed, that allows easy access to spectral data. The experiments were tested on a resolution-demanding alpha-synuclein sample. Due to superior peak dispersion in high-dimensional spectrum and availability of the sequential connectivities between four consecutive residues, the overwhelming majority of resonances could be assigned automatically using the TSAR program. PMID:25902761

  15. Sequential backbone assignment of uniformly 13C-labeled RNAs by a two-dimensional P(CC)H-TOCSY triple resonance NMR experiment.

    PubMed

    Wijmenga, S S; Heus, H A; Leeuw, H A; Hoppe, H; van der Graaf, M; Hilbers, C W

    1995-01-01

    A new 1H-13C-31P triple resonance experiment is described which allows unambiguous sequential backbone assignment in 13C-labeled oligonucleotides via through-bond coherence transfer from 31P via 13C to 1H. The approach employs INEPT to transfer coherence from 31P to 13C and homonuclear TOCSY to transfer the 13C coherence through the ribose ring, followed by 13C to 1H J-cross-polarisation. The efficiencies of the various possible transfer pathways are discussed. The most efficient route involves transfer of 31Pi coherence via C4'i and C4'i-1, because of the relatively large JPC4' couplings involved. Via the homonuclear and heteronuclear mixing periods, the C4'i and C4'i-1 coherences are subsequently transferred to, amongst others, H1'i and H1'i-1, respectively, leading to a 2D 1H-31P spectrum which allows a sequential assignment in the 31P-1H1' region of the spectrum, i.e. in the region where the proton resonances overlap least. The experiment is demonstrated on a 13C-labeled RNA hairpin with the sequence 5'(GGGC-CAAA-GCCU)3'.

  16. Atmospheric CO2 level affects plants' carbon use efficiency: insights from a 13C labeling experiment on sunflower stands

    NASA Astrophysics Data System (ADS)

    Gong, Xiaoying; Schäufele, Rudi; Schnyder, Hans

    2015-04-01

    The increase of atmospheric CO2 concentration has been shown to stimulate plant photosynthesis and (to a lesser extent) growth, thereby acting as a possible sink for the additional atmospheric CO2. However, this effect is dependent on the efficiency with which plants convert atmospheric carbon into biomass carbon, since a considerable proportion of assimilated carbon is returned to the atmosphere via plant respiration. As a core parameter for carbon cycling, carbon use efficiency of plants (CUE, the ratio of net primary production to gross primary production) quantifies the proportion of assimilated carbon that is incorporated into plant biomass. CUE has rarely been assessed based on measurements of complete carbon balance, due to methodological difficulties in measuring respiration rate of plants in light. Moreover, foliar respiration is known to be inhibited in light, thus foliar respiration rate is generally lower in light than in dark. However, this phenomenon, termed as inhibition of respiration in light (IRL), has rarely been assessed at the stand-scale and been incorporated into the calculation of CUE. Therefore, how CUE responses to atmospheric CO2 levels is still not clear. We studied CUE of sunflower stands grown at sub-ambient CO2 level (200 μmol mol-1) and elevated CO2 level (1000 μmol mol-1) using mesocosm-scale gas exchange facilities which enabled continuous measurements of 13CO2/12CO2 exchange. Appling steady-state 13C labeling, fluxes of respiration and photosynthesis in light were separated, and tracer kinetic in respiration was analyzed. This study provides the first data on CUE at a mesocosm-level including respiration in light in different CO2 environments. We found that CUE of sunflower was lower at an elevated CO2 level than at a sub-ambient CO2 level; and the ignorance of IRL lead to erroneous estimations of CUE. Variation in CUE at atmospheric CO2 levels was attributed to several mechanisms. In this study, CO2 enrichment i) affected the

  17. Proton-detected 3D 1H/13C/1H correlation experiment for structural analysis in rigid solids under ultrafast-MAS above 60 kHz

    NASA Astrophysics Data System (ADS)

    Zhang, Rongchun; Nishiyama, Yusuke; Ramamoorthy, Ayyalusamy

    2015-10-01

    A proton-detected 3D 1H/13C/1H chemical shift correlation experiment is proposed for the assignment of chemical shift resonances, identification of 13C-1H connectivities, and proximities of 13C-1H and 1H-1H nuclei under ultrafast magic-angle-spinning (ultrafast-MAS) conditions. Ultrafast-MAS is used to suppress all anisotropic interactions including 1H-1H dipolar couplings, while the finite-pulse radio frequency driven dipolar recoupling (fp-RFDR) pulse sequence is used to recouple dipolar couplings among protons and the insensitive nuclei enhanced by polarization transfer technique is used to transfer magnetization between heteronuclear spins. The 3D experiment eliminates signals from non-carbon-bonded protons and non-proton-bonded carbons to enhance spectral resolution. The 2D (F1/F3) 1H/1H and 2D 13C/1H (F2/F3) chemical shift correlation spectra extracted from the 3D spectrum enable the identification of 1H-1H proximity and 13C-1H connectivity. In addition, the 2D (F1/F2) 1H/13C chemical shift correlation spectrum, incorporated with proton magnetization exchange via the fp-RFDR recoupling of 1H-1H dipolar couplings, enables the measurement of proximities between 13C and even the remote non-carbon-bonded protons. The 3D experiment also gives three-spin proximities of 1H-1H-13C chains. Experimental results obtained from powder samples of L-alanine and L-histidine ṡ H2O ṡ HCl demonstrate the efficiency of the 3D experiment.

  18. Proton-detected 3D (1)H/(13)C/(1)H correlation experiment for structural analysis in rigid solids under ultrafast-MAS above 60 kHz.

    PubMed

    Zhang, Rongchun; Nishiyama, Yusuke; Ramamoorthy, Ayyalusamy

    2015-10-28

    A proton-detected 3D (1)H/(13)C/(1)H chemical shift correlation experiment is proposed for the assignment of chemical shift resonances, identification of (13)C-(1)H connectivities, and proximities of (13)C-(1)H and (1)H-(1)H nuclei under ultrafast magic-angle-spinning (ultrafast-MAS) conditions. Ultrafast-MAS is used to suppress all anisotropic interactions including (1)H-(1)H dipolar couplings, while the finite-pulse radio frequency driven dipolar recoupling (fp-RFDR) pulse sequence is used to recouple dipolar couplings among protons and the insensitive nuclei enhanced by polarization transfer technique is used to transfer magnetization between heteronuclear spins. The 3D experiment eliminates signals from non-carbon-bonded protons and non-proton-bonded carbons to enhance spectral resolution. The 2D (F1/F3) (1)H/(1)H and 2D (13)C/(1)H (F2/F3) chemical shift correlation spectra extracted from the 3D spectrum enable the identification of (1)H-(1)H proximity and (13)C-(1)H connectivity. In addition, the 2D (F1/F2) (1)H/(13)C chemical shift correlation spectrum, incorporated with proton magnetization exchange via the fp-RFDR recoupling of (1)H-(1)H dipolar couplings, enables the measurement of proximities between (13)C and even the remote non-carbon-bonded protons. The 3D experiment also gives three-spin proximities of (1)H-(1)H-(13)C chains. Experimental results obtained from powder samples of L-alanine and L-histidine ⋅ H2O ⋅ HCl demonstrate the efficiency of the 3D experiment.

  19. The conversion of phenylalanine to tyrosine in man. Direct measurement by continuous intravenous tracer infusions of L-(ring-/sup 2/H5)phenylalanine and L-(1-/sup 13/C) tyrosine in the postabsorptive state

    SciTech Connect

    Clarke, J.T.; Bier, D.M.

    1982-10-01

    Steady state phenylalanine and tyrosine turnover and the rate of conversion of phenylalanine of tyrosine in vivo were determined in 6 healthy postabsorptive adult volunteers. Continuous infusions of tracer amounts of L-(ring-/sup 2/H5)phenylalanine were determined intravenously for 13-14 hr. After 9-10 hr, a priming dose followed by a continuous infusion of L-(1-/sup 13/C)tyrosine was added and maintained, along with the (/sup 2/H5)phenylalanine infusion, for 4 hr. Venous plasma samples were obtained before the initiation of each infusion and every 30 min during the course of the combined (/sup 2/H5)phenylalanine and (/sup 13/C)tyrosine infusion for determination of isotopic enrichments of (/sup 2/H5)phenylalanine, (/sup 13/C)tyrosine, and (/sup 2/H4)tyrosine by gas chromatograph-mass spectrometric analysis of the N-trifluoroacetyl-, methyl ester derivatives of the amino acids. Calculated from the observed enrichments, free phenylalanine and tyrosine turnover rates were 36.1 +/- 5.1 mumole . kg-1 . h-1 and 39.8 +/- 3.5 mumole . kg-1 . h-1, respectively. Phenylalanine was converted to tyrosine at the rate of 5.83 +/- 0.59 mumole . kg-1 . h-1, accounting for approximately 16% of either the phenylalanine or the tyrosine flux. The results indicate that the normal basal steady state phenylalanine hydroxylase activity in vivo in man is lower than that obtained from phenylalanine loading studies. This supports the existence of some type of substance activation of the enzyme as reflected in the previously reported exponential relationship between phenylalanine concentration and phenylalanine hydroxylase activity in vitro. The use of continuous simultaneous infusions of tracer amounts of stable isotope-labeled phenylalanine and tyrosine provides a direct means for studying physiological regulation of phenylalanine hydroxylase activity in vivo.

  20. A roadmap for interpreting 13C metabolite labeling patterns from cells

    PubMed Central

    Buescher, Joerg M.; Antoniewicz, Maciek R.; Boros, Laszlo G.; Burgess, Shawn C.; Brunengraber, Henri; Clish, Clary B.; DeBerardinis, Ralph J.; Feron, Olivier; Frezza, Christian; Ghesquiere, Bart; Gottlieb, Eyal; Hiller, Karsten; Jones, Russell G.; Kamphorst, Jurre J.; Kibbey, Richard G.; Kimmelman, Alec C.; Locasale, Jason W.; Lunt, Sophia Y.; Maddocks, Oliver D. K.; Malloy, Craig; Metallo, Christian M.; Meuillet, Emmanuelle J.; Munger, Joshua; Nöh, Katharina; Rabinowitz, Joshua D.; Ralser, Markus; Sauer, Uwe; Stephanopoulos, Gregory; St-Pierre, Julie; Tennant, Daniel A.; Wittmann, Christoph; Vander Heiden, Matthew G.; Vazquez, Alexei; Vousden, Karen; Young, Jamey D.; Zamboni, Nicola; Fendt, Sarah-Maria

    2015-01-01

    Measuring intracellular metabolism has increasingly led to important insights in biomedical research. 13C tracer analysis, although less information-rich than quantitative 13C flux analysis that requires computational data integration, has been established as a time-efficient method to unravel relative pathway activities, qualitative changes in pathway contributions, and nutrient contributions. Here, we review selected key issues in interpreting 13C metabolite labeling patterns, with the goal of drawing accurate conclusions from steady state and dynamic stable isotopic tracer experiments. PMID:25731751

  1. 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.

  2. 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

  3. Asian Tracer Experiment and Atmospheric Modeling (TEAM) Project: Draft Field Work Plan for the Asian Long-Range Tracer Experiment

    SciTech Connect

    Allwine, K Jerry; Flaherty, Julia E.

    2007-08-01

    This report provides an experimental plan for a proposed Asian long-range tracer study as part of the international Tracer Experiment and Atmospheric Modeling (TEAM) Project. The TEAM partners are China, Japan, South Korea and the United States. Optimal times of year to conduct the study, meteorological measurements needed, proposed tracer release locations, proposed tracer sampling locations and the proposed durations of tracer releases and subsequent sampling are given. Also given are the activities necessary to prepare for the study and the schedule for completing the preparation activities leading to conducting the actual field operations. This report is intended to provide the TEAM members with the information necessary for planning and conducting the Asian long-range tracer study. The experimental plan is proposed, at this time, to describe the efforts necessary to conduct the Asian long-range tracer study, and the plan will undoubtedly be revised and refined as the planning goes forward over the next year.

  4. Soil matrix tracer contamination and canopy recycling did not impair ¹³CO₂ plant-soil pulse labelling experiments.

    PubMed

    Barthel, Matthias; Sturm, Patrick; Knohl, Alexander

    2011-09-01

    When conducting (13)CO(2) plant-soil pulse labelling experiments, tracer material might cause unwanted side effects which potentially affect δ(13)C measurements of soil respiration (δ(13)C(SR)) and the subsequent data interpretation. First, when the soil matrix is not isolated from the atmosphere, contamination of the soil matrix with tracer material occurs leading to a physical back-diffusion from soil pores. Second, when using canopy chambers continuously, (13)CO(2) is permanently re-introduced into the atmosphere due to leaf respiration which then aids re-assimilation of tracer material by the canopy. Accordingly, two climate chamber experiments on European beech saplings (Fagus sylvatica L.) were conducted to evaluate the influence of soil matrix (13)CO(2) contamination and canopy recycling on soil (13)CO(2) efflux during (13)CO(2) plant-soil pulse labelling experiments. For this purpose, a combined soil/canopy chamber system was developed which separates soil and canopy compartments in order to (a) prevent diffusion of (13)C tracer into the soil chamber during a (13)CO(2) canopy pulse labelling and (b) study stable isotope processes in soil and canopy individually and independently. In combination with laser spectrometry measuring CO(2) isotopologue mixing ratios at a rate of 1 Hz, we were able to measure δ(13)C in canopy and soil at very high temporal resolution. For the soil matrix contamination experiment, (13)CO(2) was applied to bare soil, canopy only or, simultaneously, to soil and canopy of the beech trees. The obtained δ(13)C(SR) fluxes from the different treatments were then compared with respect to label re-appearance, first peak time and magnitude. By determining the δ(13)C(SR) decay of physical (13)CO(2) back-diffusion from bare soils (contamination), it was possible to separate biological and physical components in δ(13)C(SR) of a combined flux of both. A second pulse labelling experiment, with chambers permanently enclosing the canopy

  5. An intraresidual i(HCA)CO(CA)NH experiment for the assignment of main-chain resonances in 15N, 13C labeled proteins.

    PubMed

    Mäntylahti, Sampo; Tossavainen, Helena; Hellman, Maarit; Permi, Perttu

    2009-11-01

    An improved pulse sequence, intraresidual i(HCA)CO(CA)NH, is described for establishing solely (13)C'(i), (15)N(i), (1)HN(i) connectivities in uniformly 15N/13C-labeled proteins. In comparison to the "out-and-back" style intra-HN(CA)CO experiment, the new pulse sequence offers at least two-fold higher experimental resolution in the (13)C' dimension and on average 1.6 times higher sensitivity especially for residues in alpha-helices. Performance of the new experiment was tested on a small globular protein ubiquitin and an intrinsically unfolded 110-residue cancer/testis antigen CT16/PAGE5. Use of intraresidual i(HCA)CO(CA)NH experiment in combination with the established HNCO experiment was crucial for the assignment of highly disordered CT16. PMID:19768387

  6. Interpreting tracer breakthrough tailing from different forced-gradient tracer experiment configurations in fractured bedrock

    USGS Publications Warehouse

    Becker, M.W.; Shapiro, A.M.

    2003-01-01

    Conceptual and mathematical models are presented that explain tracer breakthrough tailing in the absence of significant matrix diffusion. Model predictions are compared to field results from radially convergent, weak-dipole, and push-pull tracer experiments conducted in a saturated crystalline bedrock. The models are based upon the assumption that flow is highly channelized, that the mass of tracer in a channel is proportional to the cube of the mean channel aperture, and the mean transport time in the channel is related to the square of the mean channel aperture. These models predict the consistent -2 straight line power law slope observed in breakthrough from radially convergent and weak-dipole tracer experiments and the variable straight line power law slope observed in push-pull tracer experiments with varying injection volumes. The power law breakthrough slope is predicted in the absence of matrix diffusion. A comparison of tracer experiments in which the flow field was reversed to those in which it was not indicates that the apparent dispersion in the breakthrough curve is partially reversible. We hypothesize that the observed breakthrough tailing is due to a combination of local hydrodynamic dispersion, which always increases in the direction of fluid velocity, and heterogeneous advection, which is partially reversed when the flow field is reversed. In spite of our attempt to account for heterogeneous advection using a multipath approach, a much smaller estimate of hydrodynamic dispersivity was obtained from push-pull experiments than from radially convergent or weak dipole experiments. These results suggest that although we can explain breakthrough tailing as an advective phenomenon, we cannot ignore the relationship between hydrodynamic dispersion and flow field geometry at this site. The design of the tracer experiment can severely impact the estimation of hydrodynamic dispersion and matrix diffusion in highly heterogeneous geologic media.

  7. Sources and transfers of particulate organic matter in a tropical reservoir (Petit Saut, French Guiana): a multi-tracers analysis using δ13C, C/N ratio and pigments

    NASA Astrophysics Data System (ADS)

    de Junet, A.; Abril, G.; Guérin, F.; Billy, I.; de Wit, R.

    2005-08-01

    sedimentary POM. This material was undergoing intense degradation as revealed by high concentration of pheopigments and by an increase in C/N ratio and an increase in δ13C-OC with trap depth. Scytonemin was found in a biofilm developed on tree trunks at the reservoir surface and in all sediment traps. Other tracers showed however that the contribution of the biofilm to the sedimentary POM was minor compared to the planktonic source. In the Sinnamary downstream of the dam, POM became more 13C-depleted showing a larger contribution of methanotrophic bacteria. Chl b, Lutein and BChl c + BChl d originating from the reservoir progressively decreased downstream as the result of mineralization. At the estuarine mouth, fucoxanthin showed the presence of diatoms and the δ13C-C/N signature matched the one of POM carried by the Amazonian coastal mobile mud belt.

  8. Inferring residence time distributions from hillslope tracer experiments

    NASA Astrophysics Data System (ADS)

    McGuire, K. J.; Weiler, M.; McDonnell, J. J.

    2003-12-01

    Water residence time in catchments is typically determined by black-box modeling of environmental tracers (e.g., 18O and 2H), in which input (rainfall) and output (discharge) tracer concentrations are used to estimate parameters of an assumed distribution that represents the residence time. In this study, a different approach was taken, where the residence time distribution (RTD) was determined directly from an artificial tracer experiment. Two tracers (Amino G acid and bromide) were applied as line sources (20 and 40 m from the stream, respectively) to the soil surface of a steep forested hillslope in the H.J. Andrews Experimental Forest. Tracer concentrations and seepage flow were monitored for approximately 100 days. The resulting breakthrough curves were modeled using a simple, process-based hillslope model conditional on spatially varying soil depth and depth-varying drainable porosity and hydraulic conductivity. The model preserved internal hydrological behavior such as water table dynamics and satisfactorily reproduced seepage discharge and tracer concentrations. Using the model and assuming the same parameterization, we infer the RTD of conservative tracers applied to the entire hillslope. The derived RTDs were then compared to functions that represent RTDs used in black-box models.

  9. Visualization of enantiomers using natural abundant (13)C-filtered single and double quantum selective refocusing experiments: Application to small chiral molecules.

    PubMed

    Nath, Nilamoni; Baishya, Bikash; Suryaprakash, N

    2009-09-01

    The routine use of proton NMR for the visualization of enantiomers, aligned in the chiral liquid crystal solvent poly-gamma-benzyl-l-glutamate (PBLG), is restricted due to severe loss of resolution arising from large number of pair wise interaction of nuclear spins. In the present study, we have designed two experimental techniques for their visualization utilizing the natural abundance (13)C edited selective refocusing of single quantum (CH-SERF) and double quantum (CH-DQSERF) coherences. The methods achieve chiral discrimination and aid in the simultaneous determination of homonuclear couplings between active and passive spins and heteronuclear couplings between the excited protons and the participating (13)C spin. The CH-SERF also overcomes the problem of overlap of central transitions of the methyl selective refocusing (SERF) experiment resulting in better chiral discrimination. Theoretical description of the evolution of magnetization in both the sequences has been discussed using polarization operator formalism.

  10. Tracer experiment design for unique identification of nonlinear physiological systems.

    PubMed

    DiStefano, J J

    1976-02-01

    The design of tracer kinetic experiments, the purpose of which is to elucidate uniquely the internal couplings of a nonlinear dynamic system, is considered for a practical class of models of physiological systems. The extent of information about the real system contained in tracer kinetic data is a central issue. Criteria for determining whether nonlinear model parameters can be estimated from small-signal, "linearizing" tracer experiments are developed and illustrated by examples. The concept of "structural identifiability" is employed in this analysis to determine which model parameters can be and which cannot be determined "uniquely" from given input-output data; a step-by-step procedure based on an extension of this concept is presented for adapting the overall approach to the experimental design problem. Estimation of unmeasurable endogenous inputs and system state variables, problems that are intimately related to parameter estimation for physiological systems, are also considered. PMID:1259027

  11. High-throughput backbone resonance assignment of small 13C, 15N-labeled proteins by a triple-resonance experiment with four sequential connectivity pathways using chemical shift-dependent, apparent 1J ( 1H, 13C): HNCACB codedHAHB

    NASA Astrophysics Data System (ADS)

    Pegan, Scott; Kwiatkowski, Witek; Choe, Senyon; Riek, Roland

    2003-12-01

    The proposed three-dimensional triple-resonance experiment HNCACB codedHAHB correlates sequential 15N, 1H moieties via the chemical shifts of 13C α, 13C β, 1H α, and 1H β. The four sequential correlation pathways are achieved by the incorporation of the concept of chemical shift-coding [J. Biomol. NMR 25 (2003) 281] to the TROSY-HNCACB experiment. The monitored 1H α and 1H β chemical shifts are then coded in the line shape of the cross-peaks of 13C α, 13C β along the 13C dimension through an apparent residual scalar coupling, the size of which depends on the attached hydrogen chemical shift. The information of four sequential correlation pathways enables a rapid backbone assignment. The HNCACB codedHAHB experiment was applied to ˜85% labeled 13C, 15N-labeled amino-terminal fragment of Vaccinia virus DNA topoisomerase I comprising residues 1-77. After one day of measurement on a Bruker Avance 700 MHz spectrometer and 8 h of manual analysis of the spectrum 93% of the backbone assignment was achieved.

  12. 13C-1H dipolar-driven 13C-13C recoupling without 13C rf irradiation in nuclear magnetic resonance of rotating solids

    NASA Astrophysics Data System (ADS)

    Takegoshi, K.; Nakamura, Shinji; Terao, Takehiko

    2003-02-01

    Two recently proposed 13C-13C recoupling methods under magic angle spinning (MAS), resonant interference recoupling (RIR), and 13C-1H dipolar-assisted rotational resonance (DARR), are examined on a common theoretical foundation using the average Hamiltonian theory. In both methods, a rf field is applied on not 13C but 1H to recouple the 13C-1H dipolar interactions, and spectral overlap necessary to conserve energy for 13C-13C polarization transfer is achieved by the 13C-1H dipolar line broadening. While DARR employs time-independent 13C-1H interactions recoupled by suitable rf irradiation to 1H spins, RIR uses time-dependent 13C-1H interactions modulated appropriately by 1H rf irradiation. There are two distinct cases where 13C-1H line broadening realizes 13C-13C spectral overlap. For a pair of a carbonyl or aromatic carbon and an aliphatic carbon, spectral overlap can be achieved between one of the spinning sidebands of the former 13C resonance and the 13C-1H dipolar powder pattern of the latter. On the other hand for a pair of spins with a small chemical shift difference, the two center bands are overlapped with each other due to 13C-1H dipolar broadening. For the former, we show that both RIR and DARR occur in the first order, while for the latter, DARR recoupling is appreciable for time-independent 13C-1H interactions. We refer to the former DARR as the first-order DARR recoupling and the latter as the second-order DARR. Experimentally, we examined the following 13C-1H recoupling methods for DARR: 1H CW irradiation fulfilling a rotary-resonance condition or a modulatory-resonance condition, and 1H π pulses applied synchronously to MAS. For RIR, the FSLG-m2m¯m sequence is applied to 1H. Several one-dimensional DARR and RIR experiments were done for N-acetyl[1,2-13C, 15N] DL-valine, and [2,3-13C] L-alanine. It was found that the polarization transfer rate for RIR is larger than that for DARR except for fast spinning, while the rate for DARR is less sensitive to

  13. Conformational Analysis, Thermal Rearrangement, and EI-MS Fragmentation Mechanism of (1(10)E,4E,6S,7R)-Germacradien-6-ol by (13)C-Labeling Experiments.

    PubMed

    Rabe, Patrick; Barra, Lena; Rinkel, Jan; Riclea, Ramona; Citron, Christian A; Klapschinski, Tim A; Janusko, Aron; Dickschat, Jeroen S

    2015-11-01

    An uncharacterized terpene cyclase from Streptomyces pratensis was identified as (+)-(1(10)E,4E,6S,7R)-germacradien-6-ol synthase. The enzyme product exists as two interconvertible conformers, resulting in complex NMR spectra. For the complete assignment of NMR data, all fifteen ((13)C1)FPP isotopomers (FPP=farnesyl diphosphate) and ((13)C15)FPP were synthesized and enzymatically converted. The products were analyzed using various NMR techniques, including (13)C, (13)C COSY experiments. The ((13)C)FPP isotopomers were also used to investigate the thermal rearrangement and EI fragmentation of the enzyme product. PMID:26361082

  14. Conformational Analysis, Thermal Rearrangement, and EI-MS Fragmentation Mechanism of (1(10)E,4E,6S,7R)-Germacradien-6-ol by (13)C-Labeling Experiments.

    PubMed

    Rabe, Patrick; Barra, Lena; Rinkel, Jan; Riclea, Ramona; Citron, Christian A; Klapschinski, Tim A; Janusko, Aron; Dickschat, Jeroen S

    2015-11-01

    An uncharacterized terpene cyclase from Streptomyces pratensis was identified as (+)-(1(10)E,4E,6S,7R)-germacradien-6-ol synthase. The enzyme product exists as two interconvertible conformers, resulting in complex NMR spectra. For the complete assignment of NMR data, all fifteen ((13)C1)FPP isotopomers (FPP=farnesyl diphosphate) and ((13)C15)FPP were synthesized and enzymatically converted. The products were analyzed using various NMR techniques, including (13)C, (13)C COSY experiments. The ((13)C)FPP isotopomers were also used to investigate the thermal rearrangement and EI fragmentation of the enzyme product.

  15. Rate and δ13C values of CO2 produced during short-term online incubation experiments indicate two different processes with distinct carbon sources

    NASA Astrophysics Data System (ADS)

    Osuna, R.; Breecker, D. O.; Sharp, Z. D.

    2009-12-01

    The decomposition of organic matter in soils is a fundamental process influencing the carbon cycle. The rate of microbial oxidation of organic matter is known to increase with temperature, suggesting that many soils may turn from sinks to sources of CO2 as global temperature warms. In this study, we introduce a new, online soil incubation technique that can be used to determine the rate and carbon isotope composition (δ13C) of CO2 respired during the decomposition of organic matter. Air-dried and sieved soil samples are mechanically mixed and are rewetted and preincubated in capped 50 mL tubes. Aliquots of the soils (2 - 200 mg) taken from the storage tubes are loaded into septum-capped Labco Exetainer® vials which are themselves loaded into a thermostated sample tray maintained at a constant temperature. The vials are then flushed with synthetic CO2-free air and the soils are preincubated again for several days. The incubation experiment begins when the vials are flushed a second time with synthetic air. At the end of the incubation, all of the CO2 that was produced is flushed from the vial, cryofocused in a liquid nitrogen trap and then carried into the source of a mass spectrometer in a He stream. The CO2 from each vial is sampled automatically and directed into the mass spectrometer using a Thermo Electron Gasbench II. The number of moles and the δ13C value of CO2 produced during the incubation are determined from voltages measured by the mass spectrometer. The technique allows for rapid sample analysis and short incubation times (hours to days). Incubation of multiple aliquots of the same soil mixture allows trends to be examined. Several patterns have emerged from incubations carried out to date. An initial pulse of CO2 is emitted from the soils during the first minutes of incubation. This pulse of CO2 is emitted from dry soils (i.e. never rewetted), sterilized soils, and from soils incubated in high purity He instead of synthetic air, indicating that the

  16. The 2D {31P} Spin-Echo-Difference Constant-Time [13C, 1H]-HMQC Experiment for Simultaneous Determination of 3JH3‧P and 3JC4‧P in 13C-Labeled Nucleic Acids and Their Protein Complexes

    NASA Astrophysics Data System (ADS)

    Szyperski, Thomas; Fernández, César; Ono, Akira; Wüthrich, Kurt; Kainosho, Masatsune

    1999-10-01

    A two-dimensional {31P} spin-echo-difference constant-time [13C, 1H]-HMQC experiment (2D {31P}-sedct-[13C, 1H]-HMQC) is introduced for measurements of 3JC4‧P and 3JH3‧P scalar couplings in large 13C-labeled nucleic acids and in DNA-protein complexes. This experiment makes use of the fact that 1H-13C multiple-quantum coherences in macromolecules relax more slowly than the corresponding 13C single-quantum coherences. 3JC4‧P and 3JH3‧P are related via Karplus-type functions with the phosphodiester torsion angles β and ɛ, respectively, and their experimental assessment therefore contributes to further improved quality of NMR solution structures. Data are presented for a uniformly 13C, 15N-labeled 14-base-pair DNA duplex, both free in solution and in a 17-kDa protein-DNA complex.

  17. 13-C NMR Spectra of Styrene Derivatives: An Undergraduate Experiment Involving the Application of the Hammett Equation.

    ERIC Educational Resources Information Center

    Blunt, J. W.; Happer, D. A. R.

    1979-01-01

    In this experiment, the observation that, for meta- and para-substituted styrene derivatives, the magnitude of the C-13 shielding value for the beta-carbon is linearly related to sigma for the ring substituent is used to determine sigma. (BB)

  18. OpenMebius: An Open Source Software for Isotopically Nonstationary 13C-Based Metabolic Flux Analysis

    PubMed Central

    Furusawa, Chikara

    2014-01-01

    The in vivo measurement of metabolic flux by 13C-based metabolic flux analysis (13C-MFA) provides valuable information regarding cell physiology. Bioinformatics tools have been developed to estimate metabolic flux distributions from the results of tracer isotopic labeling experiments using a 13C-labeled carbon source. Metabolic flux is determined by nonlinear fitting of a metabolic model to the isotopic labeling enrichment of intracellular metabolites measured by mass spectrometry. Whereas 13C-MFA is conventionally performed under isotopically constant conditions, isotopically nonstationary 13C metabolic flux analysis (INST-13C-MFA) has recently been developed for flux analysis of cells with photosynthetic activity and cells at a quasi-steady metabolic state (e.g., primary cells or microorganisms under stationary phase). Here, the development of a novel open source software for INST-13C-MFA on the Windows platform is reported. OpenMebius (Open source software for Metabolic flux analysis) provides the function of autogenerating metabolic models for simulating isotopic labeling enrichment from a user-defined configuration worksheet. Analysis using simulated data demonstrated the applicability of OpenMebius for INST-13C-MFA. Confidence intervals determined by INST-13C-MFA were less than those determined by conventional methods, indicating the potential of INST-13C-MFA for precise metabolic flux analysis. OpenMebius is the open source software for the general application of INST-13C-MFA. PMID:25006579

  19. OpenMebius: an open source software for isotopically nonstationary 13C-based metabolic flux analysis.

    PubMed

    Kajihata, Shuichi; Furusawa, Chikara; Matsuda, Fumio; Shimizu, Hiroshi

    2014-01-01

    The in vivo measurement of metabolic flux by (13)C-based metabolic flux analysis ((13)C-MFA) provides valuable information regarding cell physiology. Bioinformatics tools have been developed to estimate metabolic flux distributions from the results of tracer isotopic labeling experiments using a (13)C-labeled carbon source. Metabolic flux is determined by nonlinear fitting of a metabolic model to the isotopic labeling enrichment of intracellular metabolites measured by mass spectrometry. Whereas (13)C-MFA is conventionally performed under isotopically constant conditions, isotopically nonstationary (13)C metabolic flux analysis (INST-(13)C-MFA) has recently been developed for flux analysis of cells with photosynthetic activity and cells at a quasi-steady metabolic state (e.g., primary cells or microorganisms under stationary phase). Here, the development of a novel open source software for INST-(13)C-MFA on the Windows platform is reported. OpenMebius (Open source software for Metabolic flux analysis) provides the function of autogenerating metabolic models for simulating isotopic labeling enrichment from a user-defined configuration worksheet. Analysis using simulated data demonstrated the applicability of OpenMebius for INST-(13)C-MFA. Confidence intervals determined by INST-(13)C-MFA were less than those determined by conventional methods, indicating the potential of INST-(13)C-MFA for precise metabolic flux analysis. OpenMebius is the open source software for the general application of INST-(13)C-MFA.

  20. Multi-objective experimental design for (13)C-based metabolic flux analysis.

    PubMed

    Bouvin, Jeroen; Cajot, Simon; D'Huys, Pieter-Jan; Ampofo-Asiama, Jerry; Anné, Jozef; Van Impe, Jan; Geeraerd, Annemie; Bernaerts, Kristel

    2015-10-01

    (13)C-based metabolic flux analysis is an excellent technique to resolve fluxes in the central carbon metabolism but costs can be significant when using specialized tracers. This work presents a framework for cost-effective design of (13)C-tracer experiments, illustrated on two different networks. Linear and non-linear optimal input mixtures are computed for networks for Streptomyces lividans and a carcinoma cell line. If only glucose tracers are considered as labeled substrate for a carcinoma cell line or S. lividans, the best parameter estimation accuracy is obtained by mixtures containing high amounts of 1,2-(13)C2 glucose combined with uniformly labeled glucose. Experimental designs are evaluated based on a linear (D-criterion) and non-linear approach (S-criterion). Both approaches generate almost the same input mixture, however, the linear approach is favored due to its low computational effort. The high amount of 1,2-(13)C2 glucose in the optimal designs coincides with a high experimental cost, which is further enhanced when labeling is introduced in glutamine and aspartate tracers. Multi-objective optimization gives the possibility to assess experimental quality and cost at the same time and can reveal excellent compromise experiments. For example, the combination of 100% 1,2-(13)C2 glucose with 100% position one labeled glutamine and the combination of 100% 1,2-(13)C2 glucose with 100% uniformly labeled glutamine perform equally well for the carcinoma cell line, but the first mixture offers a decrease in cost of $ 120 per ml-scale cell culture experiment. We demonstrated the validity of a multi-objective linear approach to perform optimal experimental designs for the non-linear problem of (13)C-metabolic flux analysis. Tools and a workflow are provided to perform multi-objective design. The effortless calculation of the D-criterion can be exploited to perform high-throughput screening of possible (13)C-tracers, while the illustrated benefit of multi

  1. Temporal moments in geoelectrical monitoring of salt tracer experiments

    NASA Astrophysics Data System (ADS)

    Pollock, Davina; Cirpka, Olaf A.

    2008-12-01

    Monitoring of salt tracer experiments by electrical resistivity tomography (ERT) has been shown to be a valuable tool for characterizing the hydraulics of an aquifer, but efficient approaches of determining the spatial hydraulic conductivity distribution from ERT data are still missing. Standard inversion of ERT data obtained during salt tracer tests may even lead to estimates of the concentration distribution that are in contradiction to flow and transport of conservative compounds in porous media. In order to avoid nonphysical behavior, we consider the governing equations of groundwater flow, solute transport, and geoelectrics as a coupled system. While the tracer passes through the part of the domain that is sensitive for a particular electrode configuration, the measured electrical potential differences are perturbed. We characterize these perturbations by their temporal moments and relate them to the temporal moments of concentration, which themselves depend on hydraulic conductivity. We present temporal moment-generating equations leading from the hydraulic conductivity field via heads and velocities to the temporal moments of concentration and electrical potential perturbations. The approach makes use of a linearized version of the Poisson equation. On the basis of this system of coupled steady state equations, we compute the sensitivity of electrical potential perturbations with respect to the log hydraulic conductivity distribution by the continuous adjoint state method for coupled systems. For demonstration, we simulate salt tracer experiments in a virtual quasi-two-dimensional sandbox, monitored by ERT. We show that the ratio of the first over the zeroth temporal moment of potential perturbation is less affected by the linearization of the Poisson equation than the zeroth and first moments themselves. Thus, it appears recommendable to use the ratio of first to zeroth moments also as data in inversion. We compare sensitivity patterns resulting from

  2. North beach (Nazaré) sand tracer experiment

    NASA Astrophysics Data System (ADS)

    Duarte, João; Taborda, Rui; Ribeiro, Mónica; Cascalho, João; Silva, Ana; Bosnic, Ivana

    2014-05-01

    The littoral in the vicinity of Nazaré (West Portuguese coast) is characterized by two distinct coastal stretches separated by Nazaré headland: a northern sector (Norte beach) characterized by a high energetic continuous sandy beach and a southern sector (Nazaré bay beach) that corresponds to an embayed beach, sheltered by the Nazaré headland. The bay is a geomorphological expression of the Nazaré canyon head, which acts as powerful sediment sink, capturing the large longshore net southward transport at Norte beach generated by the north Atlantic high energetic swell. The northern side of the canyon head is carved on highly resistant Cretaceous limestone sustaining an underwater vertical relief that emerges on the Nazaré headland, creating a unusual nearshore wave pattern. This wave pattern not only concentrates high energy levels at the Norte beach but also contributes to local complex longshore drift gradients capable of inducing beach seasonal cross-shore variations of more than 200 m. The main factors that influence local sediment budget are: (1) canyon head capturing and (2) headland sediment bypassing. To obtain a direct measure of the net longshore drift at Norte beach (upstream boundary of the system) a large scale fluorescent tracer experiment was performed. The data will be used to validate longshore transport formulas in a high energetic environment and to access Nazaré canyon head sediment loss. Considering the anticipation of high transport rates, approximately 10 tonnes of native sand where coated with orange fluorescent ink using a set of concrete mixers. The experiment took place on the 9th to 15th September 2013 period and followed the continuous injection method (CIM). The CIM approach was justified by the expected high energy levels that inhibits sediment sampling across the surf zone. During the tracer injection procedure (approx. 5 hours), sediment sampling was performed at 13 sites along a rectilinear coastal stretch extended through

  3. Experiments on Tracer Diffusion in Water and Aqueous Mixtures

    NASA Astrophysics Data System (ADS)

    Spiegel, Dan

    2015-03-01

    We have used forced Rayleigh scattering to measure tracer diffusion coefficients in water and other liquids. The tracer molecule is the azobenzene derivative methyl red (MR). In one set of experiments diffusion was measured at different temperatures to test the Cohen-Turnbull (CT) free volume diffusion theory. It was found that eight solvents were in reasonable agreement with CT, but the relevant energy in water was smaller by an order of magnitude. We believe this is due to the ``zero-point'' free volume that water would possess, due to its H-bonds, even at the glass transition temperature, and to the ability of MR to diffuse along its plane. In a second set of experiments we studied diffusion in aqueous and non-aqueous mixtures. The non-aqueous mixtures are in good agreement with free volume theory, but the aqueous mixtures show large disagreement. We suggest this is caused by the formation of two solvent shells in the aqueous mixtures, driven by the hydrophobicity of MR and water-amphiphile hydrogen bonding. NSF Research at Undergraduate Institutions.

  4. Proton-detected 3D {sup 1}H/{sup 13}C/{sup 1}H correlation experiment for structural analysis in rigid solids under ultrafast-MAS above 60 kHz

    SciTech Connect

    Zhang, Rongchun; Ramamoorthy, Ayyalusamy; Nishiyama, Yusuke

    2015-10-28

    A proton-detected 3D {sup 1}H/{sup 13}C/{sup 1}H chemical shift correlation experiment is proposed for the assignment of chemical shift resonances, identification of {sup 13}C-{sup 1}H connectivities, and proximities of {sup 13}C-{sup 1}H and {sup 1}H-{sup 1}H nuclei under ultrafast magic-angle-spinning (ultrafast-MAS) conditions. Ultrafast-MAS is used to suppress all anisotropic interactions including {sup 1}H-{sup 1}H dipolar couplings, while the finite-pulse radio frequency driven dipolar recoupling (fp-RFDR) pulse sequence is used to recouple dipolar couplings among protons and the insensitive nuclei enhanced by polarization transfer technique is used to transfer magnetization between heteronuclear spins. The 3D experiment eliminates signals from non-carbon-bonded protons and non-proton-bonded carbons to enhance spectral resolution. The 2D (F1/F3) {sup 1}H/{sup 1}H and 2D {sup 13}C/{sup 1}H (F2/F3) chemical shift correlation spectra extracted from the 3D spectrum enable the identification of {sup 1}H-{sup 1}H proximity and {sup 13}C-{sup 1}H connectivity. In addition, the 2D (F1/F2) {sup 1}H/{sup 13}C chemical shift correlation spectrum, incorporated with proton magnetization exchange via the fp-RFDR recoupling of {sup 1}H-{sup 1}H dipolar couplings, enables the measurement of proximities between {sup 13}C and even the remote non-carbon-bonded protons. The 3D experiment also gives three-spin proximities of {sup 1}H-{sup 1}H-{sup 13}C chains. Experimental results obtained from powder samples of L-alanine and L-histidine ⋅ H{sub 2}O ⋅ HCl demonstrate the efficiency of the 3D experiment.

  5. Impacts of proline on the central metabolism of an industrial erythromycin-producing strain Saccharopolyspora erythraea via (13)C labeling experiments.

    PubMed

    Hong, Ming; Huang, Mingzhi; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2016-08-10

    Saccharopolyspora erythraea E3 is an important industrial strain for erythromycin production and knowledge on its metabolism is limited. In the present work, (13)C labeling experiments were conducted to characterize the metabolism of S. erythraea E3. We found that S. erythraea E3 was difficult to grow on minimal medium with glucose as sole carbon source and the addition of proline remarkably improved the cell growth. The activity of EMP pathway was very low and ED pathway was alternatively the main glucose utilization pathway. The addition of proline resulted in remarkable changes in the fluxes of central metabolism. The fluxes in PP pathway, in TCA cycle and in ED pathway were 90% higher, 64% and 31% lower on Glc/Pro than on Glc, respectively. The maintenance energy on Glc/Pro was 58.4% lower than that on Glc. The energy charge was lower on Glc than on Glc/Pro, indicating that the cells on Glc suffered from energy burden. This study elucidates the impacts of proline on the central metabolism of S. erythraea and deepens the understanding of its metabolism. PMID:27215341

  6. Impacts of proline on the central metabolism of an industrial erythromycin-producing strain Saccharopolyspora erythraea via (13)C labeling experiments.

    PubMed

    Hong, Ming; Huang, Mingzhi; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2016-08-10

    Saccharopolyspora erythraea E3 is an important industrial strain for erythromycin production and knowledge on its metabolism is limited. In the present work, (13)C labeling experiments were conducted to characterize the metabolism of S. erythraea E3. We found that S. erythraea E3 was difficult to grow on minimal medium with glucose as sole carbon source and the addition of proline remarkably improved the cell growth. The activity of EMP pathway was very low and ED pathway was alternatively the main glucose utilization pathway. The addition of proline resulted in remarkable changes in the fluxes of central metabolism. The fluxes in PP pathway, in TCA cycle and in ED pathway were 90% higher, 64% and 31% lower on Glc/Pro than on Glc, respectively. The maintenance energy on Glc/Pro was 58.4% lower than that on Glc. The energy charge was lower on Glc than on Glc/Pro, indicating that the cells on Glc suffered from energy burden. This study elucidates the impacts of proline on the central metabolism of S. erythraea and deepens the understanding of its metabolism.

  7. Are δ13C values of n-alkanes affected by atmospheric CO2 concentrations? Results from a free-air CO2 enrichment (FACE) experiment.

    NASA Astrophysics Data System (ADS)

    Sandquist, D. R.; Williams, D. G.; Shuman, B. N.; Kim, S.; Chen, J.; Macdonald, C.

    2015-12-01

    Compound-specific carbon isotope (δ13C) analyses of leaf waxes (i.e., n-alkanes) can be linked to large-scale shifts in vegetation, such as dominant taxa, functional types, life-forms and photosynthetic pathways that are usually coupled with environmental changes in climate. However using these δ13C values to interpret finer-scale ecosystem properties, including climate attributes such as CO2 concentrations, is difficult owing to uncertainty in the magnitude of internal biosynthetic fractionations that determine the δ13C of waxes relative to that of bulk leaf material. We investigated the composition, abundance and δ13C of n-alkanes in the aboveground biomass of a C4 grass and a C3 grass exposed to experimentally controlled CO2 at ambient [490ppm] and elevated [630ppm] levels within natural grassland in Wyoming. The δ13C values of bulk tissues were predictably different based on the C3 and C4 photosynthetic pathways, but the difference between bulk tissue and n-alkanes (ɛlipid), for both C29 and C31, was consistently greater in the C4 grass. The magnitudes of these ɛlipid values were large (- 7‰ to -15‰) relative to those found in most other studies. CO2 concentration of the growing environment also had a significant effect on n-alkane δ13C values, with consistently higher values of ~ 2‰ under elevated CO2 found in both species and in both a wet and a dry year. These results underscore the importance of recognizing potential abiotic effects on leaf wax δ13C values, in addition to the biotic drivers their variation, when interpreting climate from leaf-wax biomarkers of terrestrial ecosystems.

  8. Stabilization of glucose-C in microbial cell membranes (PLFA) and cell walls (amino sugars) evaluated by 13C-labelling in a field experiment

    NASA Astrophysics Data System (ADS)

    Gunina, Anna; Kuzyakov, Yakov; Glaser, Bruno

    2015-04-01

    Microorganisms control carbon (C) cycle and strongly contribute to formation of soil organic matter. Strong differences in the turnover of microbial groups and cellular compounds complicate the assessment of their contribution to microbial food webs and C sequestration in soil in situ. The uptake and incorporation of 13C labeled glucose by microbial groups were traced during 50 days after the labeling under field conditions. 13C was analysed: i) in the cytosolic pool by chloroform fumigation extraction, ii) in cell membranes by phospholipid fatty acids (PLFA), iii) in cell walls by amino sugars, and iv) remaining in bulk soil. This allowed tracing C in microbial groups as well as cellular compounds. Mean residence times (MRT) of C in PLFA and the cytosol were 47 and 150 days, respectively. Such long cytosol MRT depends on its heterogeneous composition, which includes high and low molecular weight organics. Amino sugars were mainly originated from microbial residues and thus, observation periods higher than 1 year are required for estimation of their MRT. Relative 13C incorporation (13C portion in total pool C) was the highest for PLFAs (~1.5% at day 3), whereas 13C content of the cytosol and amino sugars was one and two orders of magnitude less, respectively. Relative 13C incorporation into amino sugars of living microorganisms showed only 0.57% on day 3. Therefore, the turnover of cell membrane components is two times faster than that of cell walls, even in living microorganisms. Both PLFAs and amino sugars showed that glucose C was preferentially used by bacteria. 13C incorporation into bacterial cell walls and membranes decreased with time, but increased or remained constant for fungi, reflecting faster turnover of bacteria than fungi. Consequently, bacteria contribute more to the decomposition of low molecular weight organics, whereas fungi consume bacterial products or necromass and contribute more to long-term C stabilisation. Thus, tracing of 13C in cellular

  9. Does the time of the sampling matter in 13C pulse labeling and chasing experiments? A case study on beech seedlings

    NASA Astrophysics Data System (ADS)

    Gavrichkova, Olga; Thoms, Ronny; Muhr, Jan; Karlowsky, Stefan; Keitel, Claudia; Kayler, Zachary; Calfapietra, Carlo; Gessler, Arthur; Brugnoli, Enrico; Gleixner, Gerd

    2016-04-01

    13C pulse labeling and chasing is a valuable and very popular tool for determination of the fate and turnover rates of C in plant-soil systems. Continuous isoflux measurements became an accessible reality allowing to cover completely the diurnal variation in label assimilation and respiration fluxes. Label turnover in multiple pools, especially of those located belowground, is more often assessed instead by isolated day-time samplings. By increasing the sampling frequency of belowground compartments we aimed to catch the short-term diurnal variations in label allocation and to link these processes with label dynamics in the aboveground biomass. For these purposes we labeled 3-m height soil-grown European beech seedlings with 13C enriched CO2 and traced the flow of 13C within belowground plant-soil continuum. Continuous soil isoflux measurements were accompanied by a 3-h-frequency sampling of root and soil material during the first 48 h, followed by a daily sampling in the successive 5 days. The amount of label found in microbial biomass depended partially on the amount of roots in the sample. Microbial biomass C (MBC) and microbial respiration showed very strong correlation, suggesting the possibility to use one as a proxy of the other. MBC enrichment showed a clear diurnal pattern with night-time and early morning peaks. These peaks were similar in shape and shifted by one sampling when compared to root sugars enrichment. Soil respiration showed instead a single bell-shape peak in 13C, likely due to a sequence of peaks of root and microbial origin. 13C flow into soil microbial functional groups was assessed less frequently through phospholipid fatty acid analyses (PLFA). The microorganisms were separated into two distinct groups by the time of the appearance of the label in the single PLFAs. The first group was characterized by a fast appearance of the label and higher enrichment and was composed of Gram negative bacteria and saprotrophic fungi likely living in

  10. (13)C-Breath testing in animals: theory, applications, and future directions.

    PubMed

    McCue, Marshall D; Welch, Kenneth C

    2016-04-01

    The carbon isotope values in the exhaled breath of an animal mirror the carbon isotope values of the metabolic fuels being oxidized. The measurement of stable carbon isotopes in carbon dioxide is called (13)C-breath testing and offers a minimally invasive method to study substrate oxidation in vivo. (13)C-breath testing has been broadly used to study human exercise, nutrition, and pathologies since the 1970s. Owing to reduced use of radioactive isotopes and the increased convenience and affordability of (13)C-analyzers, the past decade has witnessed a sharp increase in the use of breath testing throughout comparative physiology--especially to answer questions about how and when animals oxidize particular nutrients. Here, we review the practical aspects of (13)C-breath testing and identify the strengths and weaknesses of different methodological approaches including the use of natural abundance versus artificially-enriched (13)C tracers. We critically compare the information that can be obtained using different experimental protocols such as diet-switching versus fuel-switching. We also discuss several factors that should be considered when designing breath testing experiments including extrinsic versus intrinsic (13)C-labelling and different approaches to model nutrient oxidation. We use case studies to highlight the myriad applications of (13)C-breath testing in basic and clinical human studies as well as comparative studies of fuel use, energetics, and carbon turnover in multiple vertebrate and invertebrate groups. Lastly, we call for increased and rigorous use of (13)C-breath testing to explore a variety of new research areas and potentially answer long standing questions related to thermobiology, locomotion, and nutrition. PMID:26660654

  11. (13)C-Breath testing in animals: theory, applications, and future directions.

    PubMed

    McCue, Marshall D; Welch, Kenneth C

    2016-04-01

    The carbon isotope values in the exhaled breath of an animal mirror the carbon isotope values of the metabolic fuels being oxidized. The measurement of stable carbon isotopes in carbon dioxide is called (13)C-breath testing and offers a minimally invasive method to study substrate oxidation in vivo. (13)C-breath testing has been broadly used to study human exercise, nutrition, and pathologies since the 1970s. Owing to reduced use of radioactive isotopes and the increased convenience and affordability of (13)C-analyzers, the past decade has witnessed a sharp increase in the use of breath testing throughout comparative physiology--especially to answer questions about how and when animals oxidize particular nutrients. Here, we review the practical aspects of (13)C-breath testing and identify the strengths and weaknesses of different methodological approaches including the use of natural abundance versus artificially-enriched (13)C tracers. We critically compare the information that can be obtained using different experimental protocols such as diet-switching versus fuel-switching. We also discuss several factors that should be considered when designing breath testing experiments including extrinsic versus intrinsic (13)C-labelling and different approaches to model nutrient oxidation. We use case studies to highlight the myriad applications of (13)C-breath testing in basic and clinical human studies as well as comparative studies of fuel use, energetics, and carbon turnover in multiple vertebrate and invertebrate groups. Lastly, we call for increased and rigorous use of (13)C-breath testing to explore a variety of new research areas and potentially answer long standing questions related to thermobiology, locomotion, and nutrition.

  12. Design and implementation of the winter haze intensive tracer experiment

    SciTech Connect

    Malm, W.C.; Iyer, H.K. ); Pitchford, M. )

    1988-01-01

    Protection of vistas for certain national parks and wilderness areas as provided by the clean air act amendments of 1977 has stimulated an interest in visibility research. Methods are being developed and used to characterize atmospheric transparency, to identify the relative importance of the various particulate and gaseous atmospheric materials and to determine the role of man -made emissions. Much of the research has been conducted in the dessert southwest, in particular in northern Arizona and southern Utah. According to the authors,the juxtaposition of energy resources (especially coal) and national parks (including Grand Canyon, Bryce Canyon and Canyonlands) in an area where small changes in aerosol concentration can significantly affect visibility justifies concern by government and private organizations for visibility impacts resulting from industrial emissions.Accordingly, a cooperative effort, the subregional cooperative electric utility, national park service (NPS), Environmental Protection Agency (EPA) and Department of Defense (DOD) study, SCENES, is centered in this area. It operates on a five-year plan (1984-1989) involving continual visibility and aerosol measurements at a dozen locations, plus more in-depth intensive and special studies conducted over shorter, seasonally representative periods. In this paper, the authors discuss the winter haze intensive tracer experiment (WHITEX) which was conducted in January and February 1987 in the Colorado River area of the Colorado Plateau.

  13. 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.

  14. Feasibility of perfluorocarbon tracers (PFTs) in atmospheric source-receptor experiments

    SciTech Connect

    Dietz, R.N.; Senum, G.I.

    1984-03-01

    A brief description of the perfluorocarbon tracer (PFT) system, which includes the tracers and the release equipment, the air samplers and the analyzers, is presented along with details on the research needs to provide a viable system for MATEX-scenario experiments. The present family of 2 viable PFTs needs to be increased to 5 to 6. Given the present precision of the analysis system, a one year long tracer experiment consisting of 4 hour releases every 60 hours from 5 different sites would require nearly 150 metric tons of PFTs at a cost of $15,000,000. Shortcomings in the programmable sampler include the pump, the sampling sequence control flexibility, data storage and retrieval, and the lack of remote communication capability; sampler adsorbent studies are also needed. The analytical system, including the catalyst processing bed, the chromatography column resolution, and the linearity of the detector, is in need of significant improvement. A higher resolution analysis system could significantly reduce analysis time but, more importantly, reduce tracer requirements more than 10-fold, for a cost savings potential of more than $13,000,000. A model is presented to demonstrate the feasibility of tracer material balances. Assessment of earlier long-range tracer experiments indicates the need for possibly 400 ground sampling sites requiring $8 to $14 million worth of samplers for a one-year tracer experiment. As many as six aircraft would be needed to conduct airborne model validation and material balance studies for each tracer plume.

  15. Compartmentation of glycogen metabolism revealed from 13C isotopologue distributions

    PubMed Central

    2011-01-01

    Background Stable isotope tracers are used to assess metabolic flux profiles in living cells. The existing methods of measurement average out the isotopic isomer distribution in metabolites throughout the cell, whereas the knowledge of compartmental organization of analyzed pathways is crucial for the evaluation of true fluxes. That is why we accepted a challenge to create a software tool that allows deciphering the compartmentation of metabolites based on the analysis of average isotopic isomer distribution. Results The software Isodyn, which simulates the dynamics of isotopic isomer distribution in central metabolic pathways, was supplemented by algorithms facilitating the transition between various analyzed metabolic schemes, and by the tools for model discrimination. It simulated 13C isotope distributions in glucose, lactate, glutamate and glycogen, measured by mass spectrometry after incubation of hepatocytes in the presence of only labeled glucose or glucose and lactate together (with label either in glucose or lactate). The simulations assumed either a single intracellular hexose phosphate pool, or also channeling of hexose phosphates resulting in a different isotopic composition of glycogen. Model discrimination test was applied to check the consistency of both models with experimental data. Metabolic flux profiles, evaluated with the accepted model that assumes channeling, revealed the range of changes in metabolic fluxes in liver cells. Conclusions The analysis of compartmentation of metabolic networks based on the measured 13C distribution was included in Isodyn as a routine procedure. The advantage of this implementation is that, being a part of evaluation of metabolic fluxes, it does not require additional experiments to study metabolic compartmentation. The analysis of experimental data revealed that the distribution of measured 13C-labeled glucose metabolites is inconsistent with the idea of perfect mixing of hexose phosphates in cytosol. In contrast

  16. Active ion tracer experiments attempted in conjunction with the ion composition experiment on GEOS-2

    NASA Astrophysics Data System (ADS)

    Young, D. T.

    It is pointed out that to date six ion injection/tracer experiments have been attempted in conjunction with the GEOS-2 Ion Composition Experiment: three rocket borne Ba shaped-charge releases (Porcupine 3 and 4 and Ba-GEOS), one Li release, and two periods of operation of the Xe(+) accelerator on the SCATHA satellite. The characteristics of each of these six releases are outlined, and upper limits are placed on possible ion fluxes reaching GEOS-2. The order of magnitude of ion fluxes to be expected from each release is estimated, and it is shown that three of the experiments had no real chance of succeeding in the first place.

  17. 13C magnetic resonance spectroscopy measurements with hyperpolarized [1‐13C] pyruvate can be used to detect the expression of transgenic pyruvate decarboxylase activity in vivo

    PubMed Central

    Dzien, Piotr; Tee, Sui‐Seng; Kettunen, Mikko I.; Lyons, Scott K.; Larkin, Timothy J.; Timm, Kerstin N.; Hu, De‐En; Wright, Alan; Rodrigues, Tiago B.; Serrao, Eva M.; Marco‐Rius, Irene; Mannion, Elizabeth; D'Santos, Paula; Kennedy, Brett W. C.

    2015-01-01

    Purpose Dissolution dynamic nuclear polarization can increase the sensitivity of the 13C magnetic resonance spectroscopy experiment by at least four orders of magnitude and offers a novel approach to the development of MRI gene reporters based on enzymes that metabolize 13C‐labeled tracers. We describe here a gene reporter based on the enzyme pyruvate decarboxylase (EC 4.1.1.1), which catalyzes the decarboxylation of pyruvate to produce acetaldehyde and carbon dioxide. Methods Pyruvate decarboxylase from Zymomonas mobilis (zmPDC) and a mutant that lacked enzyme activity were expressed using an inducible promoter in human embryonic kidney (HEK293T) cells. Enzyme activity was measured in the cells and in xenografts derived from the cells using 13C MRS measurements of the conversion of hyperpolarized [1‐13C] pyruvate to H13 CO3–. Results Induction of zmPDC expression in the cells and in the xenografts derived from them resulted in an approximately two‐fold increase in the H13 CO3–/[1‐13C] pyruvate signal ratio following intravenous injection of hyperpolarized [1‐13C] pyruvate. Conclusion We have demonstrated the feasibility of using zmPDC as an in vivo reporter gene for use with hyperpolarized 13C MRS. Magn Reson Med 76:391–401, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. PMID:26388418

  18. Feasibility of Multianimal Hyperpolarized 13C MRS

    PubMed Central

    Ramirez, Marc S.; Lee, Jaehyuk; Walker, Christopher M.; Chen, Yunyun; Kingsley, Charles V.; De La Cerda, Jorge; Maldonado, Kiersten L.; Lai, Stephen Y.; Bankson, James A.

    2014-01-01

    Purpose There is great potential for real-time investigation of metabolism with MRS and hyperpolarized (HP) 13C agents. Unfortunately, HP technology has high associated costs and efficiency limitations that may constrain in vivo studies involving many animals. To improve the throughput of preclinical investigations, we evaluate the feasibility of performing HP MRS on multiple animals simultaneously. Methods Simulations helped assess the viability of a dual-coil strategy for spatially-localized multivolume MRS.A dual-mouse system was assembled and characterized based on bench- and scanner-based experiments. Enzyme phantoms mixed with HP [1-13C] pyruvate emulated real-time metabolism and offered a controlled mechanism for evaluating system performance. Finally, a normal mouse and a mouse bearing a subcutaneous xenograft of colon cancer were simultaneously scanned in vivo using an agent containing HP [1-13C] pyruvate. Results Geometric separation/rotation, active decoupling, and use of low input impedance preamplifiers permitted an encode-by-channel approach for spatially-localized MRS. A pre-calibrated shim allowed straightforward metabolite differentiation in enzyme phantom and in vivo experiments at 7 T, with performance similar to conventional acquisitions. Conclusion The initial feasibility of multi-animal HP 13C MRS was established. Throughput scales with the number of simultaneously-scanned animals, demonstrating the potential for significant improvements in study efficiency. PMID:24903532

  19. Study of stability zone influences and tracer patterns from the 1987 ANATEX (Across North America Tracer Experiment) experiment

    SciTech Connect

    Porch, W.M.; Gifford, F.A.; Hoard, D.E.

    1988-01-01

    In this paper, we will show preliminary results which appear to connect much of the hit and miss behavior of the surface tracer samples to large scale stability zones 100 to 1000 km wide. With these wintertime stability effects in mind, we have done the best we can to characterize the observed overall tracer patterns as well as individual tracer releases. This type of survey information is important to numerical model development. Diagnostic models often have difficulty reproducing surface plume concentrations where transport over stable layers have occurred. Prognostic models can, in theory, model effects of strong stable layers. However, these models would have great difficulty predicting large scale stable regions such as those observed during ANATEX. Also, though these models have ways of budging in synoptic wind fields, temperature observations are presently ignored. This is because if both wind and temperature observations are forced too strongly into the model, conflicting results may be produced. 12 refs., 5 figs.

  20. Relative contributions of rhizosphere and microbial respiration to belowground and total ecosystem respiration in arctic tussock tundra: results of a 13C pulse-chase experiment

    NASA Astrophysics Data System (ADS)

    Segal, A. D.; Sullivan, P.; Weintraub, M. N.; Darrouzet-Nardi, A.; Steltzer, H.

    2011-12-01

    Many arctic ecosystems that have historically been strong carbon (C) sinks are becoming sources of C to the atmosphere. Although ecosystem respiration is the largest C flux out of ecosystems, our ability to model respiration lags considerably behind our ability to model photosynthesis in the Arctic. Understanding the controls on respiration is especially important for an ecosystem which appears to be experiencing the greatest climate warming and also contains large stores of soil C. Partitioning respiration into its component fluxes and identifying factors controlling respiration of each component is a critical first step towards improving our ability to model changes in respiration. However, partitioning belowground constituents has proven to be challenging in most ecosystems. Therefore, to accurately estimate rhizosphere respiration and bulk soil microbial respiration in moist acidic tussock tundra, we selected an isotopic method that results in minimal disturbance of belowground processes. In mid July of 2011, we introduced a 13CO2 label into a clear ecosystem CO2 flux chamber, allowed the vegetation to incorporate the label through photosynthesis and returned 2 days and 4 days after labeling to follow the movement of the 13C signal. A smaller CO2 flux chamber was used to chase the label separately in tussock and inter-tussock areas. All above ground plant tissue was clipped immediately before the chase measurements and soil cores were taken immediately after chasing the label. Syringe samples (n=5 or 6) were collected from the small flux chamber at regular intervals as CO2 concentrations were allowed to build, and Keeling plots were used to estimate δ13C of belowground respiration. After completing the field measurements, the soil cores were sorted into live roots and root free soil. Samples of each were incubated in mason jars placed in a 10°C water bath. The jars were scrubbed free of CO2 and syringe samples were collected from each jar after CO2

  1. Tracking tracer motion in a 4-D electrical resistivity tomography experiment

    NASA Astrophysics Data System (ADS)

    Ward, W. O. C.; Wilkinson, P. B.; Chambers, J. E.; Nilsson, H.; Kuras, O.; Bai, L.

    2016-05-01

    A new framework for automatically tracking subsurface tracers in electrical resistivity tomography (ERT) monitoring images is presented. Using computer vision and Bayesian inference techniques, in the form of a Kalman filter, the trajectory of a subsurface tracer is monitored by predicting and updating a state model representing its movements. Observations for the Kalman filter are gathered using the maximally stable volumes algorithm, which is used to dynamically threshold local regions of an ERT image sequence to detect the tracer at each time step. The application of the framework to the results of 2-D and 3-D tracer monitoring experiments show that the proposed method is effective for detecting and tracking tracer plumes in ERT images in the presence of noise, without intermediate manual intervention.

  2. 13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion-isotope ratio mass spectrometry.

    PubMed

    Yuan, Yongbo; Yang, Tae Hoon; Heinzle, Elmar

    2010-07-01

    (13)C-based metabolic flux analysis ((13)CMFA) is limited to smaller scale experiments due to very high costs of labeled substrates. We measured (13)C enrichment in proteinogenic amino acid hydrolyzates using gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) from a series of parallel batch cultivations of Corynebacterium glutamicum utilizing mixtures of natural glucose and [1-(13)C] glucose, containing 0%, 0.5%, 1%, 2%, and 10% [1-(13)C] glucose. Decreasing the [1-(13)C] glucose content, kinetic isotope effects played an increasing role but could be corrected. From the corrected (13)C enrichments in vivo fluxes in the central metabolism were determined by numerical optimization. The obtained flux distribution was very similar to those obtained from parallel labeling experiments using conventional high labeling GC-MS method and to published results. The GC-C-IRMS-based method involving low labeling degree of expensive tracer substrate, e.g. 1%, is well suited for larger laboratory and industrial pilot scale fermentations.

  3. Degradation of algal lipids by deep-sea benthic foraminifera: An in situ tracer experiment

    NASA Astrophysics Data System (ADS)

    Nomaki, Hidetaka; Ohkouchi, Naohiko; Heinz, Petra; Suga, Hisami; Chikaraishi, Yoshito; Ogawa, Nanako O.; Matsumoto, Kouhei; Kitazato, Hiroshi

    2009-09-01

    We conducted an in situ feeding experiment using 13C-labeled unicellular algae in Sagami Bay, Japan (water depth, 1450 m), in order to understand the fate of lipid compounds in phytodetritus at the deep-sea floor. We examined the incorporation of excess 13C into lipid compounds extracted from bulk sediments and benthic foraminiferal cells. 13C-enriched fatty acids derived from 13C-labeled algae were exponentially degraded during 6 days of incubation in the sediment. Subsequent enrichments in 13C in sedimentary n-C 15,anteiso-C 17, and C 17 fatty acids indicated the microbial degradation of algal material and production of bacterial biomass in the sediment. We observed the incorporation of 13C-labeled algal phytol and fatty acids into foraminiferal cells. The compositions of 13C-labeled algal lipids in foraminiferal cells were different from those in the bulk sediments, indicating that foraminiferal feeding and digestion influenced the lipid distribution in the sediments. Furthermore, some sterols in Globobulimina affinis (e.g., 24-ethylcholesta-5,22-dien-3β-ol, 24-ethylcholest-5-en-3β-ol, and 23,24-dimethylcholesta-5,22E-dien-3β-ol) were newly produced via the modification of dietary algal sterols within 4-6 days. In addition to the effects of bacteria, feeding by benthic foraminifera can result in a significant reorganization of the composition of organic matter and influence benthic food webs and carbon cycling at the deep-sea floor.

  4. Identification of transport processes in Southern Indian fractured crystalline rock using forced-gradient tracer experiments

    NASA Astrophysics Data System (ADS)

    Guihéneuf, Nicolas; Bour, Olivier; Boisson, Alexandre; Le Borgne, Tanguy; Becker, Matthew R.; Nigon, Benoit; Wajiduddin, Mohammed; Ahmed, Shakeel; Maréchal, Jean-Christophe

    2015-04-01

    Understanding dominant transport processes is essential to improve prediction of contaminants transfer in fractured crystalline rocks. In such fractured media, solute transport is characterized by fast advection within open and connected fractures and sometimes by matrix diffusion that may be enhanced by chemical weathering. To investigate this phenomenon, we carried out radially convergent and push-pull tracer experiments in the fractured granite of the Experimental Hydrogeological Park of Choutuppal (Southern India). Tracer tests were performed in the same permeable fracture from few meters to several ten meters and from few hours to two weeks to check the consistency of the results at different spatial and temporal scales. These different types of forced gradient tracer experiments allow separation of the effects of advection and diffusion on transport. Breakthrough curves from radially convergent tracer tests display systematically a -2 power law slope on the late time behavior. This tailing can be adequately represented by a transport model that only takes into account heterogeneous advection caused by fluid flow channeling. The negligible impact of matrix diffusion was confirmed by the push-pull tracer tests, at least for the duration of experiments. A push-pull experiment carried out with a cocktail of two conservative tracers having different diffusion coefficients displayed similar breakthrough curves. Increasing the resting phase during the experiments did not lead to a significant decline of peak concentration. All these results suggest a negligible impact of matrix diffusion. However, increasing the scales of investigation during push-pull tracer tests led to a decrease of the power law slope on the late time behavior. This behavior that cannot be modeled with a transport model based on independent flow paths and indicate non-reversible heterogeneous advection. This process could be explained by the convergence of streamlines after a certain distance

  5. Stability and breakdown of Ca{sup 13}CO{sub 3} melt associated with formation of {sup 13}C-diamond in static high pressure experiments up to 43 GPa and 3900 K

    SciTech Connect

    Spivak, A.V.; Litvin, Yu.A.; Ovsyannikov, S.V.; Dubrovinskaia, N.A.; Dubrovinsky, L.S.

    2012-07-15

    Melting of calcium carbonate Ca{sup 13}CO{sub 3}, stability of the melt and its decomposition were studied in static high pressure experiments at pressures of 11-43 GPa and temperatures of 1600-3900 K using diamond anvil cell technique with laser heating. We observed formation of {sup 13}C-graphite (below 16 GPa) and {sup 13}C-diamond (between 16 and 43 GPa) on decomposition of the Ca{sup 13}CO{sub 3} melt at temperatures above 3400 K. At temperatures below 3400 K congruent melting of calcium carbonate was confirmed. The experimental results were applied to construction of the phase diagram of CaCO{sub 3} up to 43 GPa and 3900 K focusing at the melting curve of calcium carbonate and the decomposition phase boundary of CaCO{sub 3} melt. - Graphical abstract: Highlights: Black-Right-Pointing-Pointer Phase states of CaCO{sub 3} were studied at P=11-43 GPa and T=1600-3900 K. Black-Right-Pointing-Pointer {sup 13}C-diamond easily crystallizes in carbonate-carbon (Ca{sup 13}CO{sub 3-}{sup 13}C-graphite) melt-solutions. Black-Right-Pointing-Pointer Ca-carbonate melts congruently that was observed in experiments in DAC with laser heating. Black-Right-Pointing-Pointer Decomposition of CaCO{sub 3} melt, indicated by formation of graphite and/or diamond. Black-Right-Pointing-Pointer Decomposition of CaCO{sub 3} was observed at temperatures above 3400 K in the pressure interval studied.

  6. Highly enriched multiply-labeled stable isotopic compounds as atmospheric tracers

    DOEpatents

    Goldblatt, M.; McInteer, B.B.

    1974-01-29

    Compounds multiply-labeled with stable isotopes and highly enriched in these isotopes are readily capable of detection in tracer experiments involving high dilutions. Thus, for example, /sup 13/C/sup 18/O/sub 2/ provides a useful tracer for following atmospheric pol lution produced as a result of fossil fuel burning. (Official Gazette)

  7. Comparative evaluation of two sediment tracers in a rainfall simulation experiment

    NASA Astrophysics Data System (ADS)

    Strauss, Peter; Guzman, Gema; Mentler, Axel

    2015-04-01

    The use of sediment tracers tries to contribute to solving problems of traditional soil erosion measurements such as allocation of erosional and depositional areas. A number of different tracing approaches have already been established however, as none of available techniques are able to fully satisfy all the requirements for being an ideal tracer the search for alternative methods continues. Clays tagged with quaternary ammonium compounds are widely used in industry and are easy to incorporate into soils for sediment tracing experiments. To explore the potential of these clays a laboratory experiment was carried out in order to characterize their behaviour as compared to the well-established sediment tracer magnetic iron oxide. The experiment consisted of a simulated rainfall event of 60 mm/h in a box laboratory flume divided in two ridge-furrow subplots (200 x 57 cm). In order to evaluate transport and redistribution of soil from the ridges to the furrow and to measure the dynamic behaviour of the tracers during the rainfall a dense grid of soil samples was taken before and after the experiment (140 samples in total). Runoff and sediment were collected and all samples were analysed for both tracers. Results indicate the general suitability of organophilic clays to monitor soil redistribution by water erosion. The average relative contributions from shoulders and furrow to total sediment export determined by both tracers were similar and indicated a higher contribution from ridges. A key advantage of using organophilic clays is that the detection limit of organophilic clays is extremely low and the background concentration is zero. Despite the inherent differences between both tracers such as way of bounding, sediment enrichment or analytical technique, this experimental comparison of an established and more novel tracer method underscores the potential suitability of the latter for soil erosion studies.

  8. Two Techniques for Estimating Deglacial Mean-Ocean δ13 C Change from the Same Set of 493 Benthic δ13C Records

    NASA Astrophysics Data System (ADS)

    Peterson, C. D.; Lisiecki, L. E.; Gebbie, G.

    2013-12-01

    The crux of carbon redistribution over the deglaciation centers on the ocean, where the isotopic signature of terrestrial carbon (δ13C terrestrial carbon = -25‰) is observed as a 0.3-0.7‰ shift in benthic foraminiferal δ13C. Deglacial mean-ocean δ13C estimates vary due to different subsets of benthic δ13C data and different methods of weighting the mean δ13C by volume. Here, we present a detailed 1-to-1 comparison of two methods of calculating mean δ13C change and uncertainty estimates using the same set of 493 benthic Cibicidoides spp. δ13C measurements for the LGM and Late Holocene. The first method divides the ocean into 8 regions, and uses simple line fits to describe the distribution of δ13C data for each timeslice over 0.5-5 km depth. With these line fits, we estimate the δ13C value at 100-meter intervals and weight those estimates by the regional volume at each depth slice. The mean-ocean δ13C is the sum of these volume-weighted regional δ13C estimates and the uncertainty of these mean-ocean δ13C estimates is computed using Monte Carlo simulations. The whole-ocean δ13C change is estimated using extrapolated surface- and deep-ocean δ13C estimates, and an assumed δ13C value for the Southern Ocean. This method yields an estimated LGM-to-Holocene change of 0.38×0.07‰ for 0.5-5km and 0.35×0.16‰ for the whole ocean (Peterson et al., 2013, submitted to Paleoceanography). The second method reconstructs glacial and modern δ13C by combining the same data compilation as above with a steady-state ocean circulation model (Gebbie, 2013, submitted to Paleoceanography). The result is a tracer distribution on a 4-by-4 degree horizontal resolution grid with 23 vertical levels, and an estimate of the distribution's uncertainty that accounts for the distinct modern and glacial water-mass geometries. From both methods, we compare the regional δ13C estimates (0.5-5 km), surface δ13C estimates (0-0.5 km), deep δ13C estimates (>5 km), Southern Ocean

  9. A Cross-Hole, Multi-Year Tracer Injection Experiment in the Volcanic Ocean Crust

    NASA Astrophysics Data System (ADS)

    Fisher, A. T.; Neira, N. M.; Wheat, C. G.; Clark, J. F.; Becker, K.; Hsieh, C. C.; Rappe, M. S.

    2014-12-01

    We present preliminary results from the first cross-hole tracer injection experiment in the volcanic ocean crust. The test site is on 3.5 to 3.6 M.y. old seafloor on the eastern flank of the Juan de Fuca Ridge. Six borehole subseafloor observatories (CORKs) were installed during three scientific ocean drilling expeditions, five arrayed along a 1 km profile aligned with the strike of underlying abyssal hills (Holes 1026B, 1301A/B, and 1362A/B), and one offset 2.4 km to the east (1027C). Before installing the sixth CORK in Hole 1362B, in 2010, we injected a mixture of tracers (dissolved gas, metal salts, particles) during 24 hours into the upper ocean crust. Seafloor samplers connected CORKs, sampling from different locations in the crust, were recovered during servicing expeditions in 2011 and 2013; downhole samplers that contain records from the full four years following tracer injection will be recovered in Summer 2014. Analyses of dissolved gas tracers collected with wellhead samplers through 2013 suggest that the dominant flow direction in upper basement is south to north, as inferred from regional thermal data and the chemistry of geochemical (pore fluid and borehole) samples. The apparent tracer flow rate in upper basement is on the order of meters/day, but calculations are complicated by an incomplete CORK seal in Hole 1301A, which resulted in discharge from this system that also "pulled" water and tracer to the south. Samples were collected from the tracer injection borehole, Hole 1362B, and a sampling site 200 m to the north, Hole 1362A, beginning one year after tracer injection, after opening a large-diameter ball valve on the wellhead of Hole 1362B to initiate a long-term free flow experiment. Analyses of these samples suggest that much of the tracer injected in 2010 remained close to Hole 1362B rather than being advected and dispersed into the formation. It also appears that much of the tracer transport to Hole 1362A occurred within one or more

  10. Gaussian Modeling of Tracer Concentrations during the Joint Urban 2003 Experiment

    SciTech Connect

    Gouveia, F J

    2004-06-18

    The Joint Urban 2003 Experiment (JU2003) was conducted in Oklahoma City, Oklahoma during the summer of 2003. This extensive field experiment included over a hundred scientists measuring airflow, tracer concentration, and other variables pertinent to urban dispersion. A description of JU2003 can be found at this website: http://ju2003.pnl.gov/.

  11. (13)C metabolic flux analysis of the extremely thermophilic, fast growing, xylose-utilizing Geobacillus strain LC300.

    PubMed

    Cordova, Lauren T; Antoniewicz, Maciek R

    2016-01-01

    Thermophiles are increasingly used as versatile hosts in the biotechnology industry. One of the key advantages of thermophiles is the potential to achieve high rates of feedstock conversion at elevated temperatures. The recently isolated Geobacillus strain LC300 grows extremely fast on xylose, with a doubling time of less than 30 min. In the accompanying paper, the genome of Geobacillus LC300 was sequenced and annotated. In this work, we have experimentally validated the metabolic network model using parallel (13)C-labeling experiments and applied (13)C-metabolic flux analysis to quantify precise metabolic fluxes. Specifically, the complete set of singly labeled xylose tracers, [1-(13)C], [2-(13)C], [3-(13)C], [4-(13)C], and [5-(13)C]xylose, was used for the first time. Isotopic labeling of biomass amino acids was measured by gas chromatography mass spectrometry (GC-MS). Isotopic labeling of carbon dioxide in the off-gas was also measured by an on-line mass spectrometer. The (13)C-labeling data was then rigorously integrated for flux elucidation using the COMPLETE-MFA approach. The results provided important new insights into the metabolism of Geobacillus LC300, its efficient xylose utilization pathways, and the balance between carbon, redox and energy fluxes. The pentose phosphate pathway, glycolysis and TCA cycle were found to be highly active in Geobacillus LC300. The oxidative pentose phosphate pathway was also active and contributed significantly to NADPH production. No transhydrogenase activity was detected. Results from this work provide a solid foundation for future studies of this strain and its metabolic engineering and biotechnological applications.

  12. Heat tracer test in an alluvial aquifer: Field experiment and inverse modelling

    NASA Astrophysics Data System (ADS)

    Klepikova, Maria; Wildemeersch, Samuel; Hermans, Thomas; Jamin, Pierre; Orban, Philippe; Nguyen, Frédéric; Brouyère, Serge; Dassargues, Alain

    2016-09-01

    Using heat as an active tracer for aquifer characterization is a topic of increasing interest. In this study, we investigate the potential of using heat tracer tests for characterization of a shallow alluvial aquifer. A thermal tracer test was conducted in the alluvial aquifer of the Meuse River, Belgium. The tracing experiment consisted in simultaneously injecting heated water and a dye tracer in an injection well and monitoring the evolution of groundwater temperature and tracer concentration in the pumping well and in measurement intervals. To get insights in the 3D characteristics of the heat transport mechanisms, temperature data from a large number of observation wells closely spaced along three transects were used. Temperature breakthrough curves in observation wells are contrasted with what would be expected in an ideal layered aquifer. They reveal strongly unequal lateral and vertical components of the transport mechanisms. The observed complex behavior of the heat plume is explained by the groundwater flow gradient on the site and heterogeneities in the hydraulic conductivity field. Moreover, due to high injection temperatures during the field experiment a temperature-induced fluid density effect on heat transport occurred. By using a flow and heat transport numerical model with variable density coupled with a pilot point approach for inversion of the hydraulic conductivity field, the main preferential flow paths were delineated. The successful application of a field heat tracer test at this site suggests that heat tracer tests is a promising approach to image hydraulic conductivity field. This methodology could be applied in aquifer thermal energy storage (ATES) projects for assessing future efficiency that is strongly linked to the hydraulic conductivity variability in the considered aquifer.

  13. NMR structure analysis of uniformly 13C-labeled carbohydrates.

    PubMed

    Fontana, Carolina; Kovacs, Helena; Widmalm, Göran

    2014-06-01

    In this study, a set of nuclear magnetic resonance experiments, some of them commonly used in the study of (13)C-labeled proteins and/or nucleic acids, is applied for the structure determination of uniformly (13)C-enriched carbohydrates. Two model substances were employed: one compound of low molecular weight [(UL-(13)C)-sucrose, 342 Da] and one compound of medium molecular weight ((13)C-enriched O-antigenic polysaccharide isolated from Escherichia coli O142, ~10 kDa). The first step in this approach involves the assignment of the carbon resonances in each monosaccharide spin system using the anomeric carbon signal as the starting point. The (13)C resonances are traced using (13)C-(13)C correlations from homonuclear experiments, such as (H)CC-CT-COSY, (H)CC-NOESY, CC-CT-TOCSY and/or virtually decoupled (H)CC-TOCSY. Based on the assignment of the (13)C resonances, the (1)H chemical shifts are derived in a straightforward manner using one-bond (1)H-(13)C correlations from heteronuclear experiments (HC-CT-HSQC). In order to avoid the (1) J CC splitting of the (13)C resonances and to improve the resolution, either constant-time (CT) in the indirect dimension or virtual decoupling in the direct dimension were used. The monosaccharide sequence and linkage positions in oligosaccharides were determined using either (13)C or (1)H detected experiments, namely CC-CT-COSY, band-selective (H)CC-TOCSY, HC-CT-HSQC-NOESY or long-range HC-CT-HSQC. However, due to the short T2 relaxation time associated with larger polysaccharides, the sequential information in the O-antigen polysaccharide from E. coli O142 could only be elucidated using the (1)H-detected experiments. Exchanging protons of hydroxyl groups and N-acetyl amides in the (13)C-enriched polysaccharide were assigned by using HC-H2BC spectra. The assignment of the N-acetyl groups with (15)N at natural abundance was completed by using HN-SOFAST-HMQC, HNCA, HNCO and (13)C-detected (H)CACO spectra.

  14. NMR structure analysis of uniformly 13C-labeled carbohydrates.

    PubMed

    Fontana, Carolina; Kovacs, Helena; Widmalm, Göran

    2014-06-01

    In this study, a set of nuclear magnetic resonance experiments, some of them commonly used in the study of (13)C-labeled proteins and/or nucleic acids, is applied for the structure determination of uniformly (13)C-enriched carbohydrates. Two model substances were employed: one compound of low molecular weight [(UL-(13)C)-sucrose, 342 Da] and one compound of medium molecular weight ((13)C-enriched O-antigenic polysaccharide isolated from Escherichia coli O142, ~10 kDa). The first step in this approach involves the assignment of the carbon resonances in each monosaccharide spin system using the anomeric carbon signal as the starting point. The (13)C resonances are traced using (13)C-(13)C correlations from homonuclear experiments, such as (H)CC-CT-COSY, (H)CC-NOESY, CC-CT-TOCSY and/or virtually decoupled (H)CC-TOCSY. Based on the assignment of the (13)C resonances, the (1)H chemical shifts are derived in a straightforward manner using one-bond (1)H-(13)C correlations from heteronuclear experiments (HC-CT-HSQC). In order to avoid the (1) J CC splitting of the (13)C resonances and to improve the resolution, either constant-time (CT) in the indirect dimension or virtual decoupling in the direct dimension were used. The monosaccharide sequence and linkage positions in oligosaccharides were determined using either (13)C or (1)H detected experiments, namely CC-CT-COSY, band-selective (H)CC-TOCSY, HC-CT-HSQC-NOESY or long-range HC-CT-HSQC. However, due to the short T2 relaxation time associated with larger polysaccharides, the sequential information in the O-antigen polysaccharide from E. coli O142 could only be elucidated using the (1)H-detected experiments. Exchanging protons of hydroxyl groups and N-acetyl amides in the (13)C-enriched polysaccharide were assigned by using HC-H2BC spectra. The assignment of the N-acetyl groups with (15)N at natural abundance was completed by using HN-SOFAST-HMQC, HNCA, HNCO and (13)C-detected (H)CACO spectra. PMID:24771296

  15. Large and unexpected enrichment in stratospheric 16O13C18O and its meridional variation.

    PubMed

    Yeung, Laurence Y; Affek, Hagit P; Hoag, Katherine J; Guo, Weifu; Wiegel, Aaron A; Atlas, Elliot L; Schauffler, Sue M; Okumura, Mitchio; Boering, Kristie A; Eiler, John M

    2009-07-14

    The stratospheric CO(2) oxygen isotope budget is thought to be governed primarily by the O((1)D)+CO(2) isotope exchange reaction. However, there is increasing evidence that other important physical processes may be occurring that standard isotopic tools have been unable to identify. Measuring the distribution of the exceedingly rare CO(2) isotopologue (16)O(13)C(18)O, in concert with (18)O and (17)O abundances, provides sensitivities to these additional processes and, thus, is a valuable test of current models. We identify a large and unexpected meridional variation in stratospheric (16)O(13)C(18)O, observed as proportions in the polar vortex that are higher than in any naturally derived CO(2) sample to date. We show, through photochemical experiments, that lower (16)O(13)C(18)O proportions observed in the midlatitudes are determined primarily by the O((1)D)+CO(2) isotope exchange reaction, which promotes a stochastic isotopologue distribution. In contrast, higher (16)O(13)C(18)O proportions in the polar vortex show correlations with long-lived stratospheric tracer and bulk isotope abundances opposite to those observed at midlatitudes and, thus, opposite to those easily explained by O((1)D)+CO(2). We believe the most plausible explanation for this meridional variation is either an unrecognized isotopic fractionation associated with the mesospheric photochemistry of CO(2) or temperature-dependent isotopic exchange on polar stratospheric clouds. Unraveling the ultimate source of stratospheric (16)O(13)C(18)O enrichments may impose additional isotopic constraints on biosphere-atmosphere carbon exchange, biosphere productivity, and their respective responses to climate change.

  16. Pulsed polarization transfer for 13C NMR in solids

    NASA Astrophysics Data System (ADS)

    Bax, Ad; Szeverenyi, Nikolaus M.; Maciel, Gary E.

    A new pulsed polarization transfer experiment method is described for the polarization of 13C spins in a solid by magnetization transfer from protons. The method is directly analogous to the INEPT sequence for liquids introduced by Freeman and Morris. As polarization is transferred in PPT between individual 1H 13C pairs, rather than between spin reservoirs, different opportunities exist for structurally selective experiments. Results on p-diethoxybenzene and coronene are presented.

  17. Interpretation of injection-withdrawal tracer experiments conducted between two wells in a large single fracture.

    PubMed

    Novakowski, K S; Bickerton, G; Lapcevic, P

    2004-09-01

    Tracer experiments conducted using a flow field established by injecting water into one borehole and withdrawing water from another are often used to establish connections and investigate dispersion in fractured rock. As a result of uncertainty in the uniqueness of existing models used for interpretation, this method has not been widely used to investigate more general transport processes including matrix diffusion or advective solute exchange between mobile and immobile zones of fluid. To explore the utility of the injection-withdrawal method as a general investigative tool and with the intent to resolve the transport processes in a discrete fracture, two tracer experiments were conducted using the injection-withdrawal configuration. The experiments were conducted in a fracture which has a large aperture (>500 microm) and horizontally pervades a dolostone formation. One experiment was conducted in the direction of the hydraulic gradient and the other in the direction opposite to the natural gradient. Two tracers having significantly different values of the free-water diffusion coefficient were used. To interpret the experiments, a hybrid numerical-analytical model was developed which accounts for the arcuate shape of the flow field, advection-dispersion in the fracture, diffusion into the matrix adjacent to the fracture, and the presence of natural flow in the fracture. The model was verified by comparison to a fully analytical solution and to a well-known finite-element model. Interpretation of the tracer experiments showed that when only one tracer, advection-dispersion, and matrix diffusion are considered, non-unique results were obtained. However, by using multiple tracers and by accounting for the presence of natural flow in the fracture, unique interpretations were obtained in which a single value of matrix porosity was estimated from the results of both experiments. The estimate of porosity agrees well with independent measurements of porosity obtained from

  18. Interpretation of injection-withdrawal tracer experiments conducted between two wells in a large single fracture.

    PubMed

    Novakowski, K S; Bickerton, G; Lapcevic, P

    2004-09-01

    Tracer experiments conducted using a flow field established by injecting water into one borehole and withdrawing water from another are often used to establish connections and investigate dispersion in fractured rock. As a result of uncertainty in the uniqueness of existing models used for interpretation, this method has not been widely used to investigate more general transport processes including matrix diffusion or advective solute exchange between mobile and immobile zones of fluid. To explore the utility of the injection-withdrawal method as a general investigative tool and with the intent to resolve the transport processes in a discrete fracture, two tracer experiments were conducted using the injection-withdrawal configuration. The experiments were conducted in a fracture which has a large aperture (>500 microm) and horizontally pervades a dolostone formation. One experiment was conducted in the direction of the hydraulic gradient and the other in the direction opposite to the natural gradient. Two tracers having significantly different values of the free-water diffusion coefficient were used. To interpret the experiments, a hybrid numerical-analytical model was developed which accounts for the arcuate shape of the flow field, advection-dispersion in the fracture, diffusion into the matrix adjacent to the fracture, and the presence of natural flow in the fracture. The model was verified by comparison to a fully analytical solution and to a well-known finite-element model. Interpretation of the tracer experiments showed that when only one tracer, advection-dispersion, and matrix diffusion are considered, non-unique results were obtained. However, by using multiple tracers and by accounting for the presence of natural flow in the fracture, unique interpretations were obtained in which a single value of matrix porosity was estimated from the results of both experiments. The estimate of porosity agrees well with independent measurements of porosity obtained from

  19. Measurement of multiple psi torsion angles in uniformly 13C,15N-labeled alpha-spectrin SH3 domain using 3D 15N-13C-13C-15N MAS dipolar-chemical shift correlation spectroscopy.

    PubMed

    Ladizhansky, Vladimir; Jaroniec, Christopher P; Diehl, Annette; Oschkinat, Hartmut; Griffin, Robert G

    2003-06-01

    We demonstrate the simultaneous measurement of several backbone torsion angles psi in the uniformly (13)C,(15)N-labeled alpha-Spectrin SH3 domain using two different 3D 15N-13C-13C-15N dipolar-chemical shift magic-angle spinning (MAS) NMR experiments. The first NCCN experiment utilizes double quantum (DQ) spectroscopy combined with the INADEQUATE type 13C-13C chemical shift correlation. The decay of the DQ coherences formed between 13C'(i) and 13C(alphai) spin pairs is determined by the "correlated" dipolar field due to 15N(i)-13C(alphai) and 13C'(i)-15N(i+1) dipolar couplings and is particularly sensitive to variations of the torsion angle in the regime |psi| > 140 degrees. However, the ability of this experiment to constrain multiple psi-torsion angles is limited by the resolution of the 13C(alpha)-(13)CO correlation spectrum. This problem is partially addressed in the second approach described here, which is an NCOCA NCCN experiment. In this case the resolution is enhanced by the superior spectral dispersion of the 15N resonances present in the 15N(i+1)-13C(alphai) part of the NCOCA chemical shift correlation spectrum. For the case of the 62-residue alpha-spectrin SH3 domain, we determined 13 psi angle constraints with the INADEQUATE NCCN experiment and 22 psi constraints were measured in the NCOCA NCCN experiment.

  20. Tracer dilution measurements for two-phase geothermal production: Comparative testing and operating experience

    SciTech Connect

    Hirtz, P.; Lovekin, J.

    1995-12-31

    The tracer dilution technique for the measurement of steam and water mass flowrates and total enthalpy of two-phase geothermal fluids has been in routine use in the U.S.A. for almost three years. The tracer technique was first tested and adopted on a field-wide basis at the Coso geothermal field in California. Validation of the method was performed at the Roosevelt Hot Springs geothermal project in Utah and the Salton Sea and Heber geothermal projects in California by direct comparison to orifice-plate flowmeter measurements of the separated phases. Production well mass flowrates and total enthalpy are now regularly measured by this technique in the Coso, Salton Sea and Heber geothermal fields. Implementation of the tracer method is currently underway for the Tiwi and Bulalo geothermal fields in the Philippines. This paper presents the conceptual design of the measurement process, the results of field validations, and operating experience during field-wide testing in Coso.

  1. Measuring DNA synthesis rates with [1-13C]glycine.

    PubMed

    Chen, P; Abramson, F P

    1998-05-01

    We have devised and evaluated a stable-isotopic method for measuring DNA synthesis rates. The probe is [1-13C]-glycine that is incorporated into purines via de novo biosynthesis. The human hepatoma cell line HEP G2 was grown in medium containing [1-13C]glycine, the cells were harvested at various times, and the DNA was extracted. Following hydrolysis to the nucleosides, a reversed-phase HPLC separation was used to provide separate peaks for deoxythymidine (dT), deoxyadenosine (dA), and deoxyguanosine (dG). The HPLC effluent was continuously fed into a chemical reaction interface and an isotope ratio mass spectrometer (HPLC/CRI/IRMS). The isotope ratio of the CO2 produced in the CRI was used to monitor for enrichment. The cells were grown continuously for 5 days in labeled medium and also in a 1-day pulse labeling experiment where the washout of label was observed for the subsequent 9 days. As predicted from the role of glycine in de novo purine biosynthesis, the isotope ratio of the pyrimidine dT did not change. However, for the two purines, dA and dG, the characteristic log growth behavior of the cells was observed in their 13C/12C ratios and good agreement in the doubling time was obtained for each type of experiment. Parallel experiments that measured the HEP G2 doubling time in culture using tritiated thymidine incorporation and direct cell counts were carried out compare to our new method with established ones. We believe that the use of [1-13C]-glycine and the HPLC/CRI/IRMS is a highly sensitive and selective approach that forms the basis of a method that can measure DNA synthesis rates using a nonradioactive, nontoxic tracer. PMID:9599574

  2. Trip Report for the 2005 Sino-American SF6 Tracer Experiment

    SciTech Connect

    Bradley, M M; Sullivan, T J; Keating II, G A; Leach, M J

    2005-09-07

    The Chinese Institute for Radiation Protection (CIRP) conducted an SF6 atmospheric tracer experiment in July 2005 in the vicinity of the Qinshan Nuclear Power Company complex on the coast of the East China Sea. The experiment was partially sponsored by the US Department of Energy National Nuclear Security Administration, under the NA-23 International Emergency Management and Cooperation Program. NA-23 sent a delegation of five scientists to observe the experiment; four of the observers were from Lawrence Livermore National Laboratory (LLNL) and one was from the Japan Atomic Energy Research Institute (JAERI). CIRP's cooperation with the US-Japanese delegation was excellent, and the project was very successful from the international cooperation perspective. Although the experiment was modest in scope, it may provide one or more data sets that can be used for international dispersion model validation and intercomparison projects. Several areas for procedural improvements were noted by the US and Japanese observers, and a more concise measure of the experiment's scientific value will be available after CIRP completes and delivers the database of the experiment results by the end of the fiscal year. The consensus recommendation of the observers is that CIRP and DOE/NNSA NA-23 build on the experience and personal contacts gained during the experiment to plan and conduct an even more effective experiment in the future, perhaps as early as next year (2006). If the decision is made to conduct a follow-on experiment, we strongly recommend that the LLNL and JAERI representatives work cooperatively with CIRP throughout the entire planning phase of the experiment. As discussed in Section V, a 2006 China tracer experiment could serve as a springboard to a 2007 long-range international tracer experiment involving South Korea.

  3. Using Tracer Experiments To Study Phosphorus Transfer From Soil To Overland Flow

    NASA Astrophysics Data System (ADS)

    Vollmer, T.; Stamm, C.; Schaerer, M.; Sinaj, S.; Frossard, E.; Fluehler, H.

    Diffuse phosphorus (P) losses from agricultural land contribute to the eutrophication of surface water bodies in Switzerland. Grassland soils in areas of high animal stock densities are often prone to high P losses due to over-fertilization and a strong accumu- lation of P in the topsoil. In order to understand the effects of management practices and remediation measures on P transfer into runoff water at a small scale it is impor- tant to localize the sources of this phosphorus within the soil profile and to describe the water flows within the topsoil­overland flow system. We are studying the effects of remediation measures on P availability in the soil and on P concentrations in overland flow in a field experiment. We are using tracer exper- iments to examine the mixing behavior of water applied with a sprinkling device onto the soil surface with pre-event soil water and to trace the contribution of those two water sources to overland flow. Two plots were pre-irrigated with a solution of KBr in order to label the soil solu- tion. After a few days of equilibration, two fluorescent dyes were applied to different areas of the plots at a constant rate of 40 mm h-1. Surface runoff was analyzed for tracer concentrations. Small soil monoliths (0.35 * 0.25 *0.20 m3) were excavated and the tracer distribution within the blocks was mapped using a digital camera, optical filters, and tracer specific excitation light source. This tracing technique allowed for independent mapping of the distribution of two simultaneously applied tracers. The experiments demonstrated heterogenous infiltration of the dyes, negligible lat- eral translocation of the dyes within the soil, minimal transfer of the pre-applied Br- into overland flow, early breakthrough of the dye tracers in overland flow which was independent of the tracers sorption properties and a recovery of the dyes that corre- sponded to the runoff ratio. In all, the experiments indicate a very restricted interac- tion between

  4. Ground water pollution by roof runoff infiltration evidenced with multi-tracer experiments.

    PubMed

    Ammann, Adrian A; Hoehn, Eduard; Koch, Sabine

    2003-03-01

    The infiltration of urban roof runoff into well permeable subsurface material may have adverse effects on the ground water quality and endanger drinking water resources. Precipitation water from three different roofs of an industrial complex was channelled to a pit and infiltrated into a perialpine glaciofluvial gravel-and-sand aquifer. A shaft was constructed at the bottom of the pit and equipped with an array of TDR probes, lysimeters and suction cups that allowed measuring and sampling soil water at different depths. A fast infiltration flow was observed during natural rainfall events and during artificial infiltration experiments. For a better understanding of the behaviour of contaminants, experiments were conducted with cocktails of compounds of different reactivity (ammonium, strontium, atratone) and of non-reactive tracers (uranine, bromide, naphthionate), which represent different classes of pollutants. The experiment identified cation exchange reactions influencing the composition of the infiltrating water. These processes occurred under preferential flow conditions in macropores of the material. Measuring concentration changes under the controlled inflow of tracer experiments, the pollution potential was found to be high. Non-reactive tracers exhibited fast breakthrough and little sorption. PMID:12553990

  5. Ground water pollution by roof runoff infiltration evidenced with multi-tracer experiments.

    PubMed

    Ammann, Adrian A; Hoehn, Eduard; Koch, Sabine

    2003-03-01

    The infiltration of urban roof runoff into well permeable subsurface material may have adverse effects on the ground water quality and endanger drinking water resources. Precipitation water from three different roofs of an industrial complex was channelled to a pit and infiltrated into a perialpine glaciofluvial gravel-and-sand aquifer. A shaft was constructed at the bottom of the pit and equipped with an array of TDR probes, lysimeters and suction cups that allowed measuring and sampling soil water at different depths. A fast infiltration flow was observed during natural rainfall events and during artificial infiltration experiments. For a better understanding of the behaviour of contaminants, experiments were conducted with cocktails of compounds of different reactivity (ammonium, strontium, atratone) and of non-reactive tracers (uranine, bromide, naphthionate), which represent different classes of pollutants. The experiment identified cation exchange reactions influencing the composition of the infiltrating water. These processes occurred under preferential flow conditions in macropores of the material. Measuring concentration changes under the controlled inflow of tracer experiments, the pollution potential was found to be high. Non-reactive tracers exhibited fast breakthrough and little sorption.

  6. Hydraulic characterization of an activated sludge reactor with recycling system by tracer experiment and analytical models.

    PubMed

    Sánchez, F; Viedma, A; Kaiser, A S

    2016-09-15

    Fluid dynamic behaviour plays an important role in wastewater treatment. An efficient treatment requires the inexistence of certain hydraulic problems such as dead zones or short-circuiting flows. Residence time distribution (RTD) analysis is an excellent technique for detecting these inefficiencies. However, many wastewater treatment installations include water or sludge recycling systems, which prevent us from carrying out a conventional tracer pulse experiment to obtain the RTD curve of the installation. This paper develops an RTD analysis of an activated sludge reactor with recycling system. A tracer experiment in the reactor is carried out. Three analytical models, derived from the conventional pulse model, are proposed to obtain the RTD curve of the reactor. An analysis of the results is made, studying which model is the most suitable for each situation. This paper is useful to analyse the hydraulic efficiency of reactors with recycling systems. PMID:27288672

  7. Hydraulic characterization of an activated sludge reactor with recycling system by tracer experiment and analytical models.

    PubMed

    Sánchez, F; Viedma, A; Kaiser, A S

    2016-09-15

    Fluid dynamic behaviour plays an important role in wastewater treatment. An efficient treatment requires the inexistence of certain hydraulic problems such as dead zones or short-circuiting flows. Residence time distribution (RTD) analysis is an excellent technique for detecting these inefficiencies. However, many wastewater treatment installations include water or sludge recycling systems, which prevent us from carrying out a conventional tracer pulse experiment to obtain the RTD curve of the installation. This paper develops an RTD analysis of an activated sludge reactor with recycling system. A tracer experiment in the reactor is carried out. Three analytical models, derived from the conventional pulse model, are proposed to obtain the RTD curve of the reactor. An analysis of the results is made, studying which model is the most suitable for each situation. This paper is useful to analyse the hydraulic efficiency of reactors with recycling systems.

  8. Development of a methodology for the application of synthetic DNA in stream tracer injection experiments

    NASA Astrophysics Data System (ADS)

    Foppen, Jan Willem; Seopa, Judith; Bakobie, Noel; Bogaard, Thom

    2013-09-01

    Stream tracer injection experiments are useful for characterizing hydrological and biogeochemical processes in streams. We used nonconservative synthetic DNA and conservative NaCl in six instantaneous tracer injection experiments in streams in the Benelux. The main aim was to compare the performance of injected synthetic DNA tracer "T23" with NaCl. In all experiments, the shapes of the T23 and NaCl breakthrough curves (BTCs) were similar. Recovered T23 mass ranged from 2.9 to 52.6%, while recovered NaCl tracer mass ranged from 66.7% to complete mass recovery. In batch experiments, T23 decay was not detected. However, in those batches, we observed an unexplained initial T23 mass loss of 40-97%. In batches with sediment, T23 attachment rate coefficients ranged from close to zero to 0.2 hr-1. Advective and dispersive transport parameters of both NaCl and T23 fitted with STAMMT-L were similar. However, compared to T23, fitted storage zone areas of NaCl were 2-5 times larger, while storage zone exchange coefficients were two times larger. Fitted mass dilution factors of T23 ranged from 1.6 to 34.8. Together, these results pointed toward the disappearance of a part of the T23 mass due to both initial losses and attachment or sorption of T23 mass in those storage zone(s), while decay was not important. Our research demonstrated that artificial DNA can be a valuable tool to determine advective and dispersive transport in brooks, but not to assess solute mass exchange processes related to surface transient storage or hyporheic exchange.

  9. Development Of A Methodology For The Application Of Synthetic DNA In Stream Tracer Injection Experiments

    NASA Astrophysics Data System (ADS)

    Foppen, J.; Bogaard, T.

    2013-12-01

    Stream tracer injection experiments are useful for characterizing hydrological and biogeochemical processes in streams. We used non-conservative synthetic DNA and conservative NaCl in six instantaneous tracer injection experiments in streams in the Benelux. The main aim was to compare the performance of injected synthetic DNA tracer 'T23' with NaCl. In all experiments, the shapes of the T23 and NaCl breakthrough curves (BTCs) were similar. Recovered T23 mass ranged from 2.9-52.6%, while recovered NaCl tracer mass ranged from 66.7% to complete mass recovery. In batch experiments, T23 decay was not detected. However, in those batches, we observed an unexplained initial T23 mass loss of 40-97%. In batches with sediment, T23 attachment rate coefficients ranged from close to zero to 0.2 hr-1. Advective and dispersive transport parameters of both NaCl and T23 fitted with STAMMT-L were similar. However, compared to T23, fitted storage zone areas of NaCl were 2-5 times larger, while storage zone exchange coefficients were two times larger. Fitted mass dilution factors of T23 ranged from 1.6-34.8. Together, these results pointed towards the disappearance of a part of the T23 mass due to both initial losses and attachment or sorption of T23 mass in those storage zone(s), while decay was not important. Our research demonstrated that artificial DNA can be a valuable tool to determine advective and dispersive transport in brooks, but not to assess solute mass exchange processes related to surface transient storage or hyporheic exchange.

  10. Labeling strategies for 13C-detected aligned-sample solid-state NMR of proteins

    NASA Astrophysics Data System (ADS)

    Filipp, Fabian V.; Sinha, Neeraj; Jairam, Lena; Bradley, Joel; Opella, Stanley J.

    2009-12-01

    13C-detected solid-state NMR experiments have substantially higher sensitivity than the corresponding 15N-detected experiments on stationary, aligned samples of isotopically labeled proteins. Several methods for tailoring the isotopic labeling are described that result in spatially isolated 13C sites so that dipole-dipole couplings among the 13C are minimized, thus eliminating the need for homonuclear 13C- 13C decoupling in either indirect or direct dimensions of one- or multi-dimensional NMR experiments that employ 13C detection. The optimal percentage for random fractional 13C labeling is between 25% and 35%. Specifically labeled glycerol and glucose can be used at the carbon sources to tailor the isotopic labeling, and the choice depends on the resonances of interest for a particular study. For investigations of the protein backbone, growth of the bacteria on [2- 13C]-glucose-containing media was found to be most effective.

  11. Tomographic inversion of active thermal tracer experiments to characterize aquifer heterogeneity

    NASA Astrophysics Data System (ADS)

    Somogyvári, Márk; Bayer, Peter; Brauchler, Ralf

    2015-04-01

    The tomographical approach has been established as an efficient and robust way to characterize spatial heterogeneity of hydraulic aquifer properties. It has been successfully applied on pressure signals and solute tracers, but only little experience is available using heat tracer signals in a tomographic setup. The advantage of using a heat tracer is that temperature can be easily monitored with good temporal and spatial resolution and it provides information directly about the heat transport in the subsurface. In this study, we consider active injection of warm water into a shallow groundwater well. The concept is tested on a virtual aquifer implemented in a numerical model. The thermal evolution in the system after repeated injection at different depth levels is monitored though a tomographical observation setup. This delivers thermal tracer travel times for each combination of injection (source) and observation points (receivers). The combined inversion of all source-receiver travel times is formulated and efficiently solved as an eikonal problem. The result of the eikonal inversion is a cross section through the aquifer of mean tracer velocity and of inverted flow paths. By assuming that the heat transport is dominated by advection, a hydraulic diffusivity map can be calculated from the velocity map, similar to solute tracer tomography, introducing one new variable, which is known as the thermal retardation factor. This assumption is crucial for the inversion procedure, and it also reflects that our main interest is in the detection of preferential flow paths, where thermal diffusion plays only a minor role. For this purpose, early time diagnostics are used instead of, for example, the mean breakthrough time at the receivers. To characterize the reliability of the results, the null-space energy map is calculated based on the inverted advection flow paths. First results of the heat tracer inversion approach are comparable to those from other hydrogeological

  12. Metabolic network reconstruction, growth characterization and 13C-metabolic flux analysis of the extremophile Thermus thermophilus HB8.

    PubMed

    Swarup, Aditi; Lu, Jing; DeWoody, Kathleen C; Antoniewicz, Maciek R

    2014-07-01

    Thermus thermophilus is an extremely thermophilic bacterium with significant biotechnological potential. In this work, we have characterized aerobic growth characteristics of T. thermophilus HB8 at temperatures between 50 and 85°C, constructed a metabolic network model of its central carbon metabolism and validated the model using (13)C-metabolic flux analysis ((13)C-MFA). First, cells were grown in batch cultures in custom constructed mini-bioreactors at different temperatures to determine optimal growth conditions. The optimal temperature for T. thermophilus grown on defined medium with glucose was 81°C. The maximum growth rate was 0.25h(-1). Between 50 and 81°C the growth rate increased by 7-fold and the temperature dependence was described well by an Arrhenius model with an activation energy of 47kJ/mol. Next, we performed a (13)C-labeling experiment with [1,2-(13)C] glucose as the tracer and calculated intracellular metabolic fluxes using (13)C-MFA. The results provided support for the constructed network model and highlighted several interesting characteristics of T. thermophilus metabolism. We found that T. thermophilus largely uses glycolysis and TCA cycle to produce biosynthetic precursors, ATP and reducing equivalents needed for cells growth. Consistent with its proposed metabolic network model, we did not detect any oxidative pentose phosphate pathway flux or Entner-Doudoroff pathway activity. The biomass precursors erythrose-4-phosphate and ribose-5-phosphate were produced via the non-oxidative pentose phosphate pathway, and largely via transketolase, with little contribution from transaldolase. The high biomass yield on glucose that was measured experimentally was also confirmed independently by (13)C-MFA. The results presented here provide a solid foundation for future studies of T. thermophilus and its metabolic engineering applications.

  13. New guidelines for δ13C measurements

    USGS Publications Warehouse

    Coplen, Tyler B.; Brand, Willi A.; Gehre, Matthias; Groning, Manfred; Meijer, Harro A. J.; Toman, Blaza; Verkouteren, R. Michael

    2006-01-01

    Consistency of δ13C measurements can be improved 39−47% by anchoring the δ13C scale with two isotopic reference materials differing substantially in 13C/12C. It is recommended thatδ13C values of both organic and inorganic materials be measured and expressed relative to VPDB (Vienna Peedee belemnite) on a scale normalized by assigning consensus values of −46.6‰ to L-SVEC lithium carbonate and +1.95‰ to NBS 19 calcium carbonate. Uncertainties of other reference material values on this scale are improved by factors up to two or more, and the values of some have been notably shifted:  the δ13C of NBS 22 oil is −30.03%.

  14. Hyperpolarized (13)C Magnetic Resonance and Its Use in Metabolic Assessment of Cultured Cells and Perfused Organs.

    PubMed

    Lumata, Lloyd; Yang, Chendong; Ragavan, Mukundan; Carpenter, Nicholas; DeBerardinis, Ralph J; Merritt, Matthew E

    2015-01-01

    Diseased tissue is often characterized by abnormalities in intermediary metabolism. Observing these alterations in situ may lead to an improved understanding of pathological processes and novel ways to monitor these processes noninvasively in human patients. Although (13)C is a stable isotope safe for use in animal models of disease as well as human subjects, its utility as a metabolic tracer has largely been limited to ex vivo analyses employing analytical techniques like mass spectrometry or nuclear magnetic resonance spectroscopy. Neither of these techniques is suitable for noninvasive metabolic monitoring, and the low abundance and poor gyromagnetic ratio of conventional (13)C make it a poor nucleus for imaging. However, the recent advent of hyperpolarization methods, particularly dynamic nuclear polarization (DNP), makes it possible to enhance the spin polarization state of (13)C by many orders of magnitude, resulting in a temporary amplification of the signal sufficient for monitoring kinetics of enzyme-catalyzed reactions in living tissue through magnetic resonance spectroscopy or magnetic resonance imaging. Here, we review DNP techniques to monitor metabolism in cultured cells, perfused hearts, and perfused livers, focusing on our experiences with hyperpolarized [1-(13)C]pyruvate. We present detailed approaches to optimize the DNP procedure, streamline biological sample preparation, and maximize detection of specific metabolic activities. We also discuss practical aspects in the choice of metabolic substrates for hyperpolarization studies and outline some of the current technical and conceptual challenges in the field, including efforts to use hyperpolarization to quantify metabolic rates in vivo.

  15. Correct Characterization of Passive Tracer Dispersion in Porous Columns: Experiments vs. Theory

    NASA Astrophysics Data System (ADS)

    Cortis, A.; Scher, H.; Berkowitz, B.

    2004-12-01

    Breakthrough curves (BTC) of a passive tracer in macroscopically ``homogeneous'' granular materials (well-sorted, unconsolidated sands or glass beads) were measured in a series of column experiments. % In parallel, classical experiments on dispersion of a passive tracer in fully and partially saturated porous columns were re-examined. % All of these BTCs exhibit anomalous (non-Fickian) features: early and late arrival times are observed to differ systematically from theoretical predictions based on solution of the advective-dispersion equation (ADE) for uniform porous media. % We propose that even in these small-scale, ``homogeneous'' porous medium columns, subtle and residual pore-scale disorder effects can account for these observations. % In a Continuous Time Random Walk (CTRW) framework, we determined an ensemble-averaged distribution of particle transfer rates (based on a Master Equation for the local flux-averaged concentration) which accounts for these effects. % Solutions of the resulting CTRW transport equations yield BTCs that are in excellent agreement with the entire series of observations. % The CTRW formulation also specifies the dependence of the effective macroscopic parameters on measurable quantities. % The theoretical predictions are in excellent agreement with the observations. % It is critical to understand that as a consequence of our results, the ADE should not be taken as the starting point of any upscaling technique. % Our analyses demonstrate that existing measurements and interpretations of tracer dispersion experiments in laboratory experiments should be carefully re-considered in the framework of these recent advances in conceptual understanding and quantification. % These results have also important implications for modeling the transport of contaminants in large-scale, highly-heterogeneous, hydrogeological systems.

  16. 13C-based metabolic flux analysis: fundamentals and practice.

    PubMed

    Yang, Tae Hoon

    2013-01-01

    Isotope-based metabolic flux analysis is one of the emerging technologies applied to system level metabolic phenotype characterization in metabolic engineering. Among the developed approaches, (13)C-based metabolic flux analysis has been established as a standard tool and has been widely applied to quantitative pathway characterization of diverse biological systems. To implement (13)C-based metabolic flux analysis in practice, comprehending the underlying mathematical and computational modeling fundamentals is of importance along with carefully conducted experiments and analytical measurements. Such knowledge is also crucial when designing (13)C-labeling experiments and properly acquiring key data sets essential for in vivo flux analysis implementation. In this regard, the modeling fundamentals of (13)C-labeling systems and analytical data processing are the main topics we will deal with in this chapter. Along with this, the relevant numerical optimization techniques are addressed to help implementation of the entire computational procedures aiming at (13)C-based metabolic flux analysis in vivo.

  17. A Geoelectrically-Monitored Tracer Test At The Macrodispersion Experiment (MADE) Site In Columbus, Mississippi

    NASA Astrophysics Data System (ADS)

    Swanson, R. D.; Singha, K.; Pidlisecky, A.; Hyndman, D. W.; Butler, J. J.; Bohling, G.

    2010-12-01

    The Macrodispersion Experiment (MADE) Site in northeastern Mississippi consists of a shallow unconfined aquifer of poorly to well-sorted sand intermixed with silts and gravel. Fluvial deposits extend 12m below the surface and terminate at the low hydraulic conductivity, clay-rich Eutaw Formation. The MADE site has been the subject of numerous tracer tests due to its highly heterogeneous aquifer. Despite these tests, there is still disagreement over the dominant mechanism of transport; proposed solutions include modeling large scale hydraulic conductivity variations, allowing for poorly and well-connected flowpaths and dividing the subsurface into mobile and immobile domains. In October 2009, a doublet tracer test was performed by injecting a solution of NaCl into one well and extracting groundwater at an equal rate 6.2m down gradient. The tracer was monitored from 1 to 12m using 34 sampling points in 7 multi-level samplers for over 100 days. For 4 days after injection, over 129,000 in-well and cross-borehole electrical resistivity measurements were collected between 4 wells containing 16 electrodes each. Initial inversions of electrical resistivity data indicate two distinct regions of lower resistivity zones near the surface and the Eutaw clay with a higher resistivity zone near the middle of the aquifer. Geophysical tomograms, when compared with both gamma borehole measurements and high-resolution hydraulic conductivity measurements, suggest at this site electrical resistivity measurements are highly correlated to hydraulic conductivity and silt content. Breakthrough histories show a heterogeneous pattern of tracer arrival times and long tails in concentration at most depths. For example, measurements from a multilevel sampler 5m down gradient of the injection well indicate breakthroughs at 15 and 26 hours at a depth of 7 and 4m, respectively, but little variation in fluid conductivity was observed at 1 and 6m throughout the duration of the experiment. Both in

  18. In vivo13C spectroscopy in the rat brain using hyperpolarized [1- 13C]pyruvate and [2- 13C]pyruvate

    NASA Astrophysics Data System (ADS)

    Marjańska, Małgorzata; Iltis, Isabelle; Shestov, Alexander A.; Deelchand, Dinesh K.; Nelson, Christopher; Uğurbil, Kâmil; Henry, Pierre-Gilles

    2010-10-01

    The low sensitivity of 13C spectroscopy can be enhanced using dynamic nuclear polarization. Detection of hyperpolarized [1- 13C]pyruvate and its metabolic products has been reported in kidney, liver, and muscle. In this work, the feasibility of measuring 13C signals of hyperpolarized 13C metabolic products in the rat brain in vivo following the injection of hyperpolarized [1- 13C]pyruvate and [2- 13C]pyruvate is investigated. Injection of [2- 13C]pyruvate led to the detection of [2- 13C]lactate, but no other downstream metabolites such as TCA cycle intermediates were detected. Injection of [1- 13C]pyruvate enabled the detection of both [1- 13C]lactate and [ 13C]bicarbonate. A metabolic model was used to fit the hyperpolarized 13C time courses obtained during infusion of [1- 13C]pyruvate and to determine the values of VPDH and VLDH.

  19. Characterization of reactive tracers for C-wells field experiments 1: Electrostatic sorption mechanism, lithium

    SciTech Connect

    Fuentes, H.R.; Polzer, W.L.; Essington, E.H.; Newman, B.D.

    1989-11-01

    Lithium (Li{sup +}) was introduced as lithium bromide (LiBr), as a retarded tracer for experiments in the C-wells complex at Yucca Mountain, Nevada Test Site, Nevada. The objective was to evaluate the potential of lithium to sorb predominately by physical forces. lithium was selected as a candidate tracer on the basis of high solubility, good chemical and biological stability, and relatively low sorptivity; lack of bioaccumulation and exclusion as a priority pollutant in pertinent federal environmental regulations; good analytical detectability and low natural background concentrations; and a low cost Laboratory experiments were performed with suspensions of Prow Pass cuttings from drill hole UE-25p{number_sign}1 at depths between 549 and 594 m in J-13 water at a pH of approximately 8 and in the temperature range of 25{degree}C to 45{degree}C. Batch equilibrium and kinetics experiments were performed; estimated thermodynamic constants, relative behavior between adsorption and desorption, and potentiometric studies provided information to infer the physical nature of lithium sorption.

  20. Synthesis and applications of {sup 13}C glycerol

    SciTech Connect

    Stocking, E.; Khalsa, O.; Martinez, R.A.; Silks, L.A. III

    1994-12-01

    Due in part to the use of labeled glycerol for the {sup 13}C enrichment of biomolecules, we are currently developing new synthetic routes to various isotopomers of glycerol. Judging from our experience, traditional methods of glycerol synthesis are not easily adapted for isotopic enrichment and/or have poor overall yields (12 to 15%). Furthermore, the use of glycerol for enrichment can be prohibitively expensive and its availability is limited by the level of demand. We are presently developing a short de novo synthesis of glycerol from carbon dioxide ({approximately}53% overall yield for four steps) and are examining the feasibility of synthesizing site-specific {sup 13}C-labeled glycerol and dihydroxyacetone (DHA) from labeled methanol and carbon dioxide. One application of {sup 13}C glycerol we have examined is enzymatic conversion of glycerol to glyceraldehyde-3-monophosphate or dihydroxyacetone monophosphate (DHAP) with yields ranging from 25 to 50% (as determined by NMR spectroscopy). We are also pursuing the chemical conversion of {sup 13}C-labeled DHA to DHAP. We are especially interested in {sup 13}C-labeled DHAP because we are investigating its use as a chemo-enzymatic precursor for both labeled 2-deoxyribose and 2-deoxyribonucleic acids.

  1. Ultra-large-scale Cosmology in Next-generation Experiments with Single Tracers

    NASA Astrophysics Data System (ADS)

    Alonso, David; Bull, Philip; Ferreira, Pedro G.; Maartens, Roy; Santos, Mário G.

    2015-12-01

    Future surveys of large-scale structure will be able to measure perturbations on the scale of the cosmological horizon, and so could potentially probe a number of novel relativistic effects that are negligibly small on sub-horizon scales. These effects leave distinctive signatures in the power spectra of clustering observables and, if measurable, would open a new window on relativistic cosmology. We quantify the size and detectability of the effects for the most relevant future large-scale structure experiments: spectroscopic and photometric galaxy redshift surveys, intensity mapping surveys of neutral hydrogen, and radio continuum surveys. Our forecasts show that next-generation experiments, reaching out to redshifts z≃ 4, will not be able to detect previously undetected general-relativistic effects by using individual tracers of the density field, although the contribution of weak lensing magnification on large scales should be clearly detectable. We also perform a rigorous joint forecast for the detection of primordial non-Gaussianity through the excess power it produces in the clustering of biased tracers on large scales, finding that uncertainties of σ ({f}{{NL}})∼ 1-2 should be achievable. We study the level of degeneracy of these large-scale effects with several tracer-dependent nuisance parameters, quantifying the minimal priors on the latter that are needed for an optimal measurement of the former. Finally, we discuss the systematic effects that must be mitigated to achieve this level of sensitivity, and some alternative approaches that should help to improve the constraints. The computational tools developed to carry out this study, which requires the full-sky computation of the theoretical angular power spectra for {O}(100) redshift bins, as well as realistic models of the luminosity function, are publicly available at http://intensitymapping.physics.ox.ac.uk/codes.html.

  2. (13)C-metabolic flux analysis of lipid accumulation in the oleaginous fungus Mucor circinelloides.

    PubMed

    Zhao, Lina; Zhang, Huaiyuan; Wang, Liping; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

    2015-12-01

    The oleaginous fungus Mucor circinelloides is of industrial interest because it can produce high levels of polyunsaturated fatty acid γ-linolenic acid. M. circinelloides CBS 277.49 is able to accumulate less than 15% of cell dry weight as lipids, while M. circinelloides WJ11 can accumulate lipid up to 36%. In order to better understand the mechanisms behind the differential lipid accumulation in these two strains, tracer experiments with (13)C-glucose were performed with the growth of M. circinelloides and subsequent gas chromatography-mass spectrometric detection of (13)C-patterns in proteinogenic amino acids was carried out to identify the metabolic network topology and estimate intracellular fluxes. Our results showed that the high oleaginous strain WJ11 had higher flux of pentose phosphate pathway and malic enzyme, lower flux in tricarboxylic acid cycle, higher flux in glyoxylate cycle and ATP: citrate lyase, together, it might provide more NADPH and substrate acetyl-CoA for fatty acid synthesis.

  3. Modelling of a Tracer experiment (Bromide) at the lysimeter Wagna/Austria with MIKE-SHE

    NASA Astrophysics Data System (ADS)

    Reszler, Christian; Fank, Johann

    2015-04-01

    Data of a tracer experiment with Bromide at one of the three lysimeters in Wagna/Austria are used to test the unsaturated zone solute transport model in MIKE-SHE. On April 4th, 2005 50 mg/l of Bromide were applied on the lysimeter operated with conventional farming. At this time the lysimeter was covered with bare soil until the start of the cultivation of pumpkin one month later. Concentrations at the lysimeter bottom (180 cm depth) were measured and, after break-through, plant uptake was measured to quantify mass recovery. The model using the Richards-Van Genuchten-Mualem approach is setup by comprehensive data of vegetation and soil hydraulic properties available at the lysimeter. Water movement simulation in the unsaturated zone is tested against measured seepage rates at the lysimeter bottom and soil water contents in different soil depths in a period of five years. A sensitivity study shows that, particularly in the quaternary gravel zone two different parameter sets are necessary to represent the different dynamics of water content and seepage. With both two sets the general dynamics of the tracer experiment are simulated well. However, the early rapid rise of the measured concentrations could not be represented by either parameter set, which indicates a complex pore system consisting of different flow paths in the gravel zone, e.g., a system of matrix flow and macro-pore flow.

  4. Sulphur tracer experiments in laboratory animals using 34S-labelled yeast.

    PubMed

    Martínez-Sierra, J Giner; Moreno Sanz, F; Herrero Espílez, P; Marchante Gayón, J M; Rodríguez Fernández, J; García Alonso, J I

    2013-03-01

    We have evaluated the use of (34)S-labelled yeast to perform sulphur metabolic tracer experiments in laboratory animals. The proof of principle work included the selection of the culture conditions for the preparation of sulphur labelled yeast, the study of the suitability of this labelled yeast as sulphur source for tracer studies using in vitro gastrointestinal digestion and the administration of the (34)S-labelled yeast to laboratory animals to follow the fate and distribution of (34)S in the organism. For in vitro gastrointestinal digestion, the combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) showed that labelled methionine, cysteine and other low molecular weight sulphur-containing biomolecules were the major components in the digested extracts of the labelled yeast. Next, in vivo kinetic experiments were performed in healthy Wistar rats after the oral administration of (34)S-labelled yeast. The isotopic composition of total sulphur in tissues, urine and faeces was measured by double-focusing inductively coupled plasma mass spectrometry after microwave digestion. It was observed that measurable isotopic enrichments were detected in all samples. Finally, initial investigations on sulphur isotopic composition of serum and urine samples by HPLC-ICP-MS have been carried out. For serum samples, no conclusive data were obtained. Interestingly, chromatographic analysis of urine samples showed differential isotope enrichment for several sulphur-containing biomolecules.

  5. Tracer experiment and model evidence for macrofaunal shaping of microbial nitrogen functions along rocky shores

    NASA Astrophysics Data System (ADS)

    Pfister, Catherine A.; Altabet, Mark A.; Pather, Santhiska; Dwyer, Greg

    2016-06-01

    Seawater microbes as well as those associated with macrobiota are increasingly recognized as a key feature affecting nutrient cycling. Tidepools are ideal natural mesocosms to test macrofauna and microbe interactions, and we quantified rates of microbial nitrogen processing using tracer enrichment of ammonium (15NNH4) or nitrate (15NNO3) when tidepools were isolated from the ocean during low intertidal periods. Experiments were conducted during both day and night as well as in control tidepools and those from which mussels had been removed, allowing us to determine the role of both mussels and daylight in microbial nitrogen processing. We paired time series observations of 15N enrichment in NH4+, NO2- and NO3- with a differential equation model to quantify multiple, simultaneous nitrogen transformations. Mussel presence and daylight increased remineralization and photosynthetic nitrogen uptake. When we compared ammonium gain or loss that was attributed to any tidepool microbes vs. photosynthetic uptake, microbes accounted for 32 % of this ammonium flux on average. Microbial transformations averaged 61 % of total nitrate use; thus, microbial activity was almost 3 times that of photosynthetic nitrate uptake. Because it accounted for processes that diluted our tracer, our differential equation model assigned higher rates of nitrogen processing compared to prior source-product models. Our in situ experiments showed that animals alone elevate microbial nitrogen transformations by 2 orders of magnitude, suggesting that coastal macrobiota are key players in complex microbial nitrogen transformations.

  6. Simultaneous investigation of cardiac pyruvate dehydrogenase flux, Krebs cycle metabolism and pH, using hyperpolarized [1,2-(13)C2]pyruvate in vivo.

    PubMed

    Chen, Albert P; Hurd, Ralph E; Schroeder, Marie A; Lau, Angus Z; Gu, Yi-ping; Lam, Wilfred W; Barry, Jennifer; Tropp, James; Cunningham, Charles H

    2012-02-01

    (13)C MR spectroscopy studies performed on hearts ex vivo and in vivo following perfusion of prepolarized [1-(13)C]pyruvate have shown that changes in pyruvate dehydrogenase (PDH) flux may be monitored non-invasively. However, to allow investigation of Krebs cycle metabolism, the (13)C label must be placed on the C2 position of pyruvate. Thus, the utilization of either C1 or C2 labeled prepolarized pyruvate as a tracer can only afford a partial view of cardiac pyruvate metabolism in health and disease. If the prepolarized pyruvate molecules were labeled at both C1 and C2 positions, then it would be possible to observe the downstream metabolites that were the results of both PDH flux ((13)CO(2) and H(13)CO(3)(-)) and Krebs cycle flux ([5-(13)C]glutamate) with a single dose of the agent. Cardiac pH could also be monitored in the same experiment, but adequate SNR of the (13)CO(2) resonance may be difficult to obtain in vivo. Using an interleaved selective RF pulse acquisition scheme to improve (13)CO(2) detection, the feasibility of using dual-labeled hyperpolarized [1,2-(13)C(2)]pyruvate as a substrate for dynamic cardiac metabolic MRS studies to allow simultaneous investigation of PDH flux, Krebs cycle flux and pH, was demonstrated in vivo.

  7. (13)C-(13)c homonuclear recoupling in solid-state nuclear magnetic resonance at a moderately high magic-angle-spinning frequency.

    PubMed

    Mithu, Venus Singh; Bakthavatsalam, Subha; Madhu, Perunthiruthy K

    2013-01-01

    Two-dimensional (13)C-(13)C correlation experiments are widely employed in structure determination of protein assemblies using solid-state nuclear magnetic resonance. Here, we investigate the process of (13)C-(13)C magnetisation transfer at a moderate magic-angle-spinning frequency of 30 kHz using some of the prominent second-order dipolar recoupling schemes. The effect of isotropic chemical-shift difference and spatial distance between two carbons and amplitude of radio frequency on (1)H channel on the magnetisation transfer efficiency of these schemes is discussed in detail.

  8. Vehicle emissions of greenhouse gases and related tracers from a tunnel study: CO : CO2, N2O : CO2, CH4 : CO2, O2 : CO2 ratios, and the stable isotopes 13C and 18O in CO2 and CO

    NASA Astrophysics Data System (ADS)

    Popa, M. E.; Vollmer, M. K.; Jordan, A.; Brand, W. A.; Pathirana, S. L.; Rothe, M.; Röckmann, T.

    2014-02-01

    Measurements of CO2, CO, N2O and CH4 mole fractions, O2 / N2 ratios and the stable isotopes 13C and 18O in CO2 and CO have been performed in air samples from the Islisberg highway tunnel (Switzerland). The molar CO : CO2 ratios, with an average of (4.15 ± 0.34) ppb:ppm, are lower than reported in previous studies, pointing to a reduction in CO emissions from traffic. The 13C in CO2 reflects the isotopic composition of the fuel. 18O in CO2 is slightly depleted compared to the 18O in atmospheric O2, and shows significant variability. In contrast, the δ13C values of CO show that significant fractionation takes place during CO destruction in the catalytic converter. 13C in CO is enriched by 3‰ compared to the 13C in the fuel burnt, while the 18O content is similar to that of atmospheric O2. We compute a fractionation constant of (-2.7 ± 0.7)‰ for 13C during CO destruction. The N2O : CO2 average ratio of (1.8 ± 0.2) × 10-2 ppb:ppm is significantly lower than in past studies, showing a reduction in N2O emissions likely related to improvements in the catalytic converter technology. We also observed small CH4 emissions, with an average CH4 : CO2 ratio of (4.6 ± 0.2) × 10-2 ppb:ppm. The O2 : CO2 ratios of (-1.47 ± 0.01) ppm:ppm are very close to the expected, theoretically calculated values of O2 depletion per CO2 enhancement.

  9. Determination of 13C/ 12C ratios with (d, p) nuclear reactions

    NASA Astrophysics Data System (ADS)

    Wang, Y. Q.; Zhang, J.; Tesmer, J. R.; Li, Y. H.; Greco, R.; Grim, G. P.; Obst, A. W.; Rundberg, R. S.; Wilhelmy, J. B.

    2010-06-01

    Stable isotope ratios such as 13C/ 12C play an important role in many applications including environment and energy research. Since many surface analysis techniques are plagued with unavoidable hydrocarbon contamination issues during analysis, it is highly desirable that 13C and 12C isotopes be measured simultaneously especially in specimens with a minute amount of 13C, in order to reliably determine 13C/ 12C ratios. In this paper, we report that deuterium induced proton particle reactions, 13C(d, p) 14C and 12C(d, p) 13C, provide a convenient and reliable approach for 13C/ 12C ratio determination. Optimizations on experimental considerations and potential interferences from other common light isotopes are discussed as well as results from the application of this technique to diagnose the performance of a target debris collection in an inertial confinement fusion (ICF) experiment.

  10. Evaluation of commercial magnetic iron oxides as sediment tracers in water erosion experiments

    NASA Astrophysics Data System (ADS)

    Guzman, G.; Barron, V.; Gomez, J. A.

    2009-04-01

    Water erosion is one of the mayor concerns to sustainability of agricultural systems in Mediterranean countries, e.g. olive farming areas in Southern Spain. Despite an increase in the number of published studies on erosion rates and conservation measures, significant uncertainty persists on actual erosion rates in these areas (Gómez et al., 2008; Fleskens and Stroosnijder, 2007). Due to the limitations and cost of technologies traditionally used in erosion measurement, there is a growing interest in the use of innovative erosion tracers that could be applied to the soil and used to monitor erosion and deposition rates at experiments performed at different scales and environments. An example of these innovative traces, which could complement the potential of more traditional tracers like Cs-137, is rare earths oxides. Due to its size, D50 ranging from 1.23 to 16.38 m (Zhang et al., 2003), these rare earth oxides tagged soil aggregates more or less homogeneously and have been used in tracking sediment movement at laboratory and field scale, e.g. Polyakov and Nearing, (2004). One of the shortcomings of the use of rare earth oxides in the cost derived of the need to use Inductive Coupled Plasma Mass Spectrometry to determine its concentration in the tagged soil. The use of mineral magnetic measurements provide a less expensive alternative to complement erosion and sediment delivery in eroding landscapes (Royall, 2001), and is also an area of active research. However, most of the studies are based on measurements of magnetic properties inherent to soil materials, and little research has been done about the possibility of tagging soils with magnetic materials. Ventura et al. (2002) tagged a loamy soil with a magnetic tracer for use in rainfall simulation experiments. They concluded that the magnetic tracer used, magnetic beads of 3.2 mm of mean weight diameter, although useful in determining erosion and deposition areas presented a tracer to soil ratio that did not

  11. TRANSALP 1989 experimental campaign—II. Simulation of a tracer experiment with Lagrangian particle models

    NASA Astrophysics Data System (ADS)

    Anfossi, D.; Desiato, F.; Tinarelli, G.; Brusasca, G.; Ferrero, E.; Sacchetti, D.

    The capability of two 3D Lagrangian particle models (ARCO and SPRAY) in simulating the airborne dispersion in highly complex terrain is tested. The simulations are compared to the tracer experiment TRANSALP-I performed on the 19th of October 1989 in the Alpine region (Canton of Ticino, Switzerland). TRANSALP exercises were part of the EUROTRAC-TRACT Subproject. Tracer was released during one hour (11.00-12.00) near the ground at Iragna, a few kilometres upwind from the bifurcation of two valleys (Leventina and Blenio Valleys); about 40 samplers, located in the two valleys in the range of 40 km, measured 1/2 h average mean concentrations near the ground for five hours. The results of the simulations are presented and discussed. A statistical model evaluation aimed at quantifying the performance of the two models was carried out. The simulations reproduced with a reasonable degree of accuracy the general behaviour of the observed phenomena: the split of the plume between the two valleys, the space and time distribution of the concentration, the concentration maxima locations.

  12. Measuring (13)C-(2)D dipolar couplings with a universal REDOR dephasing curve

    PubMed

    Gullion

    2000-09-01

    A (13)C-observe REDOR experiment is described which allows (13)C-(2)D dipolar couplings to be obtained by a universal dipolar dephasing curve. Previous (13)C-observe REDOR experiments on (13)C-(2)D spin pairs generally relied on numerical simulations to obtain the dipolar coupling. The REDOR experiment described in this article is based on a deuterium composite pulse, and the data analysis eliminates the need for numerical simulations and is the same as the traditional REDOR analysis performed on pairs of spin-12 nuclei. Copyright 2000 Academic Press. PMID:10968975

  13. Coupling heat and chemical tracer experiments for estimating heat transfer parameters in shallow alluvial aquifers.

    PubMed

    Wildemeersch, S; Jamin, P; Orban, P; Hermans, T; Klepikova, M; Nguyen, F; Brouyère, S; Dassargues, A

    2014-11-15

    Geothermal energy systems, closed or open, are increasingly considered for heating and/or cooling buildings. The efficiency of such systems depends on the thermal properties of the subsurface. Therefore, feasibility and impact studies performed prior to their installation should include a field characterization of thermal properties and a heat transfer model using parameter values measured in situ. However, there is a lack of in situ experiments and methodology for performing such a field characterization, especially for open systems. This study presents an in situ experiment designed for estimating heat transfer parameters in shallow alluvial aquifers with focus on the specific heat capacity. This experiment consists in simultaneously injecting hot water and a chemical tracer into the aquifer and monitoring the evolution of groundwater temperature and concentration in the recovery well (and possibly in other piezometers located down gradient). Temperature and concentrations are then used for estimating the specific heat capacity. The first method for estimating this parameter is based on a modeling in series of the chemical tracer and temperature breakthrough curves at the recovery well. The second method is based on an energy balance. The values of specific heat capacity estimated for both methods (2.30 and 2.54MJ/m(3)/K) for the experimental site in the alluvial aquifer of the Meuse River (Belgium) are almost identical and consistent with values found in the literature. Temperature breakthrough curves in other piezometers are not required for estimating the specific heat capacity. However, they highlight that heat transfer in the alluvial aquifer of the Meuse River is complex and contrasted with different dominant process depending on the depth leading to significant vertical heat exchange between upper and lower part of the aquifer. Furthermore, these temperature breakthrough curves could be included in the calibration of a complex heat transfer model for

  14. Haze in the Grand Canyon: An evaluation of the Winter Haze Intensive Tracer Experiment

    SciTech Connect

    Not Available

    1990-10-01

    The Grand Canyon is one of the most spectacular natural sights on earth. Approximately 4 million visitors travel to Grand Canyon National Park (GCNP) each year to enjoy its majestic geological formations and intensely colored views. However, visibility in GCNP can be impaired by small increases in concentrations of fine suspended particles that scatter and absorb light; the resulting visibility degradation is perceived as haze. Sulfate particles are a major factor in visibility impairment at Grand Canyon in summer and winter. Many wintertime hazes at GCNP are believed to result from the accumulation of emissions from local sources during conditions of air stagnation, which occur more frequently in winter than in summer. In January and February 1987, the National Park Service (NPS) carried out a large-scale experiment known as the Winter Haze Intensive Tracer Experiment (WHITEX) to investigate the causes of wintertime haze in the region of GCNP and Canyonlands National Park. The overall objective of WHITEX was to assess the feasibility of attributing visibility impairment in specific geographic regions to emissions from a single point source. The experiment called for the injection of a tracer, deuterated methane (CD{sub 4}), into one of the stacks of the Navajo Generating Station (NGS), a major coal-fired power plant located 25 km from the GCNP boundary and 110 km northeast of Grand Canyon Village. A network of field stations was established in the vicinity -- mostly to the northeast of GCNP and NGS -- to measure CD{sub 4} concentrations, atmospheric aerosol and optical properties, and other chemical and physical attributes. 19 refs., 3 figs.

  15. Coupling heat and chemical tracer experiments for estimating heat transfer parameters in shallow alluvial aquifers.

    PubMed

    Wildemeersch, S; Jamin, P; Orban, P; Hermans, T; Klepikova, M; Nguyen, F; Brouyère, S; Dassargues, A

    2014-11-15

    Geothermal energy systems, closed or open, are increasingly considered for heating and/or cooling buildings. The efficiency of such systems depends on the thermal properties of the subsurface. Therefore, feasibility and impact studies performed prior to their installation should include a field characterization of thermal properties and a heat transfer model using parameter values measured in situ. However, there is a lack of in situ experiments and methodology for performing such a field characterization, especially for open systems. This study presents an in situ experiment designed for estimating heat transfer parameters in shallow alluvial aquifers with focus on the specific heat capacity. This experiment consists in simultaneously injecting hot water and a chemical tracer into the aquifer and monitoring the evolution of groundwater temperature and concentration in the recovery well (and possibly in other piezometers located down gradient). Temperature and concentrations are then used for estimating the specific heat capacity. The first method for estimating this parameter is based on a modeling in series of the chemical tracer and temperature breakthrough curves at the recovery well. The second method is based on an energy balance. The values of specific heat capacity estimated for both methods (2.30 and 2.54MJ/m(3)/K) for the experimental site in the alluvial aquifer of the Meuse River (Belgium) are almost identical and consistent with values found in the literature. Temperature breakthrough curves in other piezometers are not required for estimating the specific heat capacity. However, they highlight that heat transfer in the alluvial aquifer of the Meuse River is complex and contrasted with different dominant process depending on the depth leading to significant vertical heat exchange between upper and lower part of the aquifer. Furthermore, these temperature breakthrough curves could be included in the calibration of a complex heat transfer model for

  16. Inverse modeling of tracer experiments in FEBEX compacted Ca-bentonite

    NASA Astrophysics Data System (ADS)

    Samper, Javier; Dai, Zhenxue; Molinero, Jorge; García-Gutiérrez, M.; Missana, T.; Mingarro, M.

    Solute transport parameters of compacted Ca-bentonite used in the FEBEX Project were derived by García-Gutiérrez et al. (2001) from through- and in-diffusion experiments using analytical solutions for their interpretation. Here we expand their work and present the numerical interpretation of diffusion and permeation experiments by solving the inverse transport problem which is formulated as the minimization of a weighted least squares criterion measuring the differences between computed and measured concentration values. The inverse problem is solved with INVERSE-CORE 2 D© , a finite element code which accounts for both dissolved and sorbed concentration data, uses either the Golden section search or Gauss-Newton-Marquardt methods for minimizing the objective function and allows the estimation of transport and retardation parameters such as diffusion coefficient, total and kinematic porosity and distribution coefficients. Diffusion and permeation experiments performed on FEBEX compacted bentonite using tritium, cesium, selenium, and strontium have been effectively interpreted by inverse modeling. Estimated parameters are within the range of reported values for these tracers in bentonites. It has been found that failing to account for the role of sinters may lead to erroneous diffusion coefficients by a factor of 1.4. Possible ways to improve the design of in-diffusion and permeation experiments have been identified. The interpretation of the tritium permeation experiment requires the use of a double-porosity model with mobile porosity of 0.14 for a dry density of 1.18 g/cm 3.

  17. Synthesis Of [2h, 13c] And [2h3, 13c]Methyl Aryl Sulfides

    DOEpatents

    Martinez, Rodolfo A.; Alvarez, Marc A.; Silks, III, Louis A.; Unkefer, Clifford J.

    2004-03-30

    The present invention is directed to labeled compounds, [.sup.2 H.sub.1, .sup.13 C], [.sup.2 H.sub.2, .sup.13 C] and [.sup.2 H.sub.3, .sup.13 C]methyl aryl sulfides wherein the .sup.13 C methyl group attached to the sulfur of the sulfide includes exactly one, two or three deuterium atoms and the aryl group is selected from the group consisting of 1-naphthyl, substituted 1-naphthyl, 2-naphthyl, substituted 2-naphthyl, and phenyl groups with the structure ##STR1## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5 are each independently, hydrogen, a C.sub.1 -C.sub.4 lower alkyl, a halogen, an amino group from the group consisting of NH.sub.2, NHR and NRR' where R and R' are each a C.sub.1 -C.sub.4 lower alkyl, a phenyl, or an alkoxy group. The present invention is also directed to processes of preparing [.sup.2 H.sub.1, .sup.13 C], [.sup.2 H.sub.2,.sup.13 C] and [.sup.2 H.sub.3, .sup.13 C]methyl aryl sulfides wherein the .sup.13 C methyl group attached to the sulfur of the sulfide includes exactly one, two or three deuterium atoms. The present invention is also directed to the labeled compounds of [.sup.2 H.sub.1, .sup.13 C]methyl iodide and [.sup.2 H.sub.2, .sup.13 C]methyl iodide.

  18. Sucrose octabenzoate: assignment of 13C and 1H resonances of the sucrose moiety and the 13C resonances of the carbonyl carbons. Use of 13C-n.m.r. spectroscopy for the study of selective deacylation.

    PubMed

    Colquhoun, I J; Haines, A H; Konowicz, P A; Jones, H F

    1990-09-19

    Assignment of the 1H and 13C signals arising from the carbohydrate portion of sucrose octabenzoate has been achieved using homonuclear shift correlation experiments (COSY) and one-bond 1H-13C heteronuclear shift correlation measurements, respectively. The 13C resonances of the carbonyl carbon atoms of the eight benzoyl groups are readily distinguished for solutions in benzene-d6-pyridine-d5 (1:1), and have been assigned by means of three-bond 1H-13C shift correlation studies coupled with measurement of the 13C-n.m.r. spectrum of a sucrose octabenzoate specifically labelled with 13C in some of the carbonyl groups. With this assignment, products of partial deacylation of the octabenzoate may readily be identified by treatment with excess of benzoyl-carbonyl-13C chloride followed by measurement of the 13C-n.m.r. spectrum of the labelled sucrose octabenzoate, so prepared, in the carbonyl region. PMID:2276151

  19. Characterisation of HTO diffusion properties by an in situ tracer experiment in Opalinus clay at Mont Terri.

    PubMed

    Palut, J-M; Montarnal, Ph; Gautschi, A; Tevissen, E; Mouche, E

    2003-03-01

    A long-term single borehole diffusion experiment using tritiated water as tracer was carried out in Opalinus clay, an argillaceous rock formation that is accessible at the Mont Terri Underground Research Laboratory, situated in the Swiss Jura. The tracer was diluted in reconstituted formation water and introduced into a packed-off section of a borehole located in saturated rock. Pressure in this interval was maintained equal to the pore pressure of the surrounding rock in order to prevent any hydraulic gradient around the borehole and to avoid advective transport processes. The evolution of the tracer concentration in the injection system was monitored over time. After 1 year of diffusion, the claystone surrounding the interval was retrieved by overcoring the whole borehole and packer system, and by an adjacent oblique borehole. Compressed air was used as drilling fluid to reduce rock disturbances. The recovered overcore was sampled along profiles perpendicular to the borehole wall with a view to determining the tracer-concentration profiles in the rock. To avoid further evaporation of tritiated water, subsamples were immediately transferred into polyethylene bottles and disaggregated by adding a known amount of tracer-free water. Fifteen profiles were determined and showed a decreasing tracer concentration with distance into the rock. The pore-water contents were constant along those profiles, confirming that only very little water was lost during overcoring operations. The evolution of tritium-tracer concentration in the injection system over time and in situ profiles were interpreted with a 3-D numerical simulation of the experiment. That allowed for the identification of the transport parameters (orthotropic diffusion tensor and porosity) by minimising the relative quadratic error between the experimental and simulated data. The fitting is good and the results are consistent with data obtained on drill-core samples. The result of tritiated water is discussed

  20. Characterisation of HTO diffusion properties by an in situ tracer experiment in Opalinus clay at Mont Terri.

    PubMed

    Palut, J-M; Montarnal, Ph; Gautschi, A; Tevissen, E; Mouche, E

    2003-03-01

    A long-term single borehole diffusion experiment using tritiated water as tracer was carried out in Opalinus clay, an argillaceous rock formation that is accessible at the Mont Terri Underground Research Laboratory, situated in the Swiss Jura. The tracer was diluted in reconstituted formation water and introduced into a packed-off section of a borehole located in saturated rock. Pressure in this interval was maintained equal to the pore pressure of the surrounding rock in order to prevent any hydraulic gradient around the borehole and to avoid advective transport processes. The evolution of the tracer concentration in the injection system was monitored over time. After 1 year of diffusion, the claystone surrounding the interval was retrieved by overcoring the whole borehole and packer system, and by an adjacent oblique borehole. Compressed air was used as drilling fluid to reduce rock disturbances. The recovered overcore was sampled along profiles perpendicular to the borehole wall with a view to determining the tracer-concentration profiles in the rock. To avoid further evaporation of tritiated water, subsamples were immediately transferred into polyethylene bottles and disaggregated by adding a known amount of tracer-free water. Fifteen profiles were determined and showed a decreasing tracer concentration with distance into the rock. The pore-water contents were constant along those profiles, confirming that only very little water was lost during overcoring operations. The evolution of tritium-tracer concentration in the injection system over time and in situ profiles were interpreted with a 3-D numerical simulation of the experiment. That allowed for the identification of the transport parameters (orthotropic diffusion tensor and porosity) by minimising the relative quadratic error between the experimental and simulated data. The fitting is good and the results are consistent with data obtained on drill-core samples. The result of tritiated water is discussed

  1. Development of X-ray tracer diagnostics for radiatively-driven ablator experiments [annual report FY1998

    SciTech Connect

    J.J. MacFarlane; D.H. Cohen; P. Wang; G.A. Moses; R.R. Peterson; P.A. Jaanimagi; O.L. Langen; R.E. Olson; T.J. Murphy; G.R. Magelssen; N.D. Delamater

    1999-05-01

    This report covers fiscal year 1998 of our ongoing project to develop tracer X-ray spectroscopic diagnostics for hohlraum environments. This effort focused on an experimental campaign carried out at OMEGA on 25--27 August 1998. This phase of the project heavily emphasized experimental design, diagnostic development, and target fabrication, as well as building up numerical models for the experiments. The spectral diagnostic under development involves using two thin (few 1000 {angstrom}) mid-Z tracers in two witness plates mounted on the side of a hohlraum with the tracers' K{sub a} absorption features seen against an X-ray backlighter. The absorption data are used to sample the time-dependent, localized properties of each witness plate as a radiation wave ablates it. The experiments represented the first application of this diagnostic, in this case to side-by-side doped and undoped plastic to investigate the effects of capsule ablator dopants.

  2. The effect of entrapped nonaqueous phase liquids on tracer transport in heterogeneous porous media: Laboratory experiments at the intermediate scale

    USGS Publications Warehouse

    Barth, G.R.; Illangasekare, T.H.; Rajaram, H.

    2003-01-01

    This work considers the applicability of conservative tracers for detecting high-saturation nonaqueous-phase liquid (NAPL) entrapment in heterogeneous systems. For this purpose, a series of experiments and simulations was performed using a two-dimensional heterogeneous system (10??1.2 m), which represents an intermediate scale between laboratory and field scales. Tracer tests performed prior to injecting the NAPL provide the baseline response of the heterogeneous porous medium. Two NAPL spill experiments were performed and the entrapped-NAPL saturation distribution measured in detail using a gamma-ray attenuation system. Tracer tests following each of the NAPL spills produced breakthrough curves (BTCs) reflecting the impact of entrapped NAPL on conservative transport. To evaluate significance, the impact of NAPL entrapment on the conservative-tracer breakthrough curves was compared to simulated breakthrough curve variability for different realizations of the heterogeneous distribution. Analysis of the results reveals that the NAPL entrapment has a significant impact on the temporal moments of conservative-tracer breakthrough curves. ?? 2003 Elsevier B.V. All rights reserved.

  3. The use of synthetic colloids in tracer transport experiments in saturated rock fractures

    SciTech Connect

    Reimus, P.W.

    1995-08-01

    Studies of groundwater flow and contaminant transport in saturated, fractured geologic media are of great interest to researchers studying the potential long-term storage of hazardous wastes in or near such media. A popular technique for conducting such studies is to introduce tracers having different chemical and physical properties into a system and then observe the tracers at one or more downstream locations, inferring flow and transport mechanisms from the breakthrough characteristics of the different tracers. Many tracer studies have been conducted in saturated, fractured media to help develop and/or refine models capable of predicting contaminant transport over large scales in such media.

  4. {sup 13}C-{sup 1}H and {sup 13}C-{sup 13}C spin couplings in [2`-{sup 13}C]2`-deoxyribonucleosides: Correlations with molecular structure

    SciTech Connect

    Bandyopadhyay, T.; Stripe, W.A.; Carmichael, I.; Serianni, A.S.; Wu, J.

    1997-02-19

    2`-Deoxyribonucleosides (2`-deoxyadenosine (1), 2`-depoxycytidine (2), thymidine (3)) singly enriched with {sup 13}C at C2` have been prepared and used to obtain one-, two-, and three-bond {sup 13}C-{sup 1}H and {sup 13}C-{sup 13}C spin-coupling constants involving C2`. Spin couplings in 1-3 involving C1` and C2`are also compared to corresponding values in ribonucleosides in order to assess the effects of nucleoside structure and conformation on J values within the furanose ring. {sup 1}J{sub C2`,H2`R} and {sup 1}J{sub C2`,H2`S} in 1-3 and {sup 1}J{sub C2`,H2`} in ribonucleosides depend on C-H bond orientation: {sup 1}J{sub C1`,H1`} in 1-3 and in ribonucleosides exhibits a similar dependence. The latter couplings appear to be essentially unaffected by N-glycoside torsion. {sup 1}J{sub CC} values depend on the number and distribution of electronegative substituents on the C-C fragment. A modified projection curve is proposed to aid in the interpretation of {sup 2}J{sub C2`,H1`} values; the presence of N substitution at C1` caused a shift to more negative couplings relative to the O-substituted analog. In contrast, {sup 2}J{sub C1`,H2`} is essentially unaffected by the same change in the electronegative substituent at C1`. {sup 2}J{sub CC} values within the furanose ring are determined buy two coupling pathways; in one case (i.e., {sup 2}J{sub C1`,C3`}), the observed coupling is shown to be the algebraic sum of the two couplings arising from each pathway. 41 refs., 4 figs., 2 tabs.

  5. Use of rare earth oxides and iron oxides as soil erosion tracers in water erosion experiments at hillslope scale

    NASA Astrophysics Data System (ADS)

    Guzmán, G.; Cañasveras, J. C.; Barrón, V.; Boulal, H.; Gómez, H.; Conde, E.; Fernández, M.; Gómez, J. A.

    2010-05-01

    The characteristics of the ideal soil erosion have been defined by several authors, for example by Zhang et al. (2001). Despite intensive research on erosion tracers in the last decades there is not a single tracer fulfilling all these characteristics. That is why research on different soil erosion tracers remains as an active field. Two desirable characteristics in erosion tracers are that they should be relatively inexpensive (to purchase and analyze) and that they should be determined with high accuracy in soil or sediment. The availability of multiple tracers is another of the key requirements. In this communication we present our preliminary results on the use of two different sets of erosion tracers. One set are iron oxides with different magnetic and optical properties (Fe3O4, α-Fe2O3 and FeOOH) analyzed by NIRS and magnetic susceptibility measurements. The other set consists of five rare earth oxides (La2O3, Pr6O11, Nd2O3, Sm2O3 and Gd2O3) analyzed using inductively coupled plasma mass spectrometry (ICP-MS). These two groups were studied under controlled and natural conditions, through several water erosion experiments, in field plots with different soil management, crops and scale. In one experiment these tracers were used to determine the source of sediment within sprinkle irrigated fields planted with cotton on shoulders. For this purpose, rainfall simulations were performed under controlled conditions at two scales, one with a portable rainfall simulator at small scale (0.81m2) and with the sprinkler irrigation system in the whole cotton field (2450 m2). Furrows were tagged with both groups of tracers, keeping shoulders untagged (where cotton was planted). Soil samples before and after the rainfall simulations were collected as well as sediment samples. In another experiment four olive orchard plots (330 m2) with different soil managements (cover crop and conventional tillage) were also tagged with the two groups of tracers. Soil samples were taken at

  6. Greenland freshwater pathways in the sub-Arctic Seas from model experiments with passive tracers

    NASA Astrophysics Data System (ADS)

    Dukhovskoy, Dmitry S.; Myers, Paul G.; Platov, Gennady; Timmermans, Mary-Louise; Curry, Beth; Proshutinsky, Andrey; Bamber, Jonathan L.; Chassignet, Eric; Hu, Xianmin; Lee, Craig M.; Somavilla, Raquel

    2016-01-01

    Accelerating since the early 1990s, the Greenland Ice Sheet mass loss exerts a significant impact on thermohaline processes in the sub-Arctic seas. Surplus freshwater discharge from Greenland since the 1990s, comparable in volume to the amount of freshwater present during the Great Salinity Anomaly events, could spread and accumulate in the sub-Arctic seas, influencing convective processes there. However, hydrographic observations in the Labrador Sea and the Nordic Seas, where the Greenland freshening signal might be expected to propagate, do not show a persistent freshening in the upper ocean during last two decades. This raises the question of where the surplus Greenland freshwater has propagated. In order to investigate the fate, pathways, and propagation rate of Greenland meltwater in the sub-Arctic seas, several numerical experiments using a passive tracer to track the spreading of Greenland freshwater have been conducted as a part of the Forum for Arctic Ocean Modeling and Observational Synthesis effort. The models show that Greenland freshwater propagates and accumulates in the sub-Arctic seas, although the models disagree on the amount of tracer propagation into the convective regions. Results highlight the differences in simulated physical mechanisms at play in different models and underscore the continued importance of intercomparison studies. It is estimated that surplus Greenland freshwater flux should have caused a salinity decrease by 0.06-0.08 in the sub-Arctic seas in contradiction with the recently observed salinification (by 0.15-0.2) in the region. It is surmised that the increasing salinity of Atlantic Water has obscured the freshening signal.

  7. Coal structure at reactive sites by sup 1 H- sup 13 C- sup 19 F double cross polarization (DCP)/MAS sup 13 C NMR spectroscopy

    SciTech Connect

    Hagaman, E.W.; Woody, M.C. )

    1989-01-01

    The solid state NMR technique, {sup 1}H-{sup 13}C-{sup 31}P double cross polarization (DCP)/MAS {sup 13}C-NMR spectroscopy, uses the direct dipolar interaction between {sup 13}C-{sup 31}P spin pairs in organophosphorus substances to identify the subset of carbons within a spherical volume element of 0.4 nm radius centered on the {sup 31}P atom. In combination with chemical manipulation of coals designed to introduce phosphorus containing functionality into the organic matrix, the NMR experiment becomes a method to examine selectively the carbon bonding network at the reactive sites in the coal. This approach generates a statistical structure description of the coal at the reaction centers in contrast to bulk carbon characterization using conventional {sup 1}H-{sup 13}C CP/MAS {sup 13}C NMR spectroscopy. 3 refs.

  8. 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.

  9. The Experience of Receiving and Then Losing a Scholarship: A Tracer Study of Secondary School Scholarship Recipients in Uganda

    ERIC Educational Resources Information Center

    Watson, Cathy; Chapman, David W.; Okurut, Charles Opolot

    2014-01-01

    This study reports findings of a tracer that investigated differences in the profile and subsequent experiences of scholarship recipients in Uganda who were able to complete the lower secondary school cycle (O level) without interruption (N = 174) and those that dropped out before completing their O-level cycle (N = 51), thereby losing their…

  10. The fluorescent tracer experiment on Holiday Beach near Mugu Canyon, Southern California

    USGS Publications Warehouse

    Kinsman, Nicole; Xu, J. P.

    2012-01-01

    After revisiting sand tracer techniques originally developed in the 1960s, a range of fluorescent coating formulations were tested in the laboratory. Explicit steps are presented for the preparation of the formulation evaluated to have superior attributes, a thermoplastic pigment/dye in a colloidal mixture with a vinyl chloride/vinyl acetate copolymer. In September 2010, 0.59 cubic meters of fluorescent tracer material was injected into the littoral zone about 4 kilometers upcoast of Mugu submarine canyon in California. The movement of tracer was monitored in three dimensions over the course of 4 days using manual and automated techniques. Detailed observations of the tracer's behavior in the coastal zone indicate that this tracer successfully mimicked the native beach sand and similar methods could be used to validate models of tracer movement in this type of environment. Recommendations including how to time successful tracer studies and how to scale the field of view of automated camera systems are presented along with the advantages and disadvantages of the described tracer methodology.

  11. A sprinkling experiment using multiple tracers and ERT to conceptualize subsurface storm flow

    NASA Astrophysics Data System (ADS)

    Rothuizen, R. D.; Bogaard, T. A.; Westhoff, M. C.; Savenije, H. H. G.

    2010-05-01

    Rapid subsurface flow is one of the most important water flow processes in hillslopes. This process shows significant threshold behaviour as a certain amount of rainfall has to be added to the hillslope before the rapid subsurface flow process becomes active. Moreover, many tracer studies have shown a large fraction of ‘old' water in the rapid subsurface flow. The magnitude of the threshold and the fraction of ‘old' and ‘new' water still get significant research attention. However, this behaviour and magnitude can be explained by different hydrological concepts. In this study, we aim to identify and quantify the hydrological processes responsible for the threshold behaviour of the rapid subsurface flow and the extent of new water contribution during 5 sprinkling experiments on a 150 m2 plot, just uphill of a previously identified groundwater source. Five consecutive days we added approximately 9 mm of artificial rain on an experimental plot during one hour with salt and deuterium as artificial tracers. The analysis is based on a combination of hydrometric, hydrochemical and Electrical Resistivity Tomography (ERT) observations. The time lapse ERT observations gave most information on the saturation of and the salt content within the unsaturated soil, while the hydrometric observations quantified the storage in the saturated soil. The hydrochemical information was useful to differentiate between old and new water. We developed a conceptual model of the hydrological processes in the subsurface. The main hydrological processes that were identified and modeled are the unsaturated zone storage, the rapid subsurface flow and the slow matrix flow. By constraining the model only on the observed source outflow and not using the water quality information, we obtained a good fit between measured and modeled outflow. However, the simulated salt concentrations did not resemble the observations. This was caused by an overestimated exchange between the unsaturated zone and

  12. HYPERPOLARIZED 13C MAGNETIC RESONANCE AND ITS USE IN METABOLIC ASSESSMENT OF CULTURED CELLS AND PERFUSED ORGANS

    PubMed Central

    Lumata, Lloyd; Yang, Chendong; Ragavan, Mukundan; Carpenter, Nicholas; DeBerardinis, Ralph J.; Merritt, Matthew E.

    2016-01-01

    Diseased tissue is often characterized by abnormalities in intermediary metabolism. Observing these alterations in situ may lead to an improved understanding of pathological processes and novel ways to monitor these processes non-invasively in human patients. Although 13C is a stable isotope safe for use in animal models of disease as well as human subjects, its utility as a metabolic tracer has largely been limited to ex vivo analyses employing analytical techniques like mass spectrometry or nuclear magnetic resonance spectroscopy. Neither of these techniques is suitable for non-invasive metabolic monitoring, and the low abundance and poor gyromagnetic ratio of conventional 13C make it a poor nucleus for imaging. However, the recent advent of hyperpolarization methods, particularly dynamic nuclear polarization (DNP), make it possible to enhance the spin polarization state of 13C by many orders of magnitude, resulting in a temporary amplification of the signal sufficient for monitoring kinetics of enzyme-catalyzed reactions in living tissue through magnetic resonance spectroscopy or magnetic resonance imaging. Here we review DNP techniques to monitor metabolism in cultured cells, perfused hearts, and perfused livers, focusing on our experiences with hyperpolarized [1-13C]pyruvate. We present detailed approaches to optimize the DNP procedure, streamline biological sample preparation, and maximize detection of specific metabolic activities. We also discuss practical aspects in the choice of metabolic substrates for hyperpolarization studies, and outline some of the current technical and conceptual challenges in the field, including efforts to use hyperpolarization to quantify metabolic rates in vivo. PMID:26358902

  13. Tracing solid waste leachate in groundwater using δ13 C from dissolved inorganic carbon.

    PubMed

    Haarstad, Ketil; Mæhlum, Trond

    2013-01-01

    Tracers can be used to monitor emissions of leachate from landfills in order to detect hydrological pathways and to evaluate environmental pollution. We investigated the stable carbon isotope ratio (δ(13)C-Σ CO (2)) in dissolved inorganic carbon and tritium ((3)H) in water, in addition to the tracers of pollution commonly found in relatively high concentrations in leachate, such as chloride (Cl), organic matter (COD), nitrogen (total and NH(4)-N), iron (Fe), electrical conductivity (EC) and pH. The sampling was performed at seven landfills in the south-eastern part of Norway during a period of 5 years. The objective was to evaluate the potential for tracing leachate in the environment with emphasis on groundwater pollution. By measuring the δ(13)C-Σ CO (2) in leachates, groundwaters and surface waters, the influence of leachate can be identified. The value of δ(13)C-Σ CO (2) varied from-5.5 to 25.9 ‰ in leachate, from-25.4 to 14.7 ‰ in groundwater and from-19.7 to-13.1 ‰ in creeks. A comparison of the carbon isotope ratio with COD, EC and the concentrations of total and NH (4)-N, Cl and Fe showed that δ(13)C-Σ CO (2) is a good tracer for leachate due to higher sensitivity compared to other parameters. The mean concentrations of all the studied parameters were higher in the leachate samples; however, only the carbon isotope ratio showed significant differences between all the groups with strong and middle pollution and samples with low pollution, showing that it can be used as a convenient tracer for leachate in groundwater and surface water. The carbon isotope ratio showed strong correlation between nitrogen, EC and bicarbonate, but not with pH. Tritium was only sporadically found in measureable concentrations and is not considered as a suitable tracer at the sampled locations.

  14. Cross-hole tracer experiment reveals rapid fluid flow and low effective porosity in the upper oceanic crust

    NASA Astrophysics Data System (ADS)

    Neira, N. M.; Clark, J. F.; Fisher, A. T.; Wheat, C. G.; Haymon, R. M.; Becker, K.

    2016-09-01

    Numerous field, laboratory, and modeling studies have explored the flows of fluid, heat, and solutes during seafloor hydrothermal circulation, but it has been challenging to determine transport rates and flow directions within natural systems. Here we present results from the first cross-hole tracer experiment in the upper oceanic crust, using four subseafloor borehole observatories equipped with autonomous samplers to track the transport of a dissolved tracer (sulfur hexafluoride, SF6) injected into a ridge-flank hydrothermal system. During the first three years after tracer injection, SF6 was transported both north and south through the basaltic aquifer. The observed tracer transport rate of ∼2-3 m/day is orders of magnitude greater than bulk rates of flow inferred from thermal and chemical observations and calculated with coupled fluid-heat flow simulations. Taken together, these results suggest that the effective porosity of the upper volcanic crust through which much tracer was transported is <1%, with fluid flowing rapidly along a few well-connected channels. This is consistent with the heterogeneous (layered, faulted, and/or fractured) nature of the volcanic upper oceanic crust.

  15. The use of some ion-exchange sorbing tracer cations in in-situ experiments in high saline groundwaters

    SciTech Connect

    Byegaard, J.; Skarnemark, G.; Skaalberg, M.

    1995-12-31

    The possibility to use alkali metals and alkaline earth metals as slightly sorbing tracers in in-situ sorption experiments in high saline groundwaters has been investigated. The cation exchange characteristics of granite and some fracture minerals (chlorite and calcite) have been studied using the proposed cations as tracers. The results show low Kd`s for Na, Ca and Sr ({approximately}0.1 ml/g), while the sorption is higher for the more electropositive cations (Rb, Cs and Ba). A higher contribution of irreversible sorption can also be observed for the latter group of cations. For calcite the sorption of all the tracers, except Ca, is lower compared to the corresponding sorption to granite and chlorite. Differences in selectivity coefficients and cation exchange capacity are obtained when using different size fractions of crushed granite. The difference is even more pronounced when comparing crushed granite to intact granite.

  16. Microorganisms as tracers in groundwater injection and recovery experiments: A review

    USGS Publications Warehouse

    Harvey, R.W.

    1997-01-01

    Modern day injection and recovery techniques designed to examine the transport behavior of microorganisms in groundwater have evolved from experiments conducted in the late 1800s, in which bacteria that form red or yellow pigments were used to trace flow paths through karst and fractured- rock aquifers. A number of subsequent groundwater hydrology studies employed bacteriophage that can be injected into aquifers at very high concentrations (e g., 1013 phage ml-1) and monitored through many log units of dilution to follow groundwater flow paths for great distances, particularly in karst terrain. Starting in the 1930s, microbial indicators of fecal contamination (particularly coliform bacteria and their coliphages) were employed as tracers to determine potential migration of pathogens in groundwater. Several injection and recovery experiments performed in the 1990s employed indigenous groundwater microorganisms (both cultured and uncultured) that are better able to survive under in situ conditions. Better methods for labeling native bacteria (e.g by stable isotope labeling or inserting genetic markers; such as the ability to cause ice nucleation) are being developed that will not compromise the organisms' viability during the experimental time course.

  17. 4D ground-penetrating radar during a plot scale dye tracer experiment

    NASA Astrophysics Data System (ADS)

    Allroggen, Niklas; van Schaik, N. Loes M. B.; Tronicke, Jens

    2015-07-01

    Flow phenomena in the unsaturated zone are highly variable in time and space. Thus, it is challenging to measure and monitor such processes under field conditions. Here, we present a new setup and interpretation approach for combining a dye tracer experiment with a 4D ground-penetrating radar (GPR) survey. Therefore, we designed a rainfall experiment during which we measured three surface-based 3D GPR surveys using a pair of 500 MHz antennas. Such a survey setup requires accurate acquisition and processing techniques to extract time-lapse information supporting the interpretation of selected cross-sections photographed after excavating the site. Our results reveal patterns of traveltime changes in the measured GPR data, which are associated with soil moisture changes. As distinct horizons are present at our site, such changes can be quantified and transferred into changes in total soil moisture content. Our soil moisture estimates are similar to the amount of infiltrated water, which confirms our experimental approach and makes us confident for further developing this strategy, especially, with respect to improving the temporal and spatial resolution.

  18. Hydraulic/partitioning tracer tomography for DNAPL source zone characterization: small-scale sandbox experiments.

    PubMed

    Illman, Walter A; Berg, Steven J; Liu, Xiaoyi; Massi, Antonio

    2010-11-15

    Dense nonaqueous phase liquids (DNAPL) are prevalent at a large number of sites throughout the world. The variable release history, unstable flow, and geologic heterogeneity make the spatial distribution of DNAPLs complex. This causes difficulties in site remediation contributing to long-term groundwater contamination for decades to centuries. We present laboratory experiments to demonstrate the efficacy of Sequential Successive Linear Estimator (SSLE) algorithm that images DNAPL source zones. The algorithm relies on the fusion of hydraulic and partitioning tracer tomography (HPTT) to derive the best estimate of the K heterogeneity, DNAPL saturation (S(N)) distribution, and their uncertainty. The approach is nondestructive and can be applied repeatedly. Results from our laboratory experiments show that S(N) distributions compare favorably with DNAPL distributions observed in the sandbox but not so with local saturation estimates from core samples. We also found that the delineation of K heterogeneity can have a large impact on computed S(N) distributions emphasizing the importance of accurate delineation of hydraulic heterogeneity.

  19. Microorganisms as tracers in groundwater injection and recovery experiments: a review.

    PubMed

    Harvey, R W

    1997-07-01

    Modern day injection and recovery techniques designed to examine the transport behavior of microorganisms in groundwater have evolved from experiments conducted in the late 1800s, in which bacteria that form red or yellow pigments were used to trace flow paths through karst and fractured-rock aquifers. A number of subsequent groundwater hydrology studies employed bacteriophage that can be injected into aquifers at very high concentrations (e.g., 10(13) phage ml-1) and monitored through many log units of dilution to follow groundwater flow paths for great distances, particularly in karst terrain. Starting in the 1930s, microbial indicators of fecal contamination (particularly coliform bacteria and their coliphages) were employed as tracers to determine potential migration of pathogens in groundwater. Several injection and recovery experiments performed in the 1990s employed indigenous groundwater microorganisms (both cultured and uncultured) that are better able to survive under in situ conditions. Better methods for labeling native bacteria (e.g. by stable isotope labeling or inserting genetic markers, such as the ability to cause ice nucleation) are being developed that will not compromise the organisms' viability during the experimental time course.

  20. Effects from influent boundary conditions on tracer migration and spatial variability features in intermediate-scale experiments

    SciTech Connect

    Fuentes, H.R.; Polzer, W.L.; Springer, E.P.

    1987-04-01

    In previous unsaturated transport studies at Los Alamos dispersion coefficients were estimated to be higher close to the tracer source than at greater distances from the source. Injection of tracers through discrete influent outlets could have accounted for those higher dispersions. Also, a lack of conservation of mass of the tracers was observed and suspected to be due to spatial variability in transport. In the present study experiments were performed under uniform influent (ponded) conditions in which breakthrough of tracers was monitored at four locations at each of four depths. All other conditions were similar to those of the unsaturated transport experiments. A comparison of results from these two sets of experiments indicates differences in the parameter estimates. Estimates were made for the dispersion coefficient and the retardation factor by the one-dimensional steady flow computer code, CFITIM. Estimates were also made for mass and for velocity and the dispersion coefficient by the method of moments. The dispersion coefficient decreased with depth under discrete influent application and increased with depth under ponded influent application. Retardation was predicted better under the discrete influent application than under ponded influent application. Differences in breakthroughs and in estimated parameters among locations at the same depth were observed under ponded influent application. Those differences indicate that there is a lack of conservation of mass as well as significant spatial variability across the experimental domain. 14 refs., 9 figs., 8 tabs.

  1. Predicted fate of tritium residuum from groundwater tracer experiments in the Amargosa Desert, southern Nevada

    SciTech Connect

    Brikowski, T.

    1993-07-01

    Analytic solutions are used in this study to evaluate potential groundwater transport of tritium used in goundwater tracer tests southwest of the Nevada Test Site. Possible transport from this site is of interest because initial radionuclide concentrations were high and the site is close to goundwater discharge points (12 km). Anecdotal evidence indicates that 90 percent of these tracers were removed by pumping at the completion of the tests; this study examines the probable transport of the tracers with and without the removal. Classical dispersive transport analytic solutions are used, treating the tracer test as a point slug injection. Input parameters for the solutions were measured at the site, and consideration of parameter uncertainty is incorporated in the results. With removal of the tracer, the maximum expected region with above-Safe Drinking Water Act (40 CFR 121) concentrations of tritium extends 5 km from the injection point, and does not reach any sites of public access. Detectable tritium from the tests is likely to have reached the Ash Meadows fault zone, but flow along the fault probably diluted the tracer to below detection limits before arrival at springs along the fault. Arrival at the springs would have occurred 20 to 25 years after the tests. Without removal of the tracer, the solutions indicate that tritium concentrations just above Safe Drinking Water Act standards would have reached the Ash Meadows fault zone. In this case, detectable tritium might have been found in Devil`s Hole or Longstreet Spring, the nearest points of possible public exposure.

  2. Extreme (13)C depletion of carbonates formed during oxidation of biogenic methane in fractured granite.

    PubMed

    Drake, Henrik; Åström, Mats E; Heim, Christine; Broman, Curt; Åström, Jan; Whitehouse, Martin; Ivarsson, Magnus; Siljeström, Sandra; Sjövall, Peter

    2015-01-01

    Precipitation of exceptionally 13C-depleted authigenic carbonate is a result of, and thus a tracer for, sulphate-dependent anaerobic methane oxidation, particularly in marine sediments. Although these carbonates typically are less depleted in 13C than in the source methane, because of incorporation of C also from other sources, they are far more depleted in 13C13C as light as -69‰ V-PDB) than in carbonates formed where no methane is involved. Here we show that oxidation of biogenic methane in carbon-poor deep groundwater in fractured granitoid rocks has resulted in fracture-wall precipitation of the most extremely 13C-depleted carbonates ever reported, δ13C down to -125‰ V-PDB. A microbial consortium of sulphate reducers and methane oxidizers has been involved, as revealed by biomarker signatures in the carbonates and S-isotope compositions of co-genetic sulphide. Methane formed at shallow depths has been oxidized at several hundred metres depth at the transition to a deep-seated sulphate-rich saline water. This process is so far an unrecognized terrestrial sink of methane. PMID:25948095

  3. Extreme (13)C depletion of carbonates formed during oxidation of biogenic methane in fractured granite.

    PubMed

    Drake, Henrik; Åström, Mats E; Heim, Christine; Broman, Curt; Åström, Jan; Whitehouse, Martin; Ivarsson, Magnus; Siljeström, Sandra; Sjövall, Peter

    2015-05-07

    Precipitation of exceptionally 13C-depleted authigenic carbonate is a result of, and thus a tracer for, sulphate-dependent anaerobic methane oxidation, particularly in marine sediments. Although these carbonates typically are less depleted in 13C than in the source methane, because of incorporation of C also from other sources, they are far more depleted in 13C13C as light as -69‰ V-PDB) than in carbonates formed where no methane is involved. Here we show that oxidation of biogenic methane in carbon-poor deep groundwater in fractured granitoid rocks has resulted in fracture-wall precipitation of the most extremely 13C-depleted carbonates ever reported, δ13C down to -125‰ V-PDB. A microbial consortium of sulphate reducers and methane oxidizers has been involved, as revealed by biomarker signatures in the carbonates and S-isotope compositions of co-genetic sulphide. Methane formed at shallow depths has been oxidized at several hundred metres depth at the transition to a deep-seated sulphate-rich saline water. This process is so far an unrecognized terrestrial sink of methane.

  4. Extreme 13C depletion of carbonates formed during oxidation of biogenic methane in fractured granite

    PubMed Central

    Drake, Henrik; Åström, Mats E.; Heim, Christine; Broman, Curt; Åström, Jan; Whitehouse, Martin; Ivarsson, Magnus; Siljeström, Sandra; Sjövall, Peter

    2015-01-01

    Precipitation of exceptionally 13C-depleted authigenic carbonate is a result of, and thus a tracer for, sulphate-dependent anaerobic methane oxidation, particularly in marine sediments. Although these carbonates typically are less depleted in 13C than in the source methane, because of incorporation of C also from other sources, they are far more depleted in 13C13C as light as −69‰ V-PDB) than in carbonates formed where no methane is involved. Here we show that oxidation of biogenic methane in carbon-poor deep groundwater in fractured granitoid rocks has resulted in fracture-wall precipitation of the most extremely 13C-depleted carbonates ever reported, δ13C down to −125‰ V-PDB. A microbial consortium of sulphate reducers and methane oxidizers has been involved, as revealed by biomarker signatures in the carbonates and S-isotope compositions of co-genetic sulphide. Methane formed at shallow depths has been oxidized at several hundred metres depth at the transition to a deep-seated sulphate-rich saline water. This process is so far an unrecognized terrestrial sink of methane. PMID:25948095

  5. Optoacoustic 13C-breath test analyzer

    NASA Astrophysics Data System (ADS)

    Harde, Hermann; Helmrich, Günther; Wolff, Marcus

    2010-02-01

    The composition and concentration of exhaled volatile gases reflects the physical ability of a patient. Therefore, a breath analysis allows to recognize an infectious disease in an organ or even to identify a tumor. One of the most prominent breath tests is the 13C-urea-breath test, applied to ascertain the presence of the bacterium helicobacter pylori in the stomach wall as an indication of a gastric ulcer. In this contribution we present a new optical analyzer that employs a compact and simple set-up based on photoacoustic spectroscopy. It consists of two identical photoacoustic cells containing two breath samples, one taken before and one after capturing an isotope-marked substrate, where the most common isotope 12C is replaced to a large extent by 13C. The analyzer measures simultaneously the relative CO2 isotopologue concentrations in both samples by exciting the molecules on specially selected absorption lines with a semiconductor laser operating at a wavelength of 2.744 μm. For a reliable diagnosis changes of the 13CO2 concentration of 1% in the exhaled breath have to be detected at a concentration level of this isotope in the breath of about 500 ppm.

  6. Inversion of the Borden Tracer Experiment Data: Investigation of Stochastic Moment Models

    NASA Astrophysics Data System (ADS)

    Woodbury, Allan D.; Sudicky, E. A.

    1992-09-01

    Inversion of Dagan's two- and three-dimensional stochastic models using Freyberg's (1986), Rajaram and Gelhar's (1988, 1991), and Barry and Sposito's (1990) moment data from the Borden experiment is carried out to examine (1) the validity of the two-dimensional (Dagan, 1982) and three-dimensional (Dagan, 1988) models and (2) the reduction in uncertainty of the spatial moments over nonconditioned estimates. A direct application of Bayesian statistical inference, in conjunction with Monte Carlo integration, is used to produce posterior probability density functions for the parameters. The parameter ranges from all methods show horizontal integral scales λ between 1.85 and 4.04 m, vertical integral scales λz between 0.144 and 0.459 m, and log hydraulic conductivities In (K) between 0.120 and 0.197. These results compare well to the earlier estimates of Woodbury and Sudicky (1991) and Robin et al. (1991). We show that moment prediction uncertainty is substantially reduced when both tracer moment data and prior estimates of the In (K) geostatistical parameters are incorporated into theoretical formulae based on stochastic dispersion theory.

  7. Iterative ensemble Kalman filter for atmospheric dispersion in nuclear accidents: An application to Kincaid tracer experiment.

    PubMed

    Zhang, X L; Su, G F; Chen, J G; Raskob, W; Yuan, H Y; Huang, Q Y

    2015-10-30

    Information about atmospheric dispersion of radionuclides is vitally important for planning effective countermeasures during nuclear accidents. Results of dispersion models have high spatial and temporal resolutions, but they are not accurate enough due to the uncertain source term and the errors in meteorological data. Environmental measurements are more reliable, but they are scarce and unable to give forecasts. In this study, our newly proposed iterative ensemble Kalman filter (EnKF) data assimilation scheme is used to combine model results and environmental measurements. The system is thoroughly validated against the observations in the Kincaid tracer experiment. The initial first-guess emissions are assumed to be six magnitudes underestimated. The iterative EnKF system rapidly corrects the errors in the emission rate and wind data, thereby significantly improving the model results (>80% reduction of the normalized mean square error, r=0.71). Sensitivity tests are conducted to investigate the influence of meteorological parameters. The results indicate that the system is sensitive to boundary layer height. When the heights from the numerical weather prediction model are used, only 62.5% of reconstructed emission rates are within a factor two of the actual emissions. This increases to 87.5% when the heights derived from the on-site observations are used.

  8. Combined 13C-D and D-D clumping in methane: Methods and preliminary results

    NASA Astrophysics Data System (ADS)

    Stolper, D. A.; Sessions, A. L.; Ferreira, A. A.; Santos Neto, E. V.; Schimmelmann, A.; Shusta, S. S.; Valentine, D. L.; Eiler, J. M.

    2014-02-01

    The stable isotopic composition of methane (e.g., δD and δ13C values) is often used as a tracer for its sources and sinks. Conventional δD and δ13C measurements represent the average isotope ratios of all ten isotopologues of methane, though they are effectively controlled by the relative abundances of the three most abundant species: 12CH4, 13CH4, and 12CH3D. The precise relative abundances of the other seven isotopologues remains largely unexplored because these species contain multiple rare isotopes and are thus rare themselves. These multiply substituted (or 'clumped') isotopologues each have their own distinctive chemical and physical properties, which could provide additional constraints on the geochemistry of methane. This work focuses on quantifying the abundances of two rare isotopologues, 13CH3D and 12CH2D2, of methane in order to assess their utility as a window into methane's geochemistry. Specifically, we seek to assess whether clumped isotope distributions might be useful to quantify the temperature at which methane formed and/or equilibrated. To this end, we report the first highly precise combined measurements of the relative abundances of 13CH3D and 12CH2D2 at natural abundances (i.e., unlabeled) via the high-resolution magnetic-sector mass spectrometry of intact methane. We calibrate the use of these measurements as a geothermometer using both theory and experiment, and apply this geothermometer to representative natural samples. The method yields accurate average (i.e., bulk) isotopic ratios based on comparison with conventional techniques. We demonstrate the accuracy and precision of measurements of 13CH3D and 12CH2D2 through analyses of methane driven to high temperature (>200 °C) equilibrium in the laboratory. Application of this thermometer to natural samples yields apparent temperatures consistent with their known formation environments and appears to distinguish between biogenic and thermogenic methane.

  9. Biodegradation of the surfactant linear alkylbenzenesulfonate in sewage- contaminated groundwater: A comparison of column experiments and field tracer tests

    USGS Publications Warehouse

    Krueger, C.J.; Radakovich, K.M.; Sawyer, T.E.; Barber, L.B.; Smith, R.L.; Field, J.A.

    1998-01-01

    Transport and biodegradation of linear alkylbenzenesulfonate (LAS) in sewage-contaminated groundwater were investigated for a range of dissolved oxygen concentrations. Both laboratory column and an 80-day continuous injection tracer test field experiments were conducted. The rates of LAS biodegradation increased with increasing dissolved oxygen concentrations and indicated the preferential biodegradation of the longer alkyl chain LAS homologues (i.e., C12 and C13) and external isomers (i.e., 2-and 3- phenyl). However, for similar dissolved oxygen concentrations, mass removal rates for LAS generally were 2-3 times greater in laboratory column experiments than in the field tracer test. Under low oxygen conditions (<1 mg/L) only a fraction of the LAS mixture biodegraded in both laboratory and field experiments. Biodegradation rate constants for the continuous injection field test (0.002-0.08 day-1) were comparable to those estimated for a 3-h injection (pulsed) tracer test conducted under similar biogeochemical conditions, indicating that increasing the exposure time of aquifer sediments to LAS did not increase biodegradation rates.Transport and biodegradation of linear alkylbenzenesulfonate (LAS) in sewage-contaminated groundwater were investigated for a range of dissolved oxygen concentrations. Both laboratory column and an 80-day continuous injection tracer test field experiments were conducted. The rates of LAS biodegradation increased with increasing dissolved oxygen concentrations and indicated the preferential biodegradation of the longer alkyl chain LAS homologues (i.e., C12 and C13) and external isomers (i.e., 2- and 3-phenyl). However, for similar dissolved oxygen concentrations, mass removal rates for LAS generally were 2-3 times greater in laboratory column experiments than in the field tracer test. Under low oxygen conditions (<1 mg/L) only a fraction of the LAS mixture biodegraded in both laboratory and field experiments. Biodegradation rate constants

  10. A method for simultaneous echo planar imaging of hyperpolarized 13C pyruvate and 13C lactate

    NASA Astrophysics Data System (ADS)

    Reed, Galen D.; Larson, Peder E. Z.; von Morze, Cornelius; Bok, Robert; Lustig, Michael; Kerr, Adam B.; Pauly, John M.; Kurhanewicz, John; Vigneron, Daniel B.

    2012-04-01

    A rapid echo planar imaging sequence for dynamic imaging of [1-13C] lactate and [1-13C] pyruvate simultaneously was developed. Frequency-based separation of these metabolites was achieved by spatial shifting in the phase-encoded direction with the appropriate choice of echo spacing. Suppression of the pyruvate-hydrate and alanine resonances is achieved through an optimized spectral-spatial RF waveform. Signal sampling efficiency as a function of pyruvate and lactate excitation angle was simulated using two site exchange models. Dynamic imaging is demonstrated in a transgenic mouse model, and phantom validations of the RF pulse frequency selectivity were performed.

  11. Field-scale sulfur hexafluoride tracer experiment to understand long distance gas transport in the deep unsaturated zone

    USGS Publications Warehouse

    Walvoord, Michelle Ann; Andraski, Brian; Green, Christopher T.; Stonestrom, David A.; Striegl, Rob

    2014-01-01

    A natural gradient SF6 tracer experiment provided an unprecedented evaluation of long distance gas transport in the deep unsaturated zone (UZ) under controlled (known) conditions. The field-scale gas tracer test in the 110-m-thick UZ was conducted at the U.S. Geological Survey’s Amargosa Desert Research Site (ADRS) in southwestern Nevada. A history of anomalous (theoretically unexpected) contaminant gas transport observed at the ADRS, next to the first commercial low-level radioactive waste disposal facility in the United States, provided motivation for the SF6 tracer study. Tracer was injected into a deep UZ borehole at depths of 15 and 48 m, and plume migration was observed in a monitoring borehole 9 m away at various depths (0.5–109 m) over the course of 1 yr. Tracer results yielded useful information about gas transport as applicable to the spatial scales of interest for off-site contaminant transport in arid unsaturated zones. Modeling gas diffusion with standard empirical expressions reasonably explained SF6 plume migration, but tended to underpredict peak concentrations for the field-scale experiment given previously determined porosity information. Despite some discrepancies between observations and model results, rapid SF6 gas transport commensurate with previous contaminant migration was not observed. The results provide ancillary support for the concept that apparent anomalies in historic transport behavior at the ADRS are the result of factors other than nonreactive gas transport properties or processes currently in effect in the undisturbed UZ.

  12. Differential growth of the fungus Absidia cylindrospora on 13C/15N-labelled media.

    PubMed

    Crotty, F V; Blackshaw, R P; Murray, P J

    2011-06-15

    Many studies utilise enrichment of stable isotopes as tracers to follow the interactions occurring within soil food webs and methods have been developed to enrich bacteria, soil fauna and plant litter, Here for the first time we attempt to enrich a soil fungus to 99 atom% with (13)C and (15)N stable isotopes. In this study our objectives were to (a) assess whether the saprotrophic zygomycete fungus Absidia cylindrospora could grow on a medium enriched to 99 atom% with (13)C-glucose and (15)N-ammonium chloride, (b) to determine the level of enrichment obtained, and (c) to examine the change in growth rate of this fungus while it was growing on the dually enriched medium. To achieve this, the fungus was grown on agar enriched with (13)C and (15)N to 99 atom% and its growth rate monitored. The results showed that A. cylindrospora would grow on the highly labelled growth medium, but that its rate of growth was affected compared with the rate on either natural abundance media or media highly enriched with a single isotope ((13)C or (15)N). The implications of these results is that although the fungus is able to utilise these heavier isotopes, the biochemical processes involved in growth are affected, and consideration should be given to these differences when using stable isotope tracers in, for example, soil food web studies.

  13. Synthesis Of 2h- And 13c-Substituted Dithanes

    DOEpatents

    Martinez, Rodolfo A.; Alvarez, Marc A.; Silks, III, Louis A.; Unkefer, Clifford J.

    2004-05-04

    The present invention is directed to labeled compounds, [2-.sup.13 C]dithane wherein the .sup.13 C atom is directly bonded to one or two deuterium atoms. The present invention is also directed to processes of preparing [2-.sup.13 C]dithane wherein the .sup.13 C atom is directly bonded to one or two deuterium atoms. The present invention is also directed to labeled compounds, e.g., [.sup.2 H.sub.1-2, .sup.13 C]methanol (arylthio)-, acetates wherein the .sup.13 C atom is directly bonded to exactly one or two deuterium atoms.

  14. Synthesis of 2H- and 13C-substituted dithanes

    DOEpatents

    Martinez, Rodolfo A.; Alvarez, Marc A.; Silks, III, Louis A.; Unkefer, Clifford J.

    2003-01-01

    The present invention is directed to labeled compounds, [2-.sup.13 C]dithiane wherein the .sup.13 C atom is directly bonded to one or two deuterium atoms. The present invention is also directed to processes of preparing [2-.sup.13 C]dithiane wherein the .sup.13 C atom is directly bonded to one or two deuterium atoms. The present invention is also directed to labeled compounds, e.g., [.sup.2 H.sub.1-2, .sup.13 C]methanol (arylthio)-, acetates wherein the .sup.13 C atom is directly bonded to exactly one or two deuterium atoms.

  15. Calculation of total meal d13C from individual food d13C.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Variations in the isotopic signature of carbon in biological samples can be used to distinguish dietary patterns and monitor shifts in metabolism. But for these variations to have meaning, the isotopic signature of the diet must be known. We sought to determine if knowledge of the 13C isotopic abund...

  16. In vivo dynamic turnover of cerebral 13C isotopomers from [U- 13C]glucose

    NASA Astrophysics Data System (ADS)

    Xu, Su; Shen, Jun

    2006-10-01

    An INEPT-based 13C MRS method and a cost-effective and widely available 11.7 Tesla 89-mm bore vertical magnet were used to detect dynamic 13C isotopomer turnover from intravenously infused [U- 13C]glucose in a 211 μL voxel located in the adult rat brain. The INEPT-based 1H → 13C polarization transfer method is mostly adiabatic and therefore minimizes signal loss due to B 1 inhomogeneity of the surface coils used. High quality and reproducible data were acquired as a result of combined use of outer volume suppression, ISIS, and the single-shot three-dimensional localization scheme built in the INEPT pulse sequence. Isotopomer patterns of both glutamate C4 at 34.00 ppm and glutamine C4 at 31.38 ppm are dominated first by a doublet originated from labeling at C4 and C5 but not at C3 (with 1JC4C5 = 51 Hz) and then by a quartet originated from labeling at C3, C4, and C5 (with 1JC3C4 = 35 Hz). A lag in the transition of glutamine C4 pattern from doublet-dominance to quartet dominance as compared to glutamate C4 was observed, which provides an independent verification of the precursor-product relationship between neuronal glutamate and glial glutamine and a significant intercompartmental cerebral glutamate-glutamine cycle between neurons and glial cells.

  17. Determination of Transport Parameters in Unsaturated Zone by Tracer Experiment in the Porous Aquifer located at Ljubljana, Slovenia

    NASA Astrophysics Data System (ADS)

    Vidmar, S.; Cencur Curk, B.

    2009-04-01

    The gravel sandy aquifer of Ljubljansko polje is the source of drinking water for nearly 300.000 inhabitants of the Ljubljana city and vicinity. There are two main waterworks: Kleče and Hrastje. The plain area of Ljubljansko polje is a tectonic sink and consists of river sediments that can reach in thickness more than 100 m in the deepest part. The bedrock is the impermeable permocarbonic clayey shale, mudstones and sandstones. The hydraulic conductivity of Ljubljansko polje sediments is very good, from 10-2 m/s in the central part to 3.7•10-3 m/s on the borders of the plain. The average groundwater level is 20 m below surface. A numerical groundwater flow model was established for the wider area of the Ljubljansko polje aquifer. The fore mentioned model was not calibrated on solute transport parameters but only on water levels and this lead to unreliability in the transport model and its predictions of pollution scenarios. The transport model needs to calculate reliable scenarios of pollution dispersion, which can only be achieved with the application of real transport parameters. Human activities in the area of the Hrastje waterworks of Ljubljana threaten to degrade groundwater quality. For this reason several tracer experiments were carried out in the past. Despite a great risk, the experiments were performed on the catchment area of the Hrastje waterworks, inside the second water protection zone. During the experiments the water from Hrastje waterworks was still in use for drinking water supply. The tracer experiments were carried out in order to determine the solute transport parameters such as advection, dispersion and sorption. The research proved that the tracers could be used safely on sensitive area and that the researchers are capable and qualified to carry it out with a highest level of security. Since none of the past tracer experiments, carried out in the same area, gave us any detailed information on pollutant spreading in unsaturated zone a new

  18. Prognostic Prediction of Tracer Dispersion for the Diablo Canyon Experiments on August 31, September 2, and September 4, 1986

    SciTech Connect

    Molenkamp, C.R.

    1999-11-29

    COAMPS/LODI simulations of the tracer experiments at Diablo Canyon on August 31, September 2, and September 4, 1986 had mixed results. Simulated tracer concentrations on August 31 differed significantly from the measured concentrations. The model transported SF{sub 6} too far south and did not predict transport of SF{sub 6} north along highway 101 or into See Canyon. Early in the day the model rapidly transported SF{sub 6} away from the release point while observations suggested the tracer stayed close to Diablo Canyon for 1-2 hours. For September 2, simulations agreed very well with the measurements. The model accurately predicted the change of wind direction from north northwest to east northeast at the release point. It also predicted the advection of tracer over Mot-r-0 Bay and through the Los Osos Valley toward San Luis Obispo in excellent agreement with the observations. On September 4, the calculated transport of SF{sub 6} from Diablo Canyon had defects similar to those on August 31, a trajectory too far south and limited intrusion of tracer north along highway 101. Conversely, simulations of the Freon release from Los Osos Cemetery on September 4 corresponded well with observations. Since the simulations used only global meteorological data and no local winds for input, even the limited success of COAMPS/LODI is a favorable result. COAMPS's inability to generate southerly winds through the highway 101 corridor on August 31 and September 4 is a symptom of its underestimate of the sea breeze. The weak sea breeze correlates with a small diurnal range of air temperature possibly associated with underestimates of surface solar heating and/or overestimates of surface wetness. Improvement of COAMPS/LODI simulations requires development of new data assimilation techniques to use the local surface and low altitude wind and temperature measurements. Also, quantitative methods are needed to assess the accuracy of the models.

  19. On the use of phloem sap δ13C to estimate canopy carbon discrimination

    NASA Astrophysics Data System (ADS)

    Rascher, Katherine; Máguas, Cristina; Werner, Christiane

    2010-05-01

    Although the carbon stable isotope composition (d13C) of bulk leaf material is a good integrative parameter of photosynthetic discrimination and can be used as a reliable ecological index of plant functioning; it is not a good tracer of short-term changes in photosynthetic discrimination. In contrast, d13C of phloem sap is potentially useful as an indicator of short-term changes in canopy photosynthetic discrimination. However, recent research indicates that d13C signatures may be substantially altered by metabolic processes downstream of initial leaf-level carbon fixation (e.g. post-photosynthetic fractionation). Accordingly, before phloem sap d13C can be used as a proxy for canopy level carbon discrimination an understanding of factors influencing the degree and magnitude of post-photosynthetic fractionation and how these vary between species is of paramount importance. In this study, we measured the d13C signature along the basipetal transport pathway in two co-occurring tree species in the field - an understory invasive exotic legume, Acacia longifolia, and a native pine, Pinus pinaster. We measured d13C of bulk leaf and leaf water soluble organic matter (WSOM), phloem sap sampled at two points along the plant axis and leaf and root dark respiration. In general, species differences in photosynthetic discrimination resulted in more enriched d13C values in the water-conserving P. pinaster relative to the water-spending A. longifolia. Post-photosynthetic fractionation led to differences in d13C of carbon pools along the plant axis with progressively more depleted d13C from the canopy to the trunk (~6.5 per mil depletion in A. longifolia and ~0.8per mil depletion in P. pinaster). Leaf and root respiration, d13C, were consistently enriched relative to putative substrates. We hypothesize that the pronounced enrichment of leaf respired CO2 relative to leaf WSOM may have left behind relatively depleted carbon to be loaded into the phloem resulting in d13C depletion

  20. 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.

  1. Fate of nitrogen deposition and decomposed nitrogen from litter in a 15N-tracer mesocosm experiment

    NASA Astrophysics Data System (ADS)

    Nair, R.; Perks, M.; Mencuccini, M.

    2013-12-01

    Atmospheric deposition of anthropogenic-derived nitrogen may be a major driver of the 0.6-0.7 Pg y-1 increase in the carbon sink in historically N-limited northern and boreal forests, but the magnitude of its effect is still uncertain. A strong effect depends on the allocation of N to trees, because of their high C:N ratio in woody tissues, and isotope tracer experiments have shown that the majority of 15N tracers applied directly to the soil are lost via leeching or retained in soil pools rather than being acquired by tree root systems. However, ambient anthropogenic inputs of N to these systems are transported in the atmosphere and intercepted by foliage before they reach the soil system, while labelled fertilization experiments also can only explicitly trace the fate of the 15N-tracer from deposition, as opposed to changes in the fate of N from litter, where decomposition rates may be enhanced at low ambient levels of deposition, affecting the availability of N from this pool for tree nutrition. We present initial results from a potted Sitka Spruce mesocosm 15N-tracer experiment where ambient nitrogen deposition was supplemented with a minor (0.4 kg ha-1 y-1) input of additional N, applied to either the soil or the foliage. Either this deposition, or litter in the pots, was enriched in 15N, allowing the fate of the isotope from two different methods of deposition to be compared with that of nitrogen released from the litter under the deposition treatment.

  2. Assessment of electrical conductivity as a surrogate measurement for water samples in a tracer injection experiment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The transport behavior of solutes in streams depends on chemical, physical, biological, and hydrodynamic processes. Although it is a very complex system, it is known that this behavior is greatly influenced by surface and subsurface flows. For this reason, tracer injection in the water flows is one ...

  3. Transport of microspheres and indigenous bacteria through a sandy aquifer: Results of natural- and forced-gradient tracer experiments

    USGS Publications Warehouse

    Harvey, R.W.; George, L.H.; Smith, R.L.; LeBlanc, D.R.

    1989-01-01

    Transport of indigenous bacteria through sandy aquifer sediments was investigated in forced- and natural-gradient tracer teste. A diverse population of bacteria was collected and concentrated from groundwater at the site, stained with a DNA-specific fluorochrome, and injected back into the aquifer. Included with the injectate were a conservative tracer (Br- or Cl-) and bacteria-sized (0.2-1.3-??m) microspheres having carboxylated, carbonyl, or neutral surfaces. Transport of stained bacteria and all types and size classes of microspheres was evident. In the natural-gradient test, both surface characteristics and size of microspheres affected attenuation. Surface characteristics had the greatest effect upon retardation. Peak break-through of DAPI-stained bacteria (forced-gradient experiment) occurred well in advance of bromide at the more distal sampler. Transport behavior of bacteria was substantially different from that of carboxylated microspheres of comparable size. ?? 1988 American Chemical Society.

  4. Recover Act. Verification of Geothermal Tracer Methods in Highly Constrained Field Experiments

    SciTech Connect

    Becker, Matthew W.

    2014-05-16

    The prediction of the geothermal system efficiency is strong linked to the character of the flow system that connects injector and producer wells. If water flow develops channels or “short circuiting” between injection and extraction wells thermal sweep is poor and much of the reservoir is left untapped. The purpose of this project was to understand how channelized flow develops in fracture geothermal reservoirs and how it can be measured in the field. We explored two methods of assessing channelization: hydraulic connectivity tests and tracer tests. These methods were tested at a field site using two verification methods: ground penetrating radar (GPR) images of saline tracer and heat transfer measurements using distributed temperature sensing (DTS). The field site for these studies was the Altona Flat Fractured Rock Research Site located in northeastern New York State. Altona Flat Rock is an experimental site considered a geologic analog for some geothermal reservoirs given its low matrix porosity. Because soil overburden is thin, it provided unique access to saturated bedrock fractures and the ability image using GPR which does not effectively penetrate most soils. Five boreholes were drilled in a “five spot” pattern covering 100 m2 and hydraulically isolated in a single bedding plane fracture. This simple system allowed a complete characterization of the fracture. Nine small diameter boreholes were drilled from the surface to just above the fracture to allow the measurement of heat transfer between the fracture and the rock matrix. The focus of the hydraulic investigation was periodic hydraulic testing. In such tests, rather than pumping or injection in a well at a constant rate, flow is varied to produce an oscillating pressure signal. This pressure signal is sensed in other wells and the attenuation and phase lag between the source and receptor is an indication of hydraulic connection. We found that these tests were much more effective than constant

  5. Determination of sup 13 C labeling pattern of citric acid cycle intermediates by gas chromatography-mass spectrometry

    SciTech Connect

    Di Donato, L.; Montgomery, J.A.; Des Rosiers, C.; David, F.; Garneau, M.; Brunengraber, H. )

    1990-02-26

    Investigations of the regulation of the citric acid cycle require determination of labeling patterns of cycle intermediates. These were assayed to date, using infusion of: (i) ({sup 14}C)tracer followed by chemical degradation of intermediates and (ii) ({sup 13}C)tracer followed by NMR analysis of intermediates. The authors developed a strategy to analyze by GC-MS the ({sup 13}C) labeling pattern of {mu}mole samples of citrate (CIT), isocitrate (ICIT), 2-ketoglutarate (2-KG), glutamate (GLU) and glutamine (GLN). These are enzymatically or chemically converted to 2-KG, ICIT, 4-aminobutyrate (GABA) and 2-hydroxyglutarate (2-OHG). GC-MS analyses of TMS or TBDMS derivatives of these compounds yield the enrichment of each carbon. The authors confirmed the identity of each fragment using the spectra of (1-{sup 13}C), (5-{sup 13}C), (2,3,3,4,4-{sup 2}H{sub 5})glutamate and (1-{sup 13}C), (1,4-{sup 13}C)GABA.

  6. Determining concentration fields of tracer plumes for layered porous media in flow-tank experiments

    NASA Astrophysics Data System (ADS)

    Yu, Zhongbo; Schwartz, Franklin W.

    In the laboratory, computer-assisted image analysis provides an accurate and efficient way to monitor tracer experiments. This paper describes the determination of detailed temporal concentration distributions of tracers in a flow-tank experiment by analyzing photographs of plumes of Rhodamine dye through the glass wall of the tank. The methodology developed for this purpose consists of four steps: (1) digitally scanning black and white negatives obtained from photographs of the flow-tank experiment; (2) calibrating and normalizing each digitized image to a standard optical-density scale by determining the relation between the optical density and pixel value for each image; (3) constructing standard curves relating the concentration in an optical density from five experimental runs with predetermined concentrations (2-97mg/L) and (4) converting the optical density to concentration. The spatial distribution of concentration for two photographs was determined by applying these calibration and conversion procedures to all pixels of the digitized images. This approach provides an efficient way to study patterns of plume evolution and transport mechanisms. Résumé Au laboratoire, l'analyse d'images assistée par ordinateur est un moyen précis et efficace pour suivre certaines expériences de traçage. Ce papier présente comment sont déterminées dans le détail les distributions temporelles de la concentration en traceur au cours d'une expérience d'écoulement en réservoir au moyen de l'analyse de photographies de panaches de rhodamine à travers la paroi de verre du réservoir. La méthodologie développée dans cette expérience suit quatre étapes: (1) digitalisation par balayage des négatifs noir et blanc des prises de vue de l'expérience d'écoulement en réservoir (2) calibration et normalisation de chaque image digitalisée par rapport à une échelle étalon de densité optique en déterminant la relation entre la densité optique et la valeur des pixels

  7. 13C isotopic fractionation during biodegradation of agricultural wastes.

    PubMed

    Chalk, Phillip M; Inácio, Caio T; Urquiaga, Segundo; Chen, Deli

    2015-01-01

    Significant differences in δ(13)C signatures occur within and between plant tissues and their constituent biochemical entities, and also within and between heterotrophic bacteria and fungi and their metabolic products. Furthermore, (13)C isotopic fractionation occurs during the biodegradation of organic molecules as seen in the substrate, respired CO(2) and the microbial biomass, which could be related to substrate composition and/or microbial metabolism. The (13)C isotopic fractionation observed during the decomposition of a single defined C substrate appears to be due to the intra-molecular heterogeneity in (13)C in the substrate and to (13)C isotopic fractionation during microbial metabolism. Very limited data suggest that the latter may be quantitatively more important than the former. Studies with defined fungi in culture media have highlighted the complexities associated with the interpretation of the observed patterns of (13)C isotopic fractionation when a single defined C source is added to the culture medium which itself contains one or more C sources. Techniques involving (13)C enrichment or paired treatments involving an equivalent C(3)- and C(4)-derived substrate have been devised to overcome the problem of background C in the culture medium and (13)C isotopic fractionation during metabolism. Studies with complex substrates have shown an initial (13)C depletion phase in respired CO(2) followed by a (13)C enrichment phase which may or may not be followed by another (13)C depletion phase. Basic studies involving an integrated approach are required to gain a new insight into (13)C isotopic fractionation during organic residue decomposition, by simultaneous measurements of δ(13)C in all C moieties. New analytical tools to measure real-time changes in δ(13)CO(2) and the intra-molecular δ(13)C distribution within plant biochemical entities offer new opportunities for unravelling the complex interactions between substrate and microbial metabolism with

  8. 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.

  9. 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

  10. Gas Chromatography-Quadrupole Time-of-Flight Mass Spectrometry-Based Determination of Isotopologue and Tandem Mass Isotopomer Fractions of Primary Metabolites for (13)C-Metabolic Flux Analysis.

    PubMed

    Mairinger, Teresa; Steiger, Matthias; Nocon, Justyna; Mattanovich, Diethard; Koellensperger, Gunda; Hann, Stephan

    2015-12-01

    For the first time an analytical work flow based on accurate mass gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOFMS) with chemical ionization for analysis providing a comprehensive picture of (13)C distribution along the primary metabolism is elaborated. The method provides a powerful new toolbox for (13)C-based metabolic flux analysis, which is an emerging strategy in metabolic engineering. In this field, stable isotope tracer experiments based on, for example, (13)C are central for providing characteristic patterns of labeled metabolites, which in turn give insights into the regulation of metabolic pathway kinetics. The new method enables the analysis of isotopologue fractions of 42 free intracellular metabolites within biotechnological samples, while tandem mass isotopomer information is also accessible for a large number of analytes. Hence, the method outperforms previous approaches in terms of metabolite coverage, while also providing rich isotopomer information for a significant number of key metabolites. Moreover, the established work flow includes novel evaluation routines correcting for isotope interference of naturally distributed elements, which is crucial following derivatization of metabolites. Method validation in terms of trueness, precision, and limits of detection was performed, showing excellent analytical figures of merit with an overall maximum bias of 5.8%, very high precision for isotopologue and tandem mass isotopomer fractions representing >10% of total abundance, and absolute limits of detection in the femtomole range. The suitability of the developed method is demonstrated on a flux experiment of Pichia pastoris employing two different tracers, i.e., 1,6(13)C2-glucose and uniformly labeled (13)C-glucose. PMID:26513365

  11. In situ measurement of methane oxidation in groundwater by using natural-gradient tracer tests

    USGS Publications Warehouse

    Smith, R.L.; Howes, B.L.; Garabedian, S.P.

    1991-01-01

    Methane oxidation was measured in an unconfined sand and gravel aquifer (Cape Cod, Mass.) by using in situ natural-gradient tracer tests at both a pristine, oxygenated site and an anoxic, sewage-contaminated site. The tracer sites were equipped with multilevel sampling devices to create target grids of sampling points; the injectate was prepared with groundwater from the tracer site to maintain the same geochemical conditions. Methane oxidation was calculated from breakthrough curves of methane relative to halide and inert gas (hexafluoroethane) tracers and was confirmed by the appearance of 13C-enriched carbon dioxide in experiments in which 13C-enriched methane was used as the tracer. A V(max) for methane oxidation could be calculated when the methane concentration was sufficiently high to result in zero-order kinetics throughout the entire transport interval. Methane breakthrough curves could be simulated by modifying a one-dimensional advection-dispersion transport model to include a Michaelis-Menten-based consumption term for methane oxidation. The K(m) values for methane oxidation that gave the best match for the breakthrough curve peaks were 6.0 and 9.0 ??M for the uncontaminated and contaminated sites, respectively. Natural-gradient tracer tests are a promising approach for assessing microbial processes and for testing in situ bioremediation potential in groundwater systems.

  12. In situ measurement of methane oxidation in groundwater by using natural-gradient tracer tests.

    PubMed Central

    Smith, R L; Howes, B L; Garabedian, S P

    1991-01-01

    Methane oxidation was measured in an unconfined sand and gravel aquifer (Cape Cod, Mass.) by using in situ natural-gradient tracer tests at both a pristine, oxygenated site and an anoxic, sewage-contaminated site. The tracer sites were equipped with multilevel sampling devices to create target grids of sampling points; the injectate was prepared with groundwater from the tracer site to maintain the same geochemical conditions. Methane oxidation was calculated from breakthrough curves of methane relative to halide and inert gas (hexafluroethane) tracers and was confirmed by the appearance of 13C-enriched carbon dioxide in experiments in which 13C-enriched methane was used as the tracer. A Vmax for methane oxidation could be calculated when the methane concentration was sufficiently high to result in zero-order kinetics throughout the entire transport interval. Methane breakthrough curves could be simulated by modifying a one-dimensional adevection-dispersion transport model to include a Michaelis-Menten-based consumption term for methane oxidation. The Km values for methane oxidation that gave the best match for the breakthrough curve peaks were 6.0 and 9.0 microM for the uncontaminated and contaminated sites, respectively. Natural-gradient tracer tests are a promising approach for assessing microbial processes and for testing in situ bioremediation potential in groundwater systems. PMID:1892389

  13. Methane emission estimates using chamber and tracer release experiments for a municipal waste water treatment plant

    NASA Astrophysics Data System (ADS)

    Yver Kwok, C. E.; Müller, D.; Caldow, C.; Lebègue, B.; Mønster, J. G.; Rella, C. W.; Scheutz, C.; Schmidt, M.; Ramonet, M.; Warneke, T.; Broquet, G.; Ciais, P.

    2015-07-01

    This study presents two methods for estimating methane emissions from a waste water treatment plant (WWTP) along with results from a measurement campaign at a WWTP in Valence, France. These methods, chamber measurements and tracer release, rely on Fourier transform infrared spectroscopy and cavity ring-down spectroscopy instruments. We show that the tracer release method is suitable for quantifying facility- and some process-scale emissions, while the chamber measurements provide insight into individual process emissions. Uncertainties for the two methods are described and discussed. Applying the methods to CH4 emissions of the WWTP, we confirm that the open basins are not a major source of CH4 on the WWTP (about 10 % of the total emissions), but that the pretreatment and sludge treatment are the main emitters. Overall, the waste water treatment plant is representative of an average French WWTP.

  14. Assessing microbial utilization of free versus sorbed Alanine by using position-specific 13C labeling and 13C-PLFA analysis

    NASA Astrophysics Data System (ADS)

    Herschbach, Jennifer; Apostel, Carolin; Spielvogel, Sandra; Kuzyakov, Yakov; Dippold, Michaela

    2016-04-01

    Microbial utilization is a key transformation process of soil organic matter (SOM). Sorption of low molecular weight organic substances (LMWOS) to soil mineral surfaces blocks or delays microbial uptake and therefore mineralization of LMWOS to CO2, as well as all other biochemical transformations. We used position-specific labeling, a tool of isotope applications novel to soil science, combined with 13C-phospholipid fatty acid (PLFA) analysis, to assess microbial utilization of sorbed and non-sorbed Alanine in soil. Alanine has various functional groups enabling different sorption mechanisms via its positive charge (e.g. to clay minerals by cation exchange), as well as via its negative charge (e.g. to iron oxides by ligand exchange). To assess changes in the transformation pathways caused by sorption, we added uniformly and position-specifically 13C and 14C labeled Alanine to the Ap of a loamy Luvisol in a short-term (10 days) incubation experiment. To allow for sorption of the tracer solution to an aliquot of this soil, microbial activity was minimized in this subsample by sterilizing the soil by γ-radiation. After shaking, the remaining solutions were filtered and the non-sorbed Alanine was removed with Millipore water and then added to non-sterilized soil. For the free Alanine treatment, solutions with Alanine of similar amount and isotopic composition were prepared, added to the soil and incubated as well. The respired CO2 was trapped in NaOH and its 14C-activity was determined at increasing times intervals. Microbial utilization of Alanine's individual C positions was evaluated in distinct microbial groups classified by 13C-PLFA analysis. Sorption to soil minerals delayed respiration to CO2 and reduced initial respiration rate by 80%. Irrespective of sorption, the highest amount was respired from the carboxylic position (C-1), whereas the amino-bound (C-2) and the methylic position (C-3) were preferentially incorporated into PLFA of microorganisms due to the

  15. Practical considerations in the gas chromatography/combustion/isotope ratio monitoring mass spectrometry of 13C-enriched compounds: detection limits and carryover effects.

    PubMed

    Mottram, Hazel R; Evershed, Richard P

    2003-01-01

    This paper describes a methodological investigation of the use of gas chromatography/combustion/isotope ratio monitoring mass spectrometry (GC/C/IRMS) for the compound-specific stable isotope analysis of 13C-enriched compounds. Analysis of two 13C-enriched fatty acid methyl esters, possessing delta13C values of approximately 500 per thousand, at a range of concentrations, demonstrated that detectable responses, i.e. chromatographic peaks, could be observed in the 45/44 output even when the compound was present in such low abundance that no peak was observed in the m/z 44 ion chromatogram. A limit of detection, defined as the point at which the signal-to-background ratio was equal to 3, was calculated for two compounds and for both ion chromatograms. The limit of detection in the 45/44 chromatogram was found to be ca. 30 pg injected for methyl 13C-hexadecanoate and ca. 20 pg injected for methyl 13C-octadecanoate, whilst, in the m/z 44 ion chromatogram, detection limits were approximately 180 and approximately 200 pg, respectively. The delta13C value recorded for the analytes was found to be both inaccurate and imprecise below 5 ng of each component injected, although this would not represent a significant drawback in qualitative tracer-type experiments. In a further study of co-injected mixtures of labelled (approximately 500 per thousand) and unlabelled (natural abundance, -20 to -30 per thousand ) fatty acid methyl esters a significant within-run carryover effect was observed, where the isotope values recorded for compounds eluting immediately after enriched components were significantly affected. Whilst this would not affect qualitative results, quantitative data for mixtures containing enriched compounds should be considered with caution. The standards employed in this investigation were enriched to approximately 500 per thousand in 13C; however, these effects would probably be accentuated at higher levels of labelling and with other elements. The limit of

  16. Monitoring electron donor metabolism under variable electron acceptor conditions using 13C-labeled lactate

    NASA Astrophysics Data System (ADS)

    Bill, M.; Conrad, M. E.; Yang, L.; Beller, H. R.; Brodie, E. L.

    2010-12-01

    Three sets of flow-through columns constructed with aquifer sediment from Hanford (WA) were used to study reduction of Cr(VI) to poorly soluble Cr(III) under denitrifying, sulfate-reducing/fermentative, and iron-reducing conditions with lactate as the electron donor. In order to understand the relationship between electron donors and biomarkers, and to determine the differences in carbon isotope fractionation resulting from different microbial metabolic processes, we monitored the variation in carbon isotopes in dissolved inorganic carbon (DIC), in total organic carbon (TOC), and in lactate, acetate and propionate. The greatest enrichment in 13C in columns was observed under denitrifying conditions. The δ13C of DIC increased by ~1750 to ~2000‰ fifteen days after supplementation of natural abundance lactate with a 13C-labeled lactate tracer (for an influent δ13C of ~2250‰ for the lactate) indicating almost complete oxidation of the electron donor. The denitrifying columns were among the most active columns and had the highest cell counts and the denitrification rate was highly correlated with Cr(VI) reduction rate. δ13C values of DIC ranged from ~540 to ~1170‰ for iron-reducing conditions. The lower enrichment in iron columns was related to the lower biological activity observed with lower yields of RNA and cell numbers in the column effluents. The carbon isotope shift in the sulfate-reducing ~198 to ~1960‰ for sulfate-reducing conditions reflecting the lower levels of the lactate in these columns. Additionally, in two of the sulfate columns, almost complete fermentation of the lactate occurred, producing acetate and propionate with the labeled carbon signature, but relatively smaller amounts of inorganic carbon. For all electron-accepting conditions, TOC yielded similar δ13C values as lactate stock solutions. Differences in C use efficiency, metabolic rate or metabolic pathway contributed to the differing TOC δ13C to DIC δ13C ratios between treatments

  17. Multiresponse modeling of variably saturated flow and isotope tracer transport for a hillslope experiment at the Landscape Evolution Observatory

    NASA Astrophysics Data System (ADS)

    Scudeler, Carlotta; Pangle, Luke; Pasetto, Damiano; Niu, Guo-Yue; Volkmann, Till; Paniconi, Claudio; Putti, Mario; Troch, Peter

    2016-10-01

    This paper explores the challenges of model parameterization and process representation when simulating multiple hydrologic responses from a highly controlled unsaturated flow and transport experiment with a physically based model. The experiment, conducted at the Landscape Evolution Observatory (LEO), involved alternate injections of water and deuterium-enriched water into an initially very dry hillslope. The multivariate observations included point measures of water content and tracer concentration in the soil, total storage within the hillslope, and integrated fluxes of water and tracer through the seepage face. The simulations were performed with a three-dimensional finite element model that solves the Richards and advection-dispersion equations. Integrated flow, integrated transport, distributed flow, and distributed transport responses were successively analyzed, with parameterization choices at each step supported by standard model performance metrics. In the first steps of our analysis, where seepage face flow, water storage, and average concentration at the seepage face were the target responses, an adequate match between measured and simulated variables was obtained using a simple parameterization consistent with that from a prior flow-only experiment at LEO. When passing to the distributed responses, it was necessary to introduce complexity to additional soil hydraulic parameters to obtain an adequate match for the point-scale flow response. This also improved the match against point measures of tracer concentration, although model performance here was considerably poorer. This suggests that still greater complexity is needed in the model parameterization, or that there may be gaps in process representation for simulating solute transport phenomena in very dry soils.

  18. Stable Carbon Isotopes (δ 13C) in Coral Skeletons: Experimental Approach and Applications for Paleoceanography

    NASA Astrophysics Data System (ADS)

    Grottoli, A. G.

    2004-12-01

    Scleractinian corals obtain fixed carbon via photosynthesis by their endosymbiotic algae (zooxanthellae) and via hetertrophy (injestion of zooplankton, δ 13C ≈ -17 to -22‰ ). Carbon dioxide (CO2) used for photosynthesis is obtained from seawater (δ 13C ≈ 0%) or from respired CO2 within the coral host. The δ 13C of the carbon used in the formation of the underlying coral skeleton is fractionated as a result of both of these metabolic processes. Here I have pooled evidence from several field and tank experiments on the effect of photosynthesis and heterotrophy of coral skeletal δ 13C. In the experiments, decreases in light levels due to shading or depth resulted in a significant decrease in skeletal δ 13C in all species studied (Pavona gigantea, Pavona clavus, Porites compressa). Decreases in photosynthesis in bleached corals also resulted in a decrease in skeletal δ 13C compared to non-bleached corals growing under the same conditions and at the same location. Skeletal δ 13C also decreased at higher than normal light levels most likely due to photoinhibition. Thus, decreases in photosynthesis due to reduced light levels, due to bleaching-induced decreases in chlorophyll a concentrations, or due to photodamage-induced decreases in functional cholorphyll a, results in significant δ 13C decreases. Comprehensive interpretation of all of the data showed that changes in photosynthesis itself can drive the changes in δ 13C. In field experiments, the addition of natural concentrations of zooplankton to the diet resulted in decreases in skeletal δ 13C. Such a decrease was more pronounced with depth and in P. gigantea compared to P. clavus. In situ feeding experiments have since confirmed these findings. However under tank conditions with unaturally high feeding rates, enhanced nitrogen supply in the diet can disrupt the coral-algal symbiosis, stimlate zooxanthellae growth and photosynthesis, and cause an incrase in skeletal δ 13C. It is proposed that under

  19. sup 13 C-enriched ribonucleosides: Synthesis and application of sup 13 C- sup 1 H and sup 13 C- sup 13 C spin-coupling constants to assess furanose and N-glycoside bond conformations

    SciTech Connect

    Kline, P.C.; Serianni, A.S. )

    1990-09-26

    Adenosine (1), cytidine (2), guanosine (3), and uridine (4) have been prepared chemically with {sup 13}C enrichment (99 atom %) at C1{prime} and C2{prime} of the ribose ring. Reliable synthetic protocols have been developed to permit access to millimole quantities of labeled ribonucleosides required for structural studies of stable isotopically labeled oligonucleotides and for in vivo metabolism studies. High-resolution {sup 1}H and {sup 13}C NMR spectra of the enriched ribonucleosides have been obtained, and {sup 13}C-{sup 13}C and {sup 13}C-{sup 1}H spin-coupling constants have been measured for pathways within the {beta}-D-ribofuranose ring and across the N-glycoside bond. Related couplings were determined in methyl {alpha}- and {beta}-D-riboruanosides (5,6), and in two conformationally constrained nucleosides, 2,2{prime}-anhydro-(1-{beta}-D-arabinofuranosyl)uracil (7) and 2{prime},3{prime}-O-isopropylidene-2,5{prime}-O-cyclouridine (8). The latter data were used to construct a crude Karplus curve for the {sup 13}C-C-N-{sup 13}C coupling pathway across the N-glycoside bond in 1-4. {sup 1}H-{sup 1}H, {sup 13}C-{sup 1}H, and {sup 13}C-{sup 13}C coupling data are used to evaluate current models describing the conformational dynamics of 1-4 in aqueous solution.

  20. Synthesis Of [2h, 13c]M [2h2m 13c], And [2h3,, 13c] Methyl Aryl Sulfones And Sulfoxides

    DOEpatents

    Martinez, Rodolfo A.; Alvarez, Marc A.; Silks, III, Louis A.; Unkefer, Clifford J.; Schmidt, Jurgen G.

    2004-07-20

    The present invention is directed to labeled compounds, [.sup.2 H.sub.1, .sup.13 C], [.sup.2 H.sub.2, .sup.13 C] and [.sup.2 H.sub.3, .sup.13 C]methyl aryl sulfones and [.sup.2 H.sub.1, .sup.13 C], [.sup.2 H.sub.2, .sup.13 C] and [.sup.2 H.sub.3, .sup.13 C]methyl aryl sulfoxides, wherein the .sup.13 C methyl group attached to the sulfur of the sulfone or sulfoxide includes exactly one, two or three deuterium atoms and the aryl group is selected from the group consisting of 1-naphthyl, substituted 1-naphthyl, 2-naphthyl, substituted 2-naphthyl, and phenyl groups with the structure: ##STR1## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently, hydrogen, a C.sub.1 -C.sub.4 lower alkyl, a halogen, an amino group from the group consisting of NH.sub.2, NHR and NRR' where R and R' are each a C.sub.1 -C.sub.4 lower alkyl, a phenyl, or an alkoxy group. The present invention is also directed to processes of preparing methyl aryl sulfones and methyl aryl sulfoxides.

  1. 13C Incorporation into Signature Fatty Acids as an Assay for Carbon Allocation in Arbuscular Mycorrhiza

    PubMed Central

    Olsson, Pål Axel; van Aarle, Ingrid M.; Gavito, Mayra E.; Bengtson, Per; Bengtsson, Göran

    2005-01-01

    The ubiquitous arbuscular mycorrhizal fungi consume significant amounts of plant assimilated C, but this C flow has been difficult to quantify. The neutral lipid fatty acid 16:1ω5 is a quantitative signature for most arbuscular mycorrhizal fungi in roots and soil. We measured carbon transfer from four plant species to the arbuscular mycorrhizal fungus Glomus intraradices by estimating 13C enrichment of 16:1ω5 and compared it with 13C enrichment of total root and mycelial C. Carbon allocation to mycelia was detected within 1 day in monoxenic arbuscular mycorrhizal root cultures labeled with [13C]glucose. The 13C enrichment of neutral lipid fatty acid 16:1ω5 extracted from roots increased from 0.14% 1 day after labeling to 2.2% 7 days after labeling. The colonized roots usually were more enriched for 13C in the arbuscular mycorrhizal fungal neutral lipid fatty acid 16:1ω5 than for the root specific neutral lipid fatty acid 18:2ω6,9. We labeled plant assimilates by using 13CO2 in whole-plant experiments. The extraradical mycelium often was more enriched for 13C than was the intraradical mycelium, suggesting rapid translocation of carbon to and more active growth by the extraradical mycelium. Since there was a good correlation between 13C enrichment in neutral lipid fatty acid 16:1ω5 and total 13C in extraradical mycelia in different systems (r2 = 0.94), we propose that the total amount of labeled C in intraradical and extraradical mycelium can be calculated from the 13C enrichment of 16:1ω5. The method described enables evaluation of C flow from plants to arbuscular mycorrhizal fungi to be made without extraction, purification and identification of fungal mycelia. PMID:15870350

  2. New study of the astrophysical reaction 13C(a,n)16O via the 13C(7Li,t)17O transfer reaction

    NASA Astrophysics Data System (ADS)

    Pellegriti, Maria Grazia; Hammache, F.; Roussel, P.; Audouin, L.; Beaumel, D.; Fortier, S.; Gaudefroy, L.; Kiener, J.; Lefebvre-Schujl, A.; Stanoiu, M.; Tatischeff, V.; Vilmay, M.

    PoS(NIC-IX)161 , , [1] , L. Gaudefroy[2] , J. Kiener[3] , A. Lefebvre-Schuhl[3] , M. Stanoiu[4] , V. The cross section of the 13 C(α,n)16 O reaction is a key ingredient for the comprehension of the s-process (slow neutron captures) in stars. This reaction is considered as the main neutron source for the s-process in low-mass Asymptotic Giant Branch (AGB) stars (1-3 solar mass) [1, 2, 3]. At the α-13 C energies of astrophysical interest (Ecm around 190 keV, corresponding to a tem- perature of 108 K) the contribution of the 17 O α-decay subthreshold resonance at 6.356 MeV to the 13 C(α,n)16 O cross section should be taken into account. The effect of this resonance is controversial after the different analyses of the Kubono et al. measurement [4] of the 6.356 MeV α-spectroscopic factor (Sα ) via the transfer reaction 13 C(6 Li,d)17 O . In order to further investigate the contribution of the 6.356 MeV resonance to the 13 C(α,n)16 O cross section, we performed a new measurement of its Sα factor via a different α-transfer reac- tion, namely the 13 C(7 Li,t)17 O reaction. The experiment was performed at the Orsay Tandem by using a 7 Li beam of 28 and 34 MeV on a 13C target. The angular distribution for the transfer dif- ferential cross section was measured by detecting the tritons at the focal plane of the SPLITPOLE spectrometer. The analysis procedure used in order to extract the yield of the 6.356 MeV level will be described. Preliminary results of the angular distribution will be shown.

  3. Radioisotope tracer studies in the NASA Skylab ethothermic brazing experiment M-552

    NASA Technical Reports Server (NTRS)

    Braski, D. N.; Adair, H. L.; Kobisk, E. H.

    1974-01-01

    The first use of radioisotope tracer for mapping flow patterns during brazing of metal components in a space environment (near-zero gravity) proved successful. A nickel ferrule was brazed to a nickel tube with Lithobraze BT (71.8% Ag, 28% Cu, 0.2% Li) which contained a trace amount of radioactive Ag-110. Mapping of the flow of the braze alloy in the annulus formed between the tube and the concentric ferrule was determined by counting the radiation intensity as a function of position in the braze joint. Significant information concerning the thermal history of the braze was determined.

  4. Σ production from targets of ^4He and ^13C

    NASA Astrophysics Data System (ADS)

    Chrien, R. E.

    1996-10-01

    One of the abiding issues in hypernuclear research has been the question of the formation of nuclear bound states incorporating the Σ-hyperon. The recent increases in beam intensity at the Brookhaven AGS have enabled us to obtain a high statistics study on the production of Σ-hyperons on a ^4He target. Earlier research using stopped kaons at KEK indicated the presence of structure in the (K^-,π^-) reaction, and led to the postulate of a Σ bound state. That structure has now been definitely confirmed in the in-flight kaon experiment at the LESB2 beam line and Moby-Dick spectrometer. An improved measurement of the binding energy of the presumed state will be reported, together with a production cross section. In addition, both (K^-,π^-) and (K^-,π^+) reactions on ^13C have been studied and will be compared to similar measurements on ^9Be.

  5. Measuring long-lived 13C2 state lifetimes at natural abundance

    NASA Astrophysics Data System (ADS)

    Claytor, Kevin; Theis, Thomas; Feng, Yesu; Warren, Warren

    2014-02-01

    Long-lived disconnected eigenstates (for example, the singlet state in a system with two nearly equivalent carbons, or the singlet-singlet state in a system with two chemically equivalent carbons and two chemically equivalent hydrogens) hold the potential to drastically extend the lifetime of hyperpolarization in molecular tracers for in vivo magnetic resonance imaging (MRI). However, a first-principles calculation of the expected lifetime (and thus selection of potential imaging agents) is made very difficult because of the large variety of relevant intra- and intermolecular relaxation mechanisms. As a result, all previous measurements relied on costly and time consuming syntheses of 13C labeled compounds. Here we show that it is possible to determine 13C singlet state lifetimes by detecting the naturally abundant doubly-labeled species. This approach allows for rapid and low cost screening of potential molecular biomarkers bearing long-lived states.

  6. /sup 13/C nuclear magnetic resonance studies of cardiac metabolism

    SciTech Connect

    Seeholzer, S.H.

    1985-01-01

    The last decade has witnessed the increasing use of Nuclear Magnetic Resonance (NMR) techniques for following the metabolic fate of compounds specifically labeled with /sup 13/C. The goals of the present study are: (1) to develop reliable quantitative procedures for measuring the /sup 13/C enrichment of specific carbon sites in compounds enriched by the metabolism of /sup 13/C-labeled substrates in rat heart, and (2) to use these quantitative measurements of fractional /sup 13/C enrichment within the context of a mathematical flux model describing the carbon flow through the TCA cycle and ancillary pathways, as a means for obtaining unknown flux parameters. Rat hearts have been perfused in vitro with various combinations of glucose, acetate, pyruvate, and propionate to achieve steady state flux conditions, followed by perfusion with the same substrates labeled with /sup 13/C in specific carbon sites. The hearts were frozen at different times after addition of /sup 13/C-labeled substrates and neutralized perchloric acid extracts were used to obtain high resolution proton-decoupled /sup 13/C NMR spectra at 90.55 MHz. The fractional /sup 13/C enrichment (F.E.) of individual carbon sites in different metabolites was calculated from the area of the resolved resonances after correction for saturation and nuclear Overhauser effects. These F.E. measurements by /sup 13/C NMR were validated by the analysis of /sup 13/C-/sup 1/H scalar coupling patterns observed in /sup 1/H NMR spectra of the extracted metabolites. The results obtained from perfusion of hearts glucose plus either (2-/sup 13/C) acetate or (3-/sup 13/C) pyruvate are similar to those obtained by previous investigators using /sup 14/C-labeled substrates.

  7. Methane emission estimates using chamber and tracer release experiments for a municipal waste water treatment plant

    NASA Astrophysics Data System (ADS)

    Yver-Kwok, C. E.; Müller, D.; Caldow, C.; Lebègue, B.; Mønster, J. G.; Rella, C. W.; Scheutz, C.; Schmidt, M.; Ramonet, M.; Warneke, T.; Broquet, G.; Ciais, P.

    2015-03-01

    This study presents two methods for estimating methane emissions from a waste water treatment plant (WWTP) along with results from a measurement campaign at a WWTP in Valence, France. These methods, chamber measurements and tracer release, rely on Fourier Transform Infrared (FTIR) spectroscopy and Cavity Ring Down Spectroscopy (CRDS) instruments. We show that the tracer release method is suitable to quantify facility- and some process-scale emissions, while the chamber measurements, provide insight into individual process emissions. Uncertainties for the two methods are described and discussed. Applying the methods to CH4 emissions of the WWTP, we confirm that the open basins are not a major source of CH4 on the WWTP (about 10% of the total emissions), but that the pretreatment and sludge treatment are the main emitters. Overall, the waste water treatment plant represents a small part (about 1.5%) of the methane emissions of the city of Valence and its surroundings, which is lower than the national inventories.

  8. Chain packing in glassy polymers by natural-abundance 13C-13C spin diffusion using 2D centerband-only detection of exchange.

    PubMed

    Singh, Manmilan; Schaefer, Jacob

    2011-03-01

    The proximities of specific subgroups of nearest-neighbor chains in glassy polymers are revealed by distance-dependent (13)C-(13)C dipolar couplings and spin diffusion. The measurement of such proximities is practical even with natural-abundance levels of (13)C using a 2D version of centerband-only detection of exchange (CODEX). Two-dimensional CODEX is a relaxation-compensated experiment that avoids the problems associated with variations in T(1)(C)'s due to dynamic site heterogeneity in the glass. Isotropic chemical shifts are encoded in the t(1) preparation times before and after mixing, and variations in T(2)'s are compensated by an S(0) reference (no mixing). Data acquisition involves acquisition of an S(0) reference signal on alternate scans, and the active control of power amplifiers, to achieve stability and accuracy over long accumulation times. The model system to calibrate spin diffusion is the polymer itself. For a mixing time of 200 ms, only (13)C-(13)C pairs separated by one or two bonds (2.5 Å) show cross peaks, which therefore identify reference intrachain proximities. For a mixing time of 1200 ms, 5 Å interchain proximities appear. The resulting cross peaks are used in a simple and direct way to compare nonrandom chain packing for two commercial polycarbonates with decidedly different mechanical properties.

  9. 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).

  10. Simulation of large particle transport near the surface under stable conditions: comparison with the Hanford tracer experiments

    NASA Astrophysics Data System (ADS)

    Kim, Eugene; Larson, Timothy

    A plume model is presented describing the downwind transport of large particles (1-100 μm) under stable conditions. The model includes both vertical variations in wind speed and turbulence intensity as well as an algorithm for particle deposition at the surface. Model predictions compare favorably with the Hanford single and dual tracer experiments of crosswind integrated concentration (for particles: relative bias=-0.02 and 0.16, normalized mean square error=0.61 and 0.14, for the single and dual tracer experiments, respectively), whereas the US EPA's fugitive dust model consistently overestimates the observed concentrations at downwind distances beyond several hundred meters (for particles: relative bias=0.31 and 2.26, mean square error=0.42 and 1.71, respectively). For either plume model, the measured ratio of particle to gas concentration is consistently overestimated when using the deposition velocity algorithm of Sehmel and Hodgson (1978. DOE Report PNL-SA-6721, Pacific Northwest Laboratories, Richland, WA). In contrast, these same ratios are predicted with relatively little bias when using the algorithm of Kim et al. (2000. Atmospheric Environment 34 (15), 2387-2397).

  11. Numerical simulation of a natural gradient tracer experiment for the natural attenuation study: flow and physical transport.

    PubMed

    Julian, H E; Boggs, J M; Zheng, C; Feehley, C E

    2001-01-01

    Results are presented for numerical simulations of ground water flow and physical transport associated with a natural gradient tracer experiment conducted within a heterogeneous alluvial aquifer of the Natural Attenuation Study (NATS) site near Columbus, Mississippi. A principal goal of NATS is to evaluate biogeochemical models that predict the rate and extent of natural biodegradation under field conditions. This paper describes the initial phase in the model evaluation process, i.e., calibration of flow and physical transport models that simulate conservative bromide tracer plume evolution during NATS. An initial large-scale flow model (LSM) is developed encompassing the experimental site and surrounding region. This model is subsequently scaled down in telescopic fashion to an intermediate-scale ground water flow model (ISM) covering the tracer-monitoring network, followed by a small-scale transport model (SSM) focused on the small region of hydrocarbon plume migration observed during NATS. The LSM uses inferred depositional features of the site in conjunction with hydraulic conductivity (K) data from aquifer tests and borehole flowmeter tests to establish large-scale K and flow field trends in and around the experimental site. The subsequent ISM incorporates specified flux boundary conditions and large-scale K trends obtained from the calibrated LSM, while preserving small-scale K structure based on some 4000 flowmeter data for solute transport modeling. The configuration of the ISM-predicted potentiometric surface approximates that of the observed surface within a root mean squared error of 0.15 m. The SSM is based on the dual-domain mass-transfer approach. Despite the well-recognized difficulties in modeling solute transport in extremely heterogeneous media as found at the NATS site, the dual-domain model adequately reproduced the observed bromide concentration distributions. Differences in observed and predicted bromide concentration distributions are

  12. A bedload tracer experiment in a high-elevation mountain basin (Strimm basin, Eastern Italian Alps)

    NASA Astrophysics Data System (ADS)

    Dell'Agnese, Andrea; Brardinoni, Francesco; Mao, Luca; Comiti, Francesco

    2014-05-01

    In many applications in engineering, ecology and river management, rates and timing of bed load transport in mountain rivers are of primary importance, yet of difficult prediction. Field measurements of bedload transport rates are rare, especially in high-mountain basins, and factors controlling the sediment fluxes have not been completely understood yet. Results offered by semi-empiric transport capacity equations proposed so far are heavily dependent on the experimental setup of the flumes in which they were developed originally. Direct methods for assessing bedload transport are time-consuming and practically challenging at high flows. Therefore, indirect surrogate methods for estimating bedload transport, such as the use of tracers, represent a good alternative. This study presents results on bed sediment mobility and travel distances obtained in the Strimm Creek, a high-elevation watershed in the Eastern Italian Alps (8.5 km2). From July 2011 to June 2012 a total of 431 PIT-tagged clasts (b-axis ranging from 22.6 mm to 229.3 mm) were deployed in two contrasting channel reaches, an upstream one located within a gentle hanging valley floor and a downstream on flowing along a steep and narrow valley step. Tagged clasts were surveyed from August 2011 to October 2013 by means of a portable antenna after each main flow event, and immediately before and after the spring freshet. Motion thresholds for the different grain sizes, particle travel distances and differences in sediment mobility existing between the two reaches were hence identified. Unit stream power rather than shear stress was used to analyse particle displacement due to the extremely rough geometry of the channel which makes the estimation of water depth at different sections subject to great uncertainties. In the lower channel reach, results showed how most of the PIT-tagged clasts movement happened during snowmelt periods, with travel distances often exceeding 500 meters, and displacements during the

  13. Testing compound-specific δ13C of amino acids in mussels as a new approach to determine the average 13C values of primary production in littoral ecosystems

    NASA Astrophysics Data System (ADS)

    Vokhshoori, N. L.; Larsen, T.; McCarthy, M.

    2012-12-01

    Compound-specific isotope analysis of amino acids (CSI-AA) is a technique used to decouple trophic enrichment patterns from source changes at the base of the food web. With this new emerging tool, it is possible to precisely determine both trophic position and δ15N or δ13C source values in higher feeding organisms. While most work to date has focused on nitrogen (N) isotopic values, early work has suggested that δ13C CSI-AA has great potential as a new tracer both to a record δ13C values of primary production (unaltered by trophic transfers), and also to "fingerprint" specific carbon source organisms. Since essential amino acids (EAA) cannot be made de novo in metazoans but must be obtained from diet, the δ13C value of the primary producer is preserved through the food web. Therefore, the δ13C values of EAAs act as a unique signature of different primary producers and can be used to fingerprint the dominant carbon (C) source driving primary production at the base of the food web. In littoral ecosystems, such as the California Upwelling System (CUS), the likely dominant C sources of suspended particulate organic matter (POM) pool are kelp, upwelling phytoplankton or estuarine phytoplankton. While bulk isotopes of C and N are used extensively to resolve relative consumer hierarchy or shifting diet in a food web, we found that the δ13C bulk values in mussels cannot distinguish exact source in littoral ecosystems. Here we show 15 sites within the CUS, between Cape Blanco, OR and La Jolla, CA where mussels were sampled and analyzed for both bulk δ13C and CSI-AA. We found no latitudinal trends, but rather average bulk δ13C values for the entire coastal record were highly consistent (-15.7 ± 0.9‰). The bulk record would suggest either nutrient provisioning from kelp or upwelled phytoplankton, but 13C-AA fingerprinting confines these two sources to upwelling. This suggests that mussels are recording integrated coastal phytoplankton values, with the enriched

  14. 13C Metabolic Flux Analysis Identifies an Unusual Route for Pyruvate Dissimilation in Mycobacteria which Requires Isocitrate Lyase and Carbon Dioxide Fixation

    PubMed Central

    Beste, Dany J. V.; Bonde, Bhushan; Hawkins, Nathaniel; Ward, Jane L.; Beale, Michael H.; Noack, Stephan; Nöh, Katharina; Kruger, Nicholas J.; Ratcliffe, R. George; McFadden, Johnjoe

    2011-01-01

    Mycobacterium tuberculosis requires the enzyme isocitrate lyase (ICL) for growth and virulence in vivo. The demonstration that M. tuberculosis also requires ICL for survival during nutrient starvation and has a role during steady state growth in a glycerol limited chemostat indicates a function for this enzyme which extends beyond fat metabolism. As isocitrate lyase is a potential drug target elucidating the role of this enzyme is of importance; however, the role of isocitrate lyase has never been investigated at the level of in vivo fluxes. Here we show that deletion of one of the two icl genes impairs the replication of Mycobacterium bovis BCG at slow growth rate in a carbon limited chemostat. In order to further understand the role of isocitrate lyase in the central metabolism of mycobacteria the effect of growth rate on the in vivo fluxes was studied for the first time using 13C-metabolic flux analysis (MFA). Tracer experiments were performed with steady state chemostat cultures of BCG or M. tuberculosis supplied with 13C labeled glycerol or sodium bicarbonate. Through measurements of the 13C isotopomer labeling patterns in protein-derived amino acids and enzymatic activity assays we have identified the activity of a novel pathway for pyruvate dissimilation. We named this the GAS pathway because it utilizes the Glyoxylate shunt and Anapleurotic reactions for oxidation of pyruvate, and Succinyl CoA synthetase for the generation of succinyl CoA combined with a very low flux through the succinate – oxaloacetate segment of the tricarboxylic acid cycle. We confirm that M. tuberculosis can fix carbon from CO2 into biomass. As the human host is abundant in CO2 this finding requires further investigation in vivo as CO2 fixation may provide a point of vulnerability that could be targeted with novel drugs. This study also provides a platform for further studies into the metabolism of M. tuberculosis using 13C-MFA. PMID:21814509

  15. Perfluorocarbon Tracer Experiments on a 2 km Scale in Manchester Showing Ingress of Pollutants into a Building

    NASA Astrophysics Data System (ADS)

    Matthews, James; Wright, Matthew; Bacak, Asan; Silva, Hugo; Priestley, Michael; Martin, Damien; Percival, Carl; Shallcross, Dudley

    2016-04-01

    Cyclic perfluorocarbons (PFCs) have been used to measure the passage of air in urban and rural settings as they are chemically inert, non-toxic and have low background concentrations. The use of pre-concentrators and chemical ionisation gas chromatography enables concentrations of a few parts per quadrillion (ppq) to be measured in bag samples. Three PFC tracers were used in Manchester, UK in the summer of 2015 to map airflow in the city and ingress into buildings: perfluomethylcyclohexane (PMCH), perfluoro-2-4-dimethylcyclohexane (mPDMCH) and perfluoro-2-methyl-3-ethylpentene (PMEP). A known quantity of each PFC was released for 15 minutes from steel canisters using pre-prepared PFC mixtures. Release points were chosen to be upwind of the central sampling location (Simon Building, University of Manchester) and varied in distance up to 2.2 km. Six releases using one or three tracers in different configurations and under different conditions were undertaken in the summer. Three further experiments were conducted in the Autumn, to more closely investigate the rate of ingress and decay of tracer indoors. In each experiment, 10 litre samples were made over 30 minutes into Tedlar bags, starting at the same time the as PFC release. Samples were taken in 11 locations chosen from 15 identified areas including three in public parks, three outside within the University of Manchester area, seven inside and five outside of the Simon building and two outside a building nearby. For building measurements, receptors were placed inside the buildings on different floors; outside measurements were achieved through a sample line out of the window. Three of the sample positions inside the Simon building were paired with samplers outside to allow indoor-outdoor comparisons. PFC concentrations varied depending on location and height. The highest measured concentrations occurred when the tracer was released at sunrise; up to 330 ppq above background (11 ppq) of PMCH was measured at the 6

  16. Minnesota 1973 atmospheric boundary layer experiment: Micrometeorological and tracer data archive. Set 1 (revision 2) documentation report

    NASA Astrophysics Data System (ADS)

    Woodruff, R. K.; Droppo, J. G.; Glantz, C. S.

    1987-03-01

    An archive for micrometeorological and tracer dispersion data was developed by Battelle, Pacific Northwest Labs. for the U.S. Environmental Protection Agency. The archive was designed to make the results of extensive field tests readily accessible to EPA for model testing, development, and verification efforts. Documentation was provided for one of the archived data sets, The Minnesota 1973 Boundary Layer Experiment. The aim of the effort was to archive invaluable data sets in a timely fashion before the necessary supporting information about the data becomes lost forever. The entries are as follows: data set fact summary, a narrative description of experimental and data, special information, references, a description of archive data files, contacts (names, addresses, and phone numbers) and standard experiment summary table. Revision 2 includes previously unpublished rawinsonde profile data.

  17. A natural abundance stable isotope tracer experiment to define SO2 oxidation pathways and their fractionation during heterogeneous oxidation

    NASA Astrophysics Data System (ADS)

    Amiri, N.; Norman, A. L.

    2015-12-01

    Sulfate aerosols have crucial direct and indirect effects on climate from radiative cooling to modifying clouds by formation of cloud condensation nuclei. Secondary sulfate aerosols are formed by oxidation of SO2 and subsequent nucleation and growth and the characteristics of primary aerosol sulfate can be modified by oxidation of SO2. There are several known oxidation pathways for SO2; gaseous phase OH oxidation and aqueous phase H2O2, O3 and transition metal oxides oxidation. The SO2 oxidation pathway affects the characteristics of the aerosols formed. Stable isotope techniques are useful in determining the oxidation pathway of SO2 due to unique fractionation patterns (Harris et al., 2012). However, there are still gaps in our understanding of the oxidation pathways and fractionations affecting SO2 and secondary sulfate. A tracer experiment to investigate the oxidation of SO2 and fractionation using size segregated aerosols in the presence of different compounds is described. Two high volume samplers situated to measure background sulfate upwind, and the results of a tracer experiment, downwind, is described. After sufficient size segregated aerosol sulfate has been collected, a source of SO2 with known isotopic composition is introduced to the second high volume sampler. Changes in the isotopic composition for size segregated aerosol sulfate in comparison to the first high volume sampler are investigated. The amount of fractionation during heterogeneous oxidation of SO2 on pre-existing aerosols is calculated using the concentrations and known isotopic composition and compared to data from laboratory and field experiments. The experiment is performed downwind of sources of organic compounds such as pine forests, and characterized using co-located canister samples, to determine the effects of SO2 oxidation on secondary aerosol sulfate.

  18. 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...

  19. A three-dimensional model of atmospheric CO2 transport based on observed winds: 2. Model description and simulated tracer experiments

    NASA Astrophysics Data System (ADS)

    Heimann, Martin; Keeling, Charles D.

    The three-dimensional atmospheric transport model developed at the Goddard Institute of Space Sciences (GISS) has been modified in its coarse-grid version (7.83° × 10.00° horizontal resolution, 9 layers in vertical direction) by replacing the original model-generated wind fields with observed winds of the Global Weather Experiment, covering the period December 1978 through November 1979. The parameterization of subgridscale vertical convection was retained from the earlier model version, except that all intensities were reduced by 50 percent. To simulate atmospheric CO2 and its 13C/12C ratio, sources and sinks of carbon at the earth's surface were prescribed. The net primary productivity of the terrestrial biosphere was computed from vegetation index (NDVI) data representing the greenness of the land as recorded by the AVHRR instrument flown on satellites of the U.S. National Oceanic and Atmospheric Administration. The productivity for a given greenness was assumed to be proportional to the amount of photosynthetically active radiation reaching the plant canopy as computed from estimates of solar insolation under clear sky conditions with an allowance for attenuation caused by the presence of clouds. Estimates of cloud cover were based on satellite data of daily albedo. The respiration of terrestrial plant detritus and soils was assumed to be dependent on temperature but was globally adjusted to achieve an optimal fit of the model prediction of the seasonal cycle of atmospheric CO2 to that observed at four northern hemisphere stations. The oceanic exchange of CO2 was prescribed from rough estimates of the CO2 partial pressure of sea water, assuming a constant air-sea exchange rate. The mean annual partial pressure field was afterwards adjusted to achieve an optimal fit with respect to broad geographic features of the observed mean annual atmospheric CO2 field. The seasonal variation in partial pressure was assumed to depend on sea-surface temperature, but the

  20. SIMS measurements of intrashell δ13C in the cultured planktic foraminifer Orbulina universa

    NASA Astrophysics Data System (ADS)

    Vetter, Lael; Kozdon, Reinhard; Valley, John W.; Mora, Claudia I.; Spero, Howard J.

    2014-08-01

    In this study, we present experimental results from the planktic foraminifer Orbulina universa, cultured in the laboratory. We demonstrate that the δ13C of shell calcite precipitated in 13C-labeled seawater for 24 h can be resolved and accurately measured using Secondary Ion Mass Spectrometry (SIMS). Specimens maintained at 20 °C were transferred from ambient seawater (δ13CDIC = +1.3‰) into seawater with δ13CDIC = +51.5‰ and enriched [Ba2+] for 24 h. Specimens were then transferred into ambient seawater with elevated [87Sr] for 6-9 h of calcification, followed by a transfer back into unlabeled ambient seawater until gametogenesis. This technique produced O. universa shells with calcite layers of distinct geochemical signatures. We quantify the spatial positions of trace element labels in the shells using laser ablation ICP-MS depth profiling. Using fragments from the same shells, we quantify intrashell δ13Ccalcite using SIMS with a 6 or 8 μm spot (2 SD range ±0.5‰ to 1.7‰). Measured δ13Ccalcite values in O. universa shell layers precipitated in ambient seawater are within 2‰ of predicted δ13Ccalcite values. In 13C-labeled bands of calcite, 6 μm SIMS spot measurements are within 2‰ of predicted δ13Ccalcite values, whereas 8 μm SIMS spots yield intermediate, mixed values. The spatial agreement between trace element and carbon isotope data suggests that 13C and cation tracers are synchronously incorporated into shell calcite. These results demonstrate the ability of SIMS δ13C measurements to resolve ∼10 μm features in foraminifer shell calcite using a 6 μm spot, and highlight the potential of this technique for addressing questions about ecology, biomineralization, and paleoceanography.

  1. Directly detected 55Mn MRI: Application to phantoms for human hyperpolarized 13C MRI development

    PubMed Central

    von Morze, Cornelius; Carvajal, Lucas; Reed, Galen D.; Swisher, Christine Leon; Tropp, James; Vigneron, Daniel B.

    2014-01-01

    In this work we demonstrate for the first time directly detected manganese-55 (55Mn) MRI using a clinical 3T MRI scanner designed for human hyperpolarized 13C clinical studies with no additional hardware modifications. Due to the similar frequency of the 55Mn and 13C resonances, the use of aqueous permanganate for large, signal-dense, and cost-effective “13C” MRI phantoms was investigated, addressing the clear need for new phantoms for these studies. Due to 100% natural abundance, higher intrinsic sensitivity, and favorable relaxation properties, 55Mn MRI of aqueous permanganate demonstrates dramatically increased sensitivity over typical 13C phantom MRI, at greatly reduced cost as compared with large 13C-enriched phantoms. A large sensitivity advantage (22-fold) was demonstrated. A cylindrical phantom (d= 8 cm) containing concentrated aqueous sodium permanganate (2.7M) was scanned rapidly by 55Mn MRI in a human head coil tuned for 13C, using a balanced SSFP acquisition. The requisite penetration of RF magnetic fields into concentrated permanganate was investigated by experiments and high frequency electromagnetic simulations, and found to be sufficient for 55Mn MRI with reasonably sized phantoms. A sub-second slice-selective acquisition yielded mean image SNR of ~60 at 0.5cm3 spatial resolution, distributed with minimum central signal ~40% of the maximum edge signal. We anticipate that permanganate phantoms will be very useful for testing HP 13C coils and methods designed for human studies. PMID:25179135

  2. Kinetic isotope effects significantly influence intracellular metabolite 13C labeling patterns and flux determination

    PubMed Central

    Wasylenko, Thomas M.; Stephanopoulos, Gregory

    2014-01-01

    Rigorous mathematical modeling of carbon-labeling experiments allows estimation of fluxes through the pathways of central carbon metabolism, yielding powerful information for basic scientific studies as well as for a wide range of applications. However, the mathematical models that have been developed for flux determination from 13C labeling data have commonly neglected the influence of kinetic isotope effects on the distribution of 13C label in intracellular metabolites, as these effects have often been assumed to be inconsequential. We have used measurements of the 13C isotope effects on the pyruvate dehydrogenase enzyme from the literature to model isotopic fractionation at the pyruvate node and quantify the modeling errors expected to result from the assumption that isotope effects are negligible. We show that under some conditions kinetic isotope effects have a significant impact on the 13C labeling patterns of intracellular metabolites, and the errors associated with neglecting isotope effects in 13C-metabolic flux analysis models can be comparable in size to measurement errors associated with GC–MS. Thus, kinetic isotope effects must be considered in any rigorous assessment of errors in 13C labeling data, goodness-of-fit between model and data, confidence intervals of estimated metabolic fluxes, and statistical significance of differences between estimated metabolic flux distributions. PMID:23828762

  3. Comparison of Hydraulic Methods and Tracer Experiments as Applied to the Development of Conceptual Models for Discrete Fracture Networks

    NASA Astrophysics Data System (ADS)

    Novakowski, K. S.

    2015-12-01

    The development of conceptual models for solute migration in discrete fracture networks has typically been based on a combination of core logs, borehole geophysics, and some form of single-well hydraulic test using discrete zones. More rarely, interwell hydraulic tests and interwell tracer experiments are utilised to directly explore potential transport pathways. The latter methods are less widely employed simply due to potentially significant increases in the cost and effort in site characterization. To date however there is a paucity of literature comparing the efficacy of the standard procedure with what should be more definitive identification of transport pathways using interwell methods. In the present study, a detailed comparison is conducted by developing conceptual models from three separate data sets, the first based on core logs, geology and single-well hydraulic tests, the second based on a large suite of pulse interference tests, and the third based on a series of radially-divergent and injection-withdrawal tracer experiments. The study was conducted in an array of five HQ-sized wells, 28-32 m in depth and arranged in a five star pattern, 10 m on a side. The wells penetrate the contact between a Cambrian-aged limestone, and underlying Precambrian gneiss. The core was logged for potentially open fractures using a ranking system, and 87 contiguous hydraulic tests were conducted using a 0.85-m packer spacing. A total of 57 pulse interference tests were conducted using two wells as injection points, and 11 tracer experiments were conducted using either sample collection or in-situ detection via a submersible fluorometer. The results showed very distinct conceptual models depending on the data set, with the model based on the single-well testing significantly over-predicting the number and connection of solute transport pathways. The results of the pulse interference tests also over predict the transport pathways, but to a lesser degree. Quantification of

  4. 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.

  5. Regioselective Syntheses of [13C]4-Labelled Sodium 1-Carboxy-2-(2-ethylhexyloxycarbonyl)ethanesulfonate and Sodium 2-Carboxy-1-(2-ethylhexyloxycarbonyl)ethanesulfonate from [13C]4-Maleic Anhydride

    PubMed Central

    Barsamian, Adam L.; Perkins, Matt J.; Field, Jennifer A.; Blakemore, Paul R.

    2014-01-01

    The entitled monohydrolysis products, also known as α- and β-ethylhexyl sulfosuccinate ('EHSS'), of the surfactant diisooctyl sulfosuccinate ('DOSS') were synthesized in stable isotope labelled form from [13C]4-maleic anhydride. Sodium [13C]4-1-carboxy-2-(2-ethylhexyloxycarbonyl)ethanesulfonate (α-EHSS) was prepared by the method of Larpent by reaction of 2-ethylhexan-1-ol with [13C]4-maleic anhydride followed by regioselective conjugate addition of sodium bisulfite to the resulting monoester (38% overall yield). The regiochemical outcome of bisulfite addition was confirmed by a combination of 13C/13C (INADEQUATE) and 1H/13C (HMBC) NMR spectral correlation experiments. Sodium [13C]4-2-carboxy-1-(2-ethylhexyloxycarbonyl)ethanesulfonate (β-EHSS) was prepared in four steps by reaction of 4-methoxybenzyl alcohol (PMBOH) with [13C]4-maleic anhydride, regioselective sodium bisulfite addition, DCC mediated esterification with 2-ethylhexan-1-ol, and PMB ester deprotection with trifluoroacetic acid (13% overall yield). The regiochemical outcome of the second synthesis was confirmed by a combination of 1JCC scalar coupling constant analysis and 1H/13C (HMBC) NMR spectral correlation. The materials prepared are required as internal standards for the LC-MS/MS trace analysis of the degradation products of DOSS, the anionic surfactant found in Corexit, the oil dispersant used during emergency response efforts connected to the Deepwater Horizon oil spill of April 2010. PMID:24700711

  6. 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.

  7. Determination of the Orientation and Dynamics of Ergosterol in Model Membranes Using Uniform 13C Labeling and Dynamically Averaged 13C Chemical Shift Anisotropies as Experimental Restraints

    PubMed Central

    Soubias, O.; Jolibois, F.; Massou, S.; Milon, A.; Réat, V.

    2005-01-01

    A new strategy was established to determine the average orientation and dynamics of ergosterol in dimyristoylphosphatidylcholine model membranes. It is based on the analysis of chemical shift anisotropies (CSAs) averaged by the molecular dynamics. Static 13C CSA tensors were computed by quantum chemistry, using the gauge-including atomic-orbital approach within Hartree-Fock theory. Uniformly 13C-labeled ergosterol was purified from Pichia pastoris cells grown on labeled methanol. After reconstitution into dimyristoylphosphatidylcholine lipids, the complete 1H and 13C assignment of ergosterol's resonances was performed using a combination of magic-angle spinning two-dimensional experiments. Dynamically averaged CSAs were determined by standard side-band intensity analysis for isolated 13C resonances (C3 and ethylenic carbons) and by off-magic-angle spinning experiments for other carbons. A set of 18 constraints was thus obtained, from which the sterol's molecular order parameter and average orientation could be precisely defined. The validity of using computed CSAs in this strategy was verified on cholesterol model systems. This new method allowed us to quantify ergosterol's dynamics at three molar ratios: 16 mol % (Ld phase), 30 mol % (Lo phase), and 23 mol % (mixed phases). Contrary to cholesterol, ergosterol's molecular diffusion axis makes an important angle (14°) with the inertial axis of the rigid four-ring system. PMID:15923221

  8. Calcium-ammonium exchange experiments on clay minerals using a (45)Ca tracer technique in marine pore water.

    PubMed

    Ockert, Charlotte; Wehrmann, Laura M; Kaufhold, Stephan; Ferdelman, Tim G; Teichert, Barbara M A; Gussone, Nikolaus

    2014-01-01

    Understanding cation exchange processes is important for evaluating early diagenetic and synsedimentary processes taking place in marine sediments. To quantify calcium (Ca) exchange and Ca-ammonium exchange in a seawater environment, we performed experiments with a radioactive (45)Ca tracer on clay mineral standards (Fithian illite, montmorillonite and kaolinite) and marine sediments from the North Atlantic Integrated Ocean Drilling Program Site U1306A in artificial seawater (ASW). The results show that equilibrium during the initial attachment of Ca as well as the exchange of Ca by [Formula: see text] is attained in less than 2 min. On average 8-20% of the exchangeable sites of the clay minerals were occupied by Ca in a seawater medium. The conditional selectivity coefficient, describing the [Formula: see text] exchange in ASW is mineral specific and it was determined to be 0.07 for montmorillonite, 0.05 for a natural marine sediment and 0.013 for Fithian illite.

  9. Study of Urban environmental quality through Isotopes δ13C

    NASA Astrophysics Data System (ADS)

    González-Sosa, E.; Mastachi-Loza, C.; Becerril-Piña, R.; Ramos-Salinas, N. M.

    2012-04-01

    Usually, trees with similar pH values on their bark develop epiphytes of similar species, the acidity to be a factor for growth. The aim of the study was evaluate the air quality through isotope δ13C in order to define the levels of environmental quality in the city of Queretaro, Mexico. In this work were collected at least 4 epiphytes positioned in trees of the species Prosopis Laevigata at 25 sites of Queretaro City. The samples were analyzed for trace elements with an inductively coupled plasma atomic emission spectroscopy (ICP). The collecting took place during dry period, in May and early rain June 2011 period, and on four sectors to identify the spatial distribution of pollution, using isotopic analysis of concentration of δ 13C. According with the results there are significant differences among the species in each of the sampled areas. The 5 February Avenue presented greater diversity and richness of δ13C, followed by those who were surveyed in the proximity of the UAQ and finally in the middle-east area. An average value of δ13C-17.92%, followed by those surveyed in the vicinity of the UAQ that correspond to sector I and II with an concentration of δ13C-17.55% and δ13C-17.22%, and finally the samples collected in trees scattered in the East-Sector II and IV with a value of δ13C-17.02% and δ13C-15.62%, respectively. Also were observed differences between the dry and wet period. It is likely that these results of δ 13C in moist period reflect the drag of the isotopes due to rain events that could mark a trend in the dilution of this element, however there is a trend in terms of abundance and composition of finding more impact in those species sampled in dry period, in May and early June 2011.

  10. Central metabolic responses to the overproduction of fatty acids in Escherichia coli based on 13C-metabolic flux analysis.

    PubMed

    He, Lian; Xiao, Yi; Gebreselassie, Nikodimos; Zhang, Fuzhong; Antoniewiez, Maciek R; Tang, Yinjie J; Peng, Lifeng

    2014-03-01

    We engineered a fatty acid overproducing Escherichia coli strain through overexpressing tesA (“pull”) and fadR (“push”) and knocking out fadE (“block”). This “pull-push-block” strategy yielded 0.17 g of fatty acids (C12–C18) per gram of glucose (equivalent to 48% of the maximum theoretical yield) in batch cultures during the exponential growth phase under aerobic conditions. Metabolic fluxes were determined for the engineered E. coli and its control strain using tracer ([1,2-13C]glucose) experiments and 13C-metabolic flux analysis. Cofactor (NADPH) and energy (ATP) balances were also investigated for both strains based on estimated fluxes. Compared to the control strain, fatty acid overproduction led to significant metabolic responses in the central metabolism: (1) Acetic acid secretion flux decreased 10-fold; (2) Pentose phosphate pathway and Entner–Doudoroff pathway fluxes increased 1.5- and 2.0-fold, respectively; (3) Biomass synthesis flux was reduced 1.9-fold; (4) Anaplerotic phosphoenolpyruvate carboxylation flux decreased 1.7-fold; (5) Transhydrogenation flux converting NADH to NADPH increased by 1.7-fold. Real-time quantitative RT-PCR analysis revealed the engineered strain increased the transcription levels of pntA (encoding the membrane-bound transhydrogenase) by 2.1-fold and udhA (encoding the soluble transhydrogenase) by 1.4-fold, which is in agreement with the increased transhydrogenation flux. Cofactor and energy balances analyses showed that the fatty acid overproducing E. coli consumed significantly higher cellular maintenance energy than the control strain. We discussed the strategies to future strain development and process improvements for fatty acid production in E. coli.

  11. Direct uptake of organic carbon by grass roots and allocation in leaves and phytoliths: 13C labeling evidence

    NASA Astrophysics Data System (ADS)

    Alexandre, A.; Balesdent, J.; Cazevieille, P.; Chevassus-Rosset, C.; Signoret, P.; Mazur, J.-C.; Harutyunyan, A.; Doelsch, E.; Basile-Doelsch, I.; Miche, H.; Santos, G. M.

    2015-12-01

    In the rhizosphere, the uptake of low molecular weight carbon (C) and nitrogen (N) by plant roots has been well documented. While organic N uptake relatively to total uptake is important, organic C uptake is supposed to be low relatively to the plant's C budget. Recently, radiocarbon analyses demonstrated that a fraction of C from the soil was occluded in amorphous silica micrometric particles that precipitate in plant cells (phytoliths). Here, we investigated whether and in which extent organic C absorbed by grass roots, under the form of either intact amino acids (AAs) or microbial metabolites, can feed the organic C occluded in phytoliths. For this purpose we added 13C- and 15N-labeled AAs to the silicon-rich hydroponic solution of the grass Festuca arundinacea. The experiment was designed to prevent C leakage from the labeled nutritive solution to the chamber atmosphere. After 14 days of growth, the 13C and 15N enrichments (13C-excess and 15N-excess) in the roots, stems and leaves, and phytoliths, as well as the 13C-excess in AAs extracted from roots and stems and leaves, were quantified relatively to a control experiment in which no labelled AAs were added. The net uptake of 13C derived from the labeled AAs supplied to the nutritive solution (AA-13C) by Festuca arundinacea represented 4.5 % of the total AA-13C supply. AA-13C fixed in the plant represented only 0.13 % of total C. However, the experimental conditions may have underestimated the extent of the process under natural and field conditions. Previous studies showed that 15N and 13C can be absorbed by the roots in several organic and inorganic forms. In the present experiment, the fact that phenylalanine and methionine, that were supplied in high amount to the nutritive solution, were more 13C-enriched than other AAs in the roots and stems and leaves strongly suggested that part of AA-13C was absorbed and translocated in its original AA form. The concentration of AA-13C represented only 0.15 % of the

  12. SUMOFLUX: A Generalized Method for Targeted 13C Metabolic Flux Ratio Analysis

    PubMed Central

    Kogadeeva, Maria

    2016-01-01

    Metabolic fluxes are a cornerstone of cellular physiology that emerge from a complex interplay of enzymes, carriers, and nutrients. The experimental assessment of in vivo intracellular fluxes using stable isotopic tracers is essential if we are to understand metabolic function and regulation. Flux estimation based on 13C or 2H labeling relies on complex simulation and iterative fitting; processes that necessitate a level of expertise that ordinarily preclude the non-expert user. To overcome this, we have developed SUMOFLUX, a methodology that is broadly applicable to the targeted analysis of 13C-metabolic fluxes. By combining surrogate modeling and machine learning, we trained a predictor to specialize in estimating flux ratios from measurable 13C-data. SUMOFLUX targets specific flux features individually, which makes it fast, user-friendly, applicable to experimental design and robust in terms of experimental noise and exchange flux magnitude. Collectively, we predict that SUMOFLUX's properties realistically pave the way to high-throughput flux analyses. PMID:27626798

  13. Optimization of 13C dynamic nuclear polarization: isotopic labeling of free radicals

    NASA Astrophysics Data System (ADS)

    Niedbalski, Peter; Parish, Christopher; Kiswandi, Andhika; Lumata, Lloyd

    Dynamic nuclear polarization (DNP) is a physics technique that amplifies the nuclear magnetic resonance (NMR) signals by transferring the high polarization of the electrons to the nuclear spins. Thus, the choice of free radical is crucial in DNP as it can directly affect the NMR signal enhancement levels, typically on the order of several thousand-fold in the liquid-state. In this study, we have investigated the efficiency of four variants of the well-known 4-oxo-TEMPO radical (normal 4-oxo-TEMPO plus its 15N-enriched and/or perdeuterated variants) for use in DNP of an important metabolic tracer [1-13C]acetate. Though the variants have significant differences in electron paramagnetic resonance (EPR) spectra, we have found that changing the composition of the TEMPO radical through deuteration or 15N doping yields no significant difference in 13C DNP efficiency at 3.35 T and 1.2 K. On the other hand, deuteration of the solvent causes a significant increase of 13C polarization that is consistent over all the 4-oxo-TEMPO variants. These findings are consistent with the thermal mixing model of DNP. This work is supported by US Dept of Defense Award No. W81XWH-14-1-0048 and the Robert A. Welch Foundation Grant No. AT-1877.

  14. SUMOFLUX: A Generalized Method for Targeted 13C Metabolic Flux Ratio Analysis.

    PubMed

    Kogadeeva, Maria; Zamboni, Nicola

    2016-09-01

    Metabolic fluxes are a cornerstone of cellular physiology that emerge from a complex interplay of enzymes, carriers, and nutrients. The experimental assessment of in vivo intracellular fluxes using stable isotopic tracers is essential if we are to understand metabolic function and regulation. Flux estimation based on 13C or 2H labeling relies on complex simulation and iterative fitting; processes that necessitate a level of expertise that ordinarily preclude the non-expert user. To overcome this, we have developed SUMOFLUX, a methodology that is broadly applicable to the targeted analysis of 13C-metabolic fluxes. By combining surrogate modeling and machine learning, we trained a predictor to specialize in estimating flux ratios from measurable 13C-data. SUMOFLUX targets specific flux features individually, which makes it fast, user-friendly, applicable to experimental design and robust in terms of experimental noise and exchange flux magnitude. Collectively, we predict that SUMOFLUX's properties realistically pave the way to high-throughput flux analyses. PMID:27626798

  15. Elucidation of intrinsic biosynthesis yields using 13C-based metabolism analysis

    PubMed Central

    2014-01-01

    This paper discusses the use of 13C-based metabolism analysis for the assessment of intrinsic product yields — the actual carbon contribution from a single carbon substrate to the final product via a specific biosynthesis route — in the following four cases. First, undefined nutrients (such as yeast extract) in fermentation may contribute significantly to product synthesis, which can be quantified through an isotopic dilution method. Second, product and biomass synthesis may be dependent on the co-metabolism of multiple-carbon sources. 13C labeling experiments can track the fate of each carbon substrate in the cell metabolism and identify which substrate plays a main role in product synthesis. Third, 13C labeling can validate and quantify the contribution of the engineered pathway (versus the native pathway) to the product synthesis. Fourth, the loss of catabolic energy due to cell maintenance (energy used for functions other than production of new cell components) and low P/O ratio (Phosphate/Oxygen Ratio) significantly reduces product yields. Therefore, 13C-metabolic flux analysis is needed to assess the influence of suboptimal energy metabolism on microbial productivity, and determine how ATP/NAD(P)H are partitioned among various cellular functions. Since product yield is a major determining factor in the commercialization of a microbial cell factory, we foresee that 13C-isotopic labeling experiments, even without performing extensive flux calculations, can play a valuable role in the development and verification of microbial cell factories. PMID:24642094

  16. Elucidation of intrinsic biosynthesis yields using 13C-based metabolism analysis.

    PubMed

    Varman, Arul M; He, Lian; You, Le; Hollinshead, Whitney; Tang, Yinjie J

    2014-03-19

    This paper discusses the use of 13C-based metabolism analysis for the assessment of intrinsic product yields - the actual carbon contribution from a single carbon substrate to the final product via a specific biosynthesis route - in the following four cases. First, undefined nutrients (such as yeast extract) in fermentation may contribute significantly to product synthesis, which can be quantified through an isotopic dilution method. Second, product and biomass synthesis may be dependent on the co-metabolism of multiple-carbon sources. 13C labeling experiments can track the fate of each carbon substrate in the cell metabolism and identify which substrate plays a main role in product synthesis. Third, 13C labeling can validate and quantify the contribution of the engineered pathway (versus the native pathway) to the product synthesis. Fourth, the loss of catabolic energy due to cell maintenance (energy used for functions other than production of new cell components) and low P/O ratio (Phosphate/Oxygen Ratio) significantly reduces product yields. Therefore, 13C-metabolic flux analysis is needed to assess the influence of suboptimal energy metabolism on microbial productivity, and determine how ATP/NAD(P)H are partitioned among various cellular functions. Since product yield is a major determining factor in the commercialization of a microbial cell factory, we foresee that 13C-isotopic labeling experiments, even without performing extensive flux calculations, can play a valuable role in the development and verification of microbial cell factories.

  17. Natural-abundance sup 13 C NMR study of glycogen repletion in human liver and muscle

    SciTech Connect

    Jue, T.; Rothman, D.L.; Tavitian, B.A.; Shulman, R.G. )

    1989-03-01

    Optimizing the surface-coil design and spectral-acquisition parameters has led to the observation of the {sup 13}C NMR natural abundance glycogen signal in man at 2.1 T. Both the human muscle and hepatic glycogen signals can be detected definitively with a time resolution of {approx}13 min. A {sup 1}H/{sup 13}C concentric surface coil was used. The {sup 1}H outer coil was 11 cm in diameter; the {sup 13}C inner coil was 8 cm in diameter. The coils were tuned to 89.3 MHz and 22.4 MHz, respectively. The {sup 1}H coil was used for optimizing field homogeneity (shimming) the magnet and for single-frequency decoupling of the C{sub 1} glycogen signal. Total power deposition from both the transmitter pulse and the continuous wave decoupling did not exceed the Food and Drug Administration guideline of 8 W/kg of tissue. Experiments were done for which healthy subjects returned to the magnets at different times for {sup 13}C NMR measurement. The spectral difference between experiments was within the noise in the C{sub 1} glycogen region. Because of the spectral reproducibility and the signal sensitivity, hepatic glycogen repletion can be followed. Four hours postprandial, hepatic glycogen increases by 3.8 times from the basal fasted state. The hepatic glycogen data correspond directly to previous biopsy results and support the use of {sup 13}C NMR as a noninvasive probe of human metabolism.

  18. A Method to Constrain Genome-Scale Models with 13C Labeling Data

    PubMed Central

    García Martín, Héctor; Kumar, Vinay Satish; Weaver, Daniel; Ghosh, Amit; Chubukov, Victor; Mukhopadhyay, Aindrila; Arkin, Adam; Keasling, Jay D.

    2015-01-01

    Current limitations in quantitatively predicting biological behavior hinder our efforts to engineer biological systems to produce biofuels and other desired chemicals. Here, we present a new method for calculating metabolic fluxes, key targets in metabolic engineering, that incorporates data from 13C labeling experiments and genome-scale models. The data from 13C labeling experiments provide strong flux constraints that eliminate the need to assume an evolutionary optimization principle such as the growth rate optimization assumption used in Flux Balance Analysis (FBA). This effective constraining is achieved by making the simple but biologically relevant assumption that flux flows from core to peripheral metabolism and does not flow back. The new method is significantly more robust than FBA with respect to errors in genome-scale model reconstruction. Furthermore, it can provide a comprehensive picture of metabolite balancing and predictions for unmeasured extracellular fluxes as constrained by 13C labeling data. A comparison shows that the results of this new method are similar to those found through 13C Metabolic Flux Analysis (13C MFA) for central carbon metabolism but, additionally, it provides flux estimates for peripheral metabolism. The extra validation gained by matching 48 relative labeling measurements is used to identify where and why several existing COnstraint Based Reconstruction and Analysis (COBRA) flux prediction algorithms fail. We demonstrate how to use this knowledge to refine these methods and improve their predictive capabilities. This method provides a reliable base upon which to improve the design of biological systems. PMID:26379153

  19. Salt tracer experiments in constructed wetland ponds with emergent vegetation: laboratory study on the formation of density layers and its influence on breakthrough curve analysis.

    PubMed

    Schmid, Bernhard H; Hengl, Michael A; Stephan, Ursula

    2004-04-01

    Constructed wetlands are a rapidly expanding and intensively studied wastewater treatment system. One of the main types in use is the free water surface (FWS) wetland or wetland pond. In studies on these ponds, salt tracer experiments are a convenient tool to determine travel time distributions, which are, in turn, related to hydraulic and sedimentation (trapping) as well as nutrient removal efficiencies. Typically, flows encountered in constructed wetland ponds are characterized by low Reynolds numbers, at times even within the laminar flow regime. In such conditions the injection of salt may cause strong density effects, thereby threatening the usefulness of the recorded breakthrough curves. The processes and mechanisms governing the formation of density stratification due to salt tracer injections into wetland ponds with emergent vegetation were studied in the laboratory. The results reported are expected to be useful in the planning of future field tracer experiments.

  20. Detection of inflammatory cell function using 13C magnetic resonance spectroscopy of hyperpolarized [6-13C]-arginine

    PubMed Central

    Najac, Chloé; Chaumeil, Myriam M.; Kohanbash, Gary; Guglielmetti, Caroline; Gordon, Jeremy W.; Okada, Hideho; Ronen, Sabrina M.

    2016-01-01

    Myeloid-derived suppressor cells (MDSCs) are highly prevalent inflammatory cells that play a key role in tumor development and are considered therapeutic targets. MDSCs promote tumor growth by blocking T-cell-mediated anti-tumoral immune response through depletion of arginine that is essential for T-cell proliferation. To deplete arginine, MDSCs express high levels of arginase, which catalyzes the breakdown of arginine into urea and ornithine. Here, we developed a new hyperpolarized 13C probe, [6-13C]-arginine, to image arginase activity. We show that [6-13C]-arginine can be hyperpolarized, and hyperpolarized [13C]-urea production from [6-13C]-arginine is linearly correlated with arginase concentration in vitro. Furthermore we show that we can detect a statistically significant increase in hyperpolarized [13C]-urea production in MDSCs when compared to control bone marrow cells. This increase was associated with an increase in intracellular arginase concentration detected using a spectrophotometric assay. Hyperpolarized [6-13C]-arginine could therefore serve to image tumoral MDSC function and more broadly M2-like macrophages. PMID:27507680

  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. Structure of B sub 13 C sub 2

    SciTech Connect

    Bylander, D.M.; Kleinman, L. )

    1991-01-15

    By comparing calculated lattice constants with x-ray data as well as by comparison of calculated free energies, we find that the correct structure of B{sub 13}C{sub 2} is B{sub 12}(CBC) rather than B{sub 11}C(BBC), as had been suggested. We also show that B{sub 12}C{sub 3} is stable against 13B{sub 12}C{sub 3}{r arrow}12B{sub 13}C{sub 2}+15C as is B{sub 13}C{sub 2} against 3B{sub 13}C{sub 2}{r arrow}2B{sub 12}C{sub 3}+15B.

  3. Anomalous 13C enrichment in modern marine organic carbon

    USGS Publications Warehouse

    Arthur, M.A.; Dean, W.E.; Claypool, G.E.

    1985-01-01

    Marine organic carbon is heavier isotopically (13C enriched) than most land-plant or terrestrial organic C1. Accordingly, ??13C values of organic C in modern marine sediments are routinely interpreted in terms of the relative proportions of marine and terrestrial sources of the preserved organic matter2,3. When independent geochemical techniques are used to evaluate the source of organic matter in Cretaceous or older rocks, those rocks containing mostly marine organic C are found typically to have lighter (more-negative) ??13C values than rocks containing mostly terrestrial organic C. Here we conclude that marine photosynthesis in mid-Cretaceous and earlier oceans generally resulted in a greater fractionation of C isotopes and produced organic C having lighter ??13C values. Modern marine photosynthesis may be occurring under unusual geological conditions (higher oceanic primary production rates, lower PCO2) that limit dissolved CO2 availability and minimize carbon isotope fractionation4. ?? 1985 Nature Publishing Group.

  4. Dihydroflavanonols from Cedrus deodara, A (13)C NMR study.

    PubMed

    Agrawal, P K; Agarwal, S K; Rastogi, R P; Osterdahal, B G

    1981-09-01

    High resolution (13)C NMR study of taxifolin, cedeodarin, cedrin and their methyl ethers allowed unambiguous placement of the Me in 5,7-dihydroxyflavanonol nucleus, besides providing other valuable information on the substitution pattern in the molecule.

  5. An overview of methods using 13C for improved compound identification in metabolomics and natural products

    PubMed Central

    Clendinen, Chaevien S.; Stupp, Gregory S.; Ajredini, Ramadan; Lee-McMullen, Brittany; Beecher, Chris; Edison, Arthur S.

    2015-01-01

    Compound identification is a major bottleneck in metabolomics studies. In nuclear magnetic resonance (NMR) investigations, resonance overlap often hinders unambiguous database matching or de novo compound identification. In liquid chromatography-mass spectrometry (LC-MS), discriminating between biological signals and background artifacts and reliable determination of molecular formulae are not always straightforward. We have designed and implemented several NMR and LC-MS approaches that utilize 13C, either enriched or at natural abundance, in metabolomics applications. For LC-MS applications, we describe a technique called isotopic ratio outlier analysis (IROA), which utilizes samples that are isotopically labeled with 5% (test) and 95% (control) 13C. This labeling strategy leads to characteristic isotopic patterns that allow the differentiation of biological signals from artifacts and yield the exact number of carbons, significantly reducing possible molecular formulae. The relative abundance between the test and control samples for every IROA feature can be determined simply by integrating the peaks that arise from the 5 and 95% channels. For NMR applications, we describe two 13C-based approaches. For samples at natural abundance, we have developed a workflow to obtain 13C–13C and 13C–1H statistical correlations using 1D 13C and 1H NMR spectra. For samples that can be isotopically labeled, we describe another NMR approach to obtain direct 13C–13C spectroscopic correlations. These methods both provide extensive information about the carbon framework of compounds in the mixture for either database matching or de novo compound identification. We also discuss strategies in which 13C NMR can be used to identify unknown compounds from IROA experiments. By combining technologies with the same samples, we can identify important biomarkers and corresponding metabolites of interest. PMID:26379677

  6. Abundance anomaly of the 13C species of CCH

    NASA Astrophysics Data System (ADS)

    Sakai, N.; Saruwatari, O.; Sakai, T.; Takano, S.; Yamamoto, S.

    2010-03-01

    Aims: We have observed the N = 1-0 lines of CCH and its 13C isotopic species toward a cold dark cloud, TMC-1 and a star-forming region, L1527, to investigate the 13C abundances and formation pathways of CCH. Methods: The observations have been carried out with the IRAM 30 m telescope. Results: We have successfully detected the lines of 13CCH and C13CH toward the both sources and found a significant intensity difference between the two 13C isotopic species. The [C13CH] /[13CCH] abundance ratios are 1.6 ± 0.4 (3σ) and 1.6 ± 0.1 (3σ) for TMC-1 and L1527, respectively. The abundance difference between C13CH and 13CCH means that the two carbon atoms of CCH are not equivalent in the formation pathway. On the other hand, the [CCH]/[C13CH] and [CCH]/[13CCH] ratios are evaluated to be larger than 170 and 250 toward TMC-1, and to be larger than 80 and 135 toward L1527, respectively. Therefore, both of the 13C species are significantly diluted in comparison with the interstellar 12C/13C ratio of 60. The dilution is discussed in terms of a behavior of 13C in molecular clouds.

  7. Localized in vivo13C NMR spectroscopy of the brain

    PubMed Central

    Gruetter, Rolf; Adriany, Gregor; Choi, In-Young; Henry, Pierre-Gilles; Lei, Hongxia; Öz, Gülin

    2006-01-01

    Localized 13C NMR spectroscopy provides a new investigative tool for studying cerebral metabolism. The application of 13C NMR spectroscopy to living intact humans and animals presents the investigator with a number of unique challenges. This review provides in the first part a tutorial insight into the ingredients required for achieving a successful implementation of localized 13C NMR spectroscopy. The difficulties in establishing 13C NMR are the need for decoupling of the one-bond 13C–1H heteronuclear J coupling, the large chemical shift range, the low sensitivity and the need for localization of the signals. The methodological consequences of these technical problems are discussed, particularly with respect to (a) RF front-end considerations, (b) localization methods, (c) the low sensitivity, and (d) quantification methods. Lastly, some achievements of in vivo localized 13C NMR spectroscopy of the brain are reviewed, such as: (a) the measurement of brain glutamine synthesis and the feasibility of quantifying glutamatergic action in the brain; (b) the demonstration of significant anaplerotic fluxes in the brain; (c) the demonstration of a highly regulated malate-aspartate shuttle in brain energy metabolism and isotope flux; (d) quantification of neuronal and glial energy metabolism; and (e) brain glycogen metabolism in hypoglycemia in rats and humans. We conclude that the unique and novel insights provided by 13C NMR spectroscopy have opened many new research areas that are likely to improve the understanding of brain carbohydrate metabolism in health and disease. PMID:14679498

  8. 13C-labeled gluconate tracing as a direct and accurate method for determining the pentose phosphate pathway split ratio in Penicillium chrysogenum.

    PubMed

    Kleijn, Roelco J; van Winden, Wouter A; Ras, Cor; van Gulik, Walter M; Schipper, Dick; Heijnen, Joseph J

    2006-07-01

    In this study we developed a new method for accurately determining the pentose phosphate pathway (PPP) split ratio, an important metabolic parameter in the primary metabolism of a cell. This method is based on simultaneous feeding of unlabeled glucose and trace amounts of [U-13C]gluconate, followed by measurement of the mass isotopomers of the intracellular metabolites surrounding the 6-phosphogluconate node. The gluconate tracer method was used with a penicillin G-producing chemostat culture of the filamentous fungus Penicillium chrysogenum. For comparison, a 13C-labeling-based metabolic flux analysis (MFA) was performed for glycolysis and the PPP of P. chrysogenum. For the first time mass isotopomer measurements of 13C-labeled primary metabolites are reported for P. chrysogenum and used for a 13C-based MFA. Estimation of the PPP split ratio of P. chrysogenum at a growth rate of 0.02 h(-1) yielded comparable values for the gluconate tracer method and the 13C-based MFA method, 51.8% and 51.1%, respectively. A sensitivity analysis of the estimated PPP split ratios showed that the 95% confidence interval was almost threefold smaller for the gluconate tracer method than for the 13C-based MFA method (40.0 to 63.5% and 46.0 to 56.5%, respectively). From these results we concluded that the gluconate tracer method permits accurate determination of the PPP split ratio but provides no information about the remaining cellular metabolism, while the 13C-based MFA method permits estimation of multiple fluxes but provides a less accurate estimate of the PPP split ratio.

  9. A new tracer experiment to estimate the methane emissions from a dairy cow shed using sulfur hexafluoride (SF6)

    NASA Astrophysics Data System (ADS)

    Marik, Thomas; Levin, Ingeborg

    1996-09-01

    Methane emission from livestock and agricultural wastes contribute globally more than 30% to the anthropogenic atmospheric methane source. Estimates of this number have been derived from respiration chamber experiments. We determined methane emission rates from a tracer experiment in a modern cow shed hosting 43 dairy cows in their accustomed environment. During a 24-hour period the concentrations of CH4, CO2, and SF6, a trace gas which has been released at a constant rate into the stable air, have been measured. The ratio between SF6 release rate and measured SF6 concentration was then used to estimate the ventilation rate of the stable air during the course of the experiment. The respective ratio between CH4 or CO2 and SF6 concentration together with the known SF6 release rate allows us to calculate the CH4 (and CO2) emissions in the stable. From our experiment we derive a total daily mean CH4 emission of 441 LSTP per cow (9 cows nonlactating), which is about 15% higher than previous estimates for German cows with comparable milk production obtained during respiration chamber experiments. The higher emission in our stable experiment is attributed to the contribution of CH4 release from about 50 m3 of liquid manure present in the cow shed in underground channels. Also, considering measurements we made directly on a liquid manure tank, we obtained an estimate of the total CH4 production from manure: The normalized contribution of methane from manure amounts to 12-30% of the direct methane release of a dairy cow during rumination. The total CH4 release per dairy cow, including manure, is 521-530 LSTP CH4 per day.

  10. A Peptide-Based Method for 13C Metabolic Flux Analysis in Microbial Communities

    PubMed Central

    Ghosh, Amit; Nilmeier, Jerome; Weaver, Daniel; Adams, Paul D.; Keasling, Jay D.; Mukhopadhyay, Aindrila; Petzold, Christopher J.; Martín, Héctor García

    2014-01-01

    The study of intracellular metabolic fluxes and inter-species metabolite exchange for microbial communities is of crucial importance to understand and predict their behaviour. The most authoritative method of measuring intracellular fluxes, 13C Metabolic Flux Analysis (13C MFA), uses the labeling pattern obtained from metabolites (typically amino acids) during 13C labeling experiments to derive intracellular fluxes. However, these metabolite labeling patterns cannot easily be obtained for each of the members of the community. Here we propose a new type of 13C MFA that infers fluxes based on peptide labeling, instead of amino acid labeling. The advantage of this method resides in the fact that the peptide sequence can be used to identify the microbial species it originates from and, simultaneously, the peptide labeling can be used to infer intracellular metabolic fluxes. Peptide identity and labeling patterns can be obtained in a high-throughput manner from modern proteomics techniques. We show that, using this method, it is theoretically possible to recover intracellular metabolic fluxes in the same way as through the standard amino acid based 13C MFA, and quantify the amount of information lost as a consequence of using peptides instead of amino acids. We show that by using a relatively small number of peptides we can counter this information loss. We computationally tested this method with a well-characterized simple microbial community consisting of two species. PMID:25188426

  11. 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

  12. 1H and 13C NMR assignments for the cyanine dyes SYBR Safe and thiazole orange.

    PubMed

    Evenson, William E; Boden, Lauren M; Muzikar, Katy A; O'Leary, Daniel J

    2012-12-01

    Analysis of (1)H and (13)C NMR and mass spectral data for the fluorescent nucleic acid stain SYBR Safe indicates that it contains a cyanine-based cationic core structure identical to thiazole orange. The difference between these two compounds is the type of N-substitution on the quinolinium ring system (SYBR Safe, n-Pr; thiazole orange, Me). The (1)H and (13)C NMR resonances for both compounds were assigned on the basis of one- and two-dimensional (COSY, ROESY, HSQC, and HMBC) experiments. The preferred conformation of these compounds was computed by ab initio methods and found to be consistent with the NMR data.

  13. *d13C composition of primary producers and role of detritus in a freshwater coastal ecosystem

    USGS Publications Warehouse

    Keough, J.R.; Hagley, C.A.; Sierszen, M.

    1998-01-01

    Stable-isotope ratio signatures of primary producers in a coastal wetland and in adjacent offshore waters of western Lake Superior indicated that phytoplankton are the primary source of carbon for the grazing food web of this ecosystem. This study outlines the possible roles of other autotrophs in this regard. Isotopic signatures of macrophytes reflected their life-form-associated constraints on diffusion of inorganic carbon. Data indicated that differences between wetland and lake phytoplankton may be explained by the isotopic signatures of their dissolved inorganic carbon (DIC) sources. Results of an in situ experiment showed that respiration associated with macrophyte decomposition is capable of enriching surrounding water with significant amounts of *d13C-depleted DIC and lowering the net *d13C ratio of DIC in water in low-turbulence situations. The *d13C ratio for wetland phytoplankton may be depleted relative to pelagic algae because the fixed carbon is derived from decomposing detritus.

  14. Quantitative Analysis of Metabolic Mixtures by 2D 13C-Constant-Time TOCSY NMR Spectroscopy

    PubMed Central

    Bingol, Kerem; Zhang, Fengli; Bruschweiler-Li, Lei; Brüschweiler, Rafael

    2013-01-01

    An increasing number of organisms can be fully 13C-labeled, which has the advantage that their metabolomes can be studied by high-resolution 2D NMR 13C–13C constant-time (CT) TOCSY experiments. Individual metabolites can be identified via database searching or, in the case of novel compounds, through the reconstruction of their backbone-carbon topology. Determination of quantitative metabolite concentrations is another key task. Because significant peak overlaps in 1D NMR spectra prevents straightforward quantification through 1D peak integrals, we demonstrate here the direct use of 13C–13C CT-TOCSY spectra for metabolite quantification. This is accomplished through the quantum-mechanical treatment of the TOCSY magnetization transfer at short and long mixing times or by the use of analytical approximations, which are solely based on the knowledge of the carbon-backbone topologies. The methods are demonstrated for carbohydrate and amino-acid mixtures. PMID:23773204

  15. (1)H and (13)C magic-angle spinning nuclear magnetic resonance studies of the chicken eggshell.

    PubMed

    Pisklak, Dariusz Maciej; Szeleszczuk, Lukasz; Wawer, Iwona

    2012-12-19

    The chicken eggshell, a product of biomineralization, contains inorganic and organic substances whose content changes during the incubation process. Bloch-decay (BD) (1)H, (13)C, and cross-polarization (CP) (13)C nuclear magnetic resonance (NMR) spectra of chicken eggshells were acquired under magic-angle spinning (MAS). Variable contact time (13)C CP MAS NMR experiments revealed the signals of carbonyl groups from organic and inorganic compounds. In the (13)C BD NMR spectra, a single peak at 168.1 ppm was detected, whereas in the (1)H BD spectra, the signals from water and the bicarbonate ion were assigned. A simultaneous decrease of the water signal in the (1)H MAS NMR spectra and an increase of the carbonate ion signal in the (13)C CP MAS NMR spectra of eggshells collected during the incubation period indicate the substitution of calcium ions by hydrogen ions in the calcium carbonate crystalline phase during the incubation of an egg.

  16. Using Position-Specific 13C and 14C Labeling and 13C-PLFA Analysis to Assess Microbial Transformations of Free Versus Sorbed Alanine

    NASA Astrophysics Data System (ADS)

    Apostel, C.; Herschbach, J.; Bore, E. K.; Kuzyakov, Y.; Dippold, M. A.

    2015-12-01

    Sorption of charged or partially charged low molecular weight organic substances (LMWOS) to soil mineral surfaces delays microbial uptake and therefore mineralization of LMWOS to CO2, as well as all other biochemical transformations. We used position-specific labeling, a tool of isotope applications novel to soil sciences, to compare the transformation mechanisms of sorbed and non-sorbed alanine in soil. Alanine as an amino acid links C- and N-cycles in soil and therefore is a model substance for the pool of LMWOS. To assess transformations of sorbed alanine, we added position-specific and uniformly 13C and 14C labeled alanine tracer to soil that had previously been sterilized by γ-radiation. The labeled soil was added to non-sterilized soil from the same site and incubated. Soil labeled with the same tracers without previous sorption was prepared and incubated as well. We captured the respired CO2 and determined its 14C-activity at increasing time intervals. The incorporation of 14C into microbial biomass was determined by chloroform fumigation extraction (CFE), and utilization of individual C positions by distinct microbial groups was evaluated by 13C-phospholipid fatty acid analysis (PLFA). A dual peak in the respired CO2 revealed two sorption mechanisms. To compare the fate of individual C atoms independent of their concentration and pool size in soil, we applied the divergence index (DI). The DI reveals the convergent or divergent behavior of C from individual molecule positions during microbial utilization. Alanine C-1 position was mainly oxidized to CO2, while its C-2 and C-3 were preferentially incorporated in microbial biomass and PLFA. This indicates that sorption by the COOH group does not protect this group from preferential oxidation. Microbial metabolism was determinative for the preferential oxidation of individual molecule positions. The use of position-specific labeling revealed mechanisms and kinetics of microbial utilization of sorbed and non

  17. 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.

  18. Non invasive monitoring of water flow in the vadose zone: the issue of mass balance in controlled tracer injection experiments.

    NASA Astrophysics Data System (ADS)

    Cassiani, G.; Deiana, R.; Kemna, A.

    2006-12-01

    The non invasive characterization of the subsurface, with the goal of obtaining data for the calibration of flow and transport hydrologic models, has become very popular in recent years. However, the process of converting geophysical data into quantitative estimates of volumetric water content and/or solute concentrations is not straightforward, as it requires knowledge of (1) resolution and penetration characteristics of the geophysical methods (imaging characteristics); (2) suitable constitutive laws for the conversion of geophysical quantities into hydrologic quantities (petrophysics). In addition, the calibration of flow/transport models on the basis of geophysically-derived data requires that the space/time evolution of these data be summarized in terms that can be directly compared with simulation results. In the case of controlled injection experiments having a point source (e.g. a borehole section), an effective tool is the analysis of spatial moments of the injected slug. However, important issues are still unresolved, particularly with regard to the identifiability of second order spatial moments (spread) and, more disturbing, mass balance. Field experience demonstrates that it is rarely possible to "see" the total injected tracer mass by means of a non invasive method, be it cross-hole ERT or GPR, leading to errors of the order of 50%. The reasons of these limitations lie mostly in the imaging characteristics of the methods. A better understanding of these characteristics can, on the other hand, provide new tools for a more accurate calibration of flow/transport models.

  19. 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.

  20. Non-isothermal infiltration and tracer transport experiments on large soil columns

    NASA Astrophysics Data System (ADS)

    Sobotkova, Martina; Snehota, Michal; Cejkova, Eva; Tesar, Miroslav

    2016-04-01

    Isothermal and non-isothermal infiltration experiments were carried out in the laboratory on large undisturbed soil columns (19 cm in diameter, 25 cm high) taken at the experimental catchments Roklan (Sumava Mountains, Czech Republic) and Uhlirska (Jizera Mountains, Czech republic). The aim of the study was twofold. The first goal was to obtain water flow and heat transport data for indirect parameter estimation of thermal and hydraulic properties of soils from two sites by inverse modelling. The second aim was to investigate the extent of impact of the temperature on saturated hydraulic conductivity (Ksat) and dispersity of solute transport. The temperature of infiltrating water in isothermal experiment (20 °C) was equal to the initial temperature of the sample. For non-isothermal experiment water temperature was 5°C, while the initial temperature of the sample was 20°C as in previous case. The experiment was started by flooding the sample surface. Then water level was maintained at constant level throughout the infiltration run using the optical sensor and peristaltic pump. Concentration pulse of deuterium was applied at the top of the soil sample, during the steady state flow. Initial pressure head in the sample was close to field capacity. Two tensiometers and two temperature sensors were inserted in the soil sample in two depths (9 and 15 cm below the top of the sample). Two additional temperature sensors monitored the temperature entering and leaving the samples. Water drained freely through the perforated plate at the bottom of sample by gravity. Inflow and outflow water flux densities, water pressure heads and soil temperatures were monitored continuously during experiments. Effluent was sampled in regular time intervals and samples were analysed for deuterium concentrations by laser spectroscopy to develop breakthrough curves. The outcome of experiments are the series of measured water fluxes, pressure heads and temperatures ready for inverse modelling

  1. Transport and imaging of brute-force (13)C hyperpolarization.

    PubMed

    Hirsch, Matthew L; Smith, Bryce A; Mattingly, Mark; Goloshevsky, Artem G; Rosay, Melanie; Kempf, James G

    2015-12-01

    We demonstrate transport of hyperpolarized frozen 1-(13)C pyruvic acid from its site of production to a nearby facility, where a time series of (13)C images was acquired from the aqueous dissolution product. Transportability is tied to the hyperpolarization (HP) method we employ, which omits radical electron species used in other approaches that would otherwise relax away the HP before reaching the imaging center. In particular, we attained (13)C HP by 'brute-force', i.e., using only low temperature and high-field (e.g., T<∼2K and B∼14T) to pre-polarize protons to a large Boltzmann value (∼0.4% (1)H polarization). After polarizing the neat, frozen sample, ejection quickly (<1s) passed it through a low field (B<100G) to establish the (1)H pre-polarization spin temperature on (13)C via the process known as low-field thermal mixing (yielding ∼0.1% (13)C polarization). By avoiding polarization agents (a.k.a. relaxation agents) that are needed to hyperpolarize by the competing method of dissolution dynamic nuclear polarization (d-DNP), the (13)C relaxation time was sufficient to transport the sample for ∼10min before finally dissolving in warm water and obtaining a (13)C image of the hyperpolarized, dilute, aqueous product (∼0.01% (13)C polarization, a >100-fold gain over thermal signals in the 1T scanner). An annealing step, prior to polarizing the sample, was also key for increasing T1∼30-fold during transport. In that time, HP was maintained using only modest cryogenics and field (T∼60K and B=1.3T), for T1((13)C) near 5min. Much greater time and distance (with much smaller losses) may be covered using more-complete annealing and only slight improvements on transport conditions (e.g., yielding T1∼5h at 30K, 2T), whereas even intercity transfer is possible (T1>20h) at reasonable conditions of 6K and 2T. Finally, it is possible to increase the overall enhancement near d-DNP levels (i.e., 10(2)-fold more) by polarizing below 100mK, where nanoparticle

  2. Transport and imaging of brute-force 13C hyperpolarization

    NASA Astrophysics Data System (ADS)

    Hirsch, Matthew L.; Smith, Bryce A.; Mattingly, Mark; Goloshevsky, Artem G.; Rosay, Melanie; Kempf, James G.

    2015-12-01

    We demonstrate transport of hyperpolarized frozen 1-13C pyruvic acid from its site of production to a nearby facility, where a time series of 13C images was acquired from the aqueous dissolution product. Transportability is tied to the hyperpolarization (HP) method we employ, which omits radical electron species used in other approaches that would otherwise relax away the HP before reaching the imaging center. In particular, we attained 13C HP by 'brute-force', i.e., using only low temperature and high-field (e.g., T < ∼2 K and B ∼ 14 T) to pre-polarize protons to a large Boltzmann value (∼0.4% 1H polarization). After polarizing the neat, frozen sample, ejection quickly (<1 s) passed it through a low field (B < 100 G) to establish the 1H pre-polarization spin temperature on 13C via the process known as low-field thermal mixing (yielding ∼0.1% 13C polarization). By avoiding polarization agents (a.k.a. relaxation agents) that are needed to hyperpolarize by the competing method of dissolution dynamic nuclear polarization (d-DNP), the 13C relaxation time was sufficient to transport the sample for ∼10 min before finally dissolving in warm water and obtaining a 13C image of the hyperpolarized, dilute, aqueous product (∼0.01% 13C polarization, a >100-fold gain over thermal signals in the 1 T scanner). An annealing step, prior to polarizing the sample, was also key for increasing T1 ∼ 30-fold during transport. In that time, HP was maintained using only modest cryogenics and field (T ∼ 60 K and B = 1.3 T), for T1(13C) near 5 min. Much greater time and distance (with much smaller losses) may be covered using more-complete annealing and only slight improvements on transport conditions (e.g., yielding T1 ∼ 5 h at 30 K, 2 T), whereas even intercity transfer is possible (T1 > 20 h) at reasonable conditions of 6 K and 2 T. Finally, it is possible to increase the overall enhancement near d-DNP levels (i.e., 102-fold more) by polarizing below 100 mK, where

  3. 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

  4. A scientific workflow framework for (13)C metabolic flux analysis.

    PubMed

    Dalman, Tolga; Wiechert, Wolfgang; Nöh, Katharina

    2016-08-20

    Metabolic flux analysis (MFA) with (13)C labeling data is a high-precision technique to quantify intracellular reaction rates (fluxes). One of the major challenges of (13)C MFA is the interactivity of the computational workflow according to which the fluxes are determined from the input data (metabolic network model, labeling data, and physiological rates). Here, the workflow assembly is inevitably determined by the scientist who has to consider interacting biological, experimental, and computational aspects. Decision-making is context dependent and requires expertise, rendering an automated evaluation process hardly possible. Here, we present a scientific workflow framework (SWF) for creating, executing, and controlling on demand (13)C MFA workflows. (13)C MFA-specific tools and libraries, such as the high-performance simulation toolbox 13CFLUX2, are wrapped as web services and thereby integrated into a service-oriented architecture. Besides workflow steering, the SWF features transparent provenance collection and enables full flexibility for ad hoc scripting solutions. To handle compute-intensive tasks, cloud computing is supported. We demonstrate how the challenges posed by (13)C MFA workflows can be solved with our approach on the basis of two proof-of-concept use cases.

  5. A scientific workflow framework for (13)C metabolic flux analysis.

    PubMed

    Dalman, Tolga; Wiechert, Wolfgang; Nöh, Katharina

    2016-08-20

    Metabolic flux analysis (MFA) with (13)C labeling data is a high-precision technique to quantify intracellular reaction rates (fluxes). One of the major challenges of (13)C MFA is the interactivity of the computational workflow according to which the fluxes are determined from the input data (metabolic network model, labeling data, and physiological rates). Here, the workflow assembly is inevitably determined by the scientist who has to consider interacting biological, experimental, and computational aspects. Decision-making is context dependent and requires expertise, rendering an automated evaluation process hardly possible. Here, we present a scientific workflow framework (SWF) for creating, executing, and controlling on demand (13)C MFA workflows. (13)C MFA-specific tools and libraries, such as the high-performance simulation toolbox 13CFLUX2, are wrapped as web services and thereby integrated into a service-oriented architecture. Besides workflow steering, the SWF features transparent provenance collection and enables full flexibility for ad hoc scripting solutions. To handle compute-intensive tasks, cloud computing is supported. We demonstrate how the challenges posed by (13)C MFA workflows can be solved with our approach on the basis of two proof-of-concept use cases. PMID:26721184

  6. Implications of a Multi-well Tracer Test in the Transport of Pathogens at a Riverbank Filtration Experiment Site.

    NASA Astrophysics Data System (ADS)

    Langford, R. P.; Pillai, S.; Schulze-Makuch, D.; Widmer, K.; Abdel-Fattah, A.; Lerhner, T.

    2003-12-01

    This study tracks the transport of bromide and microspheres mimicking pathogens in an arid environment. The study site uses the Rio Grande that experiences significant annual fluctuations in both water quantity and quality. The pumping well is 17 m from the stream bank and the water table was 2 m below the stream surface. The aquifer is medium and fine-grained sand comprising two flow units. Observation wells are screened over 1 or 1.5 m intervals. The average hydraulic conductivity was about 2 x 10-3 m/s based on a test analysis, however, the responses indicated that sediment heterogeneities affected the hydraulic behavior. A 427 hour tracer test using bromide and fluorescent microspheres provides initial results that are relevant to the transport of pathogens through the subsurface under riverbank filtration conditions. Bromide was injected into an observation well at the channel margin. Differently colored fluorescent microspheres (0.25nm, 1?m, 6?m and 10?m) were injected into the stream bottom and into two observation wells. Conclusions from the tracer test are: 1) Both bromide and microspheres continued to be observed throughout the 18 days of the experiment. 2) The bromide recovery in the pumping well and in the deeper observation wells showed early and late peaks with a long tails indicating that the geological medium at the field site behaves like a double-porosity medium allowing the tracer to move relatively quickly through the higher conductivity units while being significantly retarded in the low hydraulic conductivity units. 3) Some wells showed consistently higher concentrations of bromide. 4) The 1? micospheres were abundant in the observation wells and allowed tracing of flowpaths. These showed multiple peaks similar to the bromide results. This indicates highly preferential transport paths in the sediment. 5) Microspheres from the three injection sites had distinctly different transport paths and rates. 6) Both bromide and microspheres appeared in

  7. Landmarks in the application of 13C-magnetic resonance spectroscopy to studies of neuronal/glial relationships.

    PubMed

    Bachelard, H

    1998-01-01

    The development of the use of carbon isotopes as metabolic tracers is briefly described. 13C-labelled precursors (13CO2, 13CH4) first became available in 1940 and were studied in microorganisms, but their use was limited by very low enrichments and lack of suitable analytical equipment. More success was achieved with 11C and especially 14C, as these radioactive tracers did not need to be highly enriched. Although the stable 13C isotope can be used at a low percentage enrichment in mass spectrometry, its application to magnetic resonance spectroscopy (MRS) requires very highly enriched precursors, due to its low natural abundance and low sensitivity. Despite such limitations, however, the great advantage of 13C-MRS lies in its exquisite chemical specificity, in that labelling of different carbon atoms can be distinguished within the same molecule. Effective exploitation became feasible in the early 1970s with the advent of stable instruments, Fourier transform 13C-MRS, and the availability of highly enriched precursors. Reports of its use in brain research began to appear in the mid-1980s. The applications of 13C isotopomer analysis to research on neuronal/glial relationships are reviewed. The presence of neighbouring 13C-labelled atoms affects the appearance of the resonances (splitting due to C-C coupling), and so allows for unique quantification of rates through different and possibly competing pathways. Isotopomer patterns in resonances labelled from a combination of [1-13C]glucose and [1, 2-13C2]acetate have revealed aspects of neuronal/glial metabolic trafficking on depolarization and under hypoxic conditions in vitro. This approach has now been applied to in vivo studies on inhibition of glial metabolism using fluoroacetate. The results confirm the glial specificity of the toxin and demonstrate that it does not affect entry of acetate. When the glial TCA cycle is inhibited, the ability of the glia to participate in the glutamate/glutamine cycle remains

  8. {sup 13}C relaxation in an RNA hairpin

    SciTech Connect

    King, G.C. |; Akratos, C.; Xi, Z.; Michnica, M.J.

    1994-12-01

    This initial survey of {sup 13}C relaxation in the {triangle}TAR RNA element has generated a number of interesting results that should prove generally useful for future studies. The most readily comparable study in the literature monitored {sup 13}C relaxation of the methyl groups from unusual bases in tRNA{sup Phe}. The study, which used T{sub 1} and NOE data only, reported order parameters for the methyl group axis that ranged between 0.51 and 0.97-a range similar to that observed here. However, they reported a breakdown of the standard order parameter analysis at higher (118-MHz {sup 13}C) frequencies, which should serve to emphasize the need for a thorough exploration of suitable motional models.

  9. Habitat-specific differences in plasticity of foliar δ13C in temperate steppe grasses

    PubMed Central

    Liu, Yanjie; Zhang, Lirong; Niu, Haishan; Sun, Yue; Xu, Xingliang

    2014-01-01

    A decrease in foliar δ13C with increasing precipitation is a common tendency in steppe plants. However, the rate of decrease has been reported to differ between different species or populations. We here hypothesized that plant populations in the same habitat of temperate steppes may not differ in foliar δ13C response patterns to precipitation, but could differ in the levels of plasticity of foliar δ13C across different habitats. In order to test this hypothesis, we conducted controlled watering experiments in northeast China at five sites along a west–east transect at latitude 44°N, which show substantial interannual fluctuations and intra-annual changes in precipitation among them. In 2001, watering treatment (six levels, three replicates) was assigned to 18 plots at each site. The responses of foliar δ13C to precipitation (i.e., the sum of watering and rainfall) were determined in populations of several grass species that were common across all sites. Although similar linear regression slopes were observed for populations of different species growing at the same site, significantly different slopes were obtained for populations of the same species growing at different sites. Further, the slope of the line progressively decreased from Site I to Site V for all species in this study. These results suggest habitat-specific differences in plasticity of foliar δ13C in temperate steppe grasses. This indicates that species' δ13C response to precipitation is conservative at the same site due to their long-term acclimation, but the mechanism responsible behind this needs further investigations. PMID:25035804

  10. Geochemical Approach to Archaeal Ecology: δ13C of GDGTs

    NASA Astrophysics Data System (ADS)

    Lichtin, S.; Warren, C.; Pearson, A.; Pagani, M.

    2015-12-01

    Over the last decade and a half, glycerol dialkyl glycerol tetraethers (GDGTs) have increasingly been used to reconstruct environmental temperatures; proxies like TEX86 that correlate the relative abundance of these archaeal cell membrane lipids to sea surface temperature are omnipresent in paleoclimatology literature. While it has become common to make claims about past temperatures using GDGTs, our present understanding of the organisms that synthesize the compounds is still quite limited. The generally accepted theory states that microorganisms like the Thaumarchaeota modify the structure of membrane lipids to increase intermolecular interactions, strengthening the membrane at higher temperatures. Yet to date, culture experiments have been largely restricted to a single species, Nitrosopumilus maritimes, and recent studies on oceanic archaeal rRNA have revealed that these biomarkers are produced in diverse, heterogeneous, and site-specific communities. This brings up questions as to whether different subclasses of GDGTs, and all subsequent proxies, represent adaptation within a single organismal group or a shift in community composition. To investigate whether GDGTs with different chain structures, from the simple isoprenoidal GDGT-0 to Crenarchaeol with its many cyclopentane groups, are sourced from archaea with similar or disparate metabolic pathways—and if that information is inherited in GDGTs trapped in marine sediments—this study examines the stable carbon isotope values (δ13C) of GDGTs extracted from the uppermost meters of sediment in the Orca Basin, Gulf of Mexico, using spooling-wire microcombustion isotope-ratio mass spectrometer (SWiM-IRMS), tackling a fundamental assumption of the TEX86 proxy that influences the way we perceive the veracity of existing temperature records.

  11. A Case Study of the Weather Research and Forecasting Model Applied to the Joint Urban 2003 Tracer Field Experiment. Part 2: Gas Tracer Dispersion

    NASA Astrophysics Data System (ADS)

    Nelson, Matthew A.; Brown, Michael J.; Halverson, Scot A.; Bieringer, Paul E.; Annunzio, Andrew; Bieberbach, George; Meech, Scott

    2016-07-01

    The Quick Urban & Industrial Complex (QUIC) atmospheric transport, and dispersion modelling, system was evaluated against the Joint Urban 2003 tracer-gas measurements. This was done using the wind and turbulence fields computed by the Weather Research and Forecasting (WRF) model. We compare the simulated and observed plume transport when using WRF-model-simulated wind fields, and local on-site wind measurements. Degradation of the WRF-model-based plume simulations was cased by errors in the simulated wind direction, and limitations in reproducing the small-scale wind-field variability. We explore two methods for importing turbulence from the WRF model simulations into the QUIC system. The first method uses parametrized turbulence profiles computed from WRF-model-computed boundary-layer similarity parameters; and the second method directly imports turbulent kinetic energy from the WRF model. Using the WRF model's Mellor-Yamada-Janjic boundary-layer scheme, the parametrized turbulence profiles and the direct import of turbulent kinetic energy were found to overpredict and underpredict the observed turbulence quantities, respectively. Near-source building effects were found to propagate several km downwind. These building effects and the temporal/spatial variations in the observed wind field were often found to have a stronger influence over the lateral and vertical plume spread than the intensity of turbulence. Correcting the WRF model wind directions using a single observational location improved the performance of the WRF-model-based simulations, but using the spatially-varying flow fields generated from multiple observation profiles generally provided the best performance.

  12. Calcium-ammonium exchange experiments on clay minerals using a (45)Ca tracer technique in marine pore water.

    PubMed

    Ockert, Charlotte; Wehrmann, Laura M; Kaufhold, Stephan; Ferdelman, Tim G; Teichert, Barbara M A; Gussone, Nikolaus

    2014-01-01

    Understanding cation exchange processes is important for evaluating early diagenetic and synsedimentary processes taking place in marine sediments. To quantify calcium (Ca) exchange and Ca-ammonium exchange in a seawater environment, we performed experiments with a radioactive (45)Ca tracer on clay mineral standards (Fithian illite, montmorillonite and kaolinite) and marine sediments from the North Atlantic Integrated Ocean Drilling Program Site U1306A in artificial seawater (ASW). The results show that equilibrium during the initial attachment of Ca as well as the exchange of Ca by [Formula: see text] is attained in less than 2 min. On average 8-20% of the exchangeable sites of the clay minerals were occupied by Ca in a seawater medium. The conditional selectivity coefficient, describing the [Formula: see text] exchange in ASW is mineral specific and it was determined to be 0.07 for montmorillonite, 0.05 for a natural marine sediment and 0.013 for Fithian illite. PMID:24437731

  13. Impact of Ho(3+)-doping on (13)C dynamic nuclear polarization using trityl OX063 free radical.

    PubMed

    Kiswandhi, Andhika; Niedbalski, Peter; Parish, Christopher; Kaur, Pavanjeet; Martins, André; Fidelino, Leila; Khemtong, Chalermchai; Song, Likai; Sherry, A Dean; Lumata, Lloyd

    2016-08-21

    We have investigated the effects of Ho-DOTA doping on the dynamic nuclear polarization (DNP) of [1-(13)C] sodium acetate using trityl OX063 free radical at 3.35 T and 1.2 K. Our results indicate that addition of 2 mM Ho-DOTA on 3 M [1-(13)C] sodium acetate sample in 1 : 1 v/v glycerol : water with 15 mM trityl OX063 improves the DNP-enhanced (13)C solid-state nuclear polarization by a factor of around 2.7-fold. Similar to the Gd(3+) doping effect on (13)C DNP, the locations of the positive and negative (13)C maximum polarization peaks in the (13)C microwave DNP sweep are shifted towards each other with the addition of Ho-DOTA on the DNP sample. W-band electron spin resonance (ESR) studies have revealed that while the shape and linewidth of the trityl OX063 ESR spectrum was not affected by Ho(3+)-doping, the electron spin-lattice relaxation time T1 of trityl OX063 was prominently reduced at cryogenic temperatures. The reduction of trityl OX063 electron T1 by Ho-doping is linked to the (13)C DNP improvement in light of the thermodynamic picture of DNP. Moreover, the presence of Ho-DOTA in the dissolution liquid at room temperature has negligible reduction effect on liquid-state (13)C T1, in contrast to Gd(3+)-doping which drastically reduces the (13)C T1. The results here suggest that Ho(3+)-doping is advantageous over Gd(3+) in terms of preservation of hyperpolarized state-an important aspect to consider for in vitro and in vivo NMR or imaging (MRI) experiments where a considerable preparation time is needed to administer the hyperpolarized (13)C liquid.

  14. Impact of Ho(3+)-doping on (13)C dynamic nuclear polarization using trityl OX063 free radical.

    PubMed

    Kiswandhi, Andhika; Niedbalski, Peter; Parish, Christopher; Kaur, Pavanjeet; Martins, André; Fidelino, Leila; Khemtong, Chalermchai; Song, Likai; Sherry, A Dean; Lumata, Lloyd

    2016-08-21

    We have investigated the effects of Ho-DOTA doping on the dynamic nuclear polarization (DNP) of [1-(13)C] sodium acetate using trityl OX063 free radical at 3.35 T and 1.2 K. Our results indicate that addition of 2 mM Ho-DOTA on 3 M [1-(13)C] sodium acetate sample in 1 : 1 v/v glycerol : water with 15 mM trityl OX063 improves the DNP-enhanced (13)C solid-state nuclear polarization by a factor of around 2.7-fold. Similar to the Gd(3+) doping effect on (13)C DNP, the locations of the positive and negative (13)C maximum polarization peaks in the (13)C microwave DNP sweep are shifted towards each other with the addition of Ho-DOTA on the DNP sample. W-band electron spin resonance (ESR) studies have revealed that while the shape and linewidth of the trityl OX063 ESR spectrum was not affected by Ho(3+)-doping, the electron spin-lattice relaxation time T1 of trityl OX063 was prominently reduced at cryogenic temperatures. The reduction of trityl OX063 electron T1 by Ho-doping is linked to the (13)C DNP improvement in light of the thermodynamic picture of DNP. Moreover, the presence of Ho-DOTA in the dissolution liquid at room temperature has negligible reduction effect on liquid-state (13)C T1, in contrast to Gd(3+)-doping which drastically reduces the (13)C T1. The results here suggest that Ho(3+)-doping is advantageous over Gd(3+) in terms of preservation of hyperpolarized state-an important aspect to consider for in vitro and in vivo NMR or imaging (MRI) experiments where a considerable preparation time is needed to administer the hyperpolarized (13)C liquid. PMID:27424954

  15. Where is the water going: An irrigation experiment using a natural isotopic tracer in karst SE, Australia.

    NASA Astrophysics Data System (ADS)

    Markowska, Monika; Baker, Andy; Andersen, Martin S.; Cuthbert, Mark; Rau, Gabriel; Jex, Cath; Rutlidge, Helen; Marjo, Chris; Roshan, Hamid; Treble, Pauline

    2014-05-01

    The karst unsaturated zone is a fractured rock environment associated with very heterogeneous water movement; spatial variability in the subsurface water storage; and fast preferential flow through fractures and fissures. These factors dominate the way in which water moves within the unsaturated zone in these environments, giving rise to flow path complexities less common in homogenous media. Currently there is limited research regarding karst infiltration/storage processes and potential evaporation in the unsaturated zone. Such processes may have the potential to alter the stable isotopic composition of groundwater. Caves provide a unique environment within which to examine exfiltration variability and flow dynamics in situ. In semi-arid environments evaporative processes in the unsaturated zone have been shown to directly alter the isotopic δ18O composition of cave drip waters, fractionating them towards heavier ratios, by a magnitude of 1-3 per mil relative to mean annual rainfall (Bar Matthews et al., 1996; Cuthbert et al., 2014). Here we present a novel isotopic drip water study from an artificial infiltration experiment at Wellington Caves, SE Australia. A series of four artificial infiltration events were initiated directly over Cathedral Cave, Wellington over as many days. The first event was spiked with a deuterium tracer and the subsurface response was monitored during several sampling campaigns over the following year. The infiltration study revealed: (1) isotopic break-through curves suggest a front of older water from the unsaturated zone storage arrived ahead of the infiltration water, (2) water residence times in the unsaturated zone were found to be longer than 6 months and, (3) large spatial heterogeneities existed in the proportion of exfiltrated deuterium tracer at different drip sites in the cave suggesting unique pathways and sources of water in the unsaturated zone. Implications from this study include the interpretation of paleo

  16. Accumulation of methylmercury in rice and flooded soil in experiments with an enriched isotopic Hg(II) tracer

    NASA Astrophysics Data System (ADS)

    Strickman, R. J.; Mitchell, C. P. J.

    2015-12-01

    Methylmercury (MeHg) is a neurotoxin produced in anoxic aquatic sediments. Numerous factors, including the presence of aquatic plants, alter the biogeochemistry of sediments, affecting the rate at which microorganisms transform bioavailable inorganic Hg (IHg) to MeHg. Methylmercury produced in flooded paddy soils and its transfer into rice has become an important dietary consideration. An improved understanding of how MeHg reaches the grain and the extent to which rice alters MeHg production in rhizosphere sediments could help to inform rice cultivation practices. We conducted a controlled greenhouse experiment with thirty rice plants grown in individual, flooded pots amended with enriched 200Hg. Unvegetated controls were maintained under identical conditions. At three plant growth stages (vegetative growth, flowering, and grain maturity), ten plants were sacrificed and samples collected from soil, roots, straw, panicle, and grain of vegetated and unvegetated pots, and assessed for MeHg and THg concentrations. We observed consistent ratios between ambient and tracer MeHg between soils (0.36 ±0.04 — 0.44 ± 0.09) and plant compartments (0.23 ± 0.07 -0.34 ± 0.05) indicating that plant MeHg contamination originates in the soil rather than in planta methylation. The majority of this MeHg was absorbed between the tillering (4.48 ± 2.38 ng/plant) and flowering (8.43 ± 5.12 ng/pl) phases, with a subsequent decline at maturity (2.87 ± 1.23 ng/pl) only partly explained by translocation to the developing grain, indicating that MeHg was demethylated in planta. In contrast, IHg was absorbed from both soil and air, as evidenced by the higher ambient IHg concentrations compared to tracer (3.76 ± 1.19 vs. 0.27 ± 0.40 ng/g). Surprisingly, MeHg accumulation was significantly (p= 0.042-- 0.003) lower in vegetated vs. unvegetated sediments at flowering (1.41 ± 0.26 vs. 1.57 ± 0.23) and maturity (1.27 ± 0.22 vs. 1.71 ± 0.25), suggesting that plant exudates bound Hg

  17. Nitrous oxide as a dynamical tracer in the 1987 Airborne Antarctic Ozone Experiment

    NASA Technical Reports Server (NTRS)

    Loewenstein, M.; Podolske, J. R.; Chan, K. R.; Strahan, S. E.

    1989-01-01

    In situ N2O measurements were made using an airborne tunable laser absorption spectrometer (ATLAS) on 12 flights into the Antarctic vortex, as well as on five transit flights outside the vortex region in August and September 1987, as part of the Airborne Antartic Ozone Experiment. Vertical profiles of N2O were obtained within the vortex on most of these flights and were obtained outside the vortex on several occasions. Flights into the vortex region show N2O decreasing southward between 53 and 72 S latitude on constant potential temperature surfaces in the lower stratosphere. The data lead to two important conclusions about the vortex region: (1) the lower stratosphere in August/September 1987 was occupied by 'old' air, which had subsided several kilometers during polar winter; (2) the N2O profile in the vortex was in an approximately steady state in August/September 1987, which indicates that the spring upwelling, suggested by several theories, did not occur.

  18. Measuring doubly 13C-substituted ethane by mass spectrometry

    NASA Astrophysics Data System (ADS)

    Clog, M.; Ling, C.; Eiler, J. M.

    2012-12-01

    Ethane (C2H6) is present in non-negligible amounts in most natural gas reservoirs and is used to produce ethylene for petrochemical industries. It is one of the by-products of lipid metabolism and is the arguably simplest molecule that can manifest multiple 13C substitutions. There are several plausible controls on the relative abundances of 13C2H6 in natural gases: thermodynamically controlled homogeneous isotope exchange reactions analogous to those behind carbonate clumped isotope thermometry; inheritance from larger biomolecules that under thermal degradation to produce natural gas; mixing of natural gases that differ markedly in bulk isotopic composition; or combinations of these and/or other, less expected fractionations. There is little basis for predicting which of these will dominate in natural samples. Here, we focus on an analytical techniques that will provide the avenue for exploring these phenomena. The method is based on high-resolution gas source isotope ratio mass spectrometry, using the Thermo 253-Ultra (a new prototype mass spectrometer). This instrument achieves the mass resolution (M/Δ M) up to 27,000, permitting separation of the isobaric interferences of potential contaminants and isotopologues of an analtye or its fragments which share a cardinal mass. We present techniques to analyze several isotopologues of molecular and fragment ions of C2H6. The critical isobaric separations for our purposes include: discrimination of 13C2H6 from 13C12CDH5 at mass 32 and separation of the 13CH3 fragment from 12CH4 at mass 16, both requiring at least a mass resolution of 20000 to make an adequate measurement. Other obvious interferences are either cleanly separated (e.g., O2, O) or accounted for by peak-stripping (CH3OH on mass 32 and NH2 on mass 16). We focus on a set of measurements which constrain: the doubly-substituted isotopologue, 13C2H6, and the 13CH3/12CH3 ratio of the methyl fragment, which constrains the bulk δ 13C. Similar methods can be

  19. Optimized [1-13C]glucose infusion protocol for 13C magnetic resonance spectroscopy at 3 Tesla of human brain glucose metabolism under euglycemic and hypoglycemic conditions

    PubMed Central

    van de Ven, Kim C.C.; van der Graaf, Marinette; Tack, Cees J.J.; Klomp, Dennis W.J.; Heerschap, Arend; de Galan, Bastiaan E.

    2009-01-01

    The effect of insulin-induced hypoglycemia on cerebral glucose metabolism is largely unknown. 13C MRS is a unique tool to study cerebral glucose metabolism, but the concurrent requirement for [1-13C]glucose administration limits its use under hypoglycemic conditions. To facilitate 13C MRS data analysis we designed separate [1-13C]glucose infusion protocols for hyperinsulinemic euglycemic and hypoglycemic clamps in such a way that plasma isotopic enrichment of glucose was stable and comparable under both glycemic conditions. 13C MR spectra were acquired with optimized 13C MRS measurement techniques to obtain high quality 13C MR spectra with these protocols. PMID:19913052

  20. Using Headspace Equilibration to Measure the d13C of Soil-Respired CO2

    NASA Astrophysics Data System (ADS)

    Robertson, M. A.; Powers, E.; Marshall, J.

    2007-12-01

    Soil respiration is an important component of the global carbon cycle and can account for as much as 70% of ecosystem respiration. Soil gas flux measurements have been combined with stable isotope analysis to examine ecosystem properties and processes such as water-use efficiency and the role of above ground weather in controlling soil respiration. However, current methods of measuring the δ13C of soil-respired CO2 are either inherently inaccurate or time-consuming and tedious. An alternative method of obtaining this value offers a potential solution to these problems. In this method, plastic chambers are fitted with rubber septa to allow for sample collection, then inverted and partially buried in soil. The chamber headspace is allowed to come to equilibrium with soil air. In this study we tested the viability of this method by examining whether frequent resampling of respiration chambers affected δ13C measurements, whether headspace CO2 concentration and δ13C values approached equilibrium asymptotically, and whether simulated and actual diel temperature cycles affected estimates of δ13C. All experiments were conducted on respiration chambers inverted in potting soil and placed in a Conviron growth chamber, with the exception of one field test that was conducted on respiration chambers installed in a Northern Idaho experimental forest. Samples were collected with a syringe and stored in glass vials for analysis by a ratioing mass spectrometer. We found that resampling respiration chambers as frequently as every 10 minutes had no significant effect on final δ13C values, that both chamber CO2 concentrations and δ13C values exhibited an asymptotic approach to equilibrium, and that the equilibrium value was offset from the initial flux by the amount we expected, approximately 4 ‰. However, we also found that diel temperature variation affected both headspace CO2 concentration and δ13C in the lab and in the field. We concluded that if this method is used in

  1. Nested heat tracer experiments for identifying heterogeneity of aquifer-river exchange at multiple scales

    NASA Astrophysics Data System (ADS)

    Krause, Stefan; Hannah, David; Blume, Theresa; Angermann, Lisa; Lewandowski, Joerg; Cassidy, Nigel

    2016-04-01

    This study presents the nested application of three heat tracing methods for identifying aquifer-river exchange fluxes at multiple scales ranging from centimeter to stream reach-scale. The investigations focus on a UK lowland river where hotspots of redox-reactivity were found to coincide with locations of increased streambed residence times underneath flow confining streambed peat and clay structures. In order to identify the spatial extend and patterns of reactivity hot spots associated with these streambed structures, reach-scale patterns of aquifer-river exchange fluxes have been analysed by Fibre-Optic Distributed Temperature Sensing (FO-DTS) along a cable buried in the streambed of a 250 m reach in combination with 2D thermocouple arrays in a 12 m long pool-riffle-pool sequence and small-scale heat pulse injections for tracing shallow hyporheic flow paths within the uppermost 20cm streambed sediments. FO-DTS observed streambed temperature anomalies caused by the mixing of different temperatures of GW and SW end-members were used to infer information on exchange fluxes at the aquifer-river interface. FO-DTS survey results indicate that patterns of up to 2C colder (Summer) and 3.5C warmer (Winter) temperatures in investigated streambed sediments can be attributed to fast GW up-welling in sandy and gravely sediments. Contrasting conditions were found at locations where streambed temperatures equal SW temperatures and GW-SW exchange was inhibited by the existence of peat or clay lenses within the streambed. FO-DTS observations of regional GW up-welling patterns were complemented by heat pulse injection experiments which provided essential information of the shallow aquifer- river exchange fluxes and confirmed increased SW infiltration and lateral flow in riffle crests and at locations with highly conductive streambed sediments above flow confining low conductivity structures. The propagation of diurnal temperature oscillations from the surface to streambed depths

  2. Site-specific φ- and ψ-torsion angle determination in a uniformly/extensively 13C- and 15N-labeled peptide

    NASA Astrophysics Data System (ADS)

    Wi, Sungsool; Spano, Justin

    2011-10-01

    A solid-state rotational-echo double resonance (REDOR) NMR method was introduced to identify the ϕ- and ψ-torsion angle from a 1H- 15N or 1H- 13C' spin system of alanine-like residues in a selectively, uniformly, or extensively 15N-/ 13C-labeled peptide. When a C α( i) or a 15N peak is site-specifically obtainable in the NMR spectrum of a uniformly 15N/ 13C-labeled sample system, the ψ- or ϕ-torsion angle specified by the conformational structure of peptide geometry involving 15N( i)- 1H αi - 15N( i + 1) or 13C'( i - 1)- 1H Ni- 13C'( i) spin system can be identified based on 13C α- or 15N-detected 1H α- 15N or 1H N- 13C REDOR experiment. This method will conveniently be utilized to identify major secondary motifs, such as α-helix, β-sheet, and β-turn, from a uniformly 15N-/ 13C-labled peptide sample system. When tested on a 13C-/ 15N-labeled model system of a three amino acid peptide Gly-[U- 13C, 15N]Ala-[U- 13C, 15N]Leu, the ψ-angle of alanine obtained experimentally, ψ = -40 ± 30°, agreed reasonably well with the X-ray determined angle, ψ = -39°.

  3. Neutron Backgrounds: 13C({alpha}, n) etc

    SciTech Connect

    Mitsui, Tadao

    2005-09-08

    13C({alpha}, n) reaction is the main neutron source in an underground large-volume liquid-scintillator detector KamLAND. {alpha} sources, targets, cross sections, and neutron transport are studied to estimate the backgrounds of v-bare signal.

  4. Spectral editing for in vivo 13C magnetic resonance spectroscopy

    NASA Astrophysics Data System (ADS)

    Xiang, Yun; Shen, Jun

    2012-01-01

    In vivo detection of carboxylic/amide carbons is a promising technique for studying cerebral metabolism and neurotransmission due to the very low RF power required for proton decoupling. In the carboxylic/amide region, however, there is severe spectral overlap between acetate C1 and glutamate C5, complicating studies that use acetate as an astroglia-specific substrate. There are no known in vivo MRS techniques that can spectrally resolve acetate C1 and glutamate C5 singlets. In this study, we propose to spectrally separate acetate C1 and glutamate C5 by a two-step J-editing technique after introducing homonuclear 13C- 13C scalar coupling between carboxylic/amide carbons and aliphatic carbons. By infusing [1,2- 13C 2]acetate instead of [1- 13C]acetate the acetate doublet can be spectrally edited because of the large separation between acetate C2 and glutamate C4 in the aliphatic region. This technique can be applied to studying acetate transport and metabolism in brain in the carboxylic/amide region without spectral interference.

  5. Does the Shuram δ13C excursion record Ediacaran oxygenation?

    NASA Astrophysics Data System (ADS)

    Husson, J. M.; Maloof, A. C.; Schoene, B.; Higgins, J. A.

    2013-12-01

    The most negative carbon isotope excursion in Earth history is found in carbonate rocks of the Ediacaran Period (635-542 Ma). Known colloquially as the the 'Shuram' excursion, workers have long noted its tantalizing, broad concordance with the rise of abundant macro-scale fossils in the rock record, variously interpreted as animals, giant protists, macro-algae and lichen, and known as the 'Ediacaran Biota.' Thus, the Shuram excursion has been interpreted by many in the context of a dramatically changing redox state of the Ediacaran oceans - e.g., a result of methane cycling in a low O2 atmosphere, the final destruction of a large pool of recalcitrant dissolved organic carbon (DOC), and the step-wise oxidation of the Ediacaran oceans. More recently, diagenetic interpretations of the Shuram excursion - e.g. sedimentary in-growth of very δ13C depleted authigenic carbonates, meteoric alteration of Ediacaran carbonates, late-stage burial diagenesis - have challenged the various Ediacaran redox models. A rigorous geologic context is required to discriminate between these explanatory models, and determine whether the Shuram excursion can be used to evaluate terminal Neoproterozoic oxygenation. Here, we present chemo-stratigraphic data (δ13C, δ18O, δ44/42Ca and redox sensitive trace element abundances) from 12 measured sections of the Ediacaran-aged Wonoka Formation (Fm.) of South Australia that require a syn-depositional age for the extraordinary range of δ13C values (-12 to +4‰) observed in the formation. In some locations, the Wonoka Fm. is ~700 meters (m) of mixed shelf limestones and siliclastics that record the full 16 ‰ δ13C excursion in a remarkably consistent fashion across 100s of square kilometers of basin area. Fabric-altering diagenesis, where present, occurs at the sub-meter vertical scale, only results in sub-permil offsets in δ13C and cannot be used to explain the full δ13C excursion. In other places, the Wonoka Fm. is host to deep (1 km

  6. Optical hyperpolarization of 13C nuclear spins in nanodiamond ensembles

    NASA Astrophysics Data System (ADS)

    Chen, Q.; Schwarz, I.; Jelezko, F.; Retzker, A.; Plenio, M. B.

    2015-11-01

    Dynamical nuclear polarization holds the key for orders of magnitude enhancements of nuclear magnetic resonance signals which, in turn, would enable a wide range of novel applications in biomedical sciences. However, current implementations of DNP require cryogenic temperatures and long times for achieving high polarization. Here we propose and analyze in detail protocols that can achieve rapid hyperpolarization of 13C nuclear spins in randomly oriented ensembles of nanodiamonds at room temperature. Our protocols exploit a combination of optical polarization of electron spins in nitrogen-vacancy centers and the transfer of this polarization to 13C nuclei by means of microwave control to overcome the severe challenges that are posed by the random orientation of the nanodiamonds and their nitrogen-vacancy centers. Specifically, these random orientations result in exceedingly large energy variations of the electron spin levels that render the polarization and coherent control of the nitrogen-vacancy center electron spins as well as the control of their coherent interaction with the surrounding 13C nuclear spins highly inefficient. We address these challenges by a combination of an off-resonant microwave double resonance scheme in conjunction with a realization of the integrated solid effect which, together with adiabatic rotations of external magnetic fields or rotations of nanodiamonds, leads to a protocol that achieves high levels of hyperpolarization of the entire nuclear-spin bath in a randomly oriented ensemble of nanodiamonds even at room temperature. This hyperpolarization together with the long nuclear-spin polarization lifetimes in nanodiamonds and the relatively high density of 13C nuclei has the potential to result in a major signal enhancement in 13C nuclear magnetic resonance imaging and suggests functionalized and hyperpolarized nanodiamonds as a unique probe for molecular imaging both in vitro and in vivo.

  7. TRACER: an ‘eye-opener’ to the patient experience across the transition of care in an internal medicine resident program

    PubMed Central

    Meade, Lauren B.; Hall, Susana L.; Kleppel, Reva W.; Hinchey, Kevin T.

    2015-01-01

    Background A safe patient transition requires a complex set of physician skills within the interprofessional practice. Objective To evaluate a rotation which applies self-reflection and workplace learning in a TRAnsition of CarE Rotation (TRACER) for internal medicine (IM) residents. TRACER is a 2-week required IM resident rotation where trainees join a ward team as a quality officer and follow patients into postacute care. Methods In 2010, residents participated in semistructured, one-on-one interviews as part of ongoing program evaluation. They were asked what they had learned on TRACER, the year prior, and how they used those skills in their practice. Using transcripts, the authors reviewed and coded each transcript to develop themes. Results Five themes emerged from a qualitative, grounded theory analysis: seeing things from the other side, the ‘ah ha’ moment of fragmented care, team collaboration including understanding nursing scope of practice in different settings, patient understanding, and passing the learning on. TRACER gives residents a moment to breathe and open their eyes to the interprofessional practice setting and the patient's experience of care in transition. Conclusions Residents learn about transitions of care through self-reflection. This learning is sustained over time and is valued enough to teach to their junior colleagues. PMID:25846349

  8. Measurement of position-specific 13C isotopic composition of propane at the nanomole level

    NASA Astrophysics Data System (ADS)

    Gilbert, Alexis; Yamada, Keita; Suda, Konomi; Ueno, Yuichiro; Yoshida, Naohiro

    2016-03-01

    We have developed a novel method for analyzing intramolecular carbon isotopic distribution of propane as a potential new tracer of its origin. The method is based on on-line pyrolysis of propane followed by analysis of carbon isotope ratios of the pyrolytic products methane, ethylene and ethane. Using propane samples spiked with 13C at the terminal methyl carbon, we characterize the origin of the pyrolytic fragments. We show that the exchange between C-atoms during the pyrolytic process is negligible, and thus that relative intramolecular isotope composition can be calculated. Preliminary data from 3 samples show that site-preference (SP) values, defined as the difference of δ13C values between terminal and sub-terminal C-atom positions of propane, range from -1.8‰ to -12.9‰. In addition, SP value obtained using our method for a thermogenic natural gas sample is consistent with that expected from theoretical models of thermal cracking, suggesting that the isotope fractionation associated with propane pyrolysis is negligible. The method will provide novel insights into the characterization of the origin of propane and will help better understand the biogeochemistry of natural gas deposits.

  9. Synthesis of [1-.sup.13C]pyruvic acid], [2-.sup.13C]pyruvic acid], [3-.sup.13C]pyruvic acid] and combinations thereof

    DOEpatents

    Martinez, Rodolfo A.; Unkefer, Clifford J.; Alvarez, Marc A.

    2009-09-01

    The present invention is directed to labeled compounds, of the formulae ##STR00001## wherein C* is each independently selected from the group consisting of .sup.13C and .sup.12C with the proviso that at least one C* is .sup.13C, each hydrogen of the methylene group can independently be either hydrogen or deuterium, the methyl group includes either zero or three deuterium atoms, Q is from the group of sulfide, sulfinyl, and sulfone, Z is an aryl group from the group of 1-naphthyl, substituted 1-naphthyl, 2-naphthyl, substituted 2-naphthyl, and phenyl groups with the structure ##STR00002## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently from the group of hydrogen, a C.sub.1-C.sub.4 lower alkyl, a halogen, and an amino group from the group of NH.sub.2, NHR and NRR' where R and R' are each independently from the group of a C.sub.1-C.sub.4 lower alkyl, a phenyl, and an alkoxy group, and the methyl group can include either zero or three deuterium atoms.

  10. Synthesis of [1-.sup.13C]pyruvic acid], [2-.sup.13C]pyruvic acid], [3-.sup.13C]pyruvic acid] and combinations thereof

    DOEpatents

    Martinez, Rodolfo A. , Unkefer; Clifford J. , Alvarez; Marc A.

    2012-06-12

    The present invention is directed to the labeled compounds, ##STR00001## wherein C* is each either .sup.13C and .sup.12C where at least one C* is .sup.13C, each hydrogen of the methylene group is hydrogen or deuterium, the methyl group includes either zero or three deuterium atoms, Q is sulfide, sulfinyl, or sulfone, Z is an aryl group such as 1-naphthyl, substituted 1-naphthyl, 2-naphthyl, substituted 2-naphthyl, or a phenyl group ##STR00002## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5 are each independently either hydrogen, a C.sub.1-C.sub.4 lower alkyl, a halogen, and an amino group such as NH.sub.2, NHR and NRR' where R and R' are each independently either a C.sub.1-C.sub.4 lower alkyl, a phenyl, and an alkoxy group, and the methyl group can include either zero or three deuterium atoms. The present invention is also directed to the labeled compounds ##STR00003##

  11. Strength and limits using 13C phospholipid fatty acid analysis in soil ecology

    NASA Astrophysics Data System (ADS)

    Watzinger, Andrea

    2016-04-01

    This presentation on microbial phospholipid biomarkers, their isotope analysis and their ability to reveal soil functions summarizes experiences gained by the author for more than 10 years. The amount and composition of phospholipid fatty acids (PLFAs) measured in environmental samples strongly depend on the methodology. To achieve comparable results the extraction, separation and methylation method must be kept constant. PLFAs patterns are sensitive to microbial community shifts even though the taxonomic resolution of PLFAs is low. The possibility to easily link lipid biomarkers with stable isotope techniques is identified as a major advantage when addressing soil functions. Measurement of PLFA isotopic ratios is sensitive and enables detecting isotopic fractionation. The difference between the carbon isotopic ratio of single PLFAs and their substrate (δ13C) can vary between -6 and +11‰. This difference derives from the fractionation during biosynthesis and from substrate inhomogeneity. Consequently, natural abundance studies are restricted to quantifying substrate uptake of the total microbial biomass. In contrast, artificial labelling enables quantifying carbon uptake into single PLFAs, but labelling success depends on homogeneous and undisturbed label application. Current developments in microbial ecology (e.g. 13C and 15N proteomics) and isotope techniques (online monitoring of CO2 isotope ratios) will likely improve soil functional interpretations in the future. 13C PLFA analysis will continue to contribute because it is affordable, sensitive and allows frequent sampling combined with the use of small amounts of 13C label.

  12. First airborne samples of a volcanic plume for δ13C of CO2 determinations

    NASA Astrophysics Data System (ADS)

    Fischer, Tobias P.; Lopez, Taryn M.

    2016-04-01

    Volcanic degassing is one of the main natural sources of CO2 to the atmosphere. Carbon isotopes of volcanic gases enable the determination of CO2 sources including mantle, organic or carbonate sediments, and atmosphere. Until recently, this work required sample collection from vents followed by laboratory analyses. Isotope ratio infrared analyzers now enable rapid analyses of plume δ13C-CO2, in situ and in real time. Here we report the first analyses of δ13C-CO2 from airborne samples. These data combined with plume samples from the vent area enable extrapolation to the volcanic source δ13C. We performed our experiment at the previously unsampled and remote Kanaga Volcano in the Western Aleutians. We find a δ13C source composition of -4.4‰, suggesting that CO2 from Kanaga is primarily sourced from the upper mantle with minimal contributions from subducted components. Our method is widely applicable to volcanoes where remote location or activity level precludes sampling using traditional methods.

  13. 13CFLUX2—high-performance software suite for 13C-metabolic flux analysis

    PubMed Central

    Weitzel, Michael; Nöh, Katharina; Dalman, Tolga; Niedenführ, Sebastian; Stute, Birgit; Wiechert, Wolfgang

    2013-01-01

    Summary: 13C-based metabolic flux analysis (13C-MFA) is the state-of-the-art method to quantitatively determine in vivo metabolic reaction rates in microorganisms. 13CFLUX2 contains all tools for composing flexible computational 13C-MFA workflows to design and evaluate carbon labeling experiments. A specially developed XML language, FluxML, highly efficient data structures and simulation algorithms achieve a maximum of performance and effectiveness. Support of multicore CPUs, as well as compute clusters, enables scalable investigations. 13CFLUX2 outperforms existing tools in terms of universality, flexibility and built-in features. Therewith, 13CFLUX2 paves the way for next-generation high-resolution 13C-MFA applications on the large scale. Availability and implementation: 13CFLUX2 is implemented in C++ (ISO/IEC 14882 standard) with Java and Python add-ons to run under Linux/Unix. A demo version and binaries are available at www.13cflux.net. Contact: info@13cflux.net or k.noeh@fz-juelich.de Supplementary information: Supplementary data are available at Bioinformatics online. PMID:23110970

  14. Quantitative identification of metastable magnesium carbonate minerals by solid-state 13C NMR spectroscopy.

    PubMed

    Moore, Jeremy K; Surface, J Andrew; Brenner, Allison; Wang, Louis S; Skemer, Philip; Conradi, Mark S; Hayes, Sophia E

    2015-01-01

    In the conversion of CO2 to mineral carbonates for the permanent geosequestration of CO2, there are multiple magnesium carbonate phases that are potential reaction products. Solid-state (13)C NMR is demonstrated as an effective tool for distinguishing magnesium carbonate phases and quantitatively characterizing magnesium carbonate mixtures. Several of these mineral phases include magnesite, hydromagnesite, dypingite, and nesquehonite, which differ in composition by the number of waters of hydration or the number of crystallographic hydroxyl groups. These carbonates often form in mixtures with nearly overlapping (13)C NMR resonances which makes their identification and analysis difficult. In this study, these phases have been investigated with solid-state (13)C NMR spectroscopy, including both static and magic-angle spinning (MAS) experiments. Static spectra yield chemical shift anisotropy (CSA) lineshapes that are indicative of the site-symmetry variations of the carbon environments. MAS spectra yield isotropic chemical shifts for each crystallographically inequivalent carbon and spin-lattice relaxation times, T1, yield characteristic information that assist in species discrimination. These detailed parameters, and the combination of static and MAS analyses, can aid investigations of mixed carbonates by (13)C NMR.

  15. Direct 13C-detection for carbonyl relaxation studies of protein dynamics.

    PubMed

    Pasat, Gabriela; Zintsmaster, John S; Peng, Jeffrey W

    2008-08-01

    We describe a method that uses direct 13C-detection for measuring rotating-frame carbonyl (13CO) relaxation rates to describe protein functional dynamics. Key advantages of method include the following: (i) unique access to 13CO groups that lack a scalar-coupled 15N-1H group; (ii) insensitivity to 15N/1H exchange-broadening that can derail 1H-detected 15N and HNCO methods; (iii) avoidance of artifacts caused by incomplete water suppression. We demonstrate the approach for both backbone and side-chain 13CO groups. Accuracy of the 13C-detected results is supported by their agreement with those obtained from established HNCO-based approaches. Critically, we show that the 13C-detection approach provides access to the 13CO groups of functionally important residues that are invisible via 1H-detected HNCO methods because of exchange-broadening. Hence, the 13C-based method fills gaps inherent in canonical 1H-detected relaxation experiments, and thus provides a novel complementary tool for NMR studies of biomolecular flexibility. PMID:18514001

  16. Carbon fluxes to the soil in a mature temperate forest assessed by 13C isotope tracing.

    PubMed

    Steinmann, Katharina; Siegwolf, Rolf T W; Saurer, Matthias; Körner, Christian

    2004-11-01

    Photosynthetic carbon uptake and respiratory C release from soil are major components of the global carbon balance. The use of 13C depleted CO2)(delta13C = -30 per thousand) in a free air CO2 enrichment experiment in a mature deciduous forest permitted us to trace the carbon transfer from tree crowns to the rhizosphere of 100-120 years old trees. During the first season of CO2 enrichment the CO2 released from soil originated substantially from concurrent assimilation. The small contribution of recent carbon in fine roots suggests a much slower fine root turnover than is often assumed. 13C abundance in soil air correlated best with temperature data taken from 4 to 10 days before air sampling time and is thus rapidly available for root and rhizosphere respiration. The spatial variability of delta13C in soil air showed relationships to above ground tree types such as conifers versus broad-leaved trees. Considering the complexity and strong overlap of roots from different individuals in a forest, this finding opens an exciting new possibility of associating respiration with different species. What might be seen as signal noise does in fact contain valuable information on the spatial heterogeneity of tree-soil interaction.

  17. Tracers in rainfall simulation experiments to study the onset of the wet season in Eastern Mediterranean limestone environments

    NASA Astrophysics Data System (ADS)

    Lange, Jens

    2010-05-01

    The eastern Mediterranean rainfall regime is characterized by dry and hot summers and rainy cold winters. In this climate rocky limestone environments are regarded as major recharge zones due to (a) intensively enlarged fissures by solution weathering and (b) sparse vegetation and shallow soils which limit evapotranspiration losses. However, relatively little is known on hydrological processes during high magnitude rainstorms, which, at the beginning of the rainy season, may occur on both dry and wet soils. These conditions were investigated by a series of sprinkling experiments during two successive days. Rainfall was applied on large plots (143 and 180 m2) to include the variety of different terrain elements (rocky outcrops, bare soil, different vegetation). Sprinkling units were located at each corner of the plot and supplemented by additional ones to balance wind drift. This sprinkling set-up did not guarantee a uniform distribution of applied rainfall, as overlap of sprinkling areas could not be prevented. To assess the spatial rainfall distribution, a large number of totalizers was necessary. During two days of sprinkling these totalizers were regularly measured and spatially interpolated across the plot. The temporal rainfall distribution, a series of two high intensity storms on dry and wet soil, was observed by a tipping bucket raingauge. Tracers were added to the sprinkling water to obtain additional process insights. By end member mixing analysis the contribution of different water types (pre-sprinkling, first day, second day) could be quantified. The first plot was located on a steep rocky hillslope. Significantly different concentration of chloride, nitrate and sulfate in the sprinkling waters helped to identify first day's water in second day's runoff. Surface runoff was a combination of infiltration excess runoff from rocky portions of the plot and saturation excess runoff from areas covered by soil. Soil saturation was accelerated by lateral runoff

  18. Preliminary Results from Downhole Osmotic Samplers in a Gas Tracer Injection Experiment in the Upper Oceanic Crust on the Eastern Flank of the Juan de Fuca Ridge.

    NASA Astrophysics Data System (ADS)

    de Jong, M. T.; Clark, J. F.; Neira, N. M.; Fisher, A. T.; Wheat, C. G.

    2015-12-01

    We present results from a gas tracer injection experiment in the ocean crust on the eastern flank of the Juan de Fuca Ridge, in an area of hydrothermal circulation. Sulfur hexafluoride (SF6) tracer was injected in Hole 1362B in 2010, during IODP Expedition 327. Fluid samples were subsequently collected from a borehole observatory (CORK) installed in this hole and similar CORKs in three additional holes (1026B, 1362A, and 1301A), located 300 to 500 m away. This array of holes is located on 3.5 My old seafloor, as an array oriented subparallel to the Endeavor Segment of Juan de Fuca Ridge. Borehole fluid samples were collected in copper coils using osmotic pumps. In addition to pumps at seafloor wellheads, downhole sampling pumps were installed in the perforated casing in the upper ocean crust. These downhole samplers were intended to produce a high-resolution continuous record of tracer concentrations, including records from the first year after tracer injection in Holes 1362A and 1362B. In contrast, wellhead samplers were not installed on these CORKs holes until 2011, and wellhead records from all CORKs have a record gap of up to one year, because of a delayed expedition in 2012. The downhole samples were recovered with the submersible Alvin in August 2014. SF6 concentrations in downhole samples recovered in 2014 are generally consistent with data obtained from wellhead samples. Of particular interest are the results from Hole 1362B, where a seafloor valve was opened and closed during various recovery expeditions. High resolution tracer curves produced from the 1362B downhole samples confirm that these operations produced an SF6 breakthrough curve corresponding to a classic push-pull test used to evaluate contaminant field locations in terrestrial setting. Complete analyses of downhole samples from these CORKs are expected to produce high-resolution breakthrough curves that will allow more precise analysis and modeling of hydrothermal flow in the study area.

  19. 13C NMR spectra of pyridine chalcone analogs

    NASA Astrophysics Data System (ADS)

    Jovanović, B. Ž.; Mišić-Vuković, M.; Marinković, A. D.; Csanádi, J.

    1999-05-01

    13C NMR spectra of two series of pyridine chalcone analogs were determined in deuterated dimethylsulphoxide (DMSO-d 6). It was established that these compounds were in more stable E-configurations except for the 4-pyridalacetophenone which was in Z-configuration. On the basis of the Hammett correlations of 13C NMR chemical shifts of the ethylenic bond carbon atoms and the σ values for the pyridine "aza" groups, the polarization of ethylenic bonds were discussed. It was established that the opposite effect of the pyridine substituents at the electronic density distribution in pyridalacetophenone and cinnamoylpyridines depends on its direct bonding to the ethylenic carbon or the transmission electronic effects via the carbonyl group, respectively.

  20. 13C solid-state NMR chemical shift anisotropy analysis of the anomeric carbon in carbohydrates.

    PubMed

    Chen, Ying-Ying; Luo, Shun-Yuan; Hung, Shang-Cheng; Chan, Sunney I; Tzou, Der-Lii M

    2005-03-21

    (13)C NMR solid-state structural analysis of the anomeric center in carbohydrates was performed on six monosaccharides: glucose (Glc), mannose (Man), galactose (Gal), galactosamine hydrochloride (GalN), glucosamine hydrochloride (GlcN), and N-acetyl-glucosamine (GlcNAc). In the 1D (13)C cross-polarization/magic-angle spinning (CP/MAS) spectrum, the anomeric center C-1 of these carbohydrates revealed two well resolved resonances shifted by 3-5ppm, which were readily assigned to the anomeric alpha and beta forms. From this experiment, we also extracted the (13)C chemical shift anisotropy (CSA) tensor elements of the two forms from their spinning sideband intensities, respectively. It was found out that the chemical shift tensor for the alpha anomer was more axially symmetrical than that of the beta form. A strong linear correlation was obtained when the ratio of the axial asymmetry of the (13)C chemical shift tensors of the two anomeric forms was plotted in a semilogarithmic plot against the relative population of the two anomers. Finally, we applied REDOR spectroscopy to discern whether or not there were any differences in the sugar ring conformation between the anomers. Identical two-bond distances of 2.57A (2.48A) were deduced for both the alpha and beta forms in GlcNAc (GlcN), suggesting that the two anomers have essentially identical sugar ring scaffolds in these sugars. In light of these REDOR distance measurements and the strong correlation observed between the ratio of the axial asymmetry parameters of the (13)C chemical shift tensors and the relative population between the two anomeric forms, we concluded that the anomeric effect arises principally from interaction of the electron charge clouds between the C-1-O-5 and the C-1-O-1 bonds in these monosaccharides.

  1. Metabolic Imaging in the Anesthetized Rat Brain Using Hyperpolarized [1-13C] Pyruvate and [1-13C] Ethyl Pyruvate

    PubMed Central

    Hurd, Ralph E.; Yen, Yi-Fen; Mayer, Dirk; Chen, Albert; Wilson, David; Kohler, Susan; Bok, Robert; Vigneron, Daniel; Kurhanewicz, John; Tropp, James; Spielman, Daniel; Pfefferbaum, Adolf

    2010-01-01

    Formulation, polarization, and dissolution conditions were developed to obtain a stable hyperpolarized solution of [1-13C]-ethyl pyruvate. A maximum tolerated concentration and injection rate were determined, and 13C spectroscopic imaging was used to compare the uptake of hyperpolarized [1-13C]-ethyl pyruvate relative to hyperpolarized [1-13C]-pyruvate into anesthetized rat brain. Hyperpolarized [1-13C]-ethyl pyruvate and [1-13C]-pyruvate metabolic imaging in normal brain is demonstrated and quantified in this feasibility and range-finding study. PMID:20432284

  2. Direct uptake of organically derived carbon by grass roots and allocation in leaves and phytoliths: 13C labeling evidence

    NASA Astrophysics Data System (ADS)

    Alexandre, Anne; Balesdent, Jérôme; Cazevieille, Patrick; Chevassus-Rosset, Claire; Signoret, Patrick; Mazur, Jean-Charles; Harutyunyan, Araks; Doelsch, Emmanuel; Basile-Doelsch, Isabelle; Miche, Hélène; Santos, Guaciara M.

    2016-03-01

    In the rhizosphere, the uptake of low-molecular-weight carbon (C) and nitrogen (N) by plant roots has been well documented. While organic N uptake relative to total uptake is important, organic C uptake is supposed to be low relative to the plant's C budget. Recently, radiocarbon analyses demonstrated that a fraction of C from the soil was occluded in amorphous silica micrometric particles that precipitate in plant cells (phytoliths). Here, we investigated whether and to what extent organically derived C absorbed by grass roots can feed the C occluded in phytoliths. For this purpose we added 13C- and 15N-labeled amino acids (AAs) to the silicon-rich hydroponic solution of the grass Festuca arundinacea. The experiment was designed to prevent C leakage from the labeled nutritive solution to the chamber atmosphere. After 14 days of growth, the 13C and 15N enrichments (13C excess and 15N excess) in the roots, stems and leaves as well as phytoliths were measured relative to a control experiment in which no labeled AAs were added. Additionally, the 13C excess was measured at the molecular level, in AAs extracted from roots and stems and leaves. The net uptake of labeled AA-derived 13C reached 4.5 % of the total AA 13C supply. The amount of AA-derived 13C fixed in the plant was minor but not nil (0.28 and 0.10 % of total C in roots and stems/leaves, respectively). Phenylalanine and methionine that were supplied in high amounts to the nutritive solution were more 13C-enriched than other AAs in the plant. This strongly suggested that part of AA-derived 13C was absorbed and translocated into the plant in its original AA form. In phytoliths, AA-derived 13C was detected. Its concentration was on the same order of magnitude as in bulk stems and leaves (0.15 % of the phytolith C). This finding strengthens the body of evidences showing that part of organic compounds occluded in phytoliths can be fed by C entering the plant through the roots. Although this experiment was done in

  3. 1H-detected 13C Photo-CIDNP as a Sensitivity Enhancement Tool in Solution NMR

    PubMed Central

    Lee, Jung Ho; Sekhar, Ashok; Cavagnero, Silvia

    2011-01-01

    NMR is a powerful yet intrinsically insensitive technique. The applicability of NMR to chemical and biological systems would be substantially extended by new approaches going beyond current signal-to-noise capabilities. Here, we exploit the large enhancements arising from 13C photo-chemically induced dynamic nuclear polarization (13C photo-CIDNP) in solution to improve biomolecular NMR sensitivity in the context of heteronuclear correlation spectroscopy. The 13C-PRINT pulse sequence presented here involves an initial 13C nuclear spin polarization via photo-CIDNP followed by conversion to antiphase coherence and transfer to 1H for detection. We observe substantial enhancements, up to ≫200-fold, relative to the dark (laser off) experiment. Resonances of both side-chain and backbone CH pairs are enhanced for the three aromatic residues Trp, His and Tyr and the Trp-containing σ32 peptide. The sensitivity of this experiment, defined as signal-to-noise per unit time (S/N)t, is unprecedented in the NMR polarization enhancement literature dealing with polypeptides in solution. Up to a 16-fold larger (S/N)t than the 1H-13C SE-HSQC reference sequence is achieved, for the σ32 peptide. This gain leads to a reduction in data collection time up to 256-fold, highlighting the advantages of 1H-detected 13C photo-CIDNP in solution NMR. PMID:21548581

  4. Estimation of continuous anthropogenic CO2 using CO2, CO, δ13C(CO2) and Δ14C(CO2)

    NASA Astrophysics Data System (ADS)

    Vardag, S. N.; Gerbig, C.; Janssens-Maenhout, G.; Levin, I.

    2015-07-01

    We investigate different methods for estimating anthropogenic CO2 using modelled continuous atmospheric concentrations of CO2 alone, as well as CO2 in combination with the surrogate tracers CO, δ13C(CO2) and Δ14C(CO2). These methods are applied at three hypothetical stations representing rural, urban and polluted conditions. We find that independent of the tracer used, an observation-based estimate of continuous anthropogenic CO2 is not feasible at rural measurement sites due to the low signal to noise ratio of anthropogenic CO2 estimates at such settings. At urban and polluted sites, potential future continuous Δ14C(CO2) measurements with a precision of 5 ‰ or better are most promising for anthropogenic CO2 determination (precision ca. 10-20%), but the insensitivity against CO2 contributions from biofuel emissions may reduce its accuracy in the future. Other tracers, such as δ13C(CO2) and CO could provide an accurate and already available alternative if all CO2 sources in the catchment area are well characterized with respect to their isotopic signature and CO to anthropogenic CO2 ratio. We suggest a strategy for calibrating these source characteristics on an annual basis using precise Δ14C(CO2) measurements on grab samples. The precision of anthropogenic CO2 determination using δ13C(CO2) is largely determined by the measurement precision of δ13C(CO2) and CO2. The precision when using the CO-method is mainly limited by the variation of natural CO sources and CO sinks. At present, continuous anthropogenic CO2 could be determined using the tracers δ13C(CO2) and/or CO with a precision of about 30 %, a mean bias of about 10 % and without significant diurnal discrepancies. This allows significant improvement, validation and bias reduction of highly resolved emission inventories using atmospheric observation and regional modelling.

  5. A field experiment and numerical modeling of a tracer at a gravel beach in Prince William Sound, Alaska

    NASA Astrophysics Data System (ADS)

    Guo, Qiaona; Li, Hailong; Boufadel, Michel C.; Liu, Jin

    2014-12-01

    Oil from the 1989 Exxon Valdez oil spill persists in many gravel beaches in Prince William Sound (Alaska, USA), despite great remedial efforts. A tracer study using lithium at a gravel beach on Knight Island, Prince William Sound, during the summer of 2008 is reported. The tracer injection and transport along a transect were simulated using the two-dimensional numerical model MARUN. Model results successfully reproduced the tracer concentrations observed at wells along the transect. A sensitivity analysis revealed that the estimated parameters are well determined. The simulated spatial distribution of tracer indicated that nutrients applied along the transect for bioremediation purposes would be washed to the sea very quickly (within a semi-diurnal tidal cycle) by virtue of the combination of the two-layered beach structure, the tidal fluctuation and the freshwater flow from inland. Thus, pore-water samples in the transect were found to be clean due to factors other than bioremediation. This may explain why the oil did not persist within the transect.

  6. Assessment of local hydraulic properties from electrical resistivity tomography monitoring of a three-dimensional synthetic tracer test experiment

    NASA Astrophysics Data System (ADS)

    Camporese, M.; Cassiani, G.; Deiana, R.; Salandin, P.

    2011-12-01

    In recent years geophysical methods have become increasingly popular for hydrological applications. Time-lapse electrical resistivity tomography (ERT) represents a potentially powerful tool for subsurface solute transport characterization since a full picture of the spatiotemporal evolution of the process can be obtained. However, the quantitative interpretation of tracer tests is difficult because of the uncertainty related to the geoelectrical inversion, the constitutive models linking geophysical and hydrological quantities, and the a priori unknown heterogeneous properties of natural formations. Here an approach based on the Lagrangian formulation of transport and the ensemble Kalman filter (EnKF) data assimilation technique is applied to assess the spatial distribution of hydraulic conductivity K by incorporating time-lapse cross-hole ERT data. Electrical data consist of three-dimensional cross-hole ERT images generated for a synthetic tracer test in a heterogeneous aquifer. Under the assumption that the solute spreads as a passive tracer, for high Peclet numbers the spatial moments of the evolving plume are dominated by the spatial distribution of the hydraulic conductivity. The assimilation of the electrical conductivity 4D images allows updating of the hydrological state as well as the spatial distribution of K. Thus, delineation of the tracer plume and estimation of the local aquifer heterogeneity can be achieved at the same time by means of this interpretation of time-lapse electrical images from tracer tests. We assess the impact on the performance of the hydrological inversion of (i) the uncertainty inherently affecting ERT inversions in terms of tracer concentration and (ii) the choice of the prior statistics of K. Our findings show that realistic ERT images can be integrated into a hydrological model even within an uncoupled inverse modeling framework. The reconstruction of the hydraulic conductivity spatial distribution is satisfactory in the portion

  7. Ensemble-based simultaneous emission estimates and improved forecast of radioactive pollution from nuclear power plant accidents: application to ETEX tracer experiment.

    PubMed

    Zhang, X L; Li, Q B; Su, G F; Yuan, M Q

    2015-04-01

    The accidental release of radioactive materials from nuclear power plant leads to radioactive pollution. We apply an augmented ensemble Kalman filter (EnKF) with a chemical transport model to jointly estimate the emissions of Perfluoromethylcyclohexane (PMCH), a tracer substitute for radionuclides, from a point source during the European Tracer Experiment, and to improve the forecast of its dispersion downwind. We perturb wind fields to account for meteorological uncertainties. We expand the state vector of PMCH concentrations through continuously adding an a priori emission rate for each succeeding assimilation cycle. We adopt a time-correlated red noise to simulate the temporal emission fluctuation. The improved EnKF system rapidly updates (and reduces) the excessively large initial first-guess emissions, thereby significantly improves subsequent forecasts (r = 0.83, p < 0.001). It retrieves 94% of the total PMCH released and substantially reduces transport error (>80% average reduction of the normalized mean square error).

  8. Synthesis and applications of selectively {sup 13}C-labeled RNA

    SciTech Connect

    SantaLucia, J. Jr.; Shen, L.X.; Lewis, H.; Cai, Z.; Tinoci, I. Jr.

    1994-12-01

    Spectral overlap is a substantial problem in NMR studies of RNA molecules >30 nucleotides. To overcome this difficulty, we synthesized selectively {sup 13}C-labeled RNAs and adapted several isotope-edited two- and three-dimensional NMR experiments originally developed for protein studies. We optimized protocols for synthesis of multi-gram quantities of CTP, UTp, ATP, and GTP using a combination of synthetic organic and enzymatic methods. Uracil is prepared in 40 to 50% yield from {sup 13}C-cyanide in two steps. Using acetyl- tribenzoyl-ribose and standard chemistry uracil is then attached to the sugar (90% yield). The tribenzoyl-uridine intermediate is converted into uridine or cytidine quantitatively, depending on the deblocking protocol. Labeled purines are synthesized using simple pyrimidine precursors and reacting with {sup 13}C-formic acid (80% yield). Purine nucleosides are then synthesized using uridine phosphorylase and purine nucleoside phosphorylase. The nucleosides were converted to NMPs by treatment with POC1{sub 3} in triethylphosphate. We converted NMPs to NTPs by standard enzymatic methods. Selectively labeled RNAs were synthesized by run-off transcription using {sup 13}C-labeled NTPs. Several different strategies help solve over-lap problems in larger RNAs. Isotope-edited two-dimensional NMR experiments such as {omega}1-1/2 X-filtered NOESY simplify NMR spectra by dividing the normal NOESY spectrum into two subspectra-one involving NOEs from protons bound to {sup 12}C and one from protons bound to {sup 13}C. For example, we labeled A and U residues of a 34-nucleotide pseudoknot, and the {sup 12}C subspectrum of the 1/2 X-filtered NOESY contained NOEs only from G and C residues (along with adenine 2H); the {sup 13}C subspectrum contained NOEs only from A and U residues. Each subspectrum has less overlap than the NOESY of an unlabeled sample; the editing strategy allows each resonance to be identified by residue type (A, C, G, or U).

  9. 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.

  10. Attempting to link hydro-morphology, transient storage and metabolism in streams: Insights from reactive tracer experiments

    NASA Astrophysics Data System (ADS)

    Kurz, Marie J.; Schmidt, Christian; Blaen, Phillip; Knapp, Julia L. A.; Drummond, Jennifer D.; Martí, Eugenia; Zarnetske, Jay P.; Ward, Adam S.; Krause, Stefan

    2016-04-01

    In-stream transient storage zones, including the hyporheic zone and vegetation beds, can be hotspots of biogeochemical processing in streams, enhancing ecosystem functions such as metabolism and nutrient uptake. The spatio-temporal dynamics and reactivity of these storage zones are influenced by multiple factors, including channel geomorphology, substrate composition and hydrology, and by anthropogenic modifications to flow regimes and nutrient loads. Tracer injections are a commonly employed method to evaluate solute transport and transient storage in streams; however, reactive tracers are needed to differentiate between metabolically active and inactive transient storage zones. The reactive stream tracer resazurin (Raz), a weakly fluorescent dye which irreversibly transforms to resorufin (Rru) under mildly reducing conditions, provides a proxy for aerobic respiration and an estimate of the metabolic activity associated with transient storage zones. Across a range of lotic ecosystems, we try to assess the influence of stream channel hydro-morphology, morphologic heterogeneity, and substrate type on reach (103 m) and sub-reach (102 m) scale transient storage, respiration, and nutrient uptake. To do so, we coupled injections of Raz and conservative tracers (uranine and/or salt) at each study site. The study sites included: vegetated mesocosms controlled for water depth; vegetated and un-vegetated sediment-filled mesocosms fed by waste-water effluent; a contrasting sand- vs. gravel-bedded lowland stream (Q = 0.08 m3/s); and a series of upland streams with varying size (Q = 0.1 - 1.5 m3/s) and prevalence of morphologic features. Continuous time-series of tracer concentrations were recorded using in-situ fluorometers and EC loggers. At the stream sites, time-series were recorded at multiple downstream locations in order to resolve sub-reach dynamics. Analyses yielded highly variable transport metrics and Raz-Rru transformation between study sites and between sub

  11. Detection of intracellular lactate with localized diffusion { 1H- 13C}-spectroscopy in rat glioma in vivo

    NASA Astrophysics Data System (ADS)

    Pfeuffer, Josef; Lin, Joseph C.; DelaBarre, Lance; Ugurbil, Kamil; Garwood, Michael

    2005-11-01

    The aim of this study was to compare the diffusion characteristic of lactate and alanine in a brain tumor model to that of normal brain metabolites known to be mainly intracellular such as N-acetylaspartate or creatine. The diffusion of 13C-labeled metabolites was measured in vivo with localized NMR spectroscopy at 9.4 T (400 MHz) using a previously described localization and editing pulse sequence known as ACED-STEAM ('adiabatic carbon editing and decoupling'). 13C-labeled glucose was administered and the apparent diffusion coefficients of the glycolytic products, { 1H- 13C}-lactate and { 1H- 13C}-alanine, were determined in rat intracerebral 9L glioma. To obtain insights into { 1H- 13C}-lactate compartmentation (intra- versus extracellular), the pulse sequence used very large diffusion weighting (50 ms/μm 2). Multi-exponential diffusion attenuation of the lactate metabolite signals was observed. The persistence of a lactate signal at very large diffusion weighting provided direct experimental evidence of significant intracellular lactate concentration. To investigate the spatial distribution of lactate and other metabolites, 1H spectroscopic images were also acquired. Lactate and choline-containing compounds were consistently elevated in tumor tissue, but not in necrotic regions and surrounding normal-appearing brain. Overall, these findings suggest that lactate is mainly associated with tumor tissue and that within the time-frame of these experiments at least some of the glycolytic product ([ 13C] lactate) originates from an intracellular compartment.

  12. Fossil chironomid d13C as a new proxy for past methanogenic contribution to benthic food-webs in lakes?

    NASA Astrophysics Data System (ADS)

    van Hardenbroek, M.; Heiri, O. M.; Grey, J.; Bodelier, P. L. E.; Lotter, A. F.

    2009-04-01

    Lake sediments are an important source of atmospheric methane. Methanogenic archaea in lake sediments produce 13C-depleted methane that is partly released to the water column and the atmosphere. Another part is utilized by methane oxidizing bacteria (MOB) that are an important food source for deposit-feeding chironomid larvae (Diptera: Chironomidae). If methane-derived carbon is a significant component of the chironomid diet this will lead to strongly negative d13C in the tissue and exoskeleton of chironomid larvae. Chironomid cuticles, especially the strongly sclerotized head capsules, are well preserved as fossils in lake sediments. If the relationship between modern methane fluxes in lakes and chironomid d13C can be established this would therefore provide an approach for estimating past methane fluxes based on d13C of fossil chironomid remains. Using culturing experiments we show that the stable carbon isotope signature of MOB and other food sources can be traced in chironomid muscle tissue as well as in the fossilizing exoskeleton. In addition we measured d13C in chironomid larval head capsules and other invertebrate remains from a range of surface and downcore sediment samples. Small intra-specific variability (-27.1 ± 0.08 permille) was measured in replicate samples of chironomid head capsules of Corynocera ambigua (n=7). d13C of chironomid head capsules from a several different taxa ranged from -28.0 to -25.8 permille, but in some instances we observed d13C values as low as -36.9 to -31.5 permille, suggesting that carbon from MOB can be successfully traced in fossil and subfossil chironomid remains. Our results demonstrate that the stable carbon isotope signature of MOB is incorporated into chironomid head capsules. Future research will focus on quantifying the relationship between methane fluxes, MOB, and head capsule d13C in order to reconstruct past methane fluxes based on the lake sediment record.

  13. The 13C nuclear magnetic resonance in graphite intercalation compounds

    NASA Technical Reports Server (NTRS)

    Tsang, T.; Resing, H. A.

    1985-01-01

    The (13)C NMR chemical shifts of graphite intercalation compounds were calculated. For acceptor types, the shifts come mainly from the paramagnetic (Ramsey) intra-atomic terms. They are related to the gross features of the two-dimensional band structures. The calculated anisotropy is about -140 ppm and is independent of the finer details such as charge transfer. For donor types, the carbon 2p pi orbitals are spin-polarized because of mixing with metal conduction electrons, thus there is an additional dipolar contribution which may be correlated with the electronic specific heat. The general agreement with experimental data is satisfactory.

  14. The delta 13C record of Devonian to Permian carbonates

    NASA Astrophysics Data System (ADS)

    Buggisch, W.

    2003-04-01

    A δ13Ccarb curve will be presented for samples spanning the time interval from the Silurian/Devonian to the Permian/Triassic boundary. Reliable data are usually based on analyses of brachiopod shells. Because of the huge reservoir of carbon in carbonates, also whole rock samples are suitable for stable carbon isotope analyses if they are not altered by meteoric water or by incorporation of re-oxidized organic carbon during diagenesis. There are several possibilities to test the quality of the data: (1) comparison of the δ13C record of whole rock samples with samples from brachiopod shells, (2) with the organic record, (3) analyses of the same time interval in different sections. If the same pattern of isotope data is observed in separa-ted palaeogeographic settings, it is probably caused by a change in the global carbon reservoir. Reliable δ13C data will be presented for the Devonian, Mississippian and Middle to Late Permian. During Pennsylvanian and Early Permian most carbonates were affected by meteoric diagenesis due to the large glacio-eustatic sea level changes of the Permo-Carboniferous glaciation. Long term variations (mean values for 10 Ma) are known from literature. Devonian δ13C values are about 0 to 2 ppm (V-PDB) they increase up to 5 to 6 ppm during the Mississippian - Pennsylvanian transition and drop sharply at the Permian Triassic boundary. The Devonian - Carboniferous trend is probably at least partly due to the evolution of land plants. Short term variations in the range of 0.1 to 1 Ma modify the long term trend significantly. Large positive excursions of δ13C up to 5 or 6 ppm are known from the Silurian - Devonian boundary and during the Middle Tournaisian of Laurentia and Europe. Many positive excursions of a magnitude of 2 to 3 ppm are observed, some are verified worldwide as for instance at the Frasnian - Famennian boundary which coincides with one of the largest extinction events in earth history. Short time variations in the isotopic

  15. S-Factor of radiative р 13C capture

    NASA Astrophysics Data System (ADS)

    Dubovichenko, S. B.

    2012-06-01

    The possibility of description of experimental data on the astrophysical S-factor of radiative р 13C capture within the framework of the potential cluster model with forbidden states is analyzed at energies in the range 0.03-0.8 MeV. It is demonstrated that the behavior of the astrophysical S-factor can be explained based on the Е1-transition to the bound 3 P 1 state of the 14N nucleus in the р 13С channel from the 3 S 1 wave of р 13С scattering at resonant energy of 0.55 MeV (l.s.).

  16. Design of a sup 13 C (1H) RF probe for monitoring the in vivo metabolism of (1- sup 13 C)glucose in primate brain

    SciTech Connect

    Hammer, B.E.; Sacks, W.; Bigler, R.E.; Hennessy, M.J.; Sacks, S.; Fleischer, A.; Zanzonico, P.B. )

    1990-01-01

    The design of an RF probe suitable for obtaining proton-decoupled {sup 13}C spectra from a subhuman primate brain is described. Two orthogonal saddle coils, one tuned to the resonant frequency of {sup 13}C and the other to the resonant frequency of 1H, were used to monitor the in vivo metabolism of (1-{sup 13}C)glucose in rhesus monkey brain at 2.1 T. Difference spectra showed the appearance of {sup 13}C-enriched glutamate and glutamine 30 to 40 min after a bolus injection of (1-{sup 13}C)glucose.

  17. Insights and questions raised from a multi-tracer plot-scale sprinkler experiment with time-lapse 3D GPR in a structured forested soil.

    NASA Astrophysics Data System (ADS)

    Jackisch, Conrad; Sprenger, Matthias; Allroggen, Niklas; van Schaik, Loes; Weiler, Markus; Zehe, Erwin

    2014-05-01

    Stable isotopes appear as ideal tracer commonly applied in preferential flow analyses. At the same time, central assumptions about signature mixing and propagation are founded on effective parameters merging advective and diffusive flow domains. However, in structured soils conditions are often far from well-mixed and some established assumptions may need to be reconsidered. We conducted a multi-tracer sprinkler experiment at a forested hillslope in the Attert Basin in Luxembourg with prevailing geogenic and biogenic preferential flow structures. At plot scale of 1x1 m2 we sprinkled two plots with 50 mm and one plot with 30 mm Brilliant Blue and Bromide enriched water for 1 hour. The experiments were accompanied by a high resolution 3D time-lapse GPR (Ground-Penetrating Radar) survey scanning 3x3 m2 before, directly after sprinkling and before excavation one day after sprinkling. Soil moisture was monitored with a TDR tube probe. Soil profiles were excavated and recorded for dye flow paths and for one medium resolution Bromide profile. In addition one core for pore water stable isotope analysis was taken before the sprinkling as reference and at each plot after sprinkling. We present the results with focus on the found evidence of preferential flow and the signals of the different tracers - especially the stable isotopes. While all other methods clearly show that only minor proportions of the soil took part in the infiltration process and that the sprinkler water has largely advectively propagated to the saprolite layer at about 80-100 cm depth, the stable isotopes signals from the cores indicate more intense interaction between the soil matrix and macropores, especially in the top 50 cm. This leads to the question of how the isotope signal could mix well, when most of the pore-water did not directly interact with the infiltration-water. Further questions arise to the use of tracers in general, due to the known limitations of excavation itself and rather coarse

  18. Carbon dioxide induced ocean climatic change and tracer experiment with an atmosphere-ocean general circulation model

    SciTech Connect

    Jiang, Xingjian.

    1991-01-01

    The principal objective of this study is to determine whether or not the penetration of a passive tracer is analogous to the penetration of a greenhouse-gas-induced heating. The Atmosphere Ocean General Circulation Model (A-O GCM) has been used to study CO2-induced climate change and the penetration of passive tracers into the world ocean. The present climate and a 2 x CO2 climate have been simulated. The passive tracers tritium, CFC-11, CFC-12 and a 'passive CO2- induced heating' are simulated. The CO2-induced active and passive warmings are larger in the subtropics and high latitudes than in the tropics. The largest difference between the active and passive CO2-induced heatings occur in the North Atlantic deep ocean, with maximum cooling about -1.5C for the active case in layer four of the ocean (1150m). There is no hemispherically asymmetric warming as that found by Manabe et al. (1990) and Stouffer et al. (1990). The convective overturning and large-scale sinking motion are responsible for the large penetration of CO2-induced warming in high latitudes. The CO2-induced circulation changes show that the North Atlantic thermohaline circulation is significantly weakened due to the penetration of CO2-induced heating. Associated with this change, the strength of North Atlantic conveyor belt is reduced, which results in a large warming in the upper ocean and cooling in the deep layers. The characteristic response time ranges from 40-50 years for the active CO2-induced climate change, and 70-160 years for passive CO2-induced climate change. The physical processes controlling the geochemical tracer penetration are very similar to those for the CO2-induced heating. There is not a single tracer which penetrates into the ocean exactly like the active CO2-induced heating in terms of distribution, transport or physical process. CFC's may be the best candidate as a surrogate for the CO2-induced oceanic climate study.

  19. Measuring changes in substrate utilization in the myocardium in response to fasting using hyperpolarized [1-13C]butyrate and [1-13C]pyruvate

    PubMed Central

    Bastiaansen, Jessica A. M.; Merritt, Matthew E.; Comment, Arnaud

    2016-01-01

    Cardiac dysfunction is often associated with a shift in substrate preference for ATP production. Hyperpolarized (HP) 13C magnetic resonance spectroscopy (MRS) has the unique ability to detect real-time metabolic changes in vivo due to its high sensitivity and specificity. Here a protocol using HP [1-13C]pyruvate and [1-13C]butyrate is used to measure carbohydrate versus fatty acid metabolism in vivo. Metabolic changes in fed and fasted Sprague Dawley rats (n = 36) were studied at 9.4 T after tail vein injections. Pyruvate and butyrate competed for acetyl-CoA production, as evidenced by significant changes in [13C]bicarbonate (−48%), [1-13C]acetylcarnitine (+113%), and [5-13C]glutamate (−63%), following fasting. Butyrate uptake was unaffected by fasting, as indicated by [1-13C]butyrylcarnitine. Mitochondrial pseudoketogenesis facilitated the labeling of the ketone bodies [1-13C]acetoacetate and [1-13C]β-hydroxybutyryate, without evidence of true ketogenesis. HP [1-13C]acetoacetate was increased in fasting (250%) but decreased during pyruvate co-injection (−82%). Combining HP 13C technology and co-administration of separate imaging agents enables noninvasive and simultaneous monitoring of both fatty acid and carbohydrate oxidation. This protocol illustrates a novel method for assessing metabolic flux through different enzymatic pathways simultaneously and enables mechanistic studies of the changing myocardial energetics often associated with disease. PMID:27150735

  20. Measuring changes in substrate utilization in the myocardium in response to fasting using hyperpolarized [1-(13)C]butyrate and [1-(13)C]pyruvate.

    PubMed

    Bastiaansen, Jessica A M; Merritt, Matthew E; Comment, Arnaud

    2016-01-01

    Cardiac dysfunction is often associated with a shift in substrate preference for ATP production. Hyperpolarized (HP) (13)C magnetic resonance spectroscopy (MRS) has the unique ability to detect real-time metabolic changes in vivo due to its high sensitivity and specificity. Here a protocol using HP [1-(13)C]pyruvate and [1-(13)C]butyrate is used to measure carbohydrate versus fatty acid metabolism in vivo. Metabolic changes in fed and fasted Sprague Dawley rats (n = 36) were studied at 9.4 T after tail vein injections. Pyruvate and butyrate competed for acetyl-CoA production, as evidenced by significant changes in [(13)C]bicarbonate (-48%), [1-(13)C]acetylcarnitine (+113%), and [5-(13)C]glutamate (-63%), following fasting. Butyrate uptake was unaffected by fasting, as indicated by [1-(13)C]butyrylcarnitine. Mitochondrial pseudoketogenesis facilitated the labeling of the ketone bodies [1-(13)C]acetoacetate and [1-(13)C]β-hydroxybutyryate, without evidence of true ketogenesis. HP [1-(13)C]acetoacetate was increased in fasting (250%) but decreased during pyruvate co-injection (-82%). Combining HP (13)C technology and co-administration of separate imaging agents enables noninvasive and simultaneous monitoring of both fatty acid and carbohydrate oxidation. This protocol illustrates a novel method for assessing metabolic flux through different enzymatic pathways simultaneously and enables mechanistic studies of the changing myocardial energetics often associated with disease. PMID:27150735

  1. Transmembrane Exchange of Hyperpolarized 13C-Urea in Human Erythrocytes: Subminute Timescale Kinetic Analysis

    PubMed Central

    Pagès, Guilhem; Puckeridge, Max; Liangfeng, Guo; Tan, Yee Ling; Jacob, Chacko; Garland, Marc; Kuchel, Philip W.

    2013-01-01

    The rate of exchange of urea across the membranes of human erythrocytes (red blood cells) was quantified on the 1-s to 2-min timescale. 13C-urea was hyperpolarized and subjected to rapid dissolution and the previously reported (partial) resolution of 13C NMR resonances from the molecules inside and outside red blood cells in suspensions was observed. This enabled a stopped-flow type of experiment to measure the (initially) zero-trans transport of urea with sequential single-pulse 13C NMR spectra, every second for up to ∼2 min. Data were analyzed using Bayesian reasoning and a Markov chain Monte Carlo method with a set of simultaneous nonlinear differential equations that described nuclear magnetic relaxation combined with transmembrane exchange. Our results contribute to quantitative understanding of urea-exchange kinetics in the whole body; and the methodological approach is likely to be applicable to other cellular systems and tissues in vivo. PMID:24209840

  2. NMR at cryogenic temperatures: A {sup 13}C NMR study of ferrocene

    SciTech Connect

    Orendt, A.M.; Facelli, J.C.; Jiang, Y.J.; Grant, D.M.

    1998-09-24

    A new cryogenic apparatus is described that can be used to obtain NMR spectra at temperatures down to 8--10 K. The static solid {sup 13}C NMR spectrum of ferrocene is recorded at that temperature. Spectra recorded at higher temperatures show that ferrocene is still freely rotating about its 5-fold symmetry axis on the {sup 13}C NMR time scale at 45--50 K. A comparison of the principal values of the {sup 13}C chemical-shift tensor obtained from the room- and low-temperature spectra of ferrocene indicates that the lowest frequency chemical shift principal component, {delta}{sub 33}, is tilted off this symmetry axis by approximately 12{degree}. Quantum chemical calculations of the chemical-shift tensor, completed on structures of ferrocene from the literature as well as on optimized structures with the cyclopentadienyl rings locked in both the staggered and eclipsed arrangements, predict the angle between the {delta}{sub 33} direction and the rotation axis to be between 11 and 15{degree}, depending upon the geometry used in the calculation. The calculations also predict the sign of the angular perturbation, information not obtained from the experiment. An explanation of this angular change in the {delta}{sub 33} direction is provided by the composition of the molecular orbitals.

  3. Methanogenic capabilities of ANME-archaea deduced from (13) C-labelling approaches.

    PubMed

    Bertram, Sebastian; Blumenberg, Martin; Michaelis, Walter; Siegert, Michael; Krüger, Martin; Seifert, Richard

    2013-08-01

    Anaerobic methanotrophic archaea (ANME) are ubiquitous in marine sediments where sulfate dependent anaerobic oxidation of methane (AOM) occurs. Despite considerable progress in the understanding of AOM, physiological details are still widely unresolved. We investigated two distinct microbial mat samples from the Black Sea that were dominated by either ANME-1 or ANME-2. The (13) C lipid stable isotope probing (SIP) method using labelled substances, namely methane, bicarbonate, acetate, and methanol, was applied, and the substrate-dependent methanogenic capabilities were tested. Our data provide strong evidence for a versatile physiology of both, ANME-1 and ANME-2. Considerable methane production rates (MPRs) from CO2 -reduction were observed, particularly from ANME-2 dominated samples and in the presence of methane, which supports the hypothesis of a co-occurrence of methanotrophy and methanogenesis in the AOM systems (AOM/MPR up to 2:1). The experiments also revealed strong methylotrophic capabilities through (13) C-assimilation from labelled methanol, which was independent of the presence of methane. Additionally, high MPRs from methanol were detected in both of the mat samples. As demonstrated by the (13) C-uptake into lipids, ANME-1 was found to thrive also under methane free conditions. Finally, C35 -isoprenoid hydrocarbons were identified as new lipid biomarkers for ANME-1, most likely functioning as a hydrogen sink during methanogenesis.

  4. High resolution 13C-detected solid-state NMR spectroscopy of a deuterated protein

    PubMed Central

    Tang, Ming; Comellas, Gemma; Mueller, Leonard J.

    2011-01-01

    High resolution 13C-detected solid-state NMR spectra of the deuterated beta-1 immunoglobulin binding domain of the protein G (GB1) have been collected to show that all 15N, 13C′, 13Cα and 13Cβ sites are resolved in 13C–13C and 15N–13C spectra, with significant improvement in T2 relaxation times and resolution at high magnetic field (750 MHz). The comparison of echo T2 values between deuterated and protonated GB1 at various spinning rates and under different decoupling schemes indicates that 13Cα T2′ times increase by almost a factor of two upon deuteration at all spinning rates and under moderate decoupling strength, and thus the deuteration enables application of scalar-based correlation experiments that are challenging from the standpoint of transverse relaxation, with moderate proton decoupling. Additionally, deuteration in large proteins is a useful strategy to selectively detect polar residues that are often important for protein function and protein–protein interactions. PMID:20803233

  5. Chemical Shift Separation with Controlled Aliasing for Hyperpolarized 13C Metabolic Imaging

    PubMed Central

    Shin, Peter J.; Larson, Peder E.Z.; Uecker, Martin; Reed, Galen D.; Kerr, Adam B.; Tropp, James; Ohliger, Michael A.; Nelson, Sarah J.; Pauly, John M.; Lustig, Michael; Vigneron, Daniel B.

    2014-01-01

    Purpose A chemical shift separation technique for hyperpolarized 13C metabolic imaging with high spatial and temporal resolution was developed. Specifically, a fast 3D pulse sequence and a reconstruction method were implemented to acquire signals from multiple 13C species simultaneously with subsequent separation into individual images. Methods A stack of flyback-EPI readouts and a set of multiband excitation RF pulses were designed to spatially modulate aliasing patterns of the acquired metabolite images, which translated the chemical shift separation problem into parallel imaging reconstruction problem. An eight-channel coil array was used for data acquisition and a parallel imaging method based on nonlinear inversion was developed to separate the aliased images. Results Simultaneous acquisitions of pyruvate and lactate in a phantom study and in vivo rat experiments were performed. The results demonstrated successful separation of the metabolite distributions into individual images having high spatial resolution. Conclusion This method demonstrated the ability to provide accelerated metabolite imaging in hyperpolarized 13C MR utilizing multi-channel coils, tailored readout, and specialized RF pulses. PMID:25298086

  6. Hyperpolarized 13C urea relaxation mechanism reveals renal changes in diabetic nephropathy

    PubMed Central

    Stokholm Nørlinger, Thomas; Christoffer Hansen, David; Qi, Haiyun; Mose Nielsen, Per; Bonde Bertelsen, Lotte; Henrik Ardenkjaer‐Larsen, Jan; Stødkilde Jørgensen, Hans

    2015-01-01

    Purpose Our aim was to assess a novel 13C radial fast spin echo golden ratio single shot method for interrogating early renal changes in the diabetic kidney, using hyperpolarized (HP) [13C,15N2]urea as a T2 relaxation based contrast bio‐probe. Methods A novel HP 13C MR contrast experiment was conducted in a group of streptozotocin type‐1 diabetic rat model and age matched controls. Results A significantly different relaxation time (P = 0.004) was found in the diabetic kidney (0.49 ± 0.03 s) compared with the controls (0.64 ± 0.02 s) and secondly, a strong correlation between the blood oxygen saturation level and the relaxation times were observed in the healthy controls. Conclusion HP [13C,15N2]urea apparent T2 mapping may be a useful for interrogating local renal pO2 status and renal tissue alterations. Magn Reson Med, 2015. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. Magn Reson Med 75:515–518, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. PMID:26584247

  7. A comparison of quantitative methods for clinical imaging with hyperpolarized (13)C-pyruvate.

    PubMed

    Daniels, Charlie J; McLean, Mary A; Schulte, Rolf F; Robb, Fraser J; Gill, Andrew B; McGlashan, Nicholas; Graves, Martin J; Schwaiger, Markus; Lomas, David J; Brindle, Kevin M; Gallagher, Ferdia A

    2016-04-01

    Dissolution dynamic nuclear polarization (DNP) enables the metabolism of hyperpolarized (13)C-labelled molecules, such as the conversion of [1-(13)C]pyruvate to [1-(13)C]lactate, to be dynamically and non-invasively imaged in tissue. Imaging of this exchange reaction in animal models has been shown to detect early treatment response and correlate with tumour grade. The first human DNP study has recently been completed, and, for widespread clinical translation, simple and reliable methods are necessary to accurately probe the reaction in patients. However, there is currently no consensus on the most appropriate method to quantify this exchange reaction. In this study, an in vitro system was used to compare several kinetic models, as well as simple model-free methods. Experiments were performed using a clinical hyperpolarizer, a human 3 T MR system, and spectroscopic imaging sequences. The quantitative methods were compared in vivo by using subcutaneous breast tumours in rats to examine the effect of pyruvate inflow. The two-way kinetic model was the most accurate method for characterizing the exchange reaction in vitro, and the incorporation of a Heaviside step inflow profile was best able to describe the in vivo data. The lactate time-to-peak and the lactate-to-pyruvate area under the curve ratio were simple model-free approaches that accurately represented the full reaction, with the time-to-peak method performing indistinguishably from the best kinetic model. Finally, extracting data from a single pixel was a robust and reliable surrogate of the whole region of interest. This work has identified appropriate quantitative methods for future work in the analysis of human hyperpolarized (13)C data.

  8. Vineyard weeds control practices impact on surface water transfers: using numerical tracer experiment coupled to a distributed hydrological model to manage agricultural practices spatial arrangements.

    NASA Astrophysics Data System (ADS)

    Colin, F.; Moussa, R.

    2009-04-01

    In rural basins, agricultural landscape management highly influences water and pollutants transfers. Landuse, agricultural practices and their spatial arrangements are at issue. Hydrological model are widely used to explore impacts of anthropogenic influences on experimental catchments. But planning all spatial arrangements leads to a possible cases count which cannot be considered. On the basis of the recent « numerical experiment » approach, we propose a « numerical tracer function » which had to be coupled to a distributed rainfall-runoff model. This function simulate the transfer of a virtual tracer successively spread on each distributed unit inside the catchment. It allows to rank hydrological spatial units according to their hydrological contribution to the surface flows, particularly at the catchment outlet. It was used with the distributed model MHYDAS in an agricultural context. The case study concerns the experimental Roujan vine-growing catchment (1km², south of France) studied since 1992. In this Mediterranean context, we focus on the soil hydraulic conductivity distributed parameter because it highly depends on weed control practices (chemical weeding induces a lot more runoff than mechanical weeding). We checked model sensitivity analysis to soil hydraulic conductivity spatial arrangement on runoff coefficient, peak discharge and catchment lag-time. Results show (i) the use of the tracer function is more efficient than a random approach to improve sensitivity to spatial arrangements from point of view of simulated discharge range, (ii) the first factor explaining hydrological simulations variability was practices area ratio, (iii) variability induced by practices spatial arrangements was significant on runoff coefficient and peak discharge for balanced practices area ratio and on lag-time for low area ratio of chemical weeding practices. From the actual situation on the experimental Roujan catchment (40% of tilled and 60% of non tilled vineyard

  9. Heteronuclear Cross Polarization for Enhanced Sensitivity of in Vivo13C MR Spectroscopy on a Clinical 1.5 T MR System

    NASA Astrophysics Data System (ADS)

    van den Bergh, Adrianus J.; van den Boogert, Hendrikus J.; Heerschap, Arend

    1998-11-01

    The potential of heteronuclear {1H-13C} cross polarization was studied for optimization of the signal-to-noise ratio inin vivo13C MR spectroscopy at the clinical field strength of 1.5 T. Experiments on the human calf showed a significant chemical-shift selective signal enhancement on triglyceride signals of 3.9 by heteronuclear cross polarization, compared to a standard pulse-acquire sequence. Studies on a neonatal piglet brain showed an enhancement by cross polarization of 2.2 for the detection of13C-1-glucose. This enhancement allowed a fourfold improvement in time resolution in dynamic13C MR of13C-1-glucose inflow in piglet brain. Phantom experiments demonstrated the efficiency of this technique for interleaved detection of two spectral regions. Tests with a volume coil showed the feasibility of signal enhancement by cross polarization over a large volume of interest.

  10. Probing soil C metabolism in response to temperature: results from experiments and modeling

    NASA Astrophysics Data System (ADS)

    Dijkstra, P.; Dalder, J.; Blankinship, J.; Selmants, P. C.; Schwartz, E.; Koch, G. W.; Hart, S.; Hungate, B. A.

    2010-12-01

    C use efficiency (CUE) is one of the least understood aspects of soil C cycling, has a very large effect on soil respiration and C sequestration, and decreases with elevated temperature. CUE is directly related to substrate partitioning over energy production and biosynthesis. The production of energy and metabolic precursors occurs in well-known processes such as glycolysis and Krebs cycle. We have developed a new stable isotope approach using position-specific 13C-labeled metabolic tracers to measure these fundamental metabolic processes in intact soil communities (1). We use this new approach, combined with models of soil metabolic flux patterns, to analyze the response of microbial energy production, biosynthesis, and CUE to temperature. The method consists of adding small but precise amounts of position-specific 13C -labeled metabolic tracers to parallel soil incubations, in this case 1-13C and 2,3-13C pyruvate and 1-13C and U-13C glucose. The measurement of CO2 released from the labeled tracers is used to calculate the C flux rates through various metabolic pathways. A simplified metabolic model consisting of 23 reactions is iteratively solved using results of the metabolic tracer experiments and information on microbial precursor demand under different temperatures. This new method enables direct study of fundamental aspects of microbial energy production, C use efficiency, and soil organic matter formation in response to temperature. (1) Dijkstra P, Blankinship JC, Selmants PC, Hart SC, Koch GW, Schwarz E and Hungate BA. Probing metabolic flux patterns of soil microbial communities using parallel position-specific tracer labeling. Soil Biology and Biochemistry (accepted)

  11. Millimeter and submillimeter wave spectra of 13C-glycolaldehydes

    NASA Astrophysics Data System (ADS)

    Haykal, I.; Motiyenko, R. A.; Margulès, L.; Huet, T. R.

    2013-01-01

    Context. Glycolaldehyde (CH2OHCHO) is the simplest sugar and an important intermediate in the path toward forming more complex biologically relevant molecules. Astronomical surveys of interstellar molecules, such as those available with the very sensitive ALMA telescope, require preliminary laboratory investigations of the microwave and submillimeter-wave spectra of molecular species including new isotopologs - to identify these in the interstellar media. Aims: To achieve the detection of the 13C isotopologs of glycolaldehyde in the interstellar medium, their rotational spectra in the millimeter and submillimeter-wave regions were studied. Methods: The spectra of 13CH2OHCHO and CH2OH13CHO were recorded in the 150-945 GHz spectral range in the laboratory using a solid-state submillimeter-wave spectrometer in Lille. The observed line frequencies were measured with an accuracy of 30 kHz up to 700 GHz and of 50 kHz above 700 GHz. We analyzed the spectra with a standard Watson Hamiltonian. Results: About 10 000 new lines were identified for each isotopolog. The spectroscopic parameters were determined for the ground- and the three lowest vibrational states up to 945 and 630 GHz. Previous microwave assignments of 13CH2OHCHO were not confirmed. Conclusions: The provided line-lists and sets of molecular parameters meet the needs for a first astrophysical search of 13C-glycolaldehydes. Full Tables 3 and 4 are only available at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/549/A96

  12. Approaches to studies on neuronal/glial relationships by 13C-MRS analysis.

    PubMed

    Taylor, A; McLean, M; Morris, P; Bachelard, H

    1996-01-01

    The use of different 13C-labelled precursors alone or in combination ([1-13C]glucose, [2-13C]glucose, [1-13C]acetate, [2-13C]acetate and [1,2-13C2]acetate) to study neuronal/glial metabolic relationships by MRS is discussed. Glutamine and citrate resonances represent glial metabolism if a combination of [1-13C]glucose + [2-13C]acetate is used, but only for short time periods. A combination of [2-13C]glucose + [2-13C]acetate will label -COO- groups from glucose and -CH2 groups from acetate, respectively, which distinguish well in theory. However, this approach is severely limited by the long T1S of -COO- groups and low S/N. Contributions of the anaplerotic pathway can be assessed using [2-13C]glucose, but again can be limited by the long T1S of -COO- groups. Labelling of glycerol-3-phosphate (believed to be produced in glia) from [1-13C]glucose is difficult to see under normal conditions but has proved useful in, e.g., hypoxia. We believe the most promising approach is the use of [1-13C] glucose with [1,2-13C2]acetate, by analysis of the multiplets ('isotopomers') of the amino acid resonances.

  13. 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.

  14. Hydrothermal plumes in the Gulf of Aden, as characterized by light transmission, Mn, Fe, CH4 and δ13C-CH4 anomalies

    NASA Astrophysics Data System (ADS)

    Gamo, Toshitaka; Okamura, Kei; Hatanaka, Hiroshi; Hasumoto, Hiroshi; Komatsu, Daisuke; Chinen, Masakazu; Mori, Mutsumi; Tanaka, Junya; Hirota, Akinari; Tsunogai, Urumu; Tamaki, Kensaku

    2015-11-01

    We conducted water column surveys to search for hydrothermal plumes over the spreading axes in the Gulf of Aden between 45°35‧E and 52°42‧E. We measured light transmission and chemical tracers Mn, Fe, CH4 and δ13C of CH4 in seawater taken using a CTD-Carrousel multi-sampling system at 12 locations including a control station in the Arabian Sea. We recognized three types of hydrothermal plumes at depths of ~650 to ~900 m (shallow plumes), ~1000 to ~1200 m (intermediate plumes), and >1500 m (deep plumes). The shallow plumes were apparently originated from newly discovered twin seamounts (12°03-06‧N and 45°35-41‧E) at the westernmost survey area, where two-dimensional distributions of light transmission and Mn were mapped by tow-yo observations of the CTD-sampling system with an in situ auto-analyzer GAMOS. The maximum concentrations of Mn, Fe, and CH4 of 46 nM, 251 nM, and 15 nM, respectively, were observed for collected seawater within the shallow plumes. The intermediate plumes were characterized by anomalies of light transmission, Mn, Fe, and δ13C of CH4, but by little CH4 anomalies, suggesting that CH4 had been consumed down to the background level during the aging of the plumes. Anomalies of δ3He already reported by the World Ocean Circulation Experiment (WOCE) program exhibited a hydrothermal plume-like peak at ~2000 m depth in the Gulf of Aden, which seems to coincide with the deep plumes observed in this study. The endmember δ13C-CH4 values for the shallow and the deep plumes were estimated to be in a range between -10‰ and -15‰, demonstrating that the sources of CH4 are not biogenic but magmatic as similarly observed at sediment-starved mid-oceanic ridges.

  15. Enhancing Phospholipid Fatty Acid Profiling of Soil Bacterial Communities via Substrate- Specific 13C-labelling

    NASA Astrophysics Data System (ADS)

    Evershed, R. P.; Maxfield, P. J.; Bingham, E. M.; Dildar, N.; Brennand, E. L.; Hornibrook, E.

    2008-12-01

    A range of culture-independent methods, has recently emerged to study environmental microorganisms in situ[1]. One such method is phospholipid fatty acid (PLFA) analysis, wherein these ubiquitous membrane lipids provide a powerful tool for the study of unculturable soil microorganisms. PLFA analyses have been used to investigate the impacts of a wide range of environmental factors on the soil microbial community. An acknowledged shortcoming of the PLFAs approach is the lack the chemotaxonoic specificity, which restricts the ability of the method to probe the activities of specific functional groups of the microbial community selectively. However, the selectivity of PLFAs analyses can be enhanced by incubating soils with 13C- labelled substrates followed by gas chromatography-combustion-isotope ratio mass spectrometry to reveal the specific PLFAs incorporating the 13C-label. The application of this approach will be demonstrated through our recent work on methanotrophic bacteria in soils. We applied this approach initially to mineral soils[2] and then extended chemotaxonomic assessments by using a combination of 13C-labelled PLFAs and hopanoids [3]. We have used this approach to explore the properties of high affinity methanotrophs in a range of environments, investigating the relationship between methane oxidation rates and the nature and magnitude of the methanotrophic community for the first time[4,5] More recently we extended the technique using a novel time series 13C-labelling of PLFAs[6] to estimate the rate and progression of 13C- label incorporation and turnover of methanotrophic populations. This modified approach has been used to investigate the impacts of various environmental variables, e.g. soil type, vegetation cover and land use, on the methanotrophic biomass[7.8]. The unique nature of the 13CH4 as a gaseous substate/carbon source means that can be readily introduced into soils via a specific subset of the soil microbial biomass, thereby offering many

  16. Trans and surface membrane bound zervamicin IIB: 13C-MAOSS-NMR at high spinning speed.

    PubMed

    Raap, J; Hollander, J; Ovchinnikova, T V; Swischeva, N V; Skladnev, D; Kiihne, S

    2006-08-01

    Interactions between (15)N-labelled peptides or proteins and lipids can be investigated using membranes aligned on a thin polymer film, which is rolled into a cylinder and inserted into the MAS-NMR rotor. This can be spun at high speed, which is often useful at high field strengths. Unfortuantely, substrate films like commercially available polycarbonate or PEEK produce severe overlap with peptide and protein signals in (13)C-MAOSS NMR spectra. We show that a simple house hold foil support allows clear observation of the carbonyl, aromatic and C(alpha) signals of peptides and proteins as well as the ester carbonyl and choline signals of phosphocholine lipids. The utility of the new substrate is validated in applications to the membrane active peptide zervamicin IIB. The stability and macroscopic ordering of thin PC10 bilayers was compared with that of thicker POPC bilayers, both supported on the household foil. Sidebands in the (31)P-spectra showed a high degree of alignment of both the supported POPC and PC10 lipid molecules. Compared with POPC, the PC10 lipids are slightly more disordered, most likely due to the increased mobilities of the shorter lipid molecules. This mobility prevents PC10 from forming stable vesicles for MAS studies. The (13)C-peptide peaks were selectively detected in a (13)C-detected (1)H-spin diffusion experiment. Qualitative analysis of build-up curves obtained for different mixing times allowed the transmembrane peptide in PC10 to be distinguished from the surface bound topology in POPC. The (13)C-MAOSS results thus independently confirms previous findings from (15)N spectroscopy [Bechinger, B., Skladnev, D.A., Ogrel, A., Li, X., Rogozhkina, E.V., Ovchinnikova, T.V., O'Neil, J.D.J. and Raap, J. (2001) Biochemistry, 40, 9428-9437]. In summary, application of house hold foil opens the possibility of measuring high resolution (13)C-NMR spectra of peptides and proteins in well ordered membranes, which are required to determine the secondary

  17. Reach-scale isotope tracer experiment to quantify denitrification and related processes in a nitrate-rich stream, midcontinent United States

    USGS Publications Warehouse

    Böhlke, J.K.; Harvey, J.W.; Voytek, M.A.

    2004-01-01

    We conducted an in-stream tracer experiment with Br and 15N-enriched NO3- to determine the rates of denitrification and related processes in a gaining NO3--rich stream in an agricultural watershed in the upper Mississippi basin in September 2001. We determined reach-averaged rates of N fluxes and reactions from isotopic analyses of NO3-, NO 2-, N2, and suspended particulate N in conjunction with other data in a 1.2-km reach by using a forward time-stepping numerical simulation that included groundwater discharge, denitrification, nitrification, assimilation, and air-water gas exchange with changing temperature. Denitrification was indicated by a systematic downstream increase in the ??15N values of dissolved N2. The reach-averaged rate of denitrification of surface-water NO3- indicated by the isotope tracer was approximately 120 ?? 20 ??mol m-2 h-1 (corresponding to zero- and first-order rate constants of 0.63 ??mol L-1 h-1 and 0.009 h -1, respectively). The overall rate of NO3- loss by processes other than denitrification (between O and about 200 ??mol m-2 h-1) probably was less than the denitrification rate but had a large relative uncertainty because the NO3- load was large and was increasing through the reach. The rates of denitrification and other losses would have been sufficient to reduce the stream NO 3- load substantially in the absence of NO 3- sources, but the losses were more than offset by nitrification and groundwater NO3- inputs at a combined rate of about 500-700 ??mol m-2 h-1. Despite the importance of denitrification, the overall mass fluxes of N2 were dominated by discharge of denitrified groundwater and air-water gas exchange in response to changing temperature, whereas the flux of N2 attributed to denitrification was relatively small. The in-stream isotope tracer experiment provided a sensitive direct reach-scale measurement of denitrification and related processes in a NO3--rich stream where other mass-balance methods were not suitable because

  18. Microwave spectra for the three 13C1 isotopologues of propene and new rotational constants for propene and its 13C1 isotopologues

    NASA Astrophysics Data System (ADS)

    Craig, Norman C.; Groner, Peter; Conrad, Andrew R.; Gurusinghe, Ranil; Tubergen, Michael J.

    2016-10-01

    New measurements of microwave lines (A and E) of propene and its three 13C1 isotopologues have been made in the 10-22 GHz region with FT accuracy. The revised lines for propene along with many hundreds from the literature were fitted with the ERHAM program for internal rotors to give improved rotational constants. The new constants are A0 = 46280.2904(16), B0 = 9305.24260(30), and C0 = 8134.22685(28) MHz. Lines for the 3-13C1 species were observed in a pure sample; lines for the 1-13C1 and 2-13C1 species were observed in natural abundance. In fitting the limited sets of lines for the 13C1 species, many of the centrifugal distortion constants and most of the tunneling parameters were transferred from the fit of propene itself with 27 parameters. Improved rotational constants for the 13C1 species are reported.

  19. Complete (1)H and (13)C signal assignment of prenol-10 with 3D NMR spectroscopy.

    PubMed

    Misiak, Maria; Koźmiński, Wiktor; Kwasiborska, Maria; Wójcik, Jacek; Ciepichal, Ewa; Swiezewska, Ewa

    2009-10-01

    The complete assignment of (1)H and (13)C chemical shifts of natural abundance prenol-10 is reported for the first time. It was achieved using 3D NMR experiments, which were based on random sampling of the evolution time space followed by multidimensional Fourier transform. This approach makes it possible to acquire 3D NMR spectra in a reasonable time and preserves high resolution in indirectly detected dimensions. It is shown that the interpretation of 3D COSY-HMBC and 3D TOCSY-HSQC spectra is crucial in the structural analysis of prenol-10.

  20. A high resolution δ13C record in a modern Porites lobata coral: Insights into controls on skeletal δ13C

    NASA Astrophysics Data System (ADS)

    Allison, Nicola; Finch, Adrian A.; EIMF

    2012-05-01

    δ13C was determined at a high spatial resolution by secondary ion mass spectrometry (SIMS) across a 1 year section of a modern Porites lobata coral skeleton from Hawaii. Skeletal δ13C is dominated by large oscillations of 5-7‰ that typically cover skeletal distances equivalent to periods of ˜14-40 days. These variations do not reflect seawater temperature and it is unlikely that they reflect variations in the δ13C of local seawater. We observe no correlation between skeletal δ13C and the pH of the calcification fluid (estimated from previous measurements of skeletal δ11B). We conclude that either the proportion of skeletal carbon derived from metabolic CO2 is not reflected by estimated ECF pH (as the [CO2] in the overlying coral tissue varies) and/or the δ13C composition of the metabolic CO2 is highly variable. We also observe no correlation between skeletal δ13C and previous δ18O SIMS measurements. Variations in skeletal δ13C and δ18O do not have a common timing, providing no evidence that skeletal δ13C and δ18O vary in response to a single factor. This suggests that skeletal δ13C is principally driven by variations in the δ13C composition of metabolic CO2 rather than by the abundance of metabolic CO2, which would also affect skeletal δ18O. The δ13C composition of metabolic CO2 reflects the processes of photosynthesis, heterotrophic feeding and respiration in the overlying coral tissue. Corals catabolise stored lipid reserves to meet energetic demands when photosynthesis conditions are sub-optimal. Variations in the amounts and types of reserves utilised could induce changes in the δ13C composition of metabolic CO2 and the resultant skeleton which are temporally offset from skeletal δ18O records.

  1. The 13C/12C isotopic signal of day-respired CO2 in variegated leaves of Pelargonium × hortorum.

    PubMed

    Tcherkez, Guillaume; Mauve, Caroline; Lamothe, Marlene; Le Bras, Camille; Grapin, Agnes

    2011-02-01

    In leaves, although it is accepted that CO(2) evolved by dark respiration after illumination is naturally (13) C-enriched compared to organic matter or substrate sucrose, much uncertainty remains on whether day respiration produces (13) C-depleted or (13) C-enriched CO(2). Here, we applied equations described previously for mesocosm CO(2) exchange to investigate the carbon isotope composition of CO(2) respired by autotrophic and heterotrophic tissues of Pelargonium × hortorum leaves, taking advantage of leaf variegation. Day-respired CO(2) was slightly (13) C-depleted compared to organic matter both under 21% O(2) and 2% O(2). Furthermore, most, if not all CO(2) molecules evolved in the light came from carbon atoms that had been fixed previously before the experiments, in both variegated and green leaves. We conclude that the usual definition of day respiratory fractionation, that assumes carbon fixed by current net photosynthesis is the respiratory substrate, is not valid in Pelargonium leaves under our conditions. In variegated leaves, total organic matter was slightly (13) C-depleted in white areas and so were most primary metabolites. This small isotopic difference between white and green areas probably came from the small contribution of photosynthetic CO(2) refixation and the specific nitrogen metabolism in white leaf areas. PMID:20955224

  2. The 13C/12C isotopic signal of day-respired CO2 in variegated leaves of Pelargonium × hortorum.

    PubMed

    Tcherkez, Guillaume; Mauve, Caroline; Lamothe, Marlene; Le Bras, Camille; Grapin, Agnes

    2011-02-01

    In leaves, although it is accepted that CO(2) evolved by dark respiration after illumination is naturally (13) C-enriched compared to organic matter or substrate sucrose, much uncertainty remains on whether day respiration produces (13) C-depleted or (13) C-enriched CO(2). Here, we applied equations described previously for mesocosm CO(2) exchange to investigate the carbon isotope composition of CO(2) respired by autotrophic and heterotrophic tissues of Pelargonium × hortorum leaves, taking advantage of leaf variegation. Day-respired CO(2) was slightly (13) C-depleted compared to organic matter both under 21% O(2) and 2% O(2). Furthermore, most, if not all CO(2) molecules evolved in the light came from carbon atoms that had been fixed previously before the experiments, in both variegated and green leaves. We conclude that the usual definition of day respiratory fractionation, that assumes carbon fixed by current net photosynthesis is the respiratory substrate, is not valid in Pelargonium leaves under our conditions. In variegated leaves, total organic matter was slightly (13) C-depleted in white areas and so were most primary metabolites. This small isotopic difference between white and green areas probably came from the small contribution of photosynthetic CO(2) refixation and the specific nitrogen metabolism in white leaf areas.

  3. 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

  4. Monitoring CO[subscript 2] Fixation Using GC-MS Detection of a [superscript 13]C-Label

    ERIC Educational Resources Information Center

    Hammond, Daniel G.; Bridgham, April; Reichert, Kara; Magers, Martin

    2010-01-01

    Much of our understanding of metabolic pathways has resulted from the use of chemical and isotopic labels. In this experiment, a heavy isotope of carbon, [superscript 13]C, is used to label the product of the well-known RuBisCO enzymatic reaction. This is a key reaction in photosynthesis that converts inorganic carbon to organic carbon; a process…

  5. Sorbing tracer experiments in a crystalline rock fracture at Äspö (Sweden): 3. Effect of microscale heterogeneity

    NASA Astrophysics Data System (ADS)

    Cvetkovic, V.; Cheng, H.

    2008-12-01

    We study the impact of microscale (10-3-10-2 m) heterogeneity in material and structural properties on sorbing tracer transport in a single crystalline fracture of the TRUE-1 tests (mesoscale, 5 m) at Äspö (Sweden). The analysis is based on the microscale characterization results as presented in part 1 of this series. Our main objective in this last part of the series is to provide an independent interpretation (or "prediction") of the effective parameters as estimated from calibration in part 2 by combining the data presented in part 1 with analytical and numerical transport modeling. We show here that the independent information from microscopic characterization can be used for "predicting" the effective diffusion time td reasonably well; a discrepancy is to be expected given the uncertainties of microscale retention properties, in particular of the sorption coefficient.

  6. HepatoDyn: A Dynamic Model of Hepatocyte Metabolism That Integrates 13C Isotopomer Data

    PubMed Central

    Foguet, Carles; Selivanov, Vitaly A.; Fanchon, Eric; Guinovart, Joan J.; de Atauri, Pedro; Cascante, Marta

    2016-01-01

    The liver performs many essential metabolic functions, which can be studied using computational models of hepatocytes. Here we present HepatoDyn, a highly detailed dynamic model of hepatocyte metabolism. HepatoDyn includes a large metabolic network, highly detailed kinetic laws, and is capable of dynamically simulating the redox and energy metabolism of hepatocytes. Furthermore, the model was coupled to the module for isotopic label propagation of the software package IsoDyn, allowing HepatoDyn to integrate data derived from 13C based experiments. As an example of dynamical simulations applied to hepatocytes, we studied the effects of high fructose concentrations on hepatocyte metabolism by integrating data from experiments in which rat hepatocytes were incubated with 20 mM glucose supplemented with either 3 mM or 20 mM fructose. These experiments showed that glycogen accumulation was significantly lower in hepatocytes incubated with medium supplemented with 20 mM fructose than in hepatocytes incubated with medium supplemented with 3 mM fructose. Through the integration of extracellular fluxes and 13C enrichment measurements, HepatoDyn predicted that this phenomenon can be attributed to a depletion of cytosolic ATP and phosphate induced by high fructose concentrations in the medium. PMID:27124774

  7. Acetate and Bicarbonate Assimilation and Metabolite Formation in Chlamydomonas reinhardtii: A 13C-NMR Study

    PubMed Central

    Singh, Himanshu; Shukla, Manish R.; Chary, Kandala V. R.; Rao, Basuthkar J.

    2014-01-01

    Cellular metabolite analyses by 13C-NMR showed that C. reinhardtii cells assimilate acetate at a faster rate in heterotrophy than in mixotrophy. While heterotrophic cells produced bicarbonate and CO2aq, mixotrophy cells produced bicarbonate alone as predominant metabolite. Experiments with singly 13C-labelled acetate (13CH3-COOH or CH3-13COOH) supported that both the 13C nuclei give rise to bicarbonate and CO2aq. The observed metabolite(s) upon further incubation led to the production of starch and triacylglycerol (TAG) in mixotrophy, whereas in heterotrophy the TAG production was minimal with substantial accumulation of glycerol and starch. Prolonged incubation up to eight days, without the addition of fresh acetate, led to an increased TAG production at the expense of bicarbonate, akin to that of nitrogen-starvation. However, such TAG production was substantially high in mixotrophy as compared to that in heterotrophy. Addition of mitochondrial un-coupler blocked the formation of bicarbonate and CO2aq in heterotrophic cells, even though acetate uptake ensued. Addition of PSII-inhibitor to mixotrophic cells resulted in partial conversion of bicarbonate into CO2aq, which were found to be in equilibrium. In an independent experiment, we have monitored assimilation of bicarbonate via photoautotrophy and found that the cells indeed produce starch and TAG at a much faster rate as compared to that in mixotrophy and heterotrophy. Further, we noticed that the accumulation of starch is relatively more as compared to TAG. Based on these observations, we suggest that acetate assimilation in C. reinhardtii does not directly lead to TAG formation but via bicarbonate/CO2aq pathways. Photoautotrophic mode is found to be the best growth condition for the production of starch and TAG and starch in C. reinhardtii. PMID:25207648

  8. Tracer development at ESRI

    SciTech Connect

    Adams, M.C.; Rose, P.E.; McPherson, P.

    1996-04-10

    At ESRI the Tracer Development Program is divided into three components: liquid-phase tracers, vapor-phase tracers, and pre-test modeling. The liquid-phase project has tested 40 aromatic acids and 10 fluorescent tracers for geothermal use. The vapor-phase project, which develops tracers for reservoirs such as the Geysers, is currently focused on testing SF{sub 6} at high temperatures and examining HPLC methods for the sensitive analysis of alcohol tracers. The pre-test modeling component is exploring the feasibility of using simple numerical models to lower the cost of tracer tests by providing estimates of tracer quantities, flowpaths, and arrival times.

  9. delta15N and delta13C diet-tissue discrimination factors for large sharks under semi-controlled conditions.

    PubMed

    Hussey, Nigel E; Brush, Jaclyn; McCarthy, Ian D; Fisk, Aaron T

    2010-04-01

    Stable isotopes (delta(15)N and delta(13)C) are being widely applied in ecological research but there has been a call for ecologists to determine species- and tissue-specific diet discrimination factors ((13)C and (15)N) for their study animals. For large sharks stable isotopes may provide an important tool to elucidate aspects of their ecological roles in marine systems, but laboratory based controlled feeding experiments are impractical. By utilizing commercial aquaria, we estimated (15)N and (13)C of muscle, liver, vertebral cartilage and a number of organs of three large sand tiger (Carcharias taurus) and one large lemon shark (Negaprion brevirostris) under a controlled feeding regime. For all sharks mean+/-SD for (15)N and (13)C in lipid extracted muscle using lipid extracted prey data were 2.29 per thousand+/-0.22 and 0.90 per thousand+/-0.33, respectively. The use of non-lipid extracted muscle and prey resulted in very similar (15)N and (13)C values but mixing of lipid and non-lipid extracted data produced variable estimates. Values of (15)N and (13)C in lipid extracted liver and prey were 1.50 per thousand+/-0.54 and 0.22 per thousand+/-1.18, respectively. Non-lipid extracted diet discrimination factors in liver were highly influenced by lipid content and studies that examine stable isotopes in shark liver, and likely any high lipid tissue, should strive to remove lipid effects through standardising C:N ratios, prior to isotope analysis. Mean vertebral cartilage (15)N and (13)C values were 1.45 per thousand+/-0.61 and 3.75 per thousand+/-0.44, respectively. Organ (15)N and (13)C values were more variable among individual sharks but heart tissue was consistently enriched by approximately 1-2.5 per thousand. Minimal variability in muscle and liver delta(15)N and delta(13)C sampled at different intervals along the length of individual sharks and between liver lobes suggests that stable isotope values are consistent within tissues of individual animals. To our

  10. Analysis of dynamics and mechanism of ligand binding to Artocarpus integrifolia agglutinin. A 13C and 19F NMR study.

    PubMed

    Krishna Sastry, M V; Swamy, M J; Surolia, A

    1988-10-15

    Binding of 13C-labeled N-acetylgalactosamine (13C-GalNAc) and N-trifluoroacetylgalactosamine (19F-GalNAc) to Artocarpus integrifolia agglutinin has been studied using 13C and 19F nuclear magnetic resonance spectroscopy, respectively. Binding of these saccharides resulted in broadening of the resonances, and no change in chemical shift was observed, suggesting that the alpha- and beta-anomers of 13C-GalNAc and 19F-GalNAc experience a magnetically equivalent environment in the lectin combining site. The alpha- and beta-anomers of 13C-GalNAc and 19F-GalNAc were found to be in slow exchange between free and protein bound states. Binding of 13C-GalNAc was studied as a function of temperature. From the temperature dependence of the line broadening, the thermodynamic and kinetic parameters were evaluated. The association rate constants obtained for the alpha-anomers of 13C-GalNAc and 19F-GalNAc (k+1 = 1.01 x 10(5) M-1.s-1 and 0.698 x 10(5) M-1.s-1, respectively) are in close agreement with those obtained for the corresponding beta-anomers (k+1 = 0.95 x 10(5) M-1.s-1 and 0.65 x 10(5) M-1.s-1, respectively), suggesting that the two anomers bind to the lectin by a similar mechanism. In addition these values are several orders of magnitude slower than those obtained for diffusion controlled processes. The dissociation rate constants obtained are 49.9, 56.9, 42, and 43 s-1, respectively, for the alpha- and beta-anomers of 13C-GalNAc and 19F-GalNAc. A two-step mechanism has been proposed for the interaction of 13C-GalNAc and 19F-GalNAc with A. integrifolia lectin in view of the slow association rates and high activation entropies. The thermodynamic parameters obtained for the association and dissociation reactions suggest that the binding process is entropically favored and that there is a small enthalpic contribution.

  11. Detection of Reduced GABA Synthesis Following Inhibition of GABA Transaminase Using in Vivo Magnetic Resonance Signal of [13C]GABA C1

    PubMed Central

    Yang, Jehoon; Johnson, Christopher; Shen, Jun

    2009-01-01

    Previous in vivo magnetic resonance spectroscopy (MRS) studies of gamma-aminobutyric acid (GABA) synthesis have relied on 13C label incorporation into GABA C2 from [1-13C] or [1,6-13C2]glucose. In this study, the [13C]GABA C1 signal at 182.3 ppm in the carboxylic/amide spectral region of localized in vivo 13C spectra was detected. GABA-transaminase of rat brain was inhibited by administration of gabaculine after pre-labeling of GABA C1 and its metabolic precursors with exogenous [2,5-13C2]glucose. A subsequent isotope chase experiment was performed by infusing unlabeled glucose, which revealed a markedly slow change in the labeling of GABA C1 accompanying the blockade of the GABA shunt. This slow labeling of GABA at elevated GABA concentration was attributed to the relatively small intercompartmental GABA-glutamine cycling flux that constitutes the main route of 13C label loss during the isotope chase. Because this study showed that using low RF power broadband stochastic proton decoupling is feasible at very high field strength, it has important implications for the development of carboxylic/amide 13C MRS methods to study brain metabolism and neurotransmission in human subjects at high magnetic fields. PMID:19540876

  12. 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

  13. Synthesis of 13C and 15N labeled 2,4-dinitroanisole.

    PubMed

    Jagadish, Bhumasamudram; Field, Jim A; Chorover, Jon; Sierra-Alvarez, Reyes; Abrell, Leif; Mash, Eugene A

    2014-05-30

    Syntheses of [(13)C6]-2,4-dinitroanisole (ring-(13)C6) from [(13)C6]-anisole (ring-(13)C6) and [(15)N2]-2,4-dinitroanisole from anisole using in situ generated acetyl nitrate and [(15)N]-acetyl nitrate, respectively, are described. Treatment of [(13)C6]-anisole (ring-(13)C6) with acetyl nitrate generated in 100% HNO3 gave [(13)C6]-2,4-dinitroanisole (ring-(13)C6) in 83% yield. Treatment of anisole with [(15)N]-acetyl nitrate generated in 10 N [(15)N]-HNO3 gave [(15)N2 ]-2,4-dinitroanisole in 44% yield after two cycles of nitration. Byproducts in the latter reaction included [(15)N]-2-nitroanisole and [(15)N]-4-nitroanisole.

  14. Model of decision system for 13C Isotope Separation column

    NASA Astrophysics Data System (ADS)

    Boca, M. L.

    2015-11-01

    This paper presents the model of a decisional system for 13C Isotope Separation column, which is used to detect mission critical situation. The start model was a model of one distributed control system of critical situations that may arise in the operation of the distillation column. The research work it is proposed a model of decision system which implement a temperature sensor inside of liquid nitrogen level in the condenser. The condenser is a part of column where take place the cryogenic process using nitrogen liquid. The work temperature is very low about -192oC, and because the temperature can grow or go down more than 2 degrees is a very critical location inside the column. In this way the column has a deeply monitor and supervised and it take a decision in a proper time when the temperature is grow up or getting down and became a critical situation. For monitor and supervised it was used MatLAB SimuLink. The model, the decision system gives a signal to one sensor when something is wrong in the condenser which is the most critical place of the isotopic column. In this way it creates an alarm that something is getting wrong in the isotopic column.

  15. Inelastic pion scattering by /sup 13/C at low energies

    SciTech Connect

    Mitchell, J.H.

    1987-03-01

    Angular distributions for inelastically scattered pions were obtained for several states in /sup 13/C at an incident energy of 65 MeV. The data include results from both ..pi../sup +/ and ..pi../sup -/ measurements. In addition, ..pi../sup -/ measurements were made at T/sub ..pi../ = 50 MeV at one angle to give a two point fixed-q excitation function. The data are compared to theory and the data of others. As might be expected, medium corrections are shown to be considerably more important at low energies than at resonance. This is true for inelastic transitions of multipolarity 0,2 and 3. Parameters derived from an analysis of elastic pion scattering and SCX data also provide an adequate description of the inelastic transitions. The charge asymmetry in the cross sections for the 9/2/sup +/ state that was seen at resonance persists at these energies. This result is consistent with an impulse approximation treatment of the spin-flip amplitude. This is true even though the incoming energy of the pions is far below the range where the validity of an impulse treatment is expected. 65 refs., 45 figs.

  16. New optical analyzer for 13C-breath test

    NASA Astrophysics Data System (ADS)

    Harde, Hermann; Dressler, Matthias; Helmrich, Günther; Wolff, Marcus; Groninga, Hinrich

    2008-04-01

    Medical breath tests are well established diagnostic tools, predominantly for gastroenterological inspections, but also for many other examinations. Since the composition and concentration of exhaled volatile gases reflect the physical condition of a patient, a breath analysis allows one to recognize an infectious disease in an organ or even to identify a tumor. One of the most prominent breath tests is the 13C-urea-breath test, applied to ascertain the presence of the bacterium helicobacter pylori in the stomach wall as an indication of a gastric ulcer. In this contribution we present a new optical analyzer that is based on photoacoustic spectroscopy and uses a DFB diode laser at 2.744 μm. The concentration ratio of the CO II isotopologues is determined by measuring the absorption on a 13CO II line in comparison to a 12CO II line. In the specially selected spectral range the lines have similar strengths, although the concentrations differ by a factor of 90. Therefore, the signals are well comparable. Due to an excellent signal-noise-ratio isotope variations of less than 1% can be resolved as required for the breath test.

  17. Calibration of δ13C and δ18O measurements in CO2 using Off-axis Integrated Cavity Output Spectrometer (ICOS)

    NASA Astrophysics Data System (ADS)

    Joseph, Jobin; Külls, Christoph

    2014-05-01

    The δ13C and δ18O of CO2 has enormous potential as tracers to study and quantify the interaction between the water and carbon cycles. Isotope ratio mass spectrometry (IRMS) being the conventional method for stable isotopic measurements, has many limitations making it impossible for deploying them in remote areas for online or in-situ sampling. New laser based absorption spectroscopy approaches like Cavity Ring Down Spectroscopy (CRDS) and Integrated Cavity Output Spectroscopy (ICOS) have been developed for online measurements of stable isotopes at an expense of considerably less power requirement but with precision comparable to IRMS. In this research project, we introduce a new calibration system for an Off- Axis ICOS (Los Gatos Research CCIA-36d) for a wide range of varying concentrations of CO2 (800ppm - 25,000ppm), a typical CO2 flux range at the plant-soil continuum. The calibration compensates for the concentration dependency of δ13C and δ18O measurements, and was performed using various CO2 standards with known CO2 concentration and δC13 and δO18 values. A mathematical model was developed after the calibration procedure as a correction factor for the concentration dependency of δ13C and δ18O measurements. Temperature dependency of δ13C and δ18O measurements were investigated and no significant influence was found. Simultaneous calibration of δ13C and δ18O is achieved using this calibration system with an overall accuracy of (~ 0.75±0.24 ‰ for δ13C, ~ 0.81 ±0.26‰ for δ18O). This calibration procedure is found to be appropriate for making Off-Axis ICOS suitable for measuring CO2 concentration and δ13C and δ18O measurements at atmosphere-plant-soil continuum.

  18. Suspended Sediment Moves 10 km Before Entering Storage: Re-Interpreting a 20th Century Industrial Mercury Release as a Tracer Experiment, South River, Virginia

    NASA Astrophysics Data System (ADS)

    Pizzuto, J. E.

    2014-12-01

    Recent analyses suggest that the velocity of downstream transport of suspended sediment (averaged over long timescales that include periods of transport and storage in alluvial deposits) can be represented as the ratio Ls/T, where Ls is a distance particles move before entering storage and T is the waiting time particles spend in storage before being remobilized. Sediment budget analyses suggest that Ls is 1-100 km in the mid-Atlantic region, while T may be ~103 years, such that particles move 3-5 orders of magnitude slower than the water in the channel. Given the well-known inaccuracy of sediment budgets, independent verification from a tracer study would be desirable. Here, an historic industrial release of mercury is interpreted as a decadal sediment tracer experiment, releasing sediment particles "tagged" with mercury that are deposited on floodplains. As expected, floodplain mercury inventories decrease exponentially downstream, with a characteristic decay length of 10 km (95% confidence interval: 5-25 km) that defines the distance suspended particles typically move downstream before entering storage. Floodplain mercury inventories are not significantly different above and below three colonial age mill dams (present at the time of mercury release but now breached), suggesting that these results reflect ongoing processes. Suspended sediment routing models that neglect long-term storage, and the watershed management plans based on them, may need revision.

  19. Deriving Macropore and Preferential Flow Parameters from Tracer and Time-lapse 3D GPR Experiments at the Plot-Scale

    NASA Astrophysics Data System (ADS)

    Jackisch, Conrad; Allroggen, Niklas; Tronicke, Jens; Zehe, Erwin

    2014-05-01

    "Hydrology - a science in which all processes are preferential" (Uhlenbrook, 2006) - as such preferential flow is known and discussed in hydrology since almost three decades. At the same time, preferential flow remains problematic as explicit descriptions are hard to define and upscale and implicit descriptions remain rather case sensitive. Moreover, our techniques to monitor preferential flow and especially flow structures are very limited. We conducted three multi-tracer plot-scale (1m x 1m) sprinkler experiments at a forested hillslope in the Attert Basin in Luxembourg with prevailing geogenic and biogenic preferential flow structures. It was accompanied by a 3D time-lapse GPR (Ground Penetrating Radar) survey covering an area of 3m x 3m. We present the results with special emphasis on the derivation of macropore parameters for further modelling. To do so, we developed an automated analysis of images from excavated Brilliant Blue stained profiles. Additionally, we analyse our time-lapse GPR data with respect to temporal changes and derive 3D strutural information of the preferential flow patterns. Superior to tracers, this high resolution subsurface imaging technique is non-invasive, repeatable and therefore helps to disentangle the dye stained patterns towards process observation. The results of the image analyses and the GPR surveys are compared and referenced to soil moisture monitoring, sampled Bromide profiles and stable isotope signatures. We further discuss implications for joint development of model concepts and observation methods.

  20. Two Categories of 13C/12C Ratios for Higher Plants 1

    PubMed Central

    Smith, Bruce N.; Epstein, Samuel

    1971-01-01

    13C/12C ratios have been determined for plant tissue from 104 species representing 60 families. Higher plants fall into two categories, those with low δPDBI13C values (—24 to —34‰) and those with high δ 13C values (—6 to —19‰). Algae have δ 13C values of —12 to —23‰. Photosynthetic fractionation leading to such values is discussed. PMID:16657626

  1. 40 CFR 721.6505 - Polymers of C13C15 oxoalcohol ethoxolates.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Polymers of C13C15 oxoalcohol... Specific Chemical Substances § 721.6505 Polymers of C13C15 oxoalcohol ethoxolates. (a) Chemical substance... polymers of C13C15 oxoalcohol ethoxolates (PMNs P-96-950/951) are subject to reporting under this...

  2. 40 CFR 721.6505 - Polymers of C13C15 oxoalcohol ethoxolates.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Polymers of C13C15 oxoalcohol... Specific Chemical Substances § 721.6505 Polymers of C13C15 oxoalcohol ethoxolates. (a) Chemical substance... polymers of C13C15 oxoalcohol ethoxolates (PMNs P-96-950/951) are subject to reporting under this...

  3. 40 CFR 721.6505 - Polymers of C13C15 oxoalcohol ethoxolates.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Polymers of C13C15 oxoalcohol... Specific Chemical Substances § 721.6505 Polymers of C13C15 oxoalcohol ethoxolates. (a) Chemical substance... polymers of C13C15 oxoalcohol ethoxolates (PMNs P-96-950/951) are subject to reporting under this...

  4. 40 CFR 721.6505 - Polymers of C13C15 oxoalcohol ethoxolates.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Polymers of C13C15 oxoalcohol... Specific Chemical Substances § 721.6505 Polymers of C13C15 oxoalcohol ethoxolates. (a) Chemical substance... polymers of C13C15 oxoalcohol ethoxolates (PMNs P-96-950/951) are subject to reporting under this...

  5. 40 CFR 721.6505 - Polymers of C13C15 oxoalcohol ethoxolates.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Polymers of C13C15 oxoalcohol... Specific Chemical Substances § 721.6505 Polymers of C13C15 oxoalcohol ethoxolates. (a) Chemical substance... polymers of C13C15 oxoalcohol ethoxolates (PMNs P-96-950/951) are subject to reporting under this...

  6. In situ {sup 13}C MAS NMR study of n-hexane conversion on Pt and Pd supported on basic materials. II. On the mechanism of isomerization and hydrocracking

    SciTech Connect

    Ivanova, I.I.; Seirvert, M.; Pasau-Claerbout, A.; Derouane, E.G.; Blom, N.

    1996-12-01

    {sup 13}C MAS NMR spectroscopy was performed in situ to investigate the mechanisms of n-hexane isomerization and hydrocracking on Pt and Pd supported on Al-stabilized magnesia (Pt/Mg(Al)O and Pd/Mg(Al)O), and Pt on KL zeolite (Pt/KL). All the catalysts had high metal dispersion, the metal particle sizes being 13, 11, and 18 {Angstrom}, respectively. n-Hexane 1-{sup 13}C was used for in situ label tracer experiments. {sup 13}C MAS NMR spectra were obtained during the time course of the reaction at 573 and 653 K. The NMR results were then quantified, and the reaction kinetics were studied. Identification of the primary and secondary labeled reaction products led to the conclusion that both cyclic and bond-shift isomerization mechanisms operate on the three catalysts. In the case of Pt/Mg(Al)O, the cyclic mechanism accounts for 80% of the isomerization products. In the case of Pt/KL and Pd/Mg(Al)O, the contribution of bond-shift reactions increases due to restricted formation of the methylcyclopentane intermediate on the former and to suppressed hydrogenolysis of methylcyclopentane on the latter. A nonselective cyclic isomerization mechanism operates on magnesia catalysts, while on Pt/KL selective bisecondary bond rupturing occurs. Mechanistic pathways of bond-shift and hydrocracking reactions involve both 1,3- and 2,4-metallocyclobutane intermediates in the case of magnesia-supported catalysts, while in the case of the Pt/KL catalyst a 1,3-metallocyclobutane intermediate is preferentially formed. Only terminal scission occurs on Pt/KL. The Pd catalyst demonstrates enhanced activity in demethylation. The observed differences in the mechanistic pathways are explained on the basis of the specific properties of the metal and support. 64 refs., 14 figs., 6 tabs.

  7. Preliminary Results from a Gas Tracer Injection Experiment in the Upper Oceanic Crust on the Eastern Flank of the Juan de Fuca Ridge

    NASA Astrophysics Data System (ADS)

    Neira, N. M.; Clark, J. F.; Fisher, A. T.; Wheat, C. G.

    2013-12-01

    We present the first results from a gas tracer injection experiment in the ocean crust on the eastern flank of the Juan de Fuca Ridge, in an area of vigorous hydrothermal circulation. A mixture of tracers was injected in Hole 1362B in 2010, during IODP Expedition 327, as part of a 24-hour pumping experiment. Fluid samples were subsequently collected from this hole and three additional holes (1026B, 1362A, and 1301A), located 300 to 500 m away. The array of holes is located on 3.5 M.y. old seafloor, and oriented N20°E, subparallel to the Endeavor Segment of Juan de Fuca Ridge, 100 km to the west. Sulfur hexafluoride (SF6) was injected at a concentration of 0.0192 mol/min, with fluid pumping rate of 6.7 L/s for 20.2 h, resulting in a mean concentration of 47.6 μM and 23.3 mol of SF6 being added to crustal fluids. Borehole fluid samples were collected in copper coils using osmotic pumps attached to the wellheads of several long-term, subseafloor observatories (CORKs). These samples were recovered from the seafloor using a remotely-operated vehicle in 2011 and 2013. Analyses of SF6 concentrations in samples recovered in 2011 indicate the first arrival of SF6 in Hole 1301A, 550 m south of the injection Hole 1362B, ~265 days after injection. This suggests that the most rapid lateral transport of gas (at the leading edge of the plume) occurred at ~2 m/day. Samples recovered in 2013 should provide a more complete breakthrough curve, allowing assessment of the mean lateral transport rate. Additional insights will come from analysis of metal salts and particle tracers injected contemporaneously with the SF6, the cross-hole pressure response to injection and a two-year fluid discharge experiment. Additional wellhead samples will be collected in Summer 2014, as will downhole osmosamplers deployed in perforated casing within the upper ocean crust in Holes 1362A and 1362B.

  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. 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

  10. 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.

  11. Investigating 13C +12C reaction by the activation method. Sensitivity tests

    NASA Astrophysics Data System (ADS)

    Chesneanu, Daniela; Trache, L.; Margineanu, R.; Pantelica, A.; Ghita, D.; Straticiuc, M.; Burducea, I.; Blebea-Apostu, A. M.; Gomoiu, C. M.; Tang, X.

    2015-02-01

    We have performed experiments to check the limits of sensitivity of the activation method using the new 3 MV Tandetron accelerator and the low and ultra-low background laboratories of the "Horia Hulubei" National Institute of Physics and Nuclear Engineering (IFIN-HH). We have used the 12C +13C reaction at beam energies Elab= 6, 7 and 8 MeV. The knowledge of this fusion cross section at deep sub-barrier energies is of interest for astrophysical applications, as it provides an upper limit for the fusion cross section of 12C +12C over a wide energy range. A 13C beam with intensities 0.5-2 particleμA was provided by the accelerator and used to bombard graphite targets, resulting in activation with 24Na from the 12C (13C ,p) reaction. The 1369 and 2754 keV gamma-rays from 24Na de-activation were clearly observed in the spectra obtained in two different laboratories used for measurements at low and ultralow background: one at the surface and one located underground in the Unirea salt mine from Slanic Prahova, Romania. In the underground laboratory, for Elab = 6 MeV we have measured an activity of 0.085 ± 0.011 Bq, corresponding to cross sections of 1-3 nb. This demonstrates that it is possible to measure 12C targets irradiated at lower energies for at least 10 times lower cross sections than before β-γ coincidences will lead us another factor of 10 lower, proving that this installations can be successfully used for nuclear astrophysics measurements.

  12. 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

  13. Relativistic Force Field: Parametrization of (13)C-(1)H Nuclear Spin-Spin Coupling Constants.

    PubMed

    Kutateladze, Andrei G; Mukhina, Olga A

    2015-11-01

    Previously, we reported a reliable DU8 method for natural bond orbital (NBO)-aided parametric scaling of Fermi contacts to achieve fast and accurate prediction of proton-proton spin-spin coupling constants (SSCC) in (1)H NMR. As sophisticated NMR experiments for precise measurements of carbon-proton SSCCs are becoming more user-friendly and broadly utilized by the organic chemistry community to guide and inform the process of structure determination of complex organic compounds, we have now developed a fast and accurate method for computing (13)C-(1)H SSCCs. Fermi contacts computed with the DU8 basis set are scaled using selected NBO parameters in conjunction with empirical scaling coefficients. The method is optimized for inexpensive B3LYP/6-31G(d) geometries. The parametric scaling is based on a carefully selected training set of 274 ((3)J), 193 ((2)J), and 143 ((1)J) experimental (13)C-(1)H spin-spin coupling constants reported in the literature. The DU8 basis set, optimized for computing Fermi contacts, which by design had evolved from optimization of a collection of inexpensive 3-21G*, 4-21G, and 6-31G(d) bases, offers very short computational (wall) times even for relatively large organic molecules containing 15-20 carbon atoms. The most informative SSCCs for structure determination, i.e., (3)J, were computed with an accuracy of 0.41 Hz (rmsd). The new unified approach for computing (1)H-(1)H and (13)C-(1)H SSCCs is termed "DU8c".

  14. Investigating {sup 13}C+{sup 12}C reaction by the activation method. Sensitivity tests

    SciTech Connect

    Chesneanu, Daniela Trache, L.; Margineanu, R.; Pantelica, A.; Ghita, D.; Straticiuc, M.; Burducea, I.; Blebea-Apostu, A. M.; Gomoiu, C. M.; Tang, X.

    2015-02-24

    We have performed experiments to check the limits of sensitivity of the activation method using the new 3 MV Tandetron accelerator and the low and ultra-low background laboratories of the “Horia Hulubei” National Institute of Physics and Nuclear Engineering (IFIN-HH). We have used the {sup 12}C+{sup 13}C reaction at beam energies E{sub lab}= 6, 7 and 8 MeV. The knowledge of this fusion cross section at deep sub-barrier energies is of interest for astrophysical applications, as it provides an upper limit for the fusion cross section of {sup 12}C+{sup 12}C over a wide energy range. A {sup 13}C beam with intensities 0.5–2 particleμA was provided by the accelerator and used to bombard graphite targets, resulting in activation with {sup 24}Na from the {sup 12}C({sup 13}C,p) reaction. The 1369 and 2754 keV gamma-rays from {sup 24}Na de-activation were clearly observed in the spectra obtained in two different laboratories used for measurements at low and ultralow background: one at the surface and one located underground in the Unirea salt mine from Slanic Prahova, Romania. In the underground laboratory, for E{sub lab} = 6 MeV we have measured an activity of 0.085 ± 0.011 Bq, corresponding to cross sections of 1–3 nb. This demonstrates that it is possible to measure {sup 12}C targets irradiated at lower energies for at least 10 times lower cross sections than before β–γ coincidences will lead us another factor of 10 lower, proving that this installations can be successfully used for nuclear astrophysics measurements.

  15. Relativistic Force Field: Parametrization of (13)C-(1)H Nuclear Spin-Spin Coupling Constants.

    PubMed

    Kutateladze, Andrei G; Mukhina, Olga A

    2015-11-01

    Previously, we reported a reliable DU8 method for natural bond orbital (NBO)-aided parametric scaling of Fermi contacts to achieve fast and accurate prediction of proton-proton spin-spin coupling constants (SSCC) in (1)H NMR. As sophisticated NMR experiments for precise measurements of carbon-proton SSCCs are becoming more user-friendly and broadly utilized by the organic chemistry community to guide and inform the process of structure determination of complex organic compounds, we have now developed a fast and accurate method for computing (13)C-(1)H SSCCs. Fermi contacts computed with the DU8 basis set are scaled using selected NBO parameters in conjunction with empirical scaling coefficients. The method is optimized for inexpensive B3LYP/6-31G(d) geometries. The parametric scaling is based on a carefully selected training set of 274 ((3)J), 193 ((2)J), and 143 ((1)J) experimental (13)C-(1)H spin-spin coupling constants reported in the literature. The DU8 basis set, optimized for computing Fermi contacts, which by design had evolved from optimization of a collection of inexpensive 3-21G*, 4-21G, and 6-31G(d) bases, offers very short computational (wall) times even for relatively large organic molecules containing 15-20 carbon atoms. The most informative SSCCs for structure determination, i.e., (3)J, were computed with an accuracy of 0.41 Hz (rmsd). The new unified approach for computing (1)H-(1)H and (13)C-(1)H SSCCs is termed "DU8c". PMID:26414291

  16. Enhancing the understanding of earthworm feeding behaviour via the use of fatty acid delta13C values determined by gas chromatography-combustion-isotope ratio mass spectrometry.

    PubMed

    Dungait, Jennifer A J; Briones, Maria J I; Bol, Roland; Evershed, Richard P

    2008-06-01

    Litter-dwelling (epigeic) Lumbricus rubellus and soil-dwelling (endogeic) Allolobophora chlorotica earthworms were observed aggregating under C(3) (delta(13)C = -31.3 per thousand; delta(15)N = 10.7 per thousand) and C(4) (delta(13)C = -12.6 per thousand; delta(15)N = 7.5 per thousand) synthetic dung pats applied to a temperate grassland (delta(13)C = -30.3 per thousand; delta(15)N = 5.7 per thousand) in an experiment carried out for 372 days. Bulk delta(13)C values of earthworms collected from beneath either C(3) or C(4) dung after 28, 56, 112 and 372 days demonstrated that (i) L. rubellus beneath C(4) dung were significantly (13)C-enriched after 56 days (delta(13)C = -23.8 per thousand) and 112 days (delta(13)C = -22.4 per thousand) compared with those from C(3) dung treatments (56 days, delta(13)C = -26.5 per thousand; 112 days, delta(13)C = -27.0 per thousand), and (ii) A. chlorotica were 2.1 per thousand (13)C-enriched (delta(13)C = -24.2 per thousand) relative to those from C(3) dung (delta(13)C = -26.3 per thousand) treatments after 372 days. Bulk delta(15)N values did not suggest significant uptake of dung N by either species beneath C(3) or C(4) dung, but showed that the endogeic species (total mean delta(15)N = 3.3 per thousand) had higher delta(15)N values than the epigeic species (total mean delta(15)N = 5.4 per thousand). Although the two species exhibited similar fatty acid profiles, individual fatty acid delta(13)C values revealed extensive routing of dietary C into body tissue of L. rubellus, but minor incorporation into A. chlorotica. In particular, the direct incorporation of microbial biomarker fatty acids (iC(17:0), aC(17:0)) from (13)C-labelled dung in situ, the routing of dung C into de novo synthesised compounds (iC(20:4)(omega)(6),C(20:5)(omega)(3), and the assimilation of essential fatty acids ((C(18:1)(omega)(9), C(18:1)(omega(7), C(18:2)(omega(6), C(18:3)(omega)(3)) derived from dung, were determined.

  17. 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

  18. 13C and 1H Nuclear Magnetic Resonance Study of Glycogen Futile Cycling in Strains of the Genus Fibrobacter

    PubMed Central

    Matheron, Christelle; Delort, Anne-Marie; Gaudet, Geneviève; Forano, Evelyne; Liptaj, Tibor

    1998-01-01

    We investigated the carbon metabolism of three strains of Fibrobacter succinogenes and one strain of Fibrobacter intestinalis. The four strains produced the same amounts of the metabolites succinate, acetate, and formate in approximately the same ratio (3.7/1/0.3). The four strains similarly stored glycogen during all growth phases, and the glycogen-to-protein ratio was close to 0.6 during the exponential growth phase. 13C nuclear magnetic resonance (NMR) analysis of [1-13C]glucose utilization by resting cells of the four strains revealed a reversal of glycolysis at the triose phosphate level and the same metabolic pathways. Glycogen futile cycling was demonstrated by 13C NMR by following the simultaneous metabolism of labeled [13C]glycogen and exogenous unlabeled glucose. The isotopic dilutions of the CH2 of succinate and the CH3 of acetate when the resting cells were metabolizing [1-13C]glucose and unlabeled glycogen were precisely quantified by using 13C-filtered spin-echo difference 1H NMR spectroscopy. The measured isotopic dilutions were not the same for succinate and acetate; in the case of succinate, the dilutions reflected only the contribution of glycogen futile cycling, while in the case of acetate, another mechanism was also involved. Results obtained in complementary experiments are consistent with reversal of the succinate synthesis pathway. Our results indicated that for all of the strains, from 12 to 16% of the glucose entering the metabolic pathway originated from prestored glycogen. Although genetically diverse, the four Fibrobacter strains studied had very similar carbon metabolism characteristics. PMID:12033219

  19. Isotopologues of Dense Gas Tracers in NGC 1068

    NASA Astrophysics Data System (ADS)

    Wang, Junzhi; Zhang, Zhi-Yu; Qiu, Jianjie; Shi, Yong; Zhang, Jiangshui; Fang, Min

    2014-11-01

    We present observations of isotopic lines of dense gas tracers toward the nuclear region of nearby Seyfert 2 galaxy NGC 1068 with the IRAM 30 m telescope and the Atacama Pathfinder Experiment (APEX) 12 m telescope. We detected four isotopic lines (H13CN 1-0, H13CO+ 1-0, HN13C 1-0, and HC18O+ 1-0) at the 3 mm band with the IRAM 30 m telescope and obtained upper limits of other lines. We calculated optical depths of dense gas tracers with the detected isotopic lines of HCN 1-0, HCO+ 1-0, and HNC 1-0. We find that the 14N/15N abundance ratio is greater than 420 if we adopt the upper limit of HC15N(1-0) emission. Combining this with fluxes of 1-0 lines from IRAM 30 m observations and the upper limit of 3-2 lines from APEX 12 m observations, we also estimated the excitation condition of molecular gas in the nuclear region of NGC 1068, which is less dense than that in the extreme starburst regions of galaxies.

  20. Isotopologues of dense gas tracers in NGC 1068

    SciTech Connect

    Wang, Junzhi; Qiu, Jianjie; Zhang, Zhi-Yu; Shi, Yong; Zhang, Jiangshui; Fang, Min

    2014-11-20

    We present observations of isotopic lines of dense gas tracers toward the nuclear region of nearby Seyfert 2 galaxy NGC 1068 with the IRAM 30 m telescope and the Atacama Pathfinder Experiment (APEX) 12 m telescope. We detected four isotopic lines (H{sup 13}CN 1-0, H{sup 13}CO{sup +} 1-0, HN{sup 13}C 1-0, and HC{sup 18}O{sup +} 1-0) at the 3 mm band with the IRAM 30 m telescope and obtained upper limits of other lines. We calculated optical depths of dense gas tracers with the detected isotopic lines of HCN 1-0, HCO{sup +} 1-0, and HNC 1-0. We find that the {sup 14}N/{sup 15}N abundance ratio is greater than 420 if we adopt the upper limit of HC{sup 15}N(1-0) emission. Combining this with fluxes of 1-0 lines from IRAM 30 m observations and the upper limit of 3-2 lines from APEX 12 m observations, we also estimated the excitation condition of molecular gas in the nuclear region of NGC 1068, which is less dense than that in the extreme starburst regions of galaxies.

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

    PubMed

    Kumar, Dinesh; Hosur, Ramakrishna V

    2011-09-01

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

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

    PubMed

    Kumar, Dinesh; Hosur, Ramakrishna V

    2011-09-01

    A three-dimensional nuclear magnetic resonance (NMR) pulse sequence named as hNCOcanH has been described to aid rapid sequential assignment of backbone resonances in (15)N/(13)C-labeled proteins. The experiment has been derived by a simple modification of the previously described HN(C)N pulse sequence [Panchal et al., J. Biomol. NMR 20 (2001) 135-147]; t2 evolution is used to frequency label (13)C' rather than (15)N (similar trick has also been used in the design of hNCAnH pulse sequence from hNcaNH [Frueh et al., JACS, 131 (2009) 12880-12881]). The modification results in a spectrum equivalent to HNCO, but in addition to inter-residue correlation peaks (i.e. Hi , Ci-1), the spectrum also contains additional intra-residue correlation peaks (i.e. Hi-1 , Ci-1) in the direct proton dimension which has maximum resolution. This is the main strength of the experiment and thus, even a small difference in amide (1) H chemical shifts (5-6 Hz) can be used for establishing a sequential connectivity. This experiment in combination with the HNN experiment described previously [Panchal et al., J. Biomol. NMR 20 (2001) 135-147] leads to a more robust assignment protocol for backbone resonances ((1) H(N) , (15)N) than could be derived from the combination of HNN and HN(C)N experiments [Bhavesh et al., Biochemistry, 40 (2001) 14727-14735]. Further, this new protocol enables assignment of (13)C' resonances as well. We believe that the experiment and the protocol presented here will be of immense value for structural-and functional-proteomics research by NMR. Performance of this experiment has been demonstrated using (13)C/(15)N labeled ubiquitin. PMID:21818779

  4. Pharmaceutical residues in rivers: Quantification of transient storage and travel times by tracer experiments as key parameters for the determination of mass balances

    NASA Astrophysics Data System (ADS)

    Kunkel, U.; Clemens, C.; Radke, M.

    2009-04-01

    Pharmaceutical residues are commonly detected micropollutants in the aquatic environment. To derive mass balances of these substances in river stretches and to quantify their elimination, besides highly sophisticated analytical methods precise discharge measurements are inevitable. Two additional key parameters are the travel time of water along a river stretch and the hyporheic exchange of water and solutes. Knowledge of travel times is essential to compare concentrations at the upstream and downstream end of an investigated site. As biodegradation in sediments is assumed to be a potentially significant removal process for organic micropollutants in rivers, flow of water and solutes across the sediment-water boundary and their transient storage in the hyporheic zone are also pivotal processes. To determine these river characteristics, tracer experiments using the fluorescent dye uranine were conducted at a 15 km stretch of a small river in Northern Bavaria (Roter Main) in summer 2008. Three sub-reaches were studied in separate tracer tests, and river discharge was simultaneously determined using the ultrasonic doppler technique. The travel time of uranine for the total stretch amounted to almost 1.5 days, corresponding to a mean flow velocity of about 0.4 km/h. Strong tailing of the breakthrough curves could indicate significant transient storage in the hyporheic zone. However, we tentatively attribute this tailing to the merging of two river channels having different flow characteristics and not to transient storage, and thus the hyporheic exchange is expected to be small along the river stretch. The results of the experiments are currently being analyzed by the OTIS model in order to report the exchange rates of surface and pore water on a quantitative basis.

  5. The Late Pleistocene history of surface water δ13C in the Sulu Sea: Possible relationship to Pacific Deepwater δ13C changes

    NASA Astrophysics Data System (ADS)

    Linsley, Braddock K.; Dunbar, Robert B.

    1994-04-01

    A reconstruction of late Pleistocene surface water carbon isotopic (δ13C) variability is presented from Ocean Drilling Program (ODP) site 769 in the Sulu Sea in the western tropical Pacific. The Sulu Sea is a shallowly silled back arc basin with a maximum sill depth of 420 m. Site 769 was drilled on a bathymetric high in 3643 m of water and has average late Pleistocene sedimentation rates of 8.5 cm/kyr. The oxygen isotope record (δ18O) of Globigerinoides ruber at site 769 shows a strong correlation with the SPECMAP stacked δ18O record, attesting to the continuity of sediment archive at the site. Surface δ13C displays consistent glacial-interglacial variability which averages ˜0.9‰ and has varied from 0.75 to 1.1‰ over the last 800 kyr. Comparison to surface water δ13C records in the South China Sea and western tropical Pacific suggests that the glacial-interglacial surface δ13C variability is regional in scale. Planktonic δ13C data from ODP site 677 in the eastern Pacific is also coherent with the site 769. Additionally, we have found that the site 769 surface δ13C record is coherent at periods of 100 and 41 kyr with deepwater δ13C records from the Pacific. The highest correlation occurs with the deep eastern Pacific, where benthic δ13C data from cores RC13-110 and ODP site 677 closely match the Sulu Sea surface water record. We evaluate several possible controls of surface water δ13C in the Sulu Sea that may explain the coherent timing with Pacific deepwater δ13C records. These include variations in terrestrial organic matter flux to the basin, the upwelling of subsurface water and productivity changes, and the influx of western Pacific intermediate water to the Sulu Sea. Our preferred explanation involves a region of upper intermediate water upwelling in the far western Pacific which has been shown to outgas CO2 from subsurface waters into surface waters. Upwelling also occurs in the area of Panama Basin site 677. These equatorial upwelling zones

  6. Sc3CH@C80: selective (13)C enrichment of the central carbon atom.

    PubMed

    Junghans, Katrin; Rosenkranz, Marco; Popov, Alexey A

    2016-05-01

    Sc3CH@C80 is synthesized and characterized by (1)H, (13)C, and (45)Sc NMR. A large negative chemical shift of the proton, -11.73 ppm in the Ih and -8.79 ppm in the D5h C80 cage isomers, is found. (13)C satellites in the (1)H NMR spectrum enabled indirect determination of the (13)C chemical shift for the central carbon at 173 ± 1 ppm. Intensity of the satellites allowed determination of the (13)C content for the central carbon atom. This unique possibility is applied to analyze the cluster/cage (13)C distribution in mechanistic studies employing either (13)CH4 or (13)C powder to enrich Sc3CH@C80 with (13)C. PMID:27109443

  7. Sc3CH@C80: selective 13C enrichment of the central carbon atom†

    PubMed Central

    Junghans, Katrin; Rosenkranz, Marco; Popov, Alexey A.

    2016-01-01

    Sc3CH@C80 is synthesized and characterized by 1H, 13C, and 45Sc NMR. A large negative chemical shift of the proton, −11.73 ppm in the Ih and −8.79 ppm in the D5h C80 cage isomers, is found. 13C satellites in the 1H NMR spectrum enabled indirect determination of the 13C chemical shift for the central carbon at 173 ± 1 ppm. Intensity of the satellites allowed determination of the 13C content for the central carbon atom. This unique possibility is applied to analyze the cluster/cage 13C distribution in mechanistic studies employing either 13CH4 or 13C powder to enrich Sc3CH@C80 with 13C. PMID:27109443

  8. The role of snowmelt and glacier melt on runoff in a glacierized catchment: a multi-tracer experiment

    NASA Astrophysics Data System (ADS)

    Penna, Daniele; Engel, Michael; Mao, Luca; Dell'Agnese, Andrea; Bertoldi, Giacomo; Comiti, Francesco

    2013-04-01

    The release of water as snowmelt and ice melt in high elevation catchments has significant social and economic impacts for population living in mountain areas. This is even more critical under the current conditions of glacier retreat as a consequence of global warming. Therefore, it is important to understand the role of ice and snow meltwater on runoff dynamics and groundwater recharge in glacierized environments. This task can be effectively accomplished by integrating isotopic and other tracers that are widely recognized as useful tools for the identification of the main water sources contributing to streamflow. In this work, we collected water samples from different sources in the Saldur catchment (Eastern Italian Alps). The catchment (area: 62 km², elevation range: 1600-3700 m a.s.l.) hosts a small glacier (2.8 km²) in its upper portion. Samples of rainfall, snow, snowmelt, glacier melt, stream water (main stream and tributaries) and spring water have been manually collected between April-October 2011 and April-November 2012 approximately on a monthly basis. Furthermore, 24-hour samplings with hourly collection frequency were performed at two cross sections during five melt-runoff events. The composition in stable water isotopes was determined by laser spectroscopy and mass spectrometry. Electrical conductivity (EC) and water temperature were measured in the field. Additionally, deuterium excess (DE) was computed for all samples based on the relationship between deuterium and 18-oxygen. The isotopic composition of rainfall and snow shows marked altitudinal and seasonal variations. A strong positive correlation is also evident in the relationship between DE of spring waters and elevation. Rainfall and snow samples fall perfectly on the Global Meteoric Water Line, revealing a predominant Atlantic origin of air masses producing precipitation in the study area. EC and water temperature linearly increase with the distance from the glacier snout, suggesting a

  9. Sauna, sweat and science - quantifying the proportion of condensation water versus sweat using a stable water isotope ((2)H/(1)H and (18)O/(16)O) tracer experiment.

    PubMed

    Zech, Michael; Bösel, Stefanie; Tuthorn, Mario; Benesch, Marianne; Dubbert, Maren; Cuntz, Matthias; Glaser, Bruno

    2015-01-01

    Most visitors of a sauna appreciate the heat pulse that is perceived when water is poured on the stones of a sauna stove. However, probably only few bathers are aware that this pleasant heat pulse is caused by latent heat being released onto our skin due to condensation of water vapour. In order to quantify the proportion of condensation water versus sweat to dripping water of test persons we conducted sauna experiments using isotopically labelled (δ(18)O and δ(2)H) thrown water as tracer. This allows differentiating between 'pure sweat' and 'condensation water'. Two ways of isotope mass balance calculations were applied and yielded similar results for both water isotopes. Accordingly, condensation contributed considerably to dripping water with mean proportions of 52 ± 12 and 54 ± 7% in a sauna experiment in winter semester 2011/12 and 30 ± 13 and 33 ± 6% in a sauna experiment in winter semester 2012/13, respectively, depending on the way of calculating the isotope mass balance. It can be concluded from the results of our dual isotope labelling sauna experiment that it is not all about sweat in the sauna.

  10. Sauna, sweat and science - quantifying the proportion of condensation water versus sweat using a stable water isotope ((2)H/(1)H and (18)O/(16)O) tracer experiment.

    PubMed

    Zech, Michael; Bösel, Stefanie; Tuthorn, Mario; Benesch, Marianne; Dubbert, Maren; Cuntz, Matthias; Glaser, Bruno

    2015-01-01

    Most visitors of a sauna appreciate the heat pulse that is perceived when water is poured on the stones of a sauna stove. However, probably only few bathers are aware that this pleasant heat pulse is caused by latent heat being released onto our skin due to condensation of water vapour. In order to quantify the proportion of condensation water versus sweat to dripping water of test persons we conducted sauna experiments using isotopically labelled (δ(18)O and δ(2)H) thrown water as tracer. This allows differentiating between 'pure sweat' and 'condensation water'. Two ways of isotope mass balance calculations were applied and yielded similar results for both water isotopes. Accordingly, condensation contributed considerably to dripping water with mean proportions of 52 ± 12 and 54 ± 7% in a sauna experiment in winter semester 2011/12 and 30 ± 13 and 33 ± 6% in a sauna experiment in winter semester 2012/13, respectively, depending on the way of calculating the isotope mass balance. It can be concluded from the results of our dual isotope labelling sauna experiment that it is not all about sweat in the sauna. PMID:26110629

  11. Solid-state (13)C NMR reveals effects of temperature and hydration on elastin.

    PubMed Central

    Perry, Ashlee; Stypa, Michael P; Tenn, Brandon K; Kumashiro, Kristin K

    2002-01-01

    Elastin is the principal protein component of the elastic fiber in vertebrate tissue. The waters of hydration in the elastic fiber are believed to play a critical role in the structure and function of this largely hydrophobic, amorphous protein. (13)C CPMAS NMR spectra are acquired for elastin samples with different hydration levels. The spectral intensities in the aliphatic region undergo significant changes as 70% of the water in hydrated elastin is removed. In addition, dramatic differences in the CPMAS spectra of hydrated, lyophilized, and partially dehydrated elastin samples over a relatively small temperature range (-20 degrees C to 37 degrees C) are observed. Results from other experiments, including (13)C T(1) and (1)H T(1 rho) measurements, direct polarization with magic-angle spinning, and static CP of the hydrated and lyophilized elastin preparations, also support the model that there is significant mobility in fully hydrated elastin. Our results support models in which water plays an integral role in the structure and proper function of elastin in vertebrate tissue. PMID:11806948

  12. Natural isotope correction of MS/MS measurements for metabolomics and (13)C fluxomics.

    PubMed

    Niedenführ, Sebastian; ten Pierick, Angela; van Dam, Patricia T N; Suarez-Mendez, Camilo A; Nöh, Katharina; Wahl, S Aljoscha

    2016-05-01

    Fluxomics and metabolomics are crucial tools for metabolic engineering and biomedical analysis to determine the in vivo cellular state. Especially, the application of (13)C isotopes allows comprehensive insights into the functional operation of cellular metabolism. Compared to single MS, tandem mass spectrometry (MS/MS) provides more detailed and accurate measurements of the metabolite enrichment patterns (tandem mass isotopomers), increasing the accuracy of metabolite concentration measurements and metabolic flux estimation. MS-type data from isotope labeling experiments is biased by naturally occurring stable isotopes (C, H, N, O, etc.). In particular, GC-MS(/MS) requires derivatization for the usually non-volatile intracellular metabolites introducing additional natural isotopes leading to measurements that do not directly represent the carbon labeling distribution. To make full use of LC- and GC-MS/MS mass isotopomer measurements, the influence of natural isotopes has to be eliminated (corrected). Our correction approach is analyzed for the two most common applications; (13)C fluxomics and isotope dilution mass spectrometry (IDMS) based metabolomics. Natural isotopes can have an impact on the calculated flux distribution which strongly depends on the substrate labeling and the actual flux distribution. Second, we show that in IDMS based metabolomics natural isotopes lead to underestimated concentrations that can and should be corrected with a nonlinear calibration. Our simulations indicate that the correction for natural abundance in isotope based fluxomics and quantitative metabolomics is essential for correct data interpretation.

  13. Natural isotope correction of MS/MS measurements for metabolomics and (13)C fluxomics.

    PubMed

    Niedenführ, Sebastian; ten Pierick, Angela; van Dam, Patricia T N; Suarez-Mendez, Camilo A; Nöh, Katharina; Wahl, S Aljoscha

    2016-05-01

    Fluxomics and metabolomics are crucial tools for metabolic engineering and biomedical analysis to determine the in vivo cellular state. Especially, the application of (13)C isotopes allows comprehensive insights into the functional operation of cellular metabolism. Compared to single MS, tandem mass spectrometry (MS/MS) provides more detailed and accurate measurements of the metabolite enrichment patterns (tandem mass isotopomers), increasing the accuracy of metabolite concentration measurements and metabolic flux estimation. MS-type data from isotope labeling experiments is biased by naturally occurring stable isotopes (C, H, N, O, etc.). In particular, GC-MS(/MS) requires derivatization for the usually non-volatile intracellular metabolites introducing additional natural isotopes leading to measurements that do not directly represent the carbon labeling distribution. To make full use of LC- and GC-MS/MS mass isotopomer measurements, the influence of natural isotopes has to be eliminated (corrected). Our correction approach is analyzed for the two most common applications; (13)C fluxomics and isotope dilution mass spectrometry (IDMS) based metabolomics. Natural isotopes can have an impact on the calculated flux distribution which strongly depends on the substrate labeling and the actual flux distribution. Second, we show that in IDMS based metabolomics natural isotopes lead to underestimated concentrations that can and should be corrected with a nonlinear calibration. Our simulations indicate that the correction for natural abundance in isotope based fluxomics and quantitative metabolomics is essential for correct data interpretation. PMID:26479486

  14. High-throughput hyperpolarized 13C metabolic investigations using a multi-channel acquisition system

    NASA Astrophysics Data System (ADS)

    Lee, Jaehyuk; Ramirez, Marc S.; Walker, Christopher M.; Chen, Yunyun; Yi, Stacey; Sandulache, Vlad C.; Lai, Stephen Y.; Bankson, James A.

    2015-11-01

    Magnetic resonance imaging and spectroscopy of hyperpolarized (HP) compounds such as [1-13C]-pyruvate have shown tremendous potential for offering new insight into disease and response to therapy. New applications of this technology in clinical research and care will require extensive validation in cells and animal models, a process that may be limited by the high cost and modest throughput associated with dynamic nuclear polarization. Relatively wide spectral separation between [1-13C]-pyruvate and its chemical endpoints in vivo are conducive to simultaneous multi-sample measurements, even in the presence of a suboptimal global shim. Multi-channel acquisitions could conserve costs and accelerate experiments by allowing acquisition from multiple independent samples following a single dissolution. Unfortunately, many existing preclinical MRI systems are equipped with only a single channel for broadband acquisitions. In this work, we examine the feasibility of this concept using a broadband multi-channel digital receiver extension and detector arrays that allow concurrent measurement of dynamic spectroscopic data from ex vivo enzyme phantoms, in vitro anaplastic thyroid carcinoma cells, and in vivo in tumor-bearing mice. Throughput and the cost of consumables were improved by up to a factor of four. These preliminary results demonstrate the potential for efficient multi-sample studies employing hyperpolarized agents.

  15. Incomplete fusion reactions at low energies in 13C+169Tm system

    NASA Astrophysics Data System (ADS)

    Sharma, Vijay R.; Yadav, Abhishek; Singh, Devendra P.; Singh, Pushpendra P.; Bala, Indu; Kumar, R.; Sharma, M. K.; Gupta, S.; Murlithar, S.; Singh, R. P.; Singh, B. P.; Prasad, R.

    2014-03-01

    Aiming to investigate the incomplete fusion processes at low projectile energies, experiments have been carried out for the 13C + 169Tm system at ≈ 4-7 MeV/A. Excitation functions for several heavy residues likely to be populated via complete and incomplete fusion processes have been measured using heavy recoil residue catcher technique followed by γ- ray spectroscopy. The measured cross-sections for the complete fusion (xn and pxn) channels are compared with the statistical model code PACE4, consistently using the same set of parameters. The complete fusion channels are found to be consistent with the model calculations. However, the cross-sections for all the measured α-emitting channels are found to be significantly enhanced over the calculations. Analysis of data indicate a significant fraction of incomplete fusion even at energies as low as 17% above barrier. The present results are discussed in light of the Morgenstern's systematics. Incomplete fusion strength function is found to be relatively large for alpha cluster projectile i.e. for 12C as compared to one neutron excess 13C projectile.

  16. Characterization of uniformly and atom-specifically 13C-labeled heparin and heparan sulfate polysaccharide precursors using 13C NMR spectroscopy and ESI mass spectrometry

    PubMed Central

    Nguyen, Thao K. N.; Tran, Vy M.; Victor, Xylophone V.; Skalicky, Jack J.; Kuberan, Balagurunathan

    2010-01-01

    The biological actions of heparin and heparan sulfate, two structurally related glycosaminoglycans, depend on the organization of the complex heparanome. Due to the structural complexity of the heparanome, the sequence of variably sulfonated uronic acid and glucosamine residues is usually characterized by the analysis of smaller oligosaccharide and disaccharide fragments. Even characterization of smaller heparin/heparan sulfate oligosaccharide or disaccharide fragments using simple 1D 1H NMR spectroscopy is often complicated by the extensive signal overlap. 13C NMR signals, on the other hand, overlap less and therefore, 13C NMR spectroscopy can greatly facilitate the structural elucidation of the complex heparanome and provide finer insights into the structural basis for biological functions. This is the first report of the preparation of anomeric carbon-specific 13C-labeled heparin/heparan sulfate precursors from the Escherichia coli K5 strain. Uniformly 13C- and 15N-labeled precursors were also produced and characterized by 13C NMR spectroscopy. Mass spectrometric analysis of enzymatically fragmented disaccharides revealed that anomeric carbon-specific labeling efforts resulted in a minor loss/scrambling of 13C in the precursor backbone, whereas uniform labeling efforts resulted in greater than 95% 13C isotope enrichment in the precursor backbone. These labeled precursors provided high-resolution NMR signals with great sensitivity and set the stage for studying the heparanome–proteome interactions. PMID:20832774

  17. Side-chain H and C resonance assignment in protonated/partially deuterated proteins using an improved 3D 13C-detected HCC-TOCSY

    NASA Astrophysics Data System (ADS)

    Hu, Kaifeng; Vögeli, Beat; Pervushin, Konstantin

    2005-06-01

    We propose the use of 13C-detected 3D HCC-TOCSY experiments for assignment of 1H and 13C resonances in protonated and partially deuterated proteins. The experiments extend 2D C-13-start and C-13-observe TOCSY type experiments proposed earlier [J. Biomol. NMR 26 (2) (2003) 167]. Introduction of the third 1H dimension to 2D TOCSY: (i) reduces the peak overlap and (ii) increases the sensitivity per unit time, even for highly deuterated (>85%) protein samples, which makes this improved method an attractive tool for the side-chain H and C assignment of average sized proteins with natural isotope abundance as well as large partially deuterated proteins. The experiments are demonstrated with a 16 kDa 15N, 13C-labeled non-deuterated apo-CcmE and a 48 kDa uniformly 15N, 13C-labeled and fractionally (˜90%) deuterated dimeric sFkpA. It is predicted that this method should be suitable for the assignment of methyl 13C and 1H chemical shifts of methyl protonated, highly deuterated and 13C-labeled proteins with even higher molecular weight.

  18. 13C Pathway Analysis for the Role of Formate in Electricity Generation by Shewanella Oneidensis MR-1 Using Lactate in Microbial Fuel Cells

    PubMed Central

    Luo, Shuai; Guo, Weihua; H. Nealson, Kenneth; Feng, Xueyang; He, Zhen

    2016-01-01

    Microbial fuel cell (MFC) is a promising technology for direct electricity generation from organics by microorganisms. The type of electron donors fed into MFCs affects the electrical performance, and mechanistic understanding of such effects is important to optimize the MFC performance. In this study, we used a model organism in MFCs, Shewanella oneidensis MR-1, and 13C pathway analysis to investigate the role of formate in electricity generation and the related microbial metabolism. Our results indicated a synergistic effect of formate and lactate on electricity generation, and extra formate addition on the original lactate resulted in more electrical output than using formate or lactate as a sole electron donor. Based on the 13C tracer analysis, we discovered decoupled cell growth and electricity generation in S. oneidensis MR-1 during co-utilization of lactate and formate (i.e., while the lactate was mainly metabolized to support the cell growth, the formate was oxidized to release electrons for higher electricity generation). To our best knowledge, this is the first time that 13C tracer analysis was applied to study microbial metabolism in MFCs and it was demonstrated to be a valuable tool to understand the metabolic pathways affected by electron donors in the selected electrochemically-active microorganisms. PMID:26868848

  19. FeCycle: Attempting an iron biogeochemical budget from a mesoscale SF6 tracer experiment in unperturbed low iron waters

    NASA Astrophysics Data System (ADS)

    Boyd, P. W.; Law, C. S.; Hutchins, D. A.; Abraham, E. R.; Croot, P. L.; Ellwood, M.; Frew, R. D.; Hadfield, M.; Hall, J.; Handy, S.; Hare, C.; Higgins, J.; Hill, P.; Hunter, K. A.; Leblanc, K.; Maldonado, M. T.; McKay, R. M.; Mioni, C.; Oliver, M.; Pickmere, S.; Pinkerton, M.; Safi, K.; Sander, S.; Sanudo-Wilhelmy, S. A.; Smith, M.; Strzepek, R.; Tovar-Sanchez, A.; Wilhelm, S. W.

    2005-12-01

    An improved knowledge of iron biogeochemistry is needed to better understand key controls on the functioning of high-nitrate low-chlorophyll (HNLC) oceanic regions. Iron budgets for HNLC waters have been constructed using data from disparate sources ranging from laboratory algal cultures to ocean physics. In summer 2003 we conducted FeCycle, a 10-day mesoscale tracer release in HNLC waters SE of New Zealand, and measured concurrently all sources (with the exception of aerosol deposition) to, sinks of iron from, and rates of iron recycling within, the surface mixed layer. A pelagic iron budget (timescale of days) indicated that oceanic supply terms (lateral advection and vertical diffusion) were relatively small compared to the main sink (downward particulate export). Remote sensing and terrestrial monitoring reveal 13 dust or wildfire events in Australia, prior to and during FeCycle, one of which may have deposited iron at the study location. However, iron deposition rates cannot be derived from such observations, illustrating the difficulties in closing iron budgets without quantification of episodic atmospheric supply. Despite the threefold uncertainties reported for rates of aerosol deposition (Duce et al., 1991), published atmospheric iron supply for the New Zealand region is ˜50-fold (i.e., 7- to 150-fold) greater than the oceanic iron supply measured in our budget, and thus was comparable (i.e., a third to threefold) to our estimates of downward export of particulate iron. During FeCycle, the fluxes due to short term (hours) biological iron uptake and regeneration were indicative of rapid recycling and were tenfold greater than for new iron (i.e. estimated atmospheric and measured oceanic supply), giving an "fe" ratio (uptake of new iron/uptake of new + regenerated iron) of 0.17 (i.e., a range of 0.06 to 0.51 due to uncertainties on aerosol iron supply), and an "Fe" ratio (biogenic Fe export/uptake of new + regenerated iron) of 0.09 (i.e., 0.03 to 0.24).

  20. The economical synthesis of [2'-(13)C, 1,3-(15)N2]uridine; preliminary conformational studies by solid state NMR.

    PubMed

    Patching, Simon G; Middleton, David A; Henderson, Peter J F; Herbert, Richard B

    2003-06-21

    The synthesis of [2'-(13)C, 1,3-(15)N2]uridine 11 was achieved as follows. An epimeric mixture of D-[1-(13)C]ribose 3 and D-[1-(13)C]arabinose 4 was obtained in excellent yield by condensation of K13CN with D-erythrose 2 using a modification of the Kiliani-Fischer synthesis. Efficient separation of the two aldose epimers was pivotally achieved by a novel ion-exchange (Sm3+) chromatography method. D-[2-(13)C]Ribose 5 was obtained from D-[1-(13)C]arabinose 4 using a Ni(II) diamine complex (nickel chloride plus TEMED). Combination of these procedures in a general cycling manner can lead to the very efficient preparation of specifically labelled 13C-monosaccharides of particular chirality. 15N-labelling was introduced in the preparation of [2'-(13)C, 1,3-(15)N2]uridine 11 via [15N2]urea. Cross polarisation magic angle spinning (CP-MAS) solid-state NMR experiments using rotational echo double resonance (REDOR) were carried out on crystals of the labelled uridine to show that the inter-atomic distance between C-2' and N-1 is closely similar to that calculated from X-ray crystallographic data. The REDOR method will be used now to determine the conformation of bound substrates in the bacterial nucleoside transporters NupC and NupG.

  1. Carbon isotope discrimination during litter decomposition can be explained by selective use of substrate with differing δ13C

    NASA Astrophysics Data System (ADS)

    Ngao, J.; Cotrufo, M. F.

    2011-01-01

    Temporal dynamics of C isotopic composition (δ13C) of CO2 and leaf litter was monitored during a litter decomposition experiment using Arbutus unedo L., as a slow decomposing model substrate. This allowed us (1) to quantify isotopic discrimination variation during litter decomposition, and (2) to test whether selective substrate use or kinetic fractionation could explain the observed isotopic discrimination. Total cumulative CO2-C loss (CL) comprised 27% of initial litter C. Temporal evolution of CL was simulated following a three-C-pool model. Isotopic composition of respired CO2 (δRL) was higher with respect to that of the bulk litter. The isotopic discrimination Δ(L/R) varied from -2‰ to 0‰ and it is mostly attributed to the variations of δRL. A three-pool model, with the three pools differing in their δ13C, described well the dynamic of Δ(L/R), in the intermediate stage of the process. This suggests that the observed isotopic discrimination between respired CO2 and bulk litter is in good agreement with the hypothesis of successive consumption of C compounds differing in δ13C during decomposition. However, to explain also 13C-CO2 dynamics at the beginning and end of the incubation the model had to be modified, with discrimination factors ranging from -1‰ to -4.6‰ attributed to the labile and the recalcitrance pool, respectively. We propose that this discrimination is also the result of further selective use of specific substrates within the two pools, likely being both the labile and recalcitrant pool of composite nature. In fact, the 2‰ 13C enrichment of the α-cellulose observed by the end of the experiment, and potentially attributable to kinetic fractionation, could not explain the measured Δ(L/R) dynamics.

  2. 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.

  3. The influence of fish cage culture on δ13C and δ15N of filter-feeding Bivalvia (Mollusca).

    PubMed

    Benedito, E; Figueroa, L; Takeda, A M; Manetta, G I

    2013-11-01

    The objective of this study was to evaluate the effect of Oreochromis niloticus cage culture promoted variations in the δ13C and δ15N in Corbicula fluminea (Mollusca; Bivalvia) and in the sediment of an aquatic food web. Samples were taken before and after net cage installation in the Rosana Reservoir (Paranapanema River, PR-SP). Samples of specimens of the bivalve filterer C. fluminea and samples of sediment were collected using a modified Petersen grab. All samples were dried in an oven (60 °C) for 72 hours, macerated to obtain homogenous fine powders and sent for carbon (δ13C) and nitrogen (δ15N) isotopic value analysis in a mass spectrometer. There were significant differences in the δ13C and δ15N values of the invertebrate C. fluminea between the beginning and the end of the experiment. There were no differences between the δ13C and δ15N values of sediment. These results indicate that the installation of fish cage culture promoted impacts in the isotopic composition of the aquatic food web organisms, which could exert influence over the native species and the ecosystem.

  4. δ(13)C-CH4 reveals CH4 variations over oceans from mid-latitudes to the Arctic.

    PubMed

    Yu, Juan; Xie, Zhouqing; Sun, Liguang; Kang, Hui; He, Pengzhen; Xing, Guangxi

    2015-01-01

    The biogeochemical cycles of CH4 over oceans are poorly understood, especially over the Arctic Ocean. Here we report atmospheric CH4 levels together with δ(13)C-CH4 from offshore China (31°N) to the central Arctic Ocean (up to 87°N) from July to September 2012. CH4 concentrations and δ(13)C-CH4 displayed temporal and spatial variation ranging from 1.65 to 2.63 ppm, and from -50.34% to -44.94% (mean value: -48.55 ± 0.84%), respectively. Changes in CH4 with latitude were linked to the decreasing input of enriched δ(13)C and chemical oxidation by both OH and Cl radicals as indicated by variation of δ(13)C. There were complex mixing sources outside and inside the Arctic Ocean. A keeling plot showed the dominant influence by hydrate gas in the Nordic Sea region, while the long range transport of wetland emissions were one of potentially important sources in the central Arctic Ocean. Experiments comparing sunlight and darkness indicate that microbes may also play an important role in regional variations. PMID:26323236

  5. δ13C-CH4 reveals CH4 variations over oceans from mid-latitudes to the Arctic

    PubMed Central

    Yu, Juan; Xie, Zhouqing; Sun, Liguang; Kang, Hui; He, Pengzhen; Xing, Guangxi

    2015-01-01

    The biogeochemical cycles of CH4 over oceans are poorly understood, especially over the Arctic Ocean. Here we report atmospheric CH4 levels together with δ13C-CH4 from offshore China (31°N) to the central Arctic Ocean (up to 87°N) from July to September 2012. CH4 concentrations and δ13C-CH4 displayed temporal and spatial variation ranging from 1.65 to 2.63 ppm, and from −50.34% to −44.94% (mean value: −48.55 ± 0.84%), respectively. Changes in CH4 with latitude were linked to the decreasing input of enriched δ13C and chemical oxidation by both OH and Cl radicals as indicated by variation of δ13C. There were complex mixing sources outside and inside the Arctic Ocean. A keeling plot showed the dominant influence by hydrate gas in the Nordic Sea region, while the long range transport of wetland emissions were one of potentially important sources in the central Arctic Ocean. Experiments comparing sunlight and darkness indicate that microbes may also play an important role in regional variations. PMID:26323236

  6. 13C metabolic flux analysis at a genome-scale.

    PubMed

    Gopalakrishnan, Saratram; Maranas, Costas D

    2015-11-01

    Metabolic models used in 13C metabolic flux analysis generally include a limited number of reactions primarily from central metabolism. They typically omit degradation pathways, complete cofactor balances, and atom transition contributions for reactions outside central metabolism. This study addresses the impact on prediction fidelity of scaling-up mapping models to a genome-scale. The core mapping model employed in this study accounts for (75 reactions and 65 metabolites) primarily from central metabolism. The genome-scale metabolic mapping model (GSMM) (697 reaction and 595 metabolites) is constructed using as a basis the iAF1260 model upon eliminating reactions guaranteed not to carry flux based on growth and fermentation data for a minimal glucose growth medium. Labeling data for 17 amino acid fragments obtained from cells fed with glucose labeled at the second carbon was used to obtain fluxes and ranges. Metabolic fluxes and confidence intervals are estimated, for both core and genome-scale mapping models, by minimizing the sum of square of differences between predicted and experimentally measured labeling patterns using the EMU decomposition algorithm. Overall, we find that both topology and estimated values of the metabolic fluxes remain largely consistent between core and GSM model. Stepping up to a genome-scale mapping model leads to wider flux inference ranges for 20 key reactions present in the core model. The glycolysis flux range doubles due to the possibility of active gluconeogenesis, the TCA flux range expanded by 80% due to the availability of a bypass through arginine consistent with labeling data, and the transhydrogenase reaction flux was essentially unresolved due to the presence of as many as five routes for the inter-conversion of NADPH to NADH afforded by the genome-scale model. By globally accounting for ATP demands in the GSMM model the unused ATP decreased drastically with the lower bound matching the maintenance ATP requirement. A non

  7. Determination of de novo synthesized amino acids in cellular proteins revisited by 13C NMR spectroscopy.

    PubMed

    Flögel, U; Willker, W; Leibfritz, D

    1997-04-01

    13C nuclear magnetic resonance spectroscopy was used to determine the absolute amounts to de novo synthesized amino acids in both the perchloric acid extracts and the hydrolyzed protein fractions of F98 glioma cells incubated for 2 h with 5 mmol/l [U-13C]glucose. 13C NMR spectra of the hydrolyzed protein fraction revealed a marked incorporation of 13C-labelled alanine, aspartate and glutamate into the proteins of F98 cells within the incubation period. Additionally, small amounts of 13C-labelled glycine, proline and serine could unambiguously be identified in the protein fraction. Astonishingly, approximately equal amounts of 13C-labelled glutamate and aspartate were incorporated into the cellular proteins, although the cytosolic steady-state concentration of aspartate was below 13C NMR detectability. Hypertonic stress decreased the incorporation of 13C-labelled amino acids into the total protein, albeit their cytosolic concentrations were increased, which reflects an inhibition of protein synthesis under these conditions. On the other hand, hypotonic stress increased the amount of 13C-labelled proline incorporated into the cellular proteins even though the cytosolic concentration of 13C-labelled proline was largely decreased. Apparently, hypoosmotic conditions stimulate the synthesis of proteins or peptides with a high proline content. The results show that already after 2 h of incubation with [U-13C]glucose there is a pronounced flux of 13C label into the cellular proteins, which is usually disregarded if cytosolic fluids are examined only. This means that calculations of metabolic fluxes based on 13C NMR spectroscopic data obtained from perchloric acid extracts of cells or tissues and also from in vivo measurements consider only the labelled 'NMR visible' cytosolic metabolites, which may have to be corrected for fast label flowing off into other compartments.

  8. [Distribution characteristics of soil humus fractions stable carbon isotope natural abundance (delta 13C) in paddy field under long-term ridge culture].

    PubMed

    Tang, Xiao-hong; Luo, You-jin; Ren, Zhen-jiang; Lü, Jia-ke; Wei, Chao-fu

    2011-04-01

    A 16-year field experiment was conducted in a ridge culture paddy field in the hilly region of Sichuan Basin, aimed to investigate the distribution characteristics of stable carbon isotope natural abundance (delta 13C) in soil humus fractions. The soil organic carbon (SOC) content in the paddy field under different cultivation modes ranked in the order of wide ridge culture > ridge culture > paddy and upland rotation. In soil humus substances (HS), humin (HU) was the main composition, occupying 21% - 30% of the total SOC. In the extracted soil carbon, humic acid (HA) dominated, occupying 17% - 21% of SOC and 38% - 65% of HS. The delta 13C value of SOC ranged from -27.9 per thousand to -25.6 per thousand, and the difference of the delta 13C value between 0-5 cm and 20-40 cm soil layers was about 1.9 per thousand. The delta 13C value of HA under different cultivation modes was 1 per thousand - 2 per thousand lower than that of SOC, and more approached to the delta 13C value of rapeseed and rice residues. As for fulvic acid (FA), its delta 13C value was about 2 per thousand and 4 per thousand higher than that of SOC and HA, respectively. The delta 13C value of HU in plough layer (0-20 cm) and plow layer (20-40 cm) ranged from -23.7 per thousand - -24.9 per thousand and -22.6 per thousand - -24.2 per thousand, respectively, reflecting the admixture of young and old HS. The delta 13C value in various organic carbon fractions was HU>FA>SOC>rapeseed and rice residues>HA. Long-term rice planting benefited the increase of SOC content, and cultivation mode played an important role in affecting the distribution patterns of soil humus delta 13C in plough layer and plow layer. PMID:21774322

  9. [Distribution characteristics of soil humus fractions stable carbon isotope natural abundance (delta 13C) in paddy field under long-term ridge culture].

    PubMed

    Tang, Xiao-hong; Luo, You-jin; Ren, Zhen-jiang; Lü, Jia-ke; Wei, Chao-fu

    2011-04-01

    A 16-year field experiment was conducted in a ridge culture paddy field in the hilly region of Sichuan Basin, aimed to investigate the distribution characteristics of stable carbon isotope natural abundance (delta 13C) in soil humus fractions. The soil organic carbon (SOC) content in the paddy field under different cultivation modes ranked in the order of wide ridge culture > ridge culture > paddy and upland rotation. In soil humus substances (HS), humin (HU) was the main composition, occupying 21% - 30% of the total SOC. In the extracted soil carbon, humic acid (HA) dominated, occupying 17% - 21% of SOC and 38% - 65% of HS. The delta 13C value of SOC ranged from -27.9 per thousand to -25.6 per thousand, and the difference of the delta 13C value between 0-5 cm and 20-40 cm soil layers was about 1.9 per thousand. The delta 13C value of HA under different cultivation modes was 1 per thousand - 2 per thousand lower than that of SOC, and more approached to the delta 13C value of rapeseed and rice residues. As for fulvic acid (FA), its delta 13C value was about 2 per thousand and 4 per thousand higher than that of SOC and HA, respectively. The delta 13C value of HU in plough layer (0-20 cm) and plow layer (20-40 cm) ranged from -23.7 per thousand - -24.9 per thousand and -22.6 per thousand - -24.2 per thousand, respectively, reflecting the admixture of young and old HS. The delta 13C value in various organic carbon fractions was HU>FA>SOC>rapeseed and rice residues>HA. Long-term rice planting benefited the increase of SOC content, and cultivation mode played an important role in affecting the distribution patterns of soil humus delta 13C in plough layer and plow layer.

  10. 13C-tryptophan breath test detects increased catabolic turnover of tryptophan along the kynurenine pathway in patients with major depressive disorder

    PubMed Central

    Teraishi, Toshiya; Hori, Hiroaki; Sasayama, Daimei; Matsuo, Junko; Ogawa, Shintaro; Ota, Miho; Hattori, Kotaro; Kajiwara, Masahiro; Higuchi, Teruhiko; Kunugi, Hiroshi

    2015-01-01

    Altered tryptophan–kynurenine (KYN) metabolism has been implicated in major depressive disorder (MDD). The l-[1-13C]tryptophan breath test (13C-TBT) is a noninvasive, stable-isotope tracer method in which exhaled 13CO2 is attributable to tryptophan catabolism via the KYN pathway. We included 18 patients with MDD (DSM-IV) and 24 age- and sex-matched controls. 13C-tryptophan (150 mg) was orally administered and the 13CO2/12CO2 ratio in the breath was monitored for 180 min. The cumulative recovery rate during the 180-min test (CRR0–180; %), area under the Δ13CO2-time curve (AUC; %*min), and the maximal Δ13CO2 (Cmax; %) were significantly higher in patients with MDD than in the controls (p = 0.004, p = 0.008, and p = 0.002, respectively). Plasma tryptophan concentrations correlated negatively with Cmax in both the patients and controls (p = 0.020 and p = 0.034, respectively). Our results suggest that the 13C-TBT could be a novel biomarker for detecting a subgroup of MDD with increased tryptophan–KYN metabolism. PMID:26524975

  11. 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.

  12. Internal comparison between deuterium oxide (D2O) and L-[ring-13C6] phenylalanine for acute measurement of muscle protein synthesis in humans

    PubMed Central

    Wilkinson, Daniel J; Cegielski, Jessica; Phillips, Bethan E; Boereboom, Catherine; Lund, Jonathan N; Atherton, Philip J; Smith, Kenneth

    2015-01-01

    Stable isotope tracer methodologies are becoming increasingly widespread in metabolic research; yet a number of factors restrict their implementation, such as, i.v infusions, multiple cannulae, tissue samples, and significant cost. We recently validated the sensitivity of the orally administered stable isotope tracer deuterium oxide (D2O) for quantifying day-to-day changes in muscle protein synthesis (MPS). This method is less invasive, restrictive, and more cost-effective than traditional amino acid (AA) tracer techniques. In the present study, we hypothesized the sensitivity of our analytical techniques (GC-Pyrolysis-IRMS) would permit D2O-derived measurements of MPS over much shorter periods (i.e., hours) usually only possible using AA-tracer techniques. We recruited nine males (24 ± 3 year, BMI: 25 ± 3 kg·m−²) into an internally controlled comparison of D2O versus 13C AA-tracers. The day before the acute study subjects consumed 400 mL D2O, and on the study day, received a primed (0.3 mg·kg−1) continuous (0.6 mg·kg·h−1) i.v infusion of L-[ring-13C6]-phenylalanine to quantify MPS under both: (1) basal [postabsorptive] and; (2) stimulated [postprandial] that is, consumption of 20 g EAA, conditions. Measures of MPS yielded indistinguishable technique differences with respect to EAA, 13C: 0.065 ± 0.004 to 0.089 ± 0.006%·h−1 (P < 0.05) and D2O: 0.050 ± 0.007 to 0.088 ± 0.008%·h−1 (P < 0.05) with qualitatively similar increases. Our findings reveal that acute measurement of MPS, usually only possible using AA-tracers, are feasible over shorter periods with orally administered D2O when used in tandem with GC-Pyrolysis-IRMS. We conclude that this D2O approach provides a less invasive, cost-effective, and flexible means by which to quantify MPS acutely over several hours. PMID:26149278

  13. Using regional-scale atmospheric δ13C of CO2 as an indicator of ecosystem health and function

    NASA Astrophysics Data System (ADS)

    Alden, C. B.; Miller, J. B.; White, J. W.; Yadav, V.; Michalak, A. M.

    2012-12-01

    Year to year terrestrial CO2 uptake and release is highly variable and is a result of, among other factors, weather and climate variability. One of the key ecosystem parameters that links surface-atmosphere fluxes of energy, water and carbon is stomatal conductance. By measuring and analyzing atmospheric patterns of CO2 and its 13C content over North America, we can begin to identify regional scale changes in stomatal conductance, because conductance is closely related to plant isotopic discrimination. Furthermore, 13C is a useful tracer of the differential responses of C3 and C4 plants to climate and weather anomalies, because C3 and C4 plants have very different isotopic discrimination. Both aspects of the terrestrial carbon cycle are of great interest to those seeking to understand the potential effects of global climate change on cropland and forest productivity, natural CO2 sinks, continental runoff, and continental water and energy exchange with the atmosphere. Our findings may be particularly important for parameterization of process-based models, in light of recent results suggesting that stomatal conductance models driven by vapor pressure deficit (Leuning Model) better predict atmospheric δ13C than do models driven by relative humidity (Ball-Berry Model). For the first time, spatial and temporal density of δ13C of CO2 atmospheric observations may be high enough to allow for regional inversions of δ13CO2 to optimize prior estimates of plant discrimination (and disequilibrium flux -- an isoflux resulting from the combination of a finite residence time of carbon in terrestrial biosphere pools and a changing atmospheric signature due to human burning of fossil fuels with a plant-derived δ13C signature). We perform a Bayesian synthesis inversion for 1) CO2 fluxes and 2) δ13CO2 isofluxes, over the North American region: 145-25°W longitude and 10-80°N latitude. Inversion resolution, in order to avoid aggregation errors, is 1°x1° and 3-hourly, but

  14. Computational Platform for Flux Analysis Using 13C-Label Tracing- Phase I SBIR Final Report

    SciTech Connect

    Van Dien, Stephen J.

    2005-04-12

    Isotopic label tracing is a powerful experimental technique that can be combined with metabolic models to quantify metabolic fluxes in an organism under a particular set of growth conditions. In this work we constructed a genome-scale metabolic model of Methylobacterium extorquens, a facultative methylotroph with potential application in the production of useful chemicals from methanol. A series of labeling experiments were performed using 13C-methanol, and the resulting distribution of labeled carbon in the proteinogenic amino acids was determined by mass spectrometry. Algorithms were developed to analyze this data in context of the metabolic model, yielding flux distributions for wild-type and several engineered strains of M. extorquens. These fluxes were compared to those predicted by model simulation alone, and also integrated with microarray data to give an improved understanding of the metabolic physiology of this organism.

  15. Total assignment of 1H and 13C NMR data for the sesquiterpene lactone 15-deoxygoyazensolide.

    PubMed

    Heleno, Vladimir Constantino Gomes; Crotti, Antônio Eduardo Miller; Constantino, Mauricio Gomes; Lopes, Norberto Peporine; Lopes, João Luis Callegari

    2004-03-01

    We describe a complete analysis of the 1H and 13C spectra of the anti-inflamatory, schistossomicidal and trypanosomicidal sesquiterpene lactone 15-deoxygoyazensolide. This lactone, with a structure similar to other important ones, was studied by NMR techniques such as COSY, HMQC, HMBC, Jres and NOE experiments. The comparison of the data with some computational results led to an unequivocal assignment of all hydrogen and carbon chemical shifts, even eliminating some previous ambiguities. We were able to determine all hydrogen coupling constants (J) and signal multiplicities and to confirm the stereochemistry. A new method for the determination of the relative position of the lactonization and the position of the ester group on a medium-sized ring by NMR was developed.

  16. Laccase-catalyzed reactions of 17β-estradiol in the presence of humic acid: Resolved by high-resolution mass spectrometry in combination with (13)C labeling.

    PubMed

    Sun, Kai; Luo, Qi; Gao, Yanzheng; Huang, Qingguo

    2016-02-01

    The widespread presence of estrogens in natural waters poses potential threats to the aquatic organisms and human health. It is known that estrogens undergo enzyme-catalyzed oxidative coupling (ECOC) reactions, which may impact their environmental fate and can be used in wastewater treatment to remove estrogens, but little information is available on how natural organic matter (NOM) may influence 17β-estradiol (E2) transformation in ECOC processes. A series of experiments were conducted to examine the transformation of E2 in aqueous solution containing humic acid (HA) as model NOM by laccase-mediated ECOC reactions. The impact of HA on the reaction behaviors and product distribution is systematically characterized. The presence of HA inhibited the extent of E2 self-coupling in laccase-mediated systems, while promoted cross-coupling between E2 and HA. Reconfiguration of humic molecules was also observed and characterized by changes in absorbance at 275 nm and the ratios between A250 nm/A365 nm. In particular, experiments were conducted with un-labeled E2 mixed with (13)C3-labeled E2 at a set ratio, with the products probed using high-resolution mass spectrometry (HRMS). The high m/z accuracy of HRMS enabled the use of isotope ratio as a tracer to identify possible cross-coupling products between E2 and HA. Such a method combining HRMS and isotope labeling provides a novel means for identification of products in a reaction system involving NOM or other complex matrices. These findings provide a basis for optimization of ECOC reactions for estrogen removal, and also help to understand the environmental transformation of estrogens. PMID:26692517

  17. Synthesis of isotopically labeled R- or S-[.sup.13C, .sup.2H] glycerols

    DOEpatents

    Martinez, Rodolfo A.; Unkefer, Clifford J.; Alvarez, Marc A.

    2008-01-22

    The present invention is directed to asymmetric chiral labeled glycerols including at least one chiral atom, from one to two .sup.13C atoms and from zero to four deuterium atoms bonded directly to a carbon atom, e.g., (2S) [1,2-.sup.13C.sub.2]glycerol and (2R) [1,2-.sup.13C.sub.2]glycerol, and to the use of such chiral glycerols in the preparation of labeled amino acids.

  18. 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

  19. Characterizing solute transport with transient storage across a range of flow rates: The evidence of repeated tracer experiments in Austrian and Italian streams

    NASA Astrophysics Data System (ADS)

    Schmid, Bernhard H.; Innocenti, Ilaria; Sanfilippo, Umberto

    2010-11-01

    Solute transport in rivers and streams with hyporheic zone exchange and/or in-stream storage is typically affected by the prevailing flow rate. The research reported here focuses on stream tracer experiments repeated many times along the same Austrian (Mödlingbach) and Italian (Torrente Lura) channel reaches to characterize parameter dependency on flow rate. Both groups of data sets showed an increase of storage zone area and main stream area with discharge. In either case, a strong negative correlation was obtained between storage zone residence time and flow rate. From the Mödlingbach data, no clear relationship with Q emerged for the dispersion coefficient and the dead zone ratio, whereas Torrente Lura showed a clear positive correlation of the dispersion coefficient with the flow rate and a slightly negative Q-dependency for the dead zone ratio. Mödlingbach and Torrente Lura results are presented against the background of other repeat experiments reported in literature. In practical applications, the computation of peak concentrations frequently rests on the transfer of transport parameters from one flow rate to another. Using the above Austrian and Italian data sets it was shown that the errors in simulated Mödlingbach peak concentrations remain within a 40% margin, if the ratio of flow rates (for calibration and simulation, resp.) does not exceed 2:1. With Torrente Lura, parameter transfer resulted in somewhat lower peak errors.

  20. 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.

  1. 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

  2. Quantitative (13)C Solid-State NMR Spectra by Multiple-Contact Cross-polarization for Drug Delivery: From Active Principles to Excipients and Drug Carriers.

    PubMed

    Saïdi, Fadila; Taulelle, Francis; Martineau, Charlotte

    2016-08-01

    In this contribution, we present an analysis of the main parameters influencing the efficiency of the (1)H → (13)C multiple-contact cross-polarization nuclear magnetic resonance (NMR) experiment in the context of solid pharmaceutical materials. Using the optimum experimental conditions, quantitative (13)C NMR spectra are then obtained for porous metal-organic frameworks (potential drug carriers) and for components present in drug formulations (active principle ingredient and excipients, amorphous or crystalline). Finally, we show that mixtures of components can also be quantified with this method and, hence, that it represents an ideal tool for quantification of pharmaceutical formulations by (13)C cross-polarization under magic-angle spinning NMR in the industry as it is robust and easy to set up, much faster than direct (13)C polarization and is efficient for samples at natural abundance. PMID:27372550

  3. Open-Source Automated Parahydrogen Hyperpolarizer for Molecular Imaging Using (13)C Metabolic Contrast Agents.

    PubMed

    Coffey, Aaron M; Shchepin, Roman V; Truong, Milton L; Wilkens, Ken; Pham, Wellington; Chekmenev, Eduard Y

    2016-08-16

    An open-source hyperpolarizer producing (13)C hyperpolarized contrast agents using parahydrogen induced polarization (PHIP) for biomedical and other applications is presented. This PHIP hyperpolarizer utilizes an Arduino microcontroller in conjunction with a readily modified graphical user interface written in the open-source processing software environment to completely control the PHIP hyperpolarization process including remotely triggering an NMR spectrometer for efficient production of payloads of hyperpolarized contrast agent and in situ quality assurance of the produced hyperpolarization. Key advantages of this hyperpolarizer include: (i) use of open-source software and hardware seamlessly allowing for replication and further improvement as well as readily customizable integration with other NMR spectrometers or MRI scanners (i.e., this is a multiplatform design), (ii) relatively low cost and robustness, and (iii) in situ detection capability and complete automation. The device performance is demonstrated by production of a dose (∼2-3 mL) of hyperpolarized (13)C-succinate with %P13C ∼ 28% and 30 mM concentration and (13)C-phospholactate at %P13C ∼ 15% and 25 mM concentration in aqueous medium. These contrast agents are used for ultrafast molecular imaging and spectroscopy at 4.7 and 0.0475 T. In particular, the conversion of hyperpolarized (13)C-phospholactate to (13)C-lactate in vivo is used here to demonstrate the feasibility of ultrafast multislice (13)C MRI after tail vein injection of hyperpolarized (13)C-phospholactate in mice. PMID:27478927

  4. In vivo hyperpolarized 13C MR spectroscopic imaging with 1H decoupling

    NASA Astrophysics Data System (ADS)

    Chen, Albert P.; Tropp, James; Hurd, Ralph E.; Van Criekinge, Mark; Carvajal, Lucas G.; Xu, Duan; Kurhanewicz, John; Vigneron, Daniel B.

    2009-03-01

    Application of 13C MRS in vivo on whole body MR system has been limited due to the low static field (and consequent low signal to noise ratio—SNR) of these scanners; thus there have been few reports of 1H decoupled 13C MRS in vivo using a clinical MR platform. The recent development of techniques to retain highly polarized spins in solution following DNP in a solid matrix has provided a mechanism to use endogenous pre-polarized 13C labeled substrates to study real time cellular metabolism in vivo with high SNR. In a recent in vivo hyperpolarized metabolic imaging study using 13C pyruvate, it has been demonstrated that the line shape (signal decay) of the resonances observed are greatly affected by JCH coupling in addition to inhomogeneous broadening. This study demonstrates the feasibility of improving hyperpolarized 13C metabolic imaging in vivo by incorporating 1H decoupling on a clinical whole body 3 T MR scanner. No reduction of T1 of a pre-polarized 13C substrate ([1- 13C] lactate) in solution was observed when 1H decoupling was applied with WALTZ16 sequence. Narrower linewidth for the [1- 13C] lactate resonance was observed in hyperpolarized 13C MRSI data in vivo with 1H decoupling.

  5. Comparison of different mass spectrometry techniques in the measurement of L-[ring-(13)C6]phenylalanine incorporation into mixed muscle proteins.

    PubMed

    Zabielski, Piotr; Ford, G Charles; Persson, X Mai; Jaleel, Abdul; Dewey, Jerry D; Nair, K Sreekumaran

    2013-02-01

    Precise measurement of low enrichment of stable isotope labeled amino-acid tracers in tissue samples is a prerequisite in measuring tissue protein synthesis rates. The challenge of this analysis is augmented when small sample size is a critical factor. Muscle samples from human participants following an 8 h intravenous infusion of L-[ring-(13)C(6)]phenylalanine and a bolus dose of L-[ring-(13)C(6)]phenylalanine in a mouse were utilized. Liquid chromatography tandem mass spectrometry (LC/MS/MS), gas chromatography (GC) MS/MS and GC/MS were compared to the GC-combustion-isotope ratio MS (GC/C/IRMS), to measure mixed muscle protein enrichment of [ring-(13)C(6)]phenylalanine enrichment. The sample isotope enrichment ranged from 0.0091 to 0.1312 molar percent excess. As compared with GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS showed coefficients of determination of R(2)= 0.9962 and R(2) = 0.9942, and 0.9217 respectively. However, the precision of measurements (coefficients of variation) for intra-assay are 13.0%, 1.7%, 6.3% and 13.5% and for inter-assay are 9.2%, 3.2%, 10.2% and 25% for GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS, respectively. The muscle sample sizes required to obtain these results were 8 µg, 0.8 µg, 3 µg and 3 µg for GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS, respectively. We conclude that LC/MS/MS is optimally suited for precise measurements of L-[ring-(13)C(6)]phenylalanine tracer enrichment in low abundance and in small quantity samples. PMID:23378099

  6. Comparison of different mass spectrometry techniques in the measurement of L-[ring-(13)C6]phenylalanine incorporation into mixed muscle proteins.

    PubMed

    Zabielski, Piotr; Ford, G Charles; Persson, X Mai; Jaleel, Abdul; Dewey, Jerry D; Nair, K Sreekumaran

    2013-02-01

    Precise measurement of low enrichment of stable isotope labeled amino-acid tracers in tissue samples is a prerequisite in measuring tissue protein synthesis rates. The challenge of this analysis is augmented when small sample size is a critical factor. Muscle samples from human participants following an 8 h intravenous infusion of L-[ring-(13)C(6)]phenylalanine and a bolus dose of L-[ring-(13)C(6)]phenylalanine in a mouse were utilized. Liquid chromatography tandem mass spectrometry (LC/MS/MS), gas chromatography (GC) MS/MS and GC/MS were compared to the GC-combustion-isotope ratio MS (GC/C/IRMS), to measure mixed muscle protein enrichment of [ring-(13)C(6)]phenylalanine enrichment. The sample isotope enrichment ranged from 0.0091 to 0.1312 molar percent excess. As compared with GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS showed coefficients of determination of R(2)= 0.9962 and R(2) = 0.9942, and 0.9217 respectively. However, the precision of measurements (coefficients of variation) for intra-assay are 13.0%, 1.7%, 6.3% and 13.5% and for inter-assay are 9.2%, 3.2%, 10.2% and 25% for GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS, respectively. The muscle sample sizes required to obtain these results were 8 µg, 0.8 µg, 3 µg and 3 µg for GC/C/IRMS, LC/MS/MS, GC/MS/MS and GC/MS, respectively. We conclude that LC/MS/MS is optimally suited for precise measurements of L-[ring-(13)C(6)]phenylalanine tracer enrichment in low abundance and in small quantity samples.

  7. Information content of measurements from tracer microlysimeter experiments designed for parameter identification in dual-permeability models

    NASA Astrophysics Data System (ADS)

    Larsbo, Mats; Jarvis, Nicholas

    2006-06-01

    Parameters regulating the degree of preferential flow in the dual-permeability water flow and solute transport model MACRO are difficult or impossible to derive from direct measurements. The objectives were (i) to find an improved temporal measurement scheme for identification of these parameters using laboratory microlysimeter experiments and (ii) to evaluate the possibilities of parameter identification in the MACRO model. Artificial data from laboratory microlysimeter experiments consisting of high time-resolution 'measurements' of percolation rate, effluent concentration and resident concentrations at six depths were used with PIMLI (parameter identification method using the localisation of information). The data contained enough information to successfully reduce the uncertainty in the parameter governing mass exchange between pore domains, the saturated micropore hydraulic conductivity and the dispersivity for two hypothetical soils representing one typical clay and one loam. Parameters governing water flow in the macropores were shown to be sensitive in a screening analysis with the Morris method and the uncertainty in these parameters was also reduced by PIMLI. However, some of these parameters did not converge towards their true values probably because of parameter interdependence. In all cases, 'measurements' with large information content were found early in the experiments where less than 0.2 pore volumes of water had passed through the column. For successful identification of parameters determining the degree of preferential flow, efforts should be made to perform high time-resolution measurements during the first irrigations following solute application.

  8. Effects of temperature and substrate stoichiometry on microbial specific respiration rate, carbon use efficiency, and 13C fractionation

    NASA Astrophysics Data System (ADS)

    Min, K.; Lehmeier, C.; Sellers, M.; Chen, Y.; Ballantyne, F.; Billings, S. A.

    2013-12-01

    either a C:N of 10 or 20. Nitrogen was supplied as NH4Cl and KNO3. At steady-state, the rate and δ13C of respired CO2, and biomass were measured. From 13 to 26.5°C, SRR increased and CUE decreased by 152% and 30%, respectively, and these trends were unaffected by substrate C:N. For all runs, the δ13C was highest for cellobiose, followed by microbial biomass and respired CO2. 13C fractionation between biomass and respired CO2 was positively correlated with temperature at C:N=10, while no such relationship was evident at C:N=20. Additional experiments are currently underway at a C:N of 1 to examine if reducing substrate C:N below that of microbial biomass will alter these microbial responses to temperature. This study suggests that with warming enhanced SRR linked to declining CUE drives an increase in total microbial CO2 release. Furthermore, increased total CO2 releases may be independent of substrate C:N, if substrate C:N is greater than microbial C:N. However, non-uniform 13C fractionation between biomass and respired CO2 with varying C:N suggests that different mechanisms are at play, which may confound interpretations of ecosystem-level δ13C -CO2

  9. INL Tracer Interpretation

    2007-03-27

    This spreadsheet application is for tracer test analysis. The analyses are based on the first temporal moment of a tracer. The governing equations are briefly discussed, and the individual steps required of the user are outlined. A series of Excel macros written in Visual Basic calculate mean residence time, swept pore volume, and flow-storage geometry from a tracer history.

  10. Evidence of polycyclic aromatic hydrocarbon biodegradation in a contaminated aquifer by combined application of in situ and laboratory microcosms using (13)C-labelled target compounds.

    PubMed

    Bahr, Arne; Fischer, Anko; Vogt, Carsten; Bombach, Petra

    2015-02-01

    The number of approaches to evaluate the biodegradation of polycyclic aromatic hydrocarbons (PAHs) within contaminated aquifers is limited. Here, we demonstrate the applicability of a novel method based on the combination of in situ and laboratory microcosms using (13)C-labelled PAHs as tracer compounds. The biodegradation of four PAHs (naphthalene, fluorene, phenanthrene, and acenaphthene) was investigated in an oxic aquifer at the site of a former gas plant. In situ biodegradation of naphthalene and fluorene was demonstrated using in situ microcosms (BACTRAP(®)s). BACTRAP(®)s amended with either [(13)C6]-naphthalene or [(13)C5/(13)C6]-fluorene (50:50) were incubated for a period of over two months in two groundwater wells located at the contaminant source and plume fringe, respectively. Amino acids extracted from BACTRAP(®)-grown cells showed significant (13)C-enrichments with (13)C-fractions of up to 30.4% for naphthalene and 3.8% for fluorene, thus providing evidence for the in situ biodegradation and assimilation of those PAHs at the field site. To quantify the mineralisation of PAHs, laboratory microcosms were set up with BACTRAP(®)-grown cells and groundwater. Naphthalene, fluorene, phenanthrene, or acenaphthene were added as (13)C-labelled substrates. (13)C-enrichment of the produced CO2 revealed mineralisation of between 5.9% and 19.7% for fluorene, between 11.1% and 35.1% for acenaphthene, between 14.2% and 33.1% for phenanthrene, and up to 37.0% for naphthalene over a period of 62 days. Observed PAH mineralisation rates ranged between 17 μg L(-1) d(-1) and 1639 μg L(-1) d(-1). The novel approach combining in situ and laboratory microcosms allowed a comprehensive evaluation of PAH biodegradation at the investigated field site, revealing the method's potential for the assessment of PAH degradation within contaminated aquifers.

  11. Thermal Degradation studies of nitroplasticized estane 5703 utilizing [sup 13] C enriched model polymers.

    SciTech Connect

    Wrobleski, Debra A.; Langlois, D. A.; Orler, E. B.; Schoonover, J. R.; Osborn, J. C.; Small, J. H.

    2002-01-01

    Predicting the lifetime of PBX 9501 is critical to assurance of the reliability of the nuclear weapons stockpile. Ln support of the Lifetime Prediction Model, we have been investigating the physical and chemical aging processes that can potentially affect the lifetime of PBX 9501. Surveillance data of stockpile PBX 9501 binder have shown a decrease in Estane molecular weight as a fiinction of age. Estane is a poly(ester urethane) consisting of poly(buly1ene adipate) as the flexible soft segment and 4,4{prime}-methylenediphenyl-1,1{prime}-diisocyanate (MDI) chain extended with 1,4-butanediol (BDO) as the rigid hard segment efforts have focused on chemical reactions that may cause chain scission of Estane through hydrolytic degradation of the polyester soft segments. Thermal aging of Estane in the absence of water shows no signs of hydrolytic degradation, whereas, appearance of high molecular weight species has been observed for Estane aged at elevated temperatures in the presence of nitroplasticizer. These high molecular weight species are most likely due to branching reactions that eventually leads to formation of an insoluble, cross-linked gel. While the mechanisms of hydrolytic degradation of polyesters has been addressed, the degradations reactions of poly(ester urethane) (PESU) in the presence of NP are not well understood. We have prepared a series of isotopically enriched PESU model compounds chemically similar to Estane for aging experiments in order to understand possible cross-linking mechanisms. Changes in physical properties are observed with less than 1% chemical cross-linking. Isotopic enrichment of 13C from 1.1 % natural abundance to 100% improves the sensitivity of spectroscopic techniques so that small quantities of degradation products can be observed. Because degradation reaction mechanisms of importance may occur at the hard segment portion of the polymer, we have prepared 13C PESU with 13C labeled at the methylene carbon. These model polymers

  12. A Large Metabolic Carbon Ccontribution to the δ13C Record in Marine Aragonitic Bivalve Shells

    NASA Astrophysics Data System (ADS)

    Gillikin, D. P.; Lorrain, A.; Dehairs, F.

    2006-12-01

    The stable carbon isotopic signature archived in bivalve shells was originally thought to record the δ13C of seawater dissolved inorganic carbon (δ13C-DIC). However, more recent studies have shown that the incorporation of isotopically light metabolic carbon (M) significantly affects the δ13C signal recorded in biogenic carbonates. To assess the M contribution to Mercenaria mercenaria shells collected in North Carolina, USA, we sampled seawater δ13C-DIC, tissue, hemolymph and shell δ13C. We found up to a 4‰ decrease through ontogeny in shell δ13C in a 23 year old individual. There was no correlation between shell height or age and tissue δ13C. Thus, the ontogenic decrease observed in the shell δ13C could not be attributed to changes in food sources as the animal ages leading to more negative metabolic CO2, since this would require a negative relationship between tissue δ13C and shell height. Hemolymph δ13C, on the other hand, did exhibit a negative relationship with height, but the δ13C values were more positive than expected, indicating that hemolymph may not be a good proxy of extrapallial fluid δ13C. Nevertheless, the hemolymph data indicate that respired CO2 does influence the δ13C of internal fluids and that the amount of respired CO2 is related to the age of the bivalve. The percent metabolic C incorporated into the shell (%M) was significantly higher (up to 37%) than has been found in other bivalve shells, which usually contain less than 10 %M. Attempts to use shell biometrics to predict %M could not explain more than ~60% of the observed variability. Moreover, there were large differences in the %M between different sites. Thus, the metabolic effect on shell δ13C cannot easily be accounted for to allow reliable δ13C-DIC reconstructions. However, there does seem to be a common effect of size, as all sites had indistinguishable slopes between the %M and shell height (+0.19% per mm of shell height).

  13. Preliminary studies of a canine 13C-aminopyrine demethylation blood test.

    PubMed Central

    Moeller, E M; Steiner, J M; Williams, D A; Klein, P D

    2001-01-01

    The objectives of this study were to determine whether a 13C-aminopyrine demethylation blood test is technically feasible in clinically healthy dogs, whether oral administration of 13C-aminopyrine causes a detectable increase in percent dose/min (PCD) of 13C administered as 13C-aminopyrine and recovered in gas extracted from blood, and whether gas extraction efficiency has an impact on PCD. A dose of 2 mg/kg body weight of 13C-aminopyrine dissolved in deionized water was administered orally to 6 clinically healthy dogs. Blood samples were taken from each dog 0, 30, 60, and 120 min after administration of the 13C-aminopyrine. Carbon dioxide was extracted from blood samples by addition of acid and analyzed by fractional mass spectrometry. None of the 6 dogs showed any side effects after 13C-aminopyrine administration. All 6 dogs showed a measurable increase of the PCD in gas samples extracted from blood samples at 30 min, 60 min, and 120 min after 13C-aminopyrine administration. Coefficients of variation between the triplicate samples were statistically significantly higher for the %CO2, a measure of extraction efficiency, than for PCD values (P < 0.0001). The 13C-aminopyrine demethylation blood test described here is technically feasible. Oral administration of 13C-aminopyrine did not lead to gross side effects in the 6 dogs. Clinically healthy dogs show a measurable increase of PCD in gas extracted from blood samples after oral administration of 13C-aminopyrine. Efficiency of CO2 extraction from blood samples does not have an impact on PCD determined from these blood samples. This test may prove useful to evaluate hepatic function in dogs. PMID:11227194

  14. Economical synthesis of 13C-labeled opiates, cocaine derivatives and selected urinary metabolites by derivatization of the natural products.

    PubMed

    Karlsen, Morten; Liu, Huiling; Johansen, Jon Eigill; Hoff, Bård Helge

    2015-01-01

    The illegal use of opiates and cocaine is a challenge world-wide, but some derivatives are also valuable pharmaceuticals. Reference samples of the active ingredients and their metabolites are needed both for controlling administration in the clinic and to detect drugs of abuse. Especially, (13)C-labeled compounds are useful for identification and quantification purposes by mass spectroscopic techniques, potentially increasing accuracy by minimizing ion alteration/suppression effects. Thus, t