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Sample records for 15n natural abundance

  1. 15N NATURAL ABUNDANCE AND 15N LABELLING STUDIES IN FOREST ECOSYSTEMS

    EPA Science Inventory

    The relative amounts of the two stable isotopes of Nitrogen (N), 15N, and N, vary predictably in soils and plant tissues of forests and other non-cultivated ecosystems. light fractionations, or discriminations against the heavier N isotope, that can occur as N cycles through vege...

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

    SciTech Connect

    Johnson, G.V. )

    1990-05-01

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

  3. Characteristics of the Nitrogen Cycle in Southern Everglades Marshes: 15N Tracer and Natural Abundance Experiments

    NASA Astrophysics Data System (ADS)

    Wozniak, J. R.; Anderson, W. T.; Childers, D. L.; Madden, C. J.; Rudnick, D.

    2004-12-01

    An in situ mesocosm experiment was conducted in the freshwater marshes of the Southern Everglades. The object of this experiment was to compare 15N natural abundances to N-cycling in mesocosms amended with 15N tracer. The 15N tracer technique allowed us to isolate the flows of N among various ecosystem components. Six mesocosms (2 m2) were deployed in the oligotrophic P-limited marshes of the C-111 basin. The experiment ran for a period of 21 days with sampling occurring at t=0, 5, 10, 20, 30min, 1, 3, and 6hr on the first day and subsequently on days 2, 3, 5, 9, 15, and 21. Periphyton, soil, macrophytes (Cladium jamaicense), and consumers (Gambusia holbrooki) were collected. The isotopic tracer added to the enriched mesocosms possessed a \\delta 15N value of 300\\permil. Isotopic analysis was completed on a Finnigan Delta C EA-IRMS. All ecosystem components showed tracer uptake except for the soil, which remained at natural abundance levels throughout the experiment. Periphyton showed both the most rapid and the greatest N tracer uptake with an increase of 8.7\\permil, 5 minutes after tracer addition and a maximum \\delta 15N value of 197.93\\permil at day 3. This experiment demonstrated that at study sites with low nitrate concentrations (\\approx3\\muM) a 300 permil dosing rate of 15N labled Ca(NO3)2 (\\+98% 15N) would not increase mesocosm nitrate above ambient concentrations and would provide a significant isotopic signal to track fluxes of N among ecosystem components. In addition, an experiment was conducted in the same region to determine natural abundance values for ecosystem components at both near-canal and downstream sites. All components sampled at the near-canal site had significantly heavier nitrogen isotopic values than did the downstream components (\\delta 15N 7.44\\permil \\pm2.11 and 2.01\\permil \\pm1.44 respectively); suggesting the Southern Everglades marsh is acting as a sink for canal-borne DIN and a source for "new" marsh derived DON.

  4. Regional patterns of 15N natural abundance in forest ecosystems along a large transect in eastern China

    NASA Astrophysics Data System (ADS)

    Sheng, Wenping; Yu, Guirui; Fang, Huajun; Liu, Yingchun; Wang, Qiufeng; Chen, Zhi; Zhang, Li

    2014-02-01

    The regional determining factors underlying inter- and intra-site variation of 15N natural abundance in foliage, O horizon and mineral soil were investigated in eastern China.15N natural abundance values for these forest ecosystems were in the middle of the range of values previously found for global forest ecosystems. In contrast to commonly reported global patterns, temperate forest ecosystems were significantly more15N-enriched than tropical forest ecosystems, and foliage δ15N was negatively correlated with increasing mean annual temperature and net soil N mineralisation in eastern China. Tight N cycling in forest ecosystems and the use of atmospheric N deposition by trees might underlie the δ15N distribution patterns in eastern China. The existence of mycorrhizal fungi and root distribution profiles in the soil may also influence the15N natural abundance patterns in forest ecosystems of eastern China.

  5. Regional patterns of 15N natural abundance in forest ecosystems along a large transect in eastern China

    PubMed Central

    Sheng, Wenping; Yu, Guirui; Fang, Huajun; Liu, Yingchun; Wang, Qiufeng; Chen, Zhi; Zhang, Li

    2014-01-01

    The regional determining factors underlying inter- and intra-site variation of 15N natural abundance in foliage, O horizon and mineral soil were investigated in eastern China.15N natural abundance values for these forest ecosystems were in the middle of the range of values previously found for global forest ecosystems. In contrast to commonly reported global patterns, temperate forest ecosystems were significantly more15N-enriched than tropical forest ecosystems, and foliage δ15N was negatively correlated with increasing mean annual temperature and net soil N mineralisation in eastern China. Tight N cycling in forest ecosystems and the use of atmospheric N deposition by trees might underlie the δ15N distribution patterns in eastern China. The existence of mycorrhizal fungi and root distribution profiles in the soil may also influence the15N natural abundance patterns in forest ecosystems of eastern China. PMID:24576905

  6. Rapid, storm-induced changes in the natural abundance of 15N in a planktonic ecosystem, Chesapeake Bay, USA

    NASA Astrophysics Data System (ADS)

    Montoya, J. P.; Korrigan, S. G.; McCarthy, J. J.

    1991-12-01

    Samples of dissolved inorganic nitrogen (DIN), particulate nitrogen (PN) and two species of Zooplankton were collected during two north-south transects of the Chesapeake Bay in the autumn of 1984 (27-28 September and 3-5 October). During the first transect, the natural abundance of 15N ( δ15N) in the major dissolved and planktonic pools of nitrogen suggested that the δ15N of PN was largely determined by isotopic fractionation during uptake of NH 4+ by phytoplankton. Averaged over the transect as a whole, the δ15N of the herbivorous calanoid copepod Acartia tonsa was 4.1‰ higher than that of the PN, while the δ15N of the carnivorous ctenophore Mnemiopsis leidyi was 6.4‰ higher than that of the PN. In the interval between the two transects, storm-induced mixing of the water column resulted in the injection of NH 4+ into the surface layer of the bay. This perturbation in the estuarine nitrogen cycle resulted in marked changes in the δ15N of the major dissolved and planktonic pools of nitrogen in the bay. In combination with ancillary physical, chemical and biological data, these changes in δ15N provided estimates of the isotopic fractionation factor for NH 4+ uptake by phytoplankton ( α = 1.0065 -1.0080) as well as the turnover time of nitrogen in Acartia tonsa (6.0-9.6 days). Despite the changes in δ15N observed during this cruise, the relative distribution of 15N between trophic levels was preserved: during the second transect, the difference in δ15N between Acartia tonsa and PN was 3.6‰ and the difference in δ15N between Mnemiopsis leidyi and PN was 7.3‰. These results demonstrate that the natural abundance of 15N can change dramatically on a time scale of days and that time-series studies of the natural abundance of 15N can be a useful complement to studies using tracer additions of 15N to document nitrogen transformations in planktonic ecosystems.

  7. Plant and Soil Natural Abundance delta-15N: Indicators of Nitrogen Cycling in the Catskill Mountains, New York, USA

    NASA Astrophysics Data System (ADS)

    Templer, P. H.; Lovett, G. M.; Weathers, K.; Arthur, M. A.

    2002-12-01

    We examined the potential use of natural abundance 15N of plants and soils as an indicator of forest nitrogen (N) cycling rates within the Catskill Mountains, NY. These watersheds receive among the highest rates of N deposition in the northeastern United States and are beginning to show signs of N saturation. Many studies have shown a link between increased N cycling rates and 15N enrichment of soil and plant pools. Faster rates of N cycling processes, especially nitrification, lead to fractionation of 14/15N, creating N products that are relatively depleted in 15N. This can lead to enrichment of soil pools, as lighter 14N is lost from the system via leaching or denitrification. Plant N pools can become increasingly enriched as they take up 15N-enriched soil N. Despite similar amounts of N deposition across the Catskill Mountains, forests dominated by different tree species appear to vary in the amount of N retained or lost to nearby streams. To determine if plant and soil 15N could be used as indicators of N cycling rates, we collected foliage, wood, litterfall, organic and mineral soil, and fine roots from single species stands of American beech (Fagus grandifolia), eastern hemlock (Tsuga canadensis), red oak (Quercus rubra), and sugar maple (Acer saccharum). Fine roots and soil 15N were highest within sugar maple stands (p<0.05). Sugar maple soils also had the highest rates of net nitrification and N leaching. Therefore, soil 15N appears to correlate with forest N retention and loss. However, 15N enrichment was highest within foliage, litterfall and wood of beech trees (p<0.05). The decoupling between foliage 15N and N cycling, as well as between 15N of foliage and fine roots, illustrates that it may not be possible to use a single plant pool as an indicator of N cycling rates.

  8. Determination of the natural abundance δ15N of taurine by gas chromatography-isotope ratio measurement mass spectrometry.

    PubMed

    Tea, Illa; Antheaume, Ingrid; Besnard, Jorick; Robins, Richard J

    2010-12-15

    The measurement of the nitrogen isotope ratio of taurine (2-aminoethanesulphonic acid) in biological samples has a large number of potential applications. Taurine is a small water-soluble molecule which is notoriously difficult to analyze due to its polarity and functionality. A method is described which allows the determination of the natural abundance δ(15)N values of taurine and structural analogues, such as 3-amino-1-propanesulphonic acid (APSA), by isotope ratio mass spectrometry interfaced to gas chromatography (GC-irm-MS). The one-step protocol exploits the simultaneous derivatization of both functionalities of these aminosulphonic acids by reaction with triethylorthoacetate (TEOA). Conditions have been established which ensure quantitative reaction thus avoiding any nitrogen isotope fractionation during derivatization and workup. The differences in the δ(15)N values of derivatized and non-derivatized taurine and APSA all fall within the working range of 0.4‰ (-0.02 to 0.39‰). When applied to four sources of taurine with various δ(15)N values, the method achieved excellent reproducibility and accuracy. The optimized method enables the determination of the natural abundance δ(15)N values of taurine over the concentration range 1.5-7.84 µmol.mL(-1) in samples of biological origin. PMID:21072793

  9. Estimation of biological nitrogen fixation by black locust in short-rotation forests using natural 15N abundance method

    NASA Astrophysics Data System (ADS)

    Veste, M.; Böhm, C.; Quinckenstein, A.; Freese, D.

    2012-04-01

    The importance of short rotation forests and agroforestry systems for woody biomass production for bioenergy will increase in Central Europe within the next decades. In this context, black locust (Robinia pseudoacacia) has a high growth potential especially at marginal, drought-susceptible sites such as occur in Brandenburg State (Eastern Germany). As a pioneer tree species black locust grows under a wide range of site conditions. The native range of black locust in Northern America is classified by a humid to sub-humid climate with a mean annual precipitation of 1020 to 1830 mm. In Central and Eastern Europe, this species is cultivated in a more continental climate with an annual precipitation often below 600 mm. Therefore, black locust is known to be relatively drought tolerant compared to other temperate, deciduous tree species. Because of its N2-fixation ability black locust plays generally an important role for the improvement of soil fertility. This effect is of particular interest at marginal sites in the post-mining landscapes. In order to estimate the N2-fixation potential of black locust at marginal sites leaf samples were taken from black locust trees in short rotation plantations planted between 1995 and 2007 in post-mining sites south of Cottbus (Brandenburg, NE Germany). The variation of the natural 15N abundance was measured to evaluate the biological nitrogen fixation. The nitrogen derived from the atmosphere can be calculated using a two-pool model from the quotient of the natural 15N abundances of the N2-fixing plant and the plant available soil N. Because representatively determining the plant available soil N is difficult, a non-N2-fixing reference plant growing at the same site with a similar root system and temporal N uptake pattern to the N2-fixing plant is often used. In our case we used red oak (Quercus rubra) as a reference. The average nitrogen content in the leaves of black locust ranged from 3.1% (C/N 14.8) in 15 years old trees to 3.4% (C/N 14.4) in 3 year-old trees, respectively. A higher content of nitrogen was found in leaves of re-sprouted trees with 4.3% (C/N 11.5). The estimated percentage of nitrogen derived from the atmosphere (% NdfA) in black locust was 63% - 83% compared to 56% in seabuckthorn (Hippophaë rhamnoides) and 79% in common broom (Genista scuparia). The annual leaf biomass production of black locust varied between 1325 (2 years old trees) and 2576 kg/ha a (4 years old trees). The estimated leaf nitrogen fixed by Robinia was approx. 30.5 - 59.2 kg/ha a. From the results, we can conclude that the biological nitrogen fixation by Robina is an important factor for the nitrogen balance of short-rotation plantations on nutrient poor-soils.

  10. Effects of Four Different Restoration Treatments on the Natural Abundance of 15N Stable Isotopes in Plants

    PubMed Central

    Temperton, Vicky M.; Märtin, Lea L. A.; Röder, Daniela; Lücke, Andreas; Kiehl, Kathrin

    2012-01-01

    δ15N signals in plant and soil material integrate over a number of biogeochemical processes related to nitrogen (N) and therefore provide information on net effects of multiple processes on N dynamics. In general little is known in many grassland restoration projects on soil–plant N dynamics in relation to the restoration treatments. In particular, δ15N signals may be a useful tool to assess whether abiotic restoration treatments have produced the desired result. In this study we used the range of abiotic and biotic conditions provided by a restoration experiment to assess to whether the restoration treatments and/or plant functional identity and legume neighborhood affected plant δ15N signals. The restoration treatments consisted of hay transfer and topsoil removal, thus representing increasing restoration effort, from no restoration measures, through biotic manipulation to major abiotic manipulation. We measured δ15N and %N in six different plant species (two non-legumes and four legumes) across the restoration treatments. We found that restoration treatments were clearly reflected in δ15N of the non-legume species, with very depleted δ15N associated with low soil N, and our results suggest this may be linked to uptake of ammonium (rather than nitrate). The two non-legume species differed considerably in their δ15N signals, which may be related to the two species forming different kinds of mycorrhizal symbioses. Plant δ15N signals could clearly separate legumes from non-legumes, but our results did not allow for an assessment of legume neighborhood effects on non-legume δ15N signals. We discuss our results in the light of what the δ15N signals may be telling us about plant–soil N dynamics and their potential value as an indicator for N dynamics in restoration. PMID:22645597

  11. Plant and soil natural abundance delta (15)N: indicators of relative rates of nitrogen cycling in temperate forest ecosystems.

    PubMed

    Templer, Pamela H; Arthur, Mary A; Lovett, Gary M; Weathers, Kathleen C

    2007-08-01

    Watersheds within the Catskill Mountains, New York, receive among the highest rates of nitrogen (N) deposition in the northeastern United States and are beginning to show signs of N saturation. Despite similar amounts of N deposition across watersheds within the Catskill Mountains, rates of soil N cycling and N retention vary significantly among stands of different tree species. We examined the potential use of delta (15)N of plants and soils as an indicator of relative forest soil N cycling rates. We analyzed the delta (15)N of foliage, litterfall, bole wood, surface litter layer, fine roots and organic soil from single-species stands of American beech (Fagus grandifolia), eastern hemlock (Tsuga canadensis), red oak (Quercus rubra), and sugar maple (Acer saccharum). Fine root and organic soil delta (15)N values were highest within sugar maple stands, which correlated significantly with higher rates of net mineralization and nitrification. Results from this study suggest that fine root and organic soil delta (15)N can be used as an indicator of relative rates of soil N cycling. Although not statistically significant, delta (15)N was highest within foliage, wood and litterfall of beech stands, a tree species associated with intermediate levels of soil N cycling rates and forest N retention. Our results show that belowground delta (15)N values are a better indicator of relative rates of soil N cycling than are aboveground delta (15)N values. PMID:17479293

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

    PubMed

    Lapidot, A; Gopher, A

    1997-02-01

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

  13. Natural 15N- and 13C-abundance as indicators of forest nitrogen status and soil carbon dynamics

    SciTech Connect

    Garten Jr, Charles T; Hanson, Paul J; Todd Jr, Donald E; Lu, Benwhea Bonnie; Brice, Deanne Jane

    2007-09-01

    This book highlights new and emerging uses of stable isotope analysis in a variety of ecological disciplines. While the use of natural abundance isotopes in ecological research is now relatively standard, new techniques and ways of interpreting patterns are developing rapidly. The second edition of this book provides a thorough, up-to-date examination of these methods of research. As part of the Ecological Methods and Concepts series which provides the latest information on experimental techniques in ecology, this book looks at a wide range of techniques that use natural abundance isotopes to: {sm_bullet} follow whole ecosystem element cycling {sm_bullet} understand processes of soil organic matter formation {sm_bullet} follow the movement of water in whole watersheds {sm_bullet} understand the effects of pollution in both terrestrial and aquatic environments {sm_bullet} study extreme systems such as hydrothermal vents {sm_bullet}follow migrating organisms In each case, the book explains the background to the methodology, looks at the underlying principles and assumptions, and outlines the potential limitations and pitfalls. Stable Isotopes in Ecology and Environmental Science is an ideal resource for both ecologists who are new to isotopic analysis, and more experienced isotope ecologists interested in innovative techniques and pioneering new uses.

  14. [Responses of Soil and Plant 15N Natural Abundance to Long-term N Addition in an N-Saturated Pinus massoniana Forest in Southwest China].

    PubMed

    Liu, Wen-jing; Kang, Rong-hua; Zhang, Ting; Zhu, Jing; Duan, Lei

    2015-08-01

    Increasing N deposition in China will possibly cause N saturation of forest ecosystem, further resulting in a series of serious environmental problems. In order to explore the response of forest ecosystem to N deposition in China, and further evaluate and predict the N status of ecosystem, the 15N natural abundance (delta 15N) of soil and plants was measured in a typical Masson pine (Pinus massoniana) forest in southwest China to examine the potential use of delta 15N enrichment factor (epsilon(p/s)) as an effective indicator of N status. Long-term high N addition could significantly increase delta 15N of soil and plants, which was suggested by an on-going N fertilizing experiment with NH4NO3 or NaNO3 for 7 years. Meanwhile, delta 15N of soil and plants under NH, deposition was significantly higher than that under NO- deposition, suggesting different responses of ecosystem to different N-forms of deposition. The "N enrichment factor (epsilon(p/s)) had positive correlations with N deposition, N nitrification, and N leaching in the soil water. Linear correlation between "N enrichment factor and N deposition was found for all Masson pine forests investigated in this and previous studies in China, demonstrating that 15N enrichment factor could be used as an indicator of N status. The NH3 emission control should also be carried out accompanying with NOx emission control in the future, because NH4- deposition had significantly greater impact on the forest ecosystem than NO3- deposition with the same equivalence. PMID:26592030

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

    PubMed

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

    2015-11-01

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

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

    PubMed

    Gautam, Mukesh Kumar; Lee, Kwang-Sik; Song, Byeong-Yeol; Lee, Dongho; Bong, Yeon-Sik

    2016-05-01

    Decomposition, nutrient, and isotopic (δ(13)C and δ(15)N) dynamics during 1 year were studied for leaf and twig litters of Pinus densiflora, Castanea crenata, Erigeron annuus, and Miscanthus sinensis growing on a highly weathered soil with constrained nutrient supply using litterbags in a cool temperate region of South Korea. Decay constant (k/year) ranged from 0.58 to 1.29/year, and mass loss ranged from 22.36 to 58.43 % among litter types. The results demonstrate that mass loss and nutrient dynamics of decomposing litter were influenced by the seasonality of mineralization and immobilization processes. In general, most nutrients exhibited alternate phases of rapid mineralization followed by gradual immobilization, except K, which was released throughout the field incubation. At the end of study, among all the nutrients only N and P showed net immobilization. Mobility of different nutrients from decomposing litter as the percentage of initial litter nutrient concentration was in the order of K > Mg > Ca > N ≈ P. The δ(13)C (0.32-6.70 ‰) and δ(15)N (0.74-3.90 ‰) values of residual litters showed nonlinear increase and decrease, respectively compared to initial isotopic values during decomposition. Litter of different functional types and chemical quality converged toward a conservative nutrient use strategy through mechanisms of slow decomposition and slow nutrient mobilization. Our results indicate that litter quality and season, are the most important regulators of litter decomposition in these forests. The results revealed significant relationships between litter decomposition rates and N, C:N ratio and P, and seasonality (temperature). These results and the convergence of different litters towards conservative nutrient use in these nutrient constrained ecosystems imply optimization of litter management because litter removal can have cascading effects on litter decomposition and nutrient availability in these systems. PMID:26915037

  17. Application of Natural Isotopic Abundance (1)H-(13)C- and (1)H-(15)N-Correlated Two-Dimensional NMR for Evaluation of the Structure of Protein Therapeutics.

    PubMed

    Arbogast, Luke W; Brinson, Robert G; Marino, John P

    2016-01-01

    Methods for characterizing the higher-order structure of protein therapeutics are in great demand for establishing consistency in drug manufacturing, for detecting drug product variations resulting from modifications in the manufacturing process, and for comparing a biosimilar to an innovator reference product. In principle, solution NMR can provide a robust approach for characterization of the conformation(s) of protein therapeutics in formulation at atomic resolution. However, molecular weight limitations and the perceived need for stable isotope labeling have to date limited its practical applications in the biopharmaceutical industry. Advances in NMR magnet and console technologies, cryogenically cooled probes, and new rapid acquisition methodologies, particularly selective optimized flip-angle short transient pulse schemes and nonuniform sampling, have greatly ameliorated these limitations. Here, we describe experimental methods for the collection and analysis of 2D (1)H(N)-(15)N-amide- and (1)H-(13)C-methyl-correlated spectra applied to protein drug products at natural isotopic abundance, including representatives from the rapidly growing class of monoclonal antibody (mAb) therapeutics. Practical aspects of experimental setup and data acquisition for both standard and rapid acquisition NMR techniques are described. Furthermore, strategies for the statistical comparison of 2D (1)H(N)-(15)N-amide- and (1)H-(13)C-methyl-correlated spectra are detailed. PMID:26791974

  18. Sensitive measurement of NH4+ 15N/14N (delta 15NH4+) at natural abundance levels in fresh and saltwaters.

    PubMed

    Zhang, Lin; Altabet, Mark A; Wu, Taixing; Hadas, Ora

    2007-07-15

    We report a new method for determining the 15N/14N of NH4+ at natural abundance level in both freshwater and seawater. NH4+ is first quantitatively oxidized to NO2- by hypobromite (BrO-) at pH approximately 12. After the addition of sodium arsenite to consume excess BrO-, yield is verified by colorimetric NO2- determination. NO2- is further reduced to N2O using a 1:1 sodium azide and acetic acid buffer solution using previously established procedures. The product N2O is then analyzed for isotopic composition using a continuous flow purge and cryogenic trap system coupled to an isotope ratio mass spectrometer. Reliable delta 15N values (standard deviation is 0.3 per thousand or better) are obtained over an NH4+ concentration range of 0.5-10 microM using 20 mL volumes of either freshwater or seawater samples. Higher concentration samples are readily diluted to lower concentration. Preexisting NO2- is removed by treatment with sulfanilic acid. There is no interference from any of the nitrogen-containing compounds tested except short-chain aliphatic amino acids (i.e., glycine) which typically are present at very low environmental concentrations. As compared to published methods, our approach is more robust, readily applicable at low concentrations and small sample volumes, and requires less time for preparation and analysis. PMID:17567102

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

    PubMed

    Krishnamoorthi, R; Nemmers, S; Tobias, B

    1992-06-15

    15N NMR assignments were made to the backbone amide nitrogen atoms at natural isotopic abundance of intact and reactive-site (Arg5-Ile6) hydrolyzed Cucurbita maxima trypsin inhibitor III (CMTI-III and CMTI-III*, respectively) by means of 2D proton-detected heteronuclear single bond chemical shift correlation (HSBC) spectroscopy, utilizing the previously made sequence-specific 1H NMR assignments (Krishnamoorthi et al. (1992) Biochemistry 31, 898-904). Comparison of the 15N chemical shifts of the two forms of the inhibitor molecule revealed significant changes not only for residues located near the reactive-site region, but also for those distantly located. Residues Cys3, Arg5, Leu7, Met8, Cys10, Cys16, Glu19, His25, Tyr27, Cys28 and Gly29 showed significant chemical shift changes ranging from 0.3 to 6.1 ppm, thus indicating structural perturbations that were transmitted throughout the molecule. These findings confirm the earlier conclusions based on 1H NMR investigations. PMID:1618315

  20. Adducts of rhodium(II) tetraacetate with some nitrogenous organic ligands: application of natural abundance 15N and 13C CPMAS NMR spectroscopy.

    PubMed

    Jaźwiński, Jarosław; Kamieński, Bohdan

    2007-10-01

    Adducts of rhodium(II) tetraacetate with some nitrogenous organic ligands: 1-azabicyclo[2,2,2]octane 1, 1,2-diazabicyclo[2,2,2]octane 2, pyrazine 3, pyrimidine 4, [1,3,5]triazine 5 and 1,3,5,7-tetraazatricyclo[3,3,1,1(3,7)]decane 6 have been investigated by means of natural abundance (13)C and (15)N CPMAS nuclear magnetic resonance (NMR) spectroscopy. 1-Azabicyclo[2,2,2]octane 1 having one nitrogen atom in the molecule produces either the 1:1 or 1:2-adduct depending on the reagent molar ratio; some features of its (13)C CPMAS NMR spectra suggest the dimeric structure of the 1:1-adduct. Multifunctional ligands having more than one nitrogen atom in a molecule yield the adducts insoluble in common organic solvents. Elemental analysis and NMR experiments have revealed that 1,2-diazabicyclo[2,2,2]octane, pyrazine, pyrimidine and [1,3,5]triazine produced adducts in the form of 1:1 polymeric chains. 1,3,5,7-tetraazatricyclo[3,3,1,1(3,7)]decane yields the adduct containing ligand and metal salt in the molar ratio of 3:4. The (15)N chemical shift change caused by the Rh-N bond formation (Deltadelta parameter) varies from ca. -9 ppm for aliphatic ligands to ca. -40 ppm for heteroaromatic species. The NMR findings have been supported by theoretical calculation (density functional calculation (DFT), LanLD2Z//B3LYB level) of molecular geometry, energy and chemical shieldings. PMID:17697766

  1. Leaf allocation patterns and 13C and 15N natural abundances of tropical lianas (Passiflora sp.) as dependent on external climbing support.

    PubMed

    Werth, Martin; Spiegel, Ann-Kathrin; Kazda, Marian

    2013-01-01

    The transformation from self-supporting lianas to host-supported climbing lianas is related to re-allocation of biomass and nutrients among plant organs. Therefore, first, variations in leaf mass per area (LMA), leaf carbon and nitrogen allocation and (13)C and (15)N natural abundances were analysed among three tropical Passiflora species (P. edulis, P. ligularis, and P. tripartita) in a greenhouse study. Second, the influence of a climbing support was considered for each species and parameter. P. ligularis leaves were most enriched in (13)C in both treatments when compared with the other two species. This enrichment was caused by a high LMA, which is related to a high internal resistance to CO(2) diffusion. For P. edulis and P. tripartita, δ(13)C was additionally increasing with nitrogen content per area. Generally, there were no differences when considering carbon and nitrogen allocation to leaves of host-supported and self-supporting lianas. The only hints towards increased investment into leaves after the transition from self-supporting to host-supported stages could be seen by a trend to increased leaf areas and masses. δ(13)C values of supported P. edulis or P. tripartita plants were significantly increasing faster than those of non-supported plants once the interactions of leaf mass or nitrogen content per area were accounted for. Hence, the offer of a climbing support had only a minor impact on δ(13)C or δ(15)N values in vitro, but this could be different with increasing age of lianas in vivo. PMID:23134439

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

    NASA Astrophysics Data System (ADS)

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

    2015-02-01

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

  3. Comparing the Influence of Wildfire and Prescribed Burns on Watershed Nitrogen Biogeochemistry Using 15N Natural Abundance in Terrestrial and Aquatic Ecosystem Components

    PubMed Central

    Stephan, Kirsten; Kavanagh, Kathleen L.; Koyama, Akihiro

    2015-01-01

    We evaluated differences in the effects of three low-severity spring prescribed burns and four wildfires on nitrogen (N) biogeochemistry in Rocky Mountain headwater watersheds. We compared paired (burned/unburned) watersheds of four wildfires and three spring prescribed burns for three growing seasons post-fire. To better understand fire effects on the entire watershed ecosystem, we measured N concentrations and δ15N in both the terrestrial and aquatic ecosystems components, i.e., soil, understory plants in upland and riparian areas, streamwater, and in-stream moss. In addition, we measured nitrate reductase activity in foliage of Spiraea betulifolia, a dominant understory species. We found increases of δ15N and N concentrations in both terrestrial and aquatic ecosystem N pools after wildfire, but responses were limited to terrestrial N pools after prescribed burns indicating that N transfer from terrestrial to aquatic ecosystem components did not occur in low-severity prescribed burns. Foliar δ15N differed between wildfire and prescribed burn sites; the δ15N of foliage of upland plants was enriched by 2.9 ‰ (difference between burned and unburned watersheds) in the first two years after wildfire, but only 1.3 ‰ after prescribed burns. In-stream moss δ15N in wildfire-burned watersheds was enriched by 1.3 ‰, but there was no response by moss in prescription-burned watersheds, mirroring patterns of streamwater nitrate concentrations. S. betulifolia showed significantly higher nitrate reductase activity two years after wildfires relative to corresponding unburned watersheds, but no such difference was found after prescribed burns. These responses are consistent with less altered N biogeochemistry after prescribed burns relative to wildfire. We concluded that δ15N values in terrestrial and aquatic plants and streamwater nitrate concentrations after fire can be useful indicators of the magnitude and duration of fire effects and the fate of post-fire available N. PMID:25885257

  4. Comparing the influence of wildfire and prescribed burns on watershed nitrogen biogeochemistry using 15N natural abundance in terrestrial and aquatic ecosystem components.

    PubMed

    Stephan, Kirsten; Kavanagh, Kathleen L; Koyama, Akihiro

    2015-01-01

    We evaluated differences in the effects of three low-severity spring prescribed burns and four wildfires on nitrogen (N) biogeochemistry in Rocky Mountain headwater watersheds. We compared paired (burned/unburned) watersheds of four wildfires and three spring prescribed burns for three growing seasons post-fire. To better understand fire effects on the entire watershed ecosystem, we measured N concentrations and δ15N in both the terrestrial and aquatic ecosystems components, i.e., soil, understory plants in upland and riparian areas, streamwater, and in-stream moss. In addition, we measured nitrate reductase activity in foliage of Spiraea betulifolia, a dominant understory species. We found increases of δ15N and N concentrations in both terrestrial and aquatic ecosystem N pools after wildfire, but responses were limited to terrestrial N pools after prescribed burns indicating that N transfer from terrestrial to aquatic ecosystem components did not occur in low-severity prescribed burns. Foliar δ15N differed between wildfire and prescribed burn sites; the δ15N of foliage of upland plants was enriched by 2.9 ‰ (difference between burned and unburned watersheds) in the first two years after wildfire, but only 1.3 ‰ after prescribed burns. In-stream moss δ15N in wildfire-burned watersheds was enriched by 1.3 ‰, but there was no response by moss in prescription-burned watersheds, mirroring patterns of streamwater nitrate concentrations. S. betulifolia showed significantly higher nitrate reductase activity two years after wildfires relative to corresponding unburned watersheds, but no such difference was found after prescribed burns. These responses are consistent with less altered N biogeochemistry after prescribed burns relative to wildfire. We concluded that δ15N values in terrestrial and aquatic plants and streamwater nitrate concentrations after fire can be useful indicators of the magnitude and duration of fire effects and the fate of post-fire available N. PMID:25885257

  5. Differences in animal 13C, 15N and D abundance between a polluted and an unpolluted coastal site: Likely indicators of sewage uptake by a marine food web

    NASA Astrophysics Data System (ADS)

    Rau, G. H.; Sweeney, R. E.; Kaplan, I. R.; Mearns, A. J.; Young, D. R.

    1981-12-01

    In comparing a control vs. a sewage-affected nearshore site off southern California, the natural relative abundances of 13C, 15N and D in Dover sole and Ridgeback prawn muscle were found to be significantly different. This between-site shift in animal isotope abundance is of the type expected if sewage organic materials were an important food source for animals in the latter site. Use of stable isotope natural abundance to trace the incorporation and movement of sewage-derived organics in marine food webs is suggested.

  6. 15N and 13C abundances in the Antartic Ocean with emphasis on the biogeochemical structure of the food web

    NASA Astrophysics Data System (ADS)

    Wada, Eitaro; Terazaki, Makoto; Kabaya, Yuko; Nemoto, Takahisa

    1987-06-01

    Distributions of δ 15N and δ 13C for biogenic substances in the Antarctic Ocean were investigated to construct a biogeochemical framework for assessing the Antarctic ecosystem. Phytoplankton exhibited particularly low δ 15N (0.5%) and 13C (-26.9%) values in pelagic plankton samples. High nitrate concentrations, and high PCO 2 in the surface waters on the southern side of the polar front and the resulting slow growth rate of phytoplankton under low light intensity are suggested as possible factors in causing the low isotopic compositions. Mean fractionation factors of 1.029 and 1.006 were estimated for photosynthetic carbon fixation and for the assimilation of inorganic nitrogeneous compounds (ammonium plus nitrate) during algal growth, respectively. Enrichment of 15N with increasing trophic level was confirmed for Antarctic ecosystems: δ15N animal% = 3.3 (trophic level -1) + δ5N algae, whereas 13C content did not increase in the same manner. Differences in lipid content among animals may be the main factor in causing this δ 13C anomaly. 15N and 13C abundance of sedimentary organic nitrogen differed from phytoplankton and settling particles. An exact mechanism for explaining the high δ 15N (around 5%) is not known. The very high δ 13C value of -20.5% at Sta. 3B may originate in ice algae that had grown under CO 2-limited conditions. Particles collected by sediment traps gave characteristically low δ 15N values (-3.0 to 0.9%), strongly suggesting a phytoplankton origin. The δ 15N and δ 13C values of settling material showed similar vertical profiles with depth which might arise from temporal variation of algal growth.

  7. Revision of the 15N(p, γ)16O reaction rate and oxygen abundance in H-burning zones

    NASA Astrophysics Data System (ADS)

    Caciolli, A.; Mazzocchi, C.; Capogrosso, V.; Bemmerer, D.; Broggini, C.; Corvisiero, P.; Costantini, H.; Elekes, Z.; Formicola, A.; Fülöp, Zs.; Gervino, G.; Guglielmetti, A.; Gustavino, C.; Gyürky, Gy.; Imbriani, G.; Junker, M.; Lemut, A.; Marta, M.; Menegazzo, R.; Palmerini, S.; Prati, P.; Roca, V.; Rolfs, C.; Rossi Alvarez, C.; Somorjai, E.; Straniero, O.; Strieder, F.; Terrasi, F.; Trautvetter, H. P.; Vomiero, A.

    2011-09-01

    Context. The NO cycle takes place in the deepest layer of a H-burning core or shell, when the temperature exceeds T ≃ 30 × 106 K. The O depletion observed in some globular cluster giant stars, always associated with a Na enhancement, may be due to either a deep mixing during the red giant branch (RGB) phase of the star or to the pollution of the primordial gas by an early population of massive asymptotic giant branch (AGB) stars, whose chemical composition was modified by the hot bottom burning. In both cases, the NO cycle is responsible for the O depletion. Aims: The activation of this cycle depends on the rate of the 15N(p, γ)16O reaction. A precise evaluation of this reaction rate at temperatures as low as experienced in H-burning zones in stellar interiors is mandatory to understand the observed O abundances. Methods: We present a new measurement of the 15N(p, γ)16O reaction performed at LUNA covering for the first time the center of mass energy range 70-370 keV, which corresponds to stellar temperatures between 65 × 106 K and 780 × 106 K. This range includes the 15N(p, γ)16O Gamow-peak energy of explosive H-burning taking place in the external layer of a nova and the one of the hot bottom burning (HBB) nucleosynthesis occurring in massive AGB stars. Results: With the present data, we are also able to confirm the result of the previous R-matrix extrapolation. In particular, in the temperature range of astrophysical interest, the new rate is about a factor of 2 smaller than reported in the widely adopted compilation of reaction rates (NACRE or CF88) and the uncertainty is now reduced down to the 10% level.

  8. Assessing denitrification in groundwater using natural gradient tracer tests with 15N: In situ measurement of a sequential multistep reaction

    USGS Publications Warehouse

    Smith, R.L.; Böhlke, J.K.; Garabedian, S.P.; Revesz, K.M.; Yoshinari, T.

    2004-01-01

    Denitrification was measured within a nitrate-contaminated aquifer on Cape Cod, Massachusetts, using natural gradient tracer tests with 15N nitrate. The aquifer contained zones of relatively high concentrations of nitrite (up to 77 ??M) and nitrous oxide (up to 143 ??M) and has been the site of previous studies examining ground water denitrification using the acetylene block technique. Small-scale (15-24 m travel distance) tracer tests were conducted by injecting 15N nitrate and bromide as tracers into a depth interval that contained nitrate, nitrite, nitrous oxide, and excess nitrogen gas. The timing of the bromide breakthrough curves at down-gradient wells matched peaks in 15N abundance above background for nitrate, nitrite, nitrous oxide, and nitrogen gas after more than 40 days of travel. Results were simulated with a one-dimensional transport model using linked reaction kinetics for the individual steps of the denitrification reaction pathway. It was necessary to include within the model spatial variations in background concentrations of all nitrogen oxide species. The model indicated that nitrite production (0.036-0.047 ??mol N (L aquifer)-1 d -1) was faster than the subsequent denitrification steps (0.013-0.016 ??mol N (L aquifer)-1 d-1 for nitrous oxide and 0.013-0.020 ??mol N (L aquifer)-1 d-1 for nitrogen gas) and that the total rate of reaction was slower than indicated by both acetylene block tracer tests and laboratory incubations. The rate of nitrate removal by denitrification was much slower than the rate of transport, indicating that nitrate would migrate several kilometers down-gradient before being completely consumed.

  9. 15N NMR Spectroscopy of Cyclotriphosphazenes and Polyphosphazenes Based on 31P, 15N HMQC Correlations

    NASA Astrophysics Data System (ADS)

    Peláez-Arango, Elvira; García-Alonso, Francisco J.; Carriedo, Gabino; López-Ortiz, Fernando

    A 31P and 15N NMR study of cyclotriphosphazenes 1-4 and polyphosphazene 5 has been carried out including solvent and substituent effects. 15N chemical shifts and 31P, 15N coupling constants are readily obtained through 1D and 2D 31P, 15N HMQC correlation spectroscopy on natural-abundance samples. This methodology combined with spectral simulation also yields 15N-induced isotope shifts. 31P and 15N are both relatively insensitive to changes in solvent polarity. Chemical shifts are reasonably explained in terms of electronic density variations, and 31P, 15N couplings are rationalized by considering their absolute sign, which depends on the substituent bonded to phosphorus. 15N proved to be more sensitive to structural modifications than 31P. Therefore, the availability of 15N NMR data on cyclotriphosphazenes can be considered a valuable tool in the design of model compounds for polyphosphazene synthesis.

  10. [Effects of intensive agricultural production on farmland soil carbon and nitrogen contents and their delta13C and delta15N isotope abundances].

    PubMed

    Yang, Guang-Rong; Hao, Xiying; Li, Chun-Li; Wang, Zi-Lin; Li, Yong-Mei

    2012-03-01

    Farmland soil carbon and nitrogen contents under intensive agricultural production are the important indices for the assessment of the soil fertility sustainability. This paper measured the soil pH, electrical conductivity (EC), organic carbon (SOC), total nitrogen (TN), and delta13C and delta15N isotope abundances of four types of farmland, i.e., conventional rice-broad bean rotation field, open vegetable field, 3-year plastic covered greenhouse field, and > 10-year plastic covered greenhouse field, aimed to understand the effects of intensive agricultural production degree on soil properties. In the open vegetable field, 3-year plastic covered greenhouse field, and > 10-year plastic covered greenhouse field, the soil (0-20 cm) pH decreased by 1.1, 0.8, and 0.7, and the soil EC was 4.2, 4.9, and 5.2 folds of that in conventional rice-broad bean rotation field, respectively. With the increasing year of plastic covered greenhouse production, the soil SOC and TN contents decreased after an initial increase. Comparing with those under rice-broad bean rotation, the SOC content in 0-20, 20-40, 40-60, 60-80 and 80-100 cm soil layers in >10-year plastic covered greenhouse decreased by 54%, 46%, 60%, 63%, and 59%, and the TN content decreased by 53%, 53%, 71%, 82%, and 85%, respectively. Intensive agricultural production degree had significant effects on the soil SOC and TN contents and delta13C and delta15N abundances. The delta13C abundance was significantly negatively correlated with the soil SOC, suggesting that the soil delta13C abundance could be regarded as an index for the assessment of carbon cycle in farmland soils under effects of human activities. PMID:22720621

  11. Helium isotopic abundance variation in nature

    SciTech Connect

    Holden, N.E.

    1993-08-01

    The isotopic abundance of helium in nature has been reviewed. This atomic weight value is based on the value of helium in the atmosphere, which is invariant around the world and up to a distance of 100,000 feet. Helium does vary in natural gas, volcanic rocks and gases, ocean floor sediments, waters of various types and in radioactive minerals and ores due to {alpha} particle decay of radioactive nuclides.

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

    PubMed

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

    2009-11-01

    Accurate determinations of stable isotope ratios require a calibration using at least two reference materials with different isotopic compositions to anchor the isotopic scale and compensate for differences in machine slope. Ideally, the delta values of these reference materials should bracket the isotopic range of samples with unknown delta values. While the practice of analyzing two isotopically distinct reference materials is common for water (VSMOW-SLAP) and carbonates (NBS 19 and L-SVEC), the lack of widely available organic reference materials with distinct isotopic composition has hindered the practice when analyzing organic materials by elemental analysis/isotope ratio mass spectrometry (EA-IRMS). At present only L-glutamic acids USGS40 and USGS41 satisfy these requirements for delta13C and delta15N, with the limitation that L-glutamic acid is not suitable for analysis by gas chromatography (GC). We describe the development and quality testing of (i) four nicotine laboratory reference materials for on-line (i.e. continuous flow) hydrogen reductive gas chromatography-isotope ratio mass-spectrometry (GC-IRMS), (ii) five nicotines for oxidative C, N gas chromatography-combustion-isotope ratio mass-spectrometry (GC-C-IRMS, or GC-IRMS), and (iii) also three acetanilide and three urea reference materials for on-line oxidative EA-IRMS for C and N. Isotopic off-line calibration against international stable isotope measurement standards at Indiana University adhered to the 'principle of identical treatment'. The new reference materials cover the following isotopic ranges: delta2H(nicotine) -162 to -45 per thousand, delta13C(nicotine) -30.05 to +7.72 per thousand, delta15N(nicotine) -6.03 to +33.62 per thousand; delta15N(acetanilide) +1.18 to +40.57 per thousand; delta13C(urea) -34.13 to +11.71 per thousand, delta15N(urea) +0.26 to +40.61 per thousand (recommended delta values refer to calibration with NBS 19, L-SVEC, IAEA-N-1, and IAEA-N-2). Nicotines fill a gap as the first organic nitrogen stable isotope reference materials for GC-IRMS that are available with different delta15N values. Comparative delta13C and delta15N on-line EA-IRMS data from 14 volunteering laboratories document the usefulness and reliability of acetanilides and ureas as EA-IRMS reference materials. PMID:19844968

  13. Determining the source of nitrate pollution in the Niger discontinuous aquifers using the natural {15N }/{14N } ratios

    NASA Astrophysics Data System (ADS)

    Girard, Pierre; Hillaire-Marcel, Claude

    1997-12-01

    In the semi-arid Niamey area (Niger), more than 10% of the deep wells exploiting the fracture network of the Precambrian aquifer are contaminated by nitrates, with concentrations as high as 10 meq l -1. In order to identify the source(s) of this pollution, nitrate and 15N contents in the polluted wells were monitored over a 20-month period. Potential sources of nitrate contamination were also analyzed for their 15N content. The isotopic compositions of nitrate in polluted waters were > + 12‰ and in rare cases exceeded +17‰. Latrines (˜ + 15‰) may be the major nitrate source for wells showing δ15N values above +15‰. Below this value, waters may be polluted by a combination of nitrates from both latrine and soil sources (˜ + 10‰). In some cases, the soil may account for up to 85% of the groundwater nitrate load. This mode of groundwater pollution is thought to be a consequence of deforestation. Despite their reputation as polluting agents, fertilizers ( +0.5 < δ 15N < + 3.6‰ ) which are used in rice paddies close to the contaminated areas, do not appear to be a significant source of nitrate contamination. Denitrification is probably not a significant process in the study area. Results suggest that nitrate contamination of the aquifer is a consequence of unregulated urbanization (home-made latrines) and deforestation. While latrines are limited to the urban zones, intensive cutting of the forest to meet the city dwellers' wood demand occurs in an ever increasing area around the capital, threatening the local water supply.

  14. Nitrate reductase 15N discrimination in Arabidopsis thaliana, Zea mays, Aspergillus niger, Pichea angusta, and Escherichia coli

    PubMed Central

    Carlisle, Eli; Yarnes, Chris; Toney, Michael D.; Bloom, Arnold J.

    2014-01-01

    Stable 15N isotopes have been used to examine movement of nitrogen (N) through various pools of the global N cycle. A central reaction in the cycle involves the reduction of nitrate (NO?3) to nitrite (NO?2) catalyzed by nitrate reductase (NR). Discrimination against 15N by NR is a major determinant of isotopic differences among N pools. Here, we measured in vitro 15N discrimination by several NRs purified from plants, fungi, and a bacterium to determine the intrinsic 15N discrimination by the enzyme and to evaluate the validity of measurements made using 15N-enriched NO?3. Observed NR isotope discrimination ranged from 22 to 32 (kinetic isotope effects of 1.0221.032) among the different isozymes at natural abundance 15N (0.37%). As the fractional 15N content of substrate NO?3 increased from natural abundance, the product 15N fraction deviated significantly from that expected based on substrate enrichment and 15N discrimination measured at natural abundance. Additionally, isotopic discrimination by denitrifying bacteria used to reduce NO?3 and NO?2 in some protocols became a greater source of error as 15N enrichment increased. We briefly discuss potential causes of the experimental artifacts with enriched 15N and recommend against the use of highly enriched 15N tracers to study N discrimination in plants or soils. PMID:25071800

  15. δ 15 N constraints on long-term nitrogen balances in temperate forests

    EPA Science Inventory

    Natural abundance δ15N of ecosystems integrates nitrogen (N) inputs and losses, and thus reflects factors that control the long-term development of ecosystem N balances. We here report N and carbon (C) content of forest vegetation and soils, and associated δ15N, across nine Doug...

  16. ? 15 N constraints on long-term nitrogen balances in temperate forests

    EPA Science Inventory

    Natural abundance ?15N of ecosystems integrates nitrogen (N) inputs and losses, and thus reflects factors that control the long-term development of ecosystem N balances. We here report N and carbon (C) content of forest vegetation and soils, and associated ?15N, across nine Doug...

  17. Nicotine, acetanilide and urea multi-level2H-,13C- and15N-abundance reference materials for continuous-flow isotope ratio mass spectrometry

    USGS Publications Warehouse

    Schimmelmann, A.; Albertino, A.; Sauer, P.E.; Qi, H.; Molinie, R.; Mesnard, F.

    2009-01-01

    Accurate determinations of stable isotope ratios require a calibration using at least two reference materials with different isotopic compositions to anchor the isotopic scale and compensate for differences in machine slope. Ideally, the S values of these reference materials should bracket the isotopic range of samples with unknown S values. While the practice of analyzing two isotopically distinct reference materials is common for water (VSMOW-SLAP) and carbonates (NBS 19 and L-SVEC), the lack of widely available organic reference materials with distinct isotopic composition has hindered the practice when analyzing organic materials by elemental analysis/isotope ratio mass spectrometry (EA-IRMS). At present only L-glutamic acids USGS40 and USGS41 satisfy these requirements for ??13C and ??13N, with the limitation that L-glutamic acid is not suitable for analysis by gas chromatography (GC). We describe the development and quality testing of (i) four nicotine laboratory reference materials for on-line (i.e. continuous flow) hydrogen reductive gas chromatography-isotope ratio mass-spectrometry (GC-IRMS), (ii) five nicotines for oxidative C, N gas chromatography-combustion-isotope ratio mass-spectrometry (GC-C-IRMS, or GC-IRMS), and (iii) also three acetanilide and three urea reference materials for on-line oxidative EA-IRMS for C and N. Isotopic off-line calibration against international stable isotope measurement standards at Indiana University adhered to the 'principle of identical treatment'. The new reference materials cover the following isotopic ranges: ??2Hnicotine -162 to -45%o, ??13Cnicotine -30.05 to +7.72%, ?? 15Nnicotine -6.03 to +33.62%; ??15N acetanilide +1-18 to +40.57%; ??13Curea -34.13 to +11.71%, ??15Nurea +0.26 to +40.61% (recommended ?? values refer to calibration with NBS 19, L-SVEC, IAEA-N-1, and IAEA-N-2). Nicotines fill a gap as the first organic nitrogen stable isotope reference materials for GC-IRMS that are available with different ??13N values. Comparative ??13C and ??15N on-line EA-IRMS data from 14 volunteering laboratories document the usefulness and reliability of acetanilides and ureas as EA-IRMS reference materials.

  18. Food Resources of Stream Macronivertebrates Determined by Natural-Abundance stable C and N Isotopes and a 15N Tracer Addition

    SciTech Connect

    Mulholland, P. J.

    2000-01-01

    Trophic relationships were examined using natural-abundance {sup 13}C and {sup 15}N analyses and a {sup 15}N-tracer addition experiment in Walker Branch, a 1st-order forested stream in eastern Tennessee. In the {sup 15}N-tracer addition experiment, we added {sup 15}NH{sub 4} to stream water over a 6-wk period in early spring, and measured {sup 15}N:{sup 14}N ratios in different taxa and biomass compartments over distance and time. Samples collected from a station upstream from the {sup 15}N addition provided data on natural-abundance {sup 13}C:{sup 12}C and {sup 15}N:{sup 14}N ratios. The natural-abundance {sup 15}N analysis proved to be of limited value in identifying food resources of macroinvertebrates because {sup 15}N values were not greatly different among food resources. In general, the natural-abundance stable isotope approach was most useful for determining whether epilithon or detritus were important food resources for organisms that may use both (e.g., the snail Elimia clavaeformis), and to provide corroborative evidence of food resources of taxa for which the {sup 15}N tracer results were not definitive. The {sup 15}N tracer results showed that the mayflies Stenonema spp. and Baetis spp. assimilated primarily epilithon, although Baetis appeared to assimilate a portion of the epilithon (e.g., algal cells) with more rapid N turnover than the bulk pool sampled. Although Elimia did not reach isotopic equilibrium during the tracer experiment, application of a N-turnover model to the field data suggested that it assimilated a combination of epilithon and detritus. The amphipod Gammarus minus appeared to depend mostly on fine benthic organic matter (FBOM), and the coleopteran Anchytarsus bicolor on epixylon. The caddisfly Diplectrona modesta appeared to assimilate primarily a fast N-turnover portion of the FBOM pool, and Simuliidae a fast N-turnover component of the suspended particulate organic matter pool rather than the bulk pool sampled. Together, the natural-abundance stable C and N isotope analyses and the experimental {sup 15}N tracer approach proved to be very useful tools for identifying food resources in this stream ecosystem.

  19. Application of the 15N gas-flux method for measuring in situ N2 and N2O fluxes due to denitrification in natural and semi-natural terrestrial ecosystems and comparison with the acetylene inhibition technique

    NASA Astrophysics Data System (ADS)

    Sgouridis, Fotis; Stott, Andrew; Ullah, Sami

    2016-03-01

    Soil denitrification is considered the most un-constrained process in the global N cycle due to uncertain in situ N2 flux measurements, particularly in natural and semi-natural terrestrial ecosystems. 15N tracer approaches can provide in situ measurements of both N2 and N2O simultaneously, but their use has been limited to fertilized agro-ecosystems due to the need for large 15N additions in order to detect 15N2 production against the high atmospheric N2. For 15N-N2 analyses, we have used an "in-house" laboratory designed and manufactured N2 preparation instrument which can be interfaced to any commercial continuous flow isotope ratio mass spectrometer (CF-IRMS). The N2 prep unit has gas purification steps and a copper-based reduction furnace, and allows the analysis of small gas injection volumes (4 µL) for 15N-N2 analysis. For the analysis of N2O, an automated Tracegas Preconcentrator (Isoprime Ltd) coupled to an IRMS was used to measure the 15N-N2O (4 mL gas injection volume). Consequently, the coefficient of variation for the determination of isotope ratios for N2 in air and in standard N2O (0.5 ppm) was better than 0.5 %. The 15N gas-flux method was adapted for application in natural and semi-natural land use types (peatlands, forests, and grasslands) by lowering the 15N tracer application rate to 0.04-0.5 kg 15N ha-1. The minimum detectable flux rates were 4 µg N m-2 h-1 and 0.2 ng N m-2 h-1 for the N2 and N2O fluxes respectively. Total denitrification rates measured by the acetylene inhibition technique in the same land use types correlated (r = 0.58) with the denitrification rates measured under the 15N gas-flux method, but were underestimated by a factor of 4, and this was partially attributed to the incomplete inhibition of N2O reduction to N2, under a relatively high soil moisture content, and/or the catalytic NO decomposition in the presence of acetylene. Even though relatively robust for in situ denitrification measurements, methodological uncertainties still exist in the estimation of N2 and N2O fluxes with the 15N gas-flux method due to issues related to non-homogenous distribution of the added tracer and subsoil gas diffusion using open-bottom chambers, particularly during longer incubation duration. Despite these uncertainties, the 15N gas-flux method constitutes a more reliable field technique for large-scale quantification of N2 and N2O fluxes in natural terrestrial ecosystems, thus significantly improving our ability to constrain ecosystem N budgets.

  20. Application of the 15N-Gas Flux method for measuring in situ N2 and N2O fluxes due to denitrification in natural and semi-natural terrestrial ecosystems and comparison with the acetylene inhibition technique

    NASA Astrophysics Data System (ADS)

    Sgouridis, F.; Ullah, S.; Stott, A.

    2015-08-01

    Soil denitrification is considered the most un-constrained process in the global N cycle due to uncertain in situ N2 flux measurements, particularly in natural and semi-natural terrestrial ecosystems. 15N tracer approaches can provide in situ measurements of both N2 and N2O simultaneously, but their use has been limited to fertilised agro-ecosystems due to the need for large 15N additions in order to detect 15N2 production against the high atmospheric N2. For 15N-N2 analyses, we have used an "in house" laboratory designed and manufactured N2 preparation instrument which can be interfaced to any commercial continuous flow isotope ratio mass spectrometer (CF-IRMS). The N2 prep unit has gas purification steps, a copper based reduction furnace, and allows the analysis of small gas injection volumes (4 μL) for 15N-N2 analysis. For the analysis of N2O, an automated Tracegas Pre-concentrator (Isoprime Ltd) coupled to an IRMS was used to measure the 15N-N2O (4 mL gas injection volume). Consequently, the coefficient of variation for the determination of isotope ratios for N2 in air and in standard N2O (0.5 ppm) was better than 0.5 %. The 15N Gas-Flux method was adapted for application in natural and semi-natural land use types (peatlands, forests and grasslands) by lowering the 15N tracer application rate to 0.04-0.5 kg 15N ha-1. For our chamber design (volume / surface = 8:1) and a 20 h incubation period, the minimum detectable flux rates were 4 μg N m-2 h-1 and 0.2 ng N m-2 h-1 for the N2 and N2O fluxes respectively. The N2 flux ranged between 2.4 and 416.6 μg N m-2 h-1, and the grassland soils showed on average 3 and 14 times higher denitrification rates than the woodland and organic soils respectively. The N2O flux was on average 20 to 200 times lower than the N2 flux, while the denitrification product ratio (N2O/N2 + N2O) was low, ranging between 0.03 and 13 %. Total denitrification rates measured by the acetylene inhibition technique under the same field conditions correlated (r = 0.58) with the denitrification rates measured under the 15N Gas-Flux method but were underestimated by a factor of 4 and this was attributed to the incomplete inhibition of N2O reduction to N2 under relatively high soil moisture content. The results show that the 15N Gas-Flux method can be used for quantifying N2 and N2O production rates in natural terrestrial ecosystems, thus significantly improving our ability to constrain ecosystem N budgets.

  1. Grass species influence on plant N uptake - Determination of atmospheric N deposition to a semi-natural peat bog site using a 15N labelling approach

    NASA Astrophysics Data System (ADS)

    Hurkuck, Miriam; Brümmer, Christian; Spott, Oliver; Flessa, Heinz; Kutsch, Werner L.

    2014-05-01

    Large areas of natural peat bogs in Northwestern Germany have been converted to arable land and were subjected to draining and peat cutting in the past. The few protected peatland areas remaining are affected by high nitrogen (N) deposition. Our study site - a moderately drained raised bog - is surrounded by highly fertilized agricultural land and livestock production. In this study, we used a 15N pool dilution technique called 'Integrated Total Nitrogen Input' (ITNI) to quantify annual deposition of atmospheric N into biomonitoring pots over a two-year period. Since it considers direct N uptake by plants, it was expected to result in higher N input than conventional methods for determination of N deposition (e.g. micrometeorological approaches, bulk N samplers). Using Lolium multiflorum and Eriophorum vaginatum as monitor plants and low, medium and high levels of fertilization, we aimed to simulate increasing N deposition to planted pots and to allocate airborne N after its uptake by the soil-plant system in aboveground biomass, roots and soil. Increasing N fertilization was positively correlated with biomass production of Eriophorum vaginatum, whereas atmospheric plant N uptake decreased and highest airborne N input of 899.8 ± 67.4 µg N d-1 pot-1 was found for low N fertilization. In contrast, Lolium multiflorum showed a clear dependency of N supply on plant N uptake and was highest (688.7 ± 41.4 µg N d-1 pot-1) for highly fertilized vegetation pots. Our results suggest that grass species respond differently to increasing N input. While crop grasses such as Lolium multiflorum take up N according to N availability, species adopted to nutrient-limited conditions like Eriophorum vaginatum show N saturation effects with increasing N supply. Total airborne N input ranged from about 24 to 66 kg N ha-1 yr-1 dependent on the used indicator plant and the amount of added fertilizer. Parallel determination of atmospheric N deposition using a micrometeorological approach complemented with bulk samplers was about 24 kg N ha-1 yr-1 during both years of experiments and was thus at the lower range of results obtained by the ITNI method. The low 15N recovery rate of about 50 % during some experiments indicated an underestimation of the applied ITNI approach, resulting in a maximum possible N uptake of twice as high as the determined N input. Most likely, the intensive agricultural land management of the surrounding areas leads to this high N deposition into the protected peatland area. As a result, increasing sensitivity of ombrotrophic vegetation with a subsequent change in plant species composition and a decline in bog-specific vegetation cannot be excluded.

  2. Natural abundance nitrogen-15 nuclear magnetic resonance spectral studies on selected donors

    NASA Astrophysics Data System (ADS)

    Someswara Rao, N.; Babu Rao, G.; Murthy, B. N.; Das, M. Maria; Prabhakar, T.; Lalitha, M.

    2002-10-01

    The natural abundance 15N-NMR chemical shifts of selected aliphatic amines, 2-substituted pyridine type compounds, bialicyclic tertiary amines have been measured as a function of the nature of the solvent. In the case of cyclic aliphatic amines, like piperidine, morpholine, piperazine, thiomorpholine, the nitrogen is more shielded in concentrated solution compared to that in dilute solution whereas in the hydrogen bonding and protonating solvents there is a prominent deshielding. 2-Substituted pyridines studied can be further divided into four sub groups. The site of hydrogen bonding and protonation in 2-amino, 2-hydroxy and 2-mercapto pyridines have been conclusively proved from the 15N-NMR chemical shifts and the well-known tautomeric forms of the above compounds. Similarly in the case of 2-(2-thienyl)pyridine and 2-(3-thienyl)pyridine, the site of donation has been proved as the nitrogen of the pyridine ring in both the compounds. In a similar manner, the site of hydrogen bonding and protonation in two individual compounds 2-anilinopyridine and 2-(2-pyridyl)benzimidazole have also been established. Among the bialicyclic amines, 1,2-diazabicyclo[2.2.2]octane (DABCO) behaved differently from the other two compounds. In both 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), it was possible to show that N 1-nitrogen in both the compounds is the site of donation. The effect of the second donor site on the 15N-NMR chemical shift, the site of donation in the selected compounds and some typical compounds reported in literature have been presented and discussed.

  3. Two abundant bioaccumulated halogenated compounds are natural products.

    PubMed

    Teuten, Emma L; Xu, Li; Reddy, Christopher M

    2005-02-11

    Methoxylated polybrominated diphenyl ethers (MeO-PBDEs) have been found bioaccumulated in the tissues of a variety of aquatic animals and at concentrations comparable to those of anthropogenic halogenated organic compounds, including polychlorinated biphenyls (PCBs). The origin of the MeO-PBDEs has been uncertain; circumstantial evidence supports a natural and/or an industrial source. By analyzing the natural abundance radiocarbon content of two MeO-PBDEs isolated from a True's beaked whale (Mesoplodon mirus), we show that these compounds were naturally produced. PMID:15705850

  4. Differential incorporation of natural spawners vs. artificially planted salmon carcasses in a stream food web: Evidence from delta 15N of juvenile coho salmon

    EPA Science Inventory

    Placement of salmon carcasses is a common restoration technique in Oregon and Washington streams, with the goal of improving food resources and productivity of juvenile salmon. To explore the effectiveness of this restoration technique, we measured the δ15N of juvenile coho salmo...

  5. Utility of stable isotopes ((13)C and (15)N) to demonstrate comparability between natural and experimental streams for environmental risk assessment.

    PubMed

    Morrall, Donna D; Christman, Steve C; Peterson, Bruce J; Wolheim, Wilfred M; Belanger, Scott E

    2006-09-01

    Environmental safety of contaminants sometimes requires testing at the ecosystem level. Model ecosystems can serve as a surrogate to receiving waters for the assessor. Tools to verify that model ecosystems are good surrogates for natural receiving water systems have been based on structural attributes such as taxa lists. While examining taxa lists can be useful within a geography, they are less informative when comparing across different topographical and climatic regions. This paper illustrates an alternative approach that uses natural abundance ratios of nitrogen and carbon. Comparisons were made between the Procter and Gamble Co. (P and G) Experimental Stream Facility (ESF) and the East Fork Little Miami River (EFLMR). We also conducted stable isotopes releases of nitrogen ((15)NH(4)Cl) in the ESF and EFLMR. Results from the nitrogen releases for the ESF and EFLMR were compared to each other and to stable isotope releases conducted in 9 streams as part of the Lotic Intersite Nitrogen eXperiment (LINX). Preliminary evaluations of nitrogen uptake show that the uptake rate of ammonium for the P and G experimental streams (0.019 m(-1)) is similar to uptake rates in natural streams of similar flow and fits the regression of uptake length and discharge for the LINX streams. Nutrient flow data obtained in this study in most cases verified model (SISTM) predictions. PMID:16139363

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

    PubMed

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

    2012-08-01

    Effects of anthropogenic nitrogen (N) deposition and the ability of terrestrial ecosystems to store carbon (C) depend in part on the amount of N retained in the system and its partitioning among plant and soil pools. We conducted a meta-analysis of studies at 48 sites across four continents that used enriched 15N isotope tracers in order to synthesize information about total ecosystem N retention (i.e., total ecosystem 15N recovery in plant and soil pools) across natural systems and N partitioning among ecosystem pools. The greatest recoveries of ecosystem 15N tracer occurred in shrublands (mean, 89.5%) and wetlands (84.8%) followed by forests (74.9%) and grasslands (51.8%). In the short term (< 1 week after 15N tracer application), total ecosystem 15N recovery was negatively correlated with fine-root and soil 15N natural abundance, and organic soil C and N concentration but was positively correlated with mean annual temperature and mineral soil C:N. In the longer term (3-18 months after 15N tracer application), total ecosystem 15N retention was negatively correlated with foliar natural-abundance 15N but was positively correlated with mineral soil C and N concentration and C:N, showing that plant and soil natural-abundance 15N and soil C:N are good indicators of total ecosystem N retention. Foliar N concentration was not significantly related to ecosystem 15N tracer recovery, suggesting that plant N status is not a good predictor of total ecosystem N retention. Because the largest ecosystem sinks for 15N tracer were below ground in forests, shrublands, and grasslands, we conclude that growth enhancement and potential for increased C storage in aboveground biomass from atmospheric N deposition is likely to be modest in these ecosystems. Total ecosystem 15N recovery decreased with N fertilization, with an apparent threshold fertilization rate of 46 kg N x ha(-1) x yr(-1) above which most ecosystems showed net losses of applied 15N tracer in response to N fertilizer addition. PMID:22928411

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

    USGS Publications Warehouse

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

    2012-01-01

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

  8. Assessment of 15N15N16O as a tracer of stratospheric processes

    NASA Astrophysics Data System (ADS)

    Kaiser, Jan; Röckmann, Thomas; Brenninkmeijer, Carl A. M.

    2003-01-01

    Isotopic fractionation of mono-substituted N2O species by the stratospheric sinks, photolysis and reaction with O(1D), has been investigated previously by atmospheric and laboratory measurements. Here, we report on photolysis of the most abundant poly-substituted isotopologue, 15N15N16O. The associated fractionation constant equals the sum of individual fractionation constants at central and terminal nitrogen sites. Based on statistical considerations, a more general ``additivity rule'' is proposed that extends to the remaining poly-substituted N2O species and is compared to predictions from two theories. Atmospheric mixing effects do not lead to measurable deviations from random isotope distributions since variations of isotope ratios are too small. Fractionation by the O(1D) sink likely also follows the ``additivity rule''. Consequently, the stratospheric 15N15N16O abundance is expected to reflect its statistically dictated value. The value of 15N15N16O as a potential new tracer of stratospheric processes may thus be limited, but this remains to be verified by actual measurements.

  9. Chlorine-36 abundance in natural and synthetic perchlorate

    SciTech Connect

    Heikoop, Jeffrey M; Dale, M; Sturchio, Neil C; Caffee, M; Belosa, A D; Heraty, Jr., L J; Bohike, J K; Hatzinger, P B; Jackson, W A; Gu, B

    2009-01-01

    Perchlorate (ClO{sub 4}{sup -}) is ubiquitous in the environment. It occurs naturally as a product of atmospheric photochemical reactions, and is synthesized for military, aerospace, and industrial applications. Nitrate-enriched soils of the Atacama Desert (Chile) contain high concentrations of natural ClO{sub 4}{sup -}; nitrate produced from these soils has been exported worldwide since the mid-1800's for use in agriculture. The widespread introduction of synthetic and agricultural ClO{sub 4}{sup -} into the environment has complicated attempts to understand the geochemical cycle of ClO{sub 4}{sup -}. Natural ClO{sub 4}{sup -} samples from the southwestern United States have relatively high {sup 36}Cl abundances ({sup 36}Cl/Cl = 3,100 x 10{sup -15} to 28,800 x 10{sup -15}), compared with samples of synthetic ({sup 36}Cl/Cl = 0.0 x 10{sup -15} to 40 x 10{sup -15}) and Atacama Desert ({sup 36}Cl/Cl = 0.9 x 10{sup -15} to 590 x 10{sup -15}) ClO{sub 4}{sup -}. These data give a lower limit for the initial {sup 36}Cl abundance of natural ClO{sub 4}{sup -} and provide temporal and other constraints on its geochemical cycle.

  10. QUANTITATIVE 15N NMR SPECTROSCOPY

    EPA Science Inventory

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

  11. A robust method for ammonium nitrogen isotopic analysis in freshwater and seawater at natural abundance levels

    NASA Astrophysics Data System (ADS)

    Zhang, L.; Altabet, M. A.; Wu, T.; Hadas, O.

    2006-12-01

    Natural ammonium N isotopic abundance has been increasingly used in studies of marine and freshwater biogeochemistry. However, current methods are time-consuming, subject to interference from DON, and not reliable at low concentrations. Our new method for determining the δ15N of ammonium overcomes these difficulties by employing the oxidation of ammonium to nitrite followed by conversion of nitrite to nitrous oxide. In the first step, ammonium is quantitatively oxidized by hypobromite at pH~12. After the addition of sodium arsenite to consume excess hypobromite, yield is verified by colorimetric NO2-measurement using sulfanilamide and naphthyl ethylenediamine (NED). Nitrite is further reduced to N2O by a 1:1 sodium azide and acetic acid buffer solution using previously established procedures. Buffer concentration can be varied according to sample matrix to ensure that a reaction pH between 2 and 4 is reached. The product nitrous oxide is then isotopically analyzed using a continuous flow purge and cryogenic trap system coupled to an isotope ratio mass spectrometer. Reliable δ15N values (±0.31‰) are obtained over a concentration range of 0.5 μM to 20 μM using 20 ml volumes of either fresh or seawater samples. Reagent blanks are very low, about 0.05 μM. There is no interference from any of the nitrogen containing compounds tested except short chain aliphatic amino acid (i.e. glycine) which typically are not present at sufficiently high environmental concentrations to pose a problem.

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

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

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

  13. N-15 NMR spectra of naturally abundant nitrogen in soil and aquatic natural organic matter samples of the International Humic Substances Society

    USGS Publications Warehouse

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

    2009-01-01

    The naturally abundant nitrogen in soil and aquatic NOM samples from the International Humic Substances Society has been characterized by solid state CP/MAS 15N NMR. Soil samples include humic and fulvic acids from the Elliot soil, Minnesota Waskish peat and Florida Pahokee peat, as well as the Summit Hill soil humic acid and the Leonardite humic acid. Aquatic samples include Suwannee River humic, fulvic and reverse osmosis isolates, Nordic humic and fulvic acids and Pony Lake fulvic acid. Additionally, Nordic and Suwannee River XAD-4 acids and Suwannee River hydrophobic neutral fractions were analyzed. Similar to literature reports, amide/aminoquinone nitrogens comprised the major peaks in the solid state spectra of the soil humic and fulvic acids, along with heterocyclic and amino sugar/terminal amino acid nitrogens. Spectra of aquatic samples, including the XAD-4 acids, contain resolved heterocyclic nitrogen peaks in addition to the amide nitrogens. The spectrum of the nitrogen enriched, microbially derived Pony Lake, Antarctica fulvic acid, appeared to contain resonances in the region of pyrazine, imine and/or pyridine nitrogens, which have not been observed previously in soil or aquatic humic substances by 15N NMR. Liquid state 15N NMR experiments were also recorded on the Elliot soil humic acid and Pony Lake fulvic acid, both to examine the feasibility of the techniques, and to determine whether improvements in resolution over the solid state could be realized. For both samples, polarization transfer (DEPT) and indirect detection (1H-15N gHSQC) spectra revealed greater resolution among nitrogens directly bonded to protons. The amide/aminoquinone nitrogens could also be observed by direct detection experiments.

  14. Diversity and abundance of phosphonate biosynthetic genes in nature

    PubMed Central

    Yu, Xiaomin; Doroghazi, James R.; Janga, Sarath C.; Zhang, Jun Kai; Circello, Benjamin; Griffin, Benjamin M.; Labeda, David P.; Metcalf, William W.

    2013-01-01

    Phosphonates, molecules containing direct carbon–phosphorus bonds, compose a structurally diverse class of natural products with interesting and useful biological properties. Although their synthesis in protozoa was discovered more than 50 y ago, the extent and diversity of phosphonate production in nature remains poorly characterized. The rearrangement of phosphoenolpyruvate (PEP) to phosphonopyruvate, catalyzed by the enzyme PEP mutase (PepM), is shared by the vast majority of known phosphonate biosynthetic pathways. Thus, the pepM gene can be used as a molecular marker to examine the occurrence and abundance of phosphonate-producing organisms. Based on the presence of this gene, phosphonate biosynthesis is common in microbes, with ∼5% of sequenced bacterial genomes and 7% of genome equivalents in metagenomic datasets carrying pepM homologs. Similarly, we detected the pepM gene in ∼5% of random actinomycete isolates. The pepM-containing gene neighborhoods from 25 of these isolates were cloned, sequenced, and compared with those found in sequenced genomes. PEP mutase sequence conservation is strongly correlated with conservation of other nearby genes, suggesting that the diversity of phosphonate biosynthetic pathways can be predicted by examining PEP mutase diversity. We used this approach to estimate the range of phosphonate biosynthetic pathways in nature, revealing dozens of discrete groups in pepM amplicons from local soils, whereas hundreds were observed in metagenomic datasets. Collectively, our analyses show that phosphonate biosynthesis is both diverse and relatively common in nature, suggesting that the role of phosphonate molecules in the biosphere may be more important than is often recognized. PMID:24297932

  15. Practical recommendations for the reduction of memory effects in compound-specific 15N/14N-ratio analysis of enriched amino acids by gas chromatography/combustion/isotope ratio mass spectrometry.

    PubMed

    Petzke, Klaus J; Metges, Cornelia C

    2012-01-30

    Gas chromatography/combustion/isotope ratio mass spectrometry (GC-C-IRMS) is a highly sensitive approach which allows the analysis of the (13)C/(12)C and (15)N/(14)N isotope composition of amino acids in the range of natural abundance or in slightly (13)C- and (15)N-enriched samples. However, the accuracy of measurements remains a permanent challenge. Here we show the effect of the presence of slightly (15)N-enriched compounds in physiological samples on the accuracy and reproducibility of (15)N-abundances of amino acids within or between analytical runs. We spiked several individual amino acids with the respective (15)N-labelled isotopomer and measured the (15)N/(14)N ratios of other amino acids in the same sample or in the following analytical runs. Intra- and inter-run memory effects can be observed in (15)N/(14)N ratios of amino acids. Sample throughput is reduced when cleaning runs using standard mixtures are required to restore initial conditions after runs of samples with (15)N-enriched analytes. Possible reasons for the observed phenomenon and its implications for work in the lower (15)N-enrichment range (<0.5 APE) are discussed and include different aspects of gas chromatography, derivatisation, and hot catalytic metal surface effects. Results need to be interpreted with caution if complex physiological samples contain (15)N-enriched amino acids beyond 500 ?(15)N (~0.18 APE). PMID:22173808

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

    PubMed

    Schmidt, S; Stewart, G R

    2003-03-01

    A large number of herbaceous and woody plants from tropical woodland, savanna, and monsoon forest were analysed to determine the impact of environmental factors (nutrient and water availability, fire) and biological factors (microbial associations, systematics) on plant delta(15)N values. Foliar delta(15)N values of herbaceous and woody species were not related to growth form or phenology, but a strong relationship existed between mycorrhizal status and plant delta(15)N. In woodland and savanna, woody species with ectomycorrhizal (ECM) associations and putative N(2)-fixing species with ECM/arbuscular (AM) associations had lowest foliar delta(15)N values (1.0-0.6 per thousand ), AM species had mostly intermediate delta(15)N values (average +0.6 per thousand ), while non-mycorrhizal Proteaceae had highest delta(15)N values (+2.9 to +4.1 per thousand ). Similar differences in foliar delta(15)N were observed between AM (average 0.1 and 0.2 per thousand ) and non-mycorrhizal (average +0.8 and +0.3 per thousand ) herbaceous species in woodland and savanna. Leguminous savanna species had significantly higher leaf N contents (1.8-2.5% N) than non-fixing species (0.9-1.2% N) indicating substantial N acquisition via N(2) fixation. Monsoon forest species had similar leaf N contents (average 2.4% N) and positive delta(15)N values (+0.9 to +2.4 per thousand ). Soil nitrification and plant NO(3)(-) use was substantially higher in monsoon forest than in woodland or savanna. In the studied communities, higher soil N content and nitrification rates were associated with more positive soil delta(15)N and plant delta(15)N. In support of this notion, Ficus, a high NO(3)(-) using taxa associated with NO(3)(-) rich sites in the savanna, had the highest delta(15)N values of all AM species in the savanna. delta(15)N of xylem sap was examined as a tool for studying plant delta(15)N relations. delta(15)N of xylem sap varied seasonally and between differently aged Acacia and other savanna species. Plants from annually burnt savanna had significantly higher delta(15)N values compared to plants from less frequently burnt savanna, suggesting that foliar (15)N natural abundance could be used as marker for assessing historic fire regimes. Australian woodland and savanna species had low leaf delta(15)N and N content compared to species from equivalent African communities indicating that Australian biota are the more N depauperate. The largest differences in leaf delta(15)N occurred between the dominant ECM Australian and African savanna (miombo) species, which were depleted and enriched in (15)N, respectively. While the depleted delta(15)N of Australian ECM species are similar to those of previous reports on ECM species in natural plant communities, the (15)N-enriched delta(15)N of African ECM species represent an anomaly. PMID:12647130

  17. Highly 15N-Enriched Chondritic Clasts in the Isheyevo Meteorite

    SciTech Connect

    Bonal, L; Huss, G R; Krot, A N; Nagashima, K; Ishii, H A; Bradley, J P; Hutcheon, I D

    2009-01-14

    The metal-rich carbonaceous chondrites (CB and CH) have the highest whole-rock {sup 15}N enrichment ({delta}{sup 15}N up to +1500{per_thousand}), similar to {delta}{sup 15}N values reported in micron-sized regions (hotspots) of Interplanetary Dust Particles (IDPs) of possibly cometary origin and fine-grained matrices of unmetamorphosed chondrites. These {sup 15}N-rich hotspots are commonly attributed to low-temperature ion-molecule reactions in the protosolar molecular cloud or in the outer part of the protoplanetary disk. The nature of the whole-rock {sup 15}N enrichment of the metal-rich chondrites is not understood. We report a discovery of a unique type of primitive chondritic clasts in the CH/CB-like meteorite Isheyevo, which provides important constraints on the origin of {sup 15}N anomaly in metal-rich chondrites and nitrogen-isotope fractionation in the Solar System. These clasts contain tiny chondrules and refractory inclusions (5-15 {micro}m in size), and abundant ferromagnesian chondrule fragments (1-50 {micro}m in size) embedded in the partly hydrated, fine-grained matrix material composed of olivines, pyroxenes, poorly-organized aromatic organics, phyllosilicates and other hydrous phases. The mineralogy and oxygen isotope compositions of chondrules and refractory inclusions in the clasts are similar to those in the Isheyevo host, suggesting formation at similar heliocentric distances. In contrast to the previously known extraterrestrial samples, the fine-grained material in the clasts is highly and rather uniformly enriched in {sup 15}N, with bulk {delta}{sup 15}N values ranging between +1000 and +1300{per_thousand}; the {delta}{sup 15}N values in rare hotspots range from +1400 to +4000{per_thousand}. Since fine-grained matrices in the lithic clasts are the only component containing thermally unprocessed (during CAI and chondrule formation or during impact melting) materials that accreted into the metal rich chondrite parent body(ies), the {sup 15}N-enriched clasts as present in Isheyevo probably represent the major primordial carrier of {sup 15}N anomaly in these meteorites. The rather uniform and very high bulk {sup 15}N enrichment of the Isheyevo chondritic clasts, in contrast to rare hotspots of possibly molecular cloud origin in cometary IDPs, indicates that the nitrogen-isotope fractionation recorded by these clasts, containing abundant solar system materials, could have resulted from processes in the protoplanetary disk (e.g., ultraviolet photodissociation of {sup 15}N{sub 2} followed by trapping {sup 15}N atoms into NH-bearing ices) rather than solely by inheritance from the protosolar molecular cloud. If this is the case, the unique {sup 15}N-rich signature of the clasts could have resulted from nitrogen-isotope fractionation in cold and optically thin parts of the protoplanetary disk.

  18. CHANGE IN NATURAL ABUNDANCE OF 15N AND ESTIMATION OF N LOSSES FROM DAIRY MANURE DURING STORAGE BY MASS BALANCE AND NITROGEN-TO-PHOSPHORUS RATIO

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The main objective was to evaluate methodologies to estimate N losses from stored dairy manure. Manure with high N (HN) and low N (LN) content was obtained from two groups of cows assigned diets of 17 and 15% CP (DM), respectively. Manure collected from the barn floor was diluted with water to 10% ...

  19. Bodily variability of zinc natural isotope abundances in sheep.

    PubMed

    Balter, Vincent; Zazzo, Antoine; Moloney, Aidan P; Moynier, Fred; Schmidt, Olaf; Monahan, Frank J; Albarède, Francis

    2010-03-15

    Evidence is growing that the range of zinc stable isotope compositions, represented by the deviation of (66)Zn in permil units relative to a standard and expressed as delta(66)Zn, is larger in organic matter than in inorganic material. This study reports the variations of delta(66)Zn in various organs of sheep raised on a controlled diet. Zinc was purified by anion-exchange chromatography. The Zn concentrations and Zn stable isotope compositions were determined by quadrupole inductively coupled plasma mass spectrometry and multi-collector inductively coupled plasma mass spectrometry, respectively. The data show that delta(66)Zn variability exceeds 1 per thousand, with bone, muscle, serum and urine enriched in the heavy isotopes, and feces, red blood cells, kidney and liver enriched in light isotopes, all relative to the diet value. The (66)Zn enrichment of the circulating serum reservoir is likely to take place in the digestive tract, probably through the preferential binding of lighter isotopes with phytic acid, which is known to control the uptake of metallic elements. Mass balance calculations suggest that the (66)Zn depletion between diet and feces, which is not balanced by any other outward flux, leads to a secular isotopic drift in serum. A simple time-dependent two-box model, involving the gastro-intestinal tract on the one hand and the muscle and bone on the other, predicts that the maximum (66)Zn enrichment, which equals the difference in delta(66)Zn between diet and bulk (approximately 0.25 per thousand), is reached after about ten years. Therefore, a better understanding of the variations of natural abundance of Zn isotopes in animals and humans will probably bring new perspectives for the assessment of their Zn status. PMID:20155761

  20. Stable isotopes (?13C and ?15N) of organic matrix from coral skeleton

    PubMed Central

    Muscatine, Leonard; Goiran, Claire; Land, Lynton; Jaubert, Jean; Cuif, Jean-Pierre; Allemand, Denis

    2005-01-01

    The evolutionary success of reef-building corals in nutrient-poor tropical waters is attributed to endosymbiotic dinoflagellates. The algae release photosynthetic products to the coral animal cells, augment nutrient flux, and enhance the rate of coral calcification. Natural abundance of stable isotopes (?13C and ?18O) provides answers to modern and paleobiological questions about the effect of photosymbiosis on sources of carbon and oxygen in coral skeletal calcium carbonate. Here we compare 17 species of symbiotic and nonsymbiotic corals to determine whether evidence for photosymbiosis appears in stable isotopes (?13C and ?15N) of an organic skeletal compartment, the coral skeletal organic matrix (OM). Mean OM ?13C in symbiotic and nonsymbiotic corals was similar (-26.08 vs. -24.31), but mean OM ?15N was significantly depleted in 15N in the former (4.09) relative to the latter (12.28), indicating an effect of the algae on OM synthesis and revealing OM ?15N as a proxy for photosymbiosis. To answer an important paleobiological question about the origin of photosymbiosis in reef-building corals, we applied this proxy test to a fossil coral (Pachythecalis major) from the Triassic (240 million years ago) in which OM is preserved. Mean OM ?15N was 4.66, suggesting that P. major was photosymbiotic. The results show that symbiotic algae augment coral calcification by contributing to the synthesis of skeletal OM and that they may have done so as early as the Triassic. PMID:15671164

  1. Natural-abundance measurement of spin-spin couplings the nitrogen-15 in 1D and 2D NMR spectra by HEED pulse sequences

    NASA Astrophysics Data System (ADS)

    Kupče, Ērik; Wrackmeyer, Bernd

    Standard pulse sequences frequently employed in NMR studies, such as INEPT, DEPT, HETCOR, phase-sensitive HETCOR, and HETCOR with nongeminal proton decoupling in the F1 dimension, have been extended by Hahn spin echoes. This enables measurement of 15NX and long-range 15N 1H couplings (together with the comparison of their relative signs) at the natural-abundance level of isotopes. The sequences were optimized and verified for X  13C, 29Si, 31P, 119Sn, 207Pb, using a wide variety of nitrogen compounds (e.g., pyrroles, nitro compounds, cyanides, cyanates, isocyanates, isothiocyanates, carbodiimides, silylamines, PN, SiN, SnN, and PbN compounds). Both positive and negative 1J( 15N13C) couplings were observed. The trends were reproduced by SCF INDO FPT calculations. Reduced coupling constants 1K119 Sn15 N and 1 K207, 15Pb 15N were all negative. Two-and four-bond 15N 1H couplings were of either sign, whereas vicinal 3J( 15N1H) couplings were always negative, showing a crude linear relationship with V 1J( 15N13C) in the compounds studied. Since the intensity of the residual signal in the HEED experiments is readily adjustable, the measurement of {15}/{14}N isotope shifts, 1Δ {15}/{14}N(X) , is straightforward. The 1|gD {151}/{14}N( 13C) values determined for rather different bonding situations show a complex behavior and there is no simple relation between 1Δ values and bond order, s character, or hybridization. A previously proposed classification of {15}/{14} ( NC) isotope shifts is inadequate in the light of the present data. The 1Δ {15}/{14}N values for 3P(III) chemical shifts are much larger than those for P(V) derivatives. The {15}/{14} isotope effects on 13C and 29Si chemical shifts are similar in magnitude. Unexpectedly, several PbN compounds showed 1Δ {15}/{14}N( 207Pb) values close to zero.

  2. Carbon and nitrogen biogeochemistry in the ocean: A study using stable isotope natural abundance

    NASA Technical Reports Server (NTRS)

    Rau, G. H.; Desmarais, David J.

    1985-01-01

    Determining the biogeochemical pathways traveled by carbon and nitrogen in the ocean is fundamental to the understanding of how the ocean participates in the cycling of these elements within the biosphere. Because biological production, metabolism, and respiration can significantly alter the natural abundance of C-13 and N-15, these abundances can provide important information about the nature of these biological processes and their variability in the marine environment. The research initially seeks to characterize the spatial and temporal patterns of stable isotope abundances in organic matter, and to relate these abundances to C and N biogeochemical processes within selected areas of the northeastern Pacific Ocean.

  3. Experimental evidence for diel δ15N-patterns in different tissues, xylem and phloem saps of castor bean (Ricinus communis L.).

    PubMed

    Peuke, A D; Gessler, A; Tcherkez, G

    2013-12-01

    Nitrogen isotope signatures in plants might give insights in the metabolism and allocation of nitrogen. To obtain a deeper understanding of the modifications of the nitrogen isotope signatures, we determined δ(15)N in transport saps and in different fractions of leaves, axes and roots during a diel course along the plant axis. The most significant diel variations were observed in xylem and phloem saps where δ(15)N was significantly higher during the day compared with during the night. However in xylem saps, this was observed only in the canopy, but not at the hypocotyl positions. In the canopy, δ(15)N was correlated fairly well between phloem and xylem saps. These variations in δ(15)N in transport saps can be attributed to nitrate reduction in leaves during the photoperiod as well as to (15)N-enriched glutamine acting as transport form of N. δ(15)N of the water soluble fraction of roots and leaves partially affected δ(15)N of phloem and xylems saps. δ(15)N patterns are likely the result of a complex set of interactions and N-fluxes between plant organs. Furthermore, the natural nitrogen isotope abundance in plant tissue is not constant during the diel course - a fact that needs to be taken into account when sampling for isotopic studies. PMID:23663089

  4. Mycorrhizal Fungi Provide Most of the Nitrogen for Symbiotic Arctic Plants: 15N Evidence

    NASA Astrophysics Data System (ADS)

    Hobbie, J. E.; Hobbie, E. A.

    2004-12-01

    When soil nitrogen is in short supply, most terrestrial plants form symbioses with fungi (mycorrhizae) in which fine hyphal threads take up soil nitrogen, transport it into plant roots, and in return receive plant sugars. Because the transfer rates are very difficult to measure in nature, ecologists need new tools by which to assess the role of mycorrhizal fungi in carbon and nitrogen cycling. Recent studies indicate that the natural abundance of 15N taken up from the soil by hyphae is changed during transfer of nitrogen to roots; the result is large differences among the natural abundance of 15N in soil, symbiotic plants, and symbiotic fungi that depend on the mass balance of nitrogen in the mycorrhizal symbiosis. Measurements were carried out in acidic tussock tundra at the Toolik Lake LTER site in Arctic Alaska (68\\deg N 149\\deg W). The \\delta15N of soil N was 1.5%, of soil ammonium was 1.5%, of ericoid and ectomycorrhizal plants was -5.0%, and of ectomycorrhizal fungi was 7.0 parts per mille%. The mass balance of the 15N shows that the plants received 61-86% of their nitrogen from the fungal hyphae. These values, when combined with known plant growth rates, reveal that the plants provided 7-16% of their photosynthetic carbon to the fungi for growth and respiration, or about 25% of all carbon allocated to belowground processes. This analytical technique could be readily applied to other nitrogen-limited ecosystems such as many temperate and boreal forests to quantify the importance for terrestrial carbon and nitrogen cycling of mycorrhizally mediated transfers at the plant-soil interface.

  5. Natural Abundance 14C Content of Dibutyl Phthalate (DBP) from Three Marine Algae

    PubMed Central

    Namikoshi, Michio; Fujiwara, Takeshi; Nishikawa, Teruaki; Ukai, Kazuyo

    2006-01-01

    Analysis of the natural abundance 14C content of dibutyl phthalate (DBP) from two edible brown algae, Undaria pinnatifida and Laminaria japonica, and a green alga, Ulva sp., revealed that the DBP was naturally produced. The natural abundance 14C content of di-(2-ethylhexyl) phthalate (DEHP) obtained from the same algae was about 50–80% of the standard sample and the 14C content of the petrochemical (industrial) products of DBP and DEHP were below the detection limit.

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

    NASA Astrophysics Data System (ADS)

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

    2015-11-01

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

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

    PubMed

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

    2015-11-21

    The (15)N chemical shift tensor is shown to be extremely sensitive to lattice structure and a powerful metric for monitoring density functional theory refinements of crystal structures. These refinements include lattice effects and are applied here to five crystal structures. All structures improve based on a better agreement between experimental and calculated (15)N tensors, with an average improvement of 47.0 ppm. Structural improvement is further indicated by a decrease in forces on the atoms by 2-3 orders of magnitude and a greater similarity in atom positions to neutron diffraction structures. These refinements change bond lengths by more than the diffraction errors including adjustments to X-Y and X-H bonds (X, Y = C, N, and O) of 0.028 0.002 and 0.144 0.036 , respectively. The acquisition of (15)N tensors at natural abundance is challenging and this limitation is overcome by improved (1)H decoupling in the FIREMAT method. This decoupling dramatically narrows linewidths, improves signal-to-noise by up to 317%, and significantly improves the accuracy of measured tensors. A total of 39 tensors are measured with shifts distributed over a range of more than 400 ppm. Overall, experimental (15)N tensors are at least 5 times more sensitive to crystal structure than (13)C tensors due to nitrogen's greater polarizability and larger range of chemical shifts. PMID:26590548

  8. THE EFFECTS OF PARAMAGNETIC RELAXATION REAGENTS ON 15N SPIN RELAXATION AND THE USE OF GD(DPM)3 AS A NITROGEN-15 NUCLEAR MAGNETIC RESONANCE SPIN LABEL

    EPA Science Inventory

    Electron-nuclear relaxation times (T(1) sup e's) for (15)N and (13)C in natural abundance are measured for a series of amines of a wide range of pK(a)s using four paramagnetic relaxation reagents that are soluable in organic solutions. Cr(acac)3 and Cr(dpm)3 are seen to affect th...

  9. Vertical δ13C and δ15N changes during pedogenesis

    NASA Astrophysics Data System (ADS)

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

    2015-04-01

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

  10. Natural variability in abundance of prevalent soybean proteins.

    PubMed

    Natarajan, Savithiry S

    2010-12-01

    Soybean is an inexpensive source of protein for humans and animals. Genetic modifications (GMO) to soybean have become inevitable on two fronts, both quality and yield will need to improve to meet increasing global demand. To ensure the safety of the crop for consumers it is important to determine the natural variation in seed protein constituents as well as any unintended changes that may occur in the GMO as a result of genetic modification. Understanding the natural variation of seed proteins in wild and cultivated soybeans that have been used in conventional soybean breeding programs is critical for determining unintended protein expression in GMO soybeans. In recent years, proteomic technologies have been used as an effective analytical tool for examining modifications of protein profiles. We have standardized and applied these technologies to determine and quantify the spectrum of proteins present in soybean seed. We used two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and liquid chromatography mass spectrometry (LC-MS) for the separation, quantification, and identification of different classes of soybean seed proteins. We have observed significant variations in different classes of proteins, including storage, allergen and anti-nutritional protein profiles, between non-GMO cultivated and wild soybean varieties. This information is useful for scientists and regulatory agencies to determine whether the unintended expression of proteins found in transgenic soybean is within the range of natural variation. PMID:20709130

  11. Increasing plant use of organic nitrogen with elevation is reflected in nitrogen uptake rates and ecosystem delta15N.

    PubMed

    Averill, Colin; Finzi, Adrien

    2011-04-01

    It is hypothesized that decreasing mean annual temperature and rates of nitrogen (N) cycling causes plants to switch from inorganic to organic forms of N as the primary mode of N nutrition. To test this hypothesis, we conducted field experiments and collected natural-abundance delta15N signatures of foliage, soils, and ectomycorrhizal sporocarps along a steep elevation-climate gradient in the White Mountains, New Hampshire, USA. Here we show that with increasing elevation organic forms of N became the dominant source of N taken up by hardwood and coniferous tree species based on dual-labeled glycine uptake analysis, an important confirmation of an emerging theory for the biogeochemistry of the N cycle. Variation in natural abundance foliar delta15N with elevation was also consistent with increasing organic N uptake, though a simple, mass balance model demonstrated that the uptake of delta15N depleted inorganic N, rather than fractionation upon transfer of N from mycorrhizal fungi, best explains variations in foliar delta15N with elevation. PMID:21661551

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

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

    We used 15N2 gas to trace nitrogen (N) from biological N2-fixation to vascular plant uptake in an Alberta bog in order to determine if neighboring bog plants acquire recently fixed N from diazotrophs associating with Sphagnum mosses. Recent evidence indicates high rates of N2-fixation in Sphagnum mosses of Alberta bogs (Vile et al. 2013). Our previous work has shown that mosses can assimilate fixed N from associated diazotrophs as evidenced by the high N content of mosses despite minimal inputs from atmospheric deposition, retranslocation, and N mineralization. Therefore, the potential exists for vascular plants to obtain N from ';leaky' tissues of live mosses, however, this phenomenon has not been tested previously. Here we document the potential for relatively rapid transfer to vascular plants of N fixed by Sphagnum moss-associated diazotrophs. We utilized the novel approach of incubating mosses in 15N2 to allow the process of diazotrophic N2-fixation to mechanistically provide the 15N label, which is subsequently transferred to Sphagnum mosses. The potential for vascular bog natives to tap this N was assessed by planting the vascular plants in the labeled moss. Sphagnum mosses (upper 3 cm of live plants) were incubated in the presence of 98 atom % 15N2 gas for 48 hours. Two vascular plants common to Alberta bogs; Picea mariana and Vaccinium oxycoccus were then placed in the labeled mosses, where the mosses served as the substrate. Tissue samples from these plants were collected at three time points during the incubation; prior to 15N2 exposure (to determine natural abundance 15N), and at one and two months after 15N2 exposure. Roots and leaves were separated and run separately on a mass spectrometer to determine 15N concentrations. Sphagnum moss capitula obtained N from N2-fixation (δ15N of -2.43 × 0.40, 122.76 × 23.78, 224.92 × 68.37, 143.74 × 54.38 prior to, immediately after, and at 1 and 2 months after exposure to 15N2, respectively). Nitrogen was transferred to P. mariana roots (mean δ15N at 1 and 2 months of 15.26 × 3.30 and 16.19 × 1.21) more than shoots (mean δ15N at 1 and 2 months of 6.57 × 0.52 and 4.67×0.17) (initial δ15N values of roots and shoots of 2.16 × 0.37 and 5.54 × 0.35, respectively). Nitrogen also was transferred to V. oxycoccos roots (δ15N at 2 months of 21.46 × 3.61) more than shoots (δ15N 2 months of -2.17 × 0.23) (initial δ15N values of roots and shoots of -6.41 × 0.21 and -6.85 × 0.15, respectively). A two-way ANOVA and Tukey's HSD verified that both vascular plants' roots were significantly enriched with 15N (P. mariana roots; p < 0.0001, V. oxycoccus roots; p < 0.0001) after 1 month. These results indicate that bog vascular plants may derive considerable nitrogen from biological N2-fixation taking place in Sphagnum moss capitula. The experiment was subsequently repeated in-situ.

  13. PARTITIONING THE RELATIVE INFLUENCE OF SOIL N, MYCORRHIZAE, AND FOLIAR N UPTAKE ON FOLIAR δ15N PATTERNS: CAN WE DETECT FOLIAR UPTAKE OF REACTIVE N?

    NASA Astrophysics Data System (ADS)

    Vallano, D.; Sparks, J. P.

    2009-12-01

    Vegetation is an important sink for atmospheric reactive N in N-limited systems and may be capable of incorporating reactive N compounds directly into leaves through the foliar uptake pathway. A proxy for atmospheric reactive N entering vegetation would be useful to estimate the impact of direct foliar N uptake on plant metabolism. Natural abundance foliar N isotopic composition (δ15N) is a practical tool for this purpose because plant-available N sources often have different isotopic compositions. Current understanding of foliar δ15N suggests these values primarily represent the integration of soil δ15N, direct foliar N uptake, mycorrhizal fractionation, and within-plant fractionations. Using a potted plant mesocosm system, we estimated the influence of mycorrhizae on foliar δ15N patterns in red maple (Acer rubrum) seedlings along an N deposition gradient in New York State. We found that mycorrhizal associations altered foliar δ15N in red maple seedlings from 0.03 - 1.01‰ across sites. Along the same temporal and spatial scales, we examined the influence of soil δ15N, foliar N uptake, and mycorrhizae on foliar δ15N in adult stands of American beech (Fagus grandifolia), black birch (Betula lenta), red maple (A. rubrum), and red oak (Quercus rubra). Using multiple regression models, atmospheric NO2 concentration explained 0%, 69%, 23%, and 45% of the residual variation in foliar δ15N remaining in American beech, red maple, red oak, and black birch, respectively, after accounting for soil δ15N. Our results suggest that foliar δ15N may be used to estimate pollution-derived atmospheric reactive N entering vegetation via the foliar N uptake pathway.

  14. Protein retention assessment of four levels of poultry by-product substitution of fishmeal in rainbow trout (Oncorhynchus mykiss) diets using stable isotopes of nitrogen (δ15N) as natural tracers.

    PubMed

    Badillo, Daniel; Herzka, Sharon Z; Viana, Maria Teresa

    2014-01-01

    This is second part from an experiment where the nitrogen retention of poultry by-product meal (PBM) compared to fishmeal (FM) was evaluated using traditional indices. Here a quantitative method using stable isotope ratios of nitrogen (δ(15)N values) as natural tracers of nitrogen incorporation into fish biomass is assessed. Juvenile rainbow trout (Oncorhynchus mykiss) were fed for 80 days on isotopically distinct diets in which 0, 33, 66 and 100% of FM as main protein source was replaced by PBM. The diets were isonitrogenous, isolipidic and similar in gross energy content. Fish in all treatments reached isotopic equilibrium by the end of the experiment. Two-source isotope mixing models that incorporated the isotopic composition of FM and PBM as well as that of formulated feeds, empirically derived trophic discrimination factors and the isotopic composition of fish that had reached isotopic equilibrium to the diets were used to obtain a quantitative estimate of the retention of each source of nitrogen. Fish fed the diets with 33 and 66% replacement of FM by PBM retained poultry by-product meal roughly in proportion to its level of inclusion in the diets, whereas no differences were detected in the protein efficiency ratio. Coupled with the similar biomass gain of fishes fed the different diets, our results support the inclusion of PBM as replacement for fishmeal in aquaculture feeds. A re-feeding experiment in which all fish were fed a diet of 100% FM for 28 days indicated isotopic turnover occurred very fast, providing further support for the potential of isotopic ratios as tracers of the retention of specific protein sources into fish tissues. Stable isotope analysis is a useful tool for studies that seek to obtain quantitative estimates of the retention of different protein sources. PMID:25226392

  15. Protein Retention Assessment of Four Levels of Poultry By-Product Substitution of Fishmeal in Rainbow Trout (Oncorhynchus mykiss) Diets Using Stable Isotopes of Nitrogen (δ15N) as Natural Tracers

    PubMed Central

    Badillo, Daniel; Herzka, Sharon Z.; Viana, Maria Teresa

    2014-01-01

    This is second part from an experiment where the nitrogen retention of poultry by-product meal (PBM) compared to fishmeal (FM) was evaluated using traditional indices. Here a quantitative method using stable isotope ratios of nitrogen (δ15N values) as natural tracers of nitrogen incorporation into fish biomass is assessed. Juvenile rainbow trout (Oncorhynchus mykiss) were fed for 80 days on isotopically distinct diets in which 0, 33, 66 and 100% of FM as main protein source was replaced by PBM. The diets were isonitrogenous, isolipidic and similar in gross energy content. Fish in all treatments reached isotopic equilibrium by the end of the experiment. Two-source isotope mixing models that incorporated the isotopic composition of FM and PBM as well as that of formulated feeds, empirically derived trophic discrimination factors and the isotopic composition of fish that had reached isotopic equilibrium to the diets were used to obtain a quantitative estimate of the retention of each source of nitrogen. Fish fed the diets with 33 and 66% replacement of FM by PBM retained poultry by-product meal roughly in proportion to its level of inclusion in the diets, whereas no differences were detected in the protein efficiency ratio. Coupled with the similar biomass gain of fishes fed the different diets, our results support the inclusion of PBM as replacement for fishmeal in aquaculture feeds. A re-feeding experiment in which all fish were fed a diet of 100% FM for 28 days indicated isotopic turnover occurred very fast, providing further support for the potential of isotopic ratios as tracers of the retention of specific protein sources into fish tissues. Stable isotope analysis is a useful tool for studies that seek to obtain quantitative estimates of the retention of different protein sources. PMID:25226392

  16. Diversity, abundance and natural products of marine sponge-associated actinomycetes.

    PubMed

    Abdelmohsen, Usama Ramadan; Bayer, Kristina; Hentschel, Ute

    2014-03-01

    Actinomycetes are known for their unprecedented ability to produce novel lead compounds of clinical and pharmaceutical importance. This review focuses on the diversity, abundance and methodological approaches targeting marine sponge-associated actinomycetes. Additionally, novel qPCR data on actinomycete abundances in different sponge species and other environmental sources are presented. The natural products literature is covered, and we are here reporting on their chemical structures, their biological activities, as well as the source organisms from which they were isolated. PMID:24496105

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

    NASA Astrophysics Data System (ADS)

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

    2007-11-01

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

  18. 15N Hyperpolarization by Reversible Exchange Using SABRE-SHEATH

    PubMed Central

    2016-01-01

    NMR signal amplification by reversible exchange (SABRE) is a NMR hyperpolarization technique that enables nuclear spin polarization enhancement of molecules via concurrent chemical exchange of a target substrate and parahydrogen (the source of spin order) on an iridium catalyst. Recently, we demonstrated that conducting SABRE in microtesla fields provided by a magnetic shield enables up to 10% 15N-polarization (Theis, T.; et al. J. Am. Chem. Soc.2015, 137, 1404). Hyperpolarization on 15N (and heteronuclei in general) may be advantageous because of the long-lived nature of the hyperpolarization on 15N relative to the short-lived hyperpolarization of protons conventionally hyperpolarized by SABRE, in addition to wider chemical shift dispersion and absence of background signal. Here we show that these unprecedented polarization levels enable 15N magnetic resonance imaging. We also present a theoretical model for the hyperpolarization transfer to heteronuclei, and detail key parameters that should be optimized for efficient 15N-hyperpolarization. The effects of parahydrogen pressure, flow rate, sample temperature, catalyst-to-substrate ratio, relaxation time (T1), and reversible oxygen quenching are studied on a test system of 15N-pyridine in methanol-d4. Moreover, we demonstrate the first proof-of-principle 13C-hyperpolarization using this method. This simple hyperpolarization scheme only requires access to parahydrogen and a magnetic shield, and it provides large enough signal gains to enable one of the first 15N images (2 × 2 mm2 resolution). Importantly, this method enables hyperpolarization of molecular sites with NMR T1 relaxation times suitable for biomedical imaging and spectroscopy. PMID:25960823

  19. Natural Abundance 17O NMR Spectroscopy of Rat Brain In Vivo

    PubMed Central

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

    2008-01-01

    Oxygen is an abundant element that is present in almost all biologically relevant molecules. NMR observation of oxygen has been relatively limited since the NMR-active isotope, oxygen-17, is only present at a 0.037% natural abundance. Furthermore, as a spin 5/2 nucleus oxygen-17 has a moderately strong quadrupole moment which leads to fairly broad resonances (T2* = 1 - 4 ms). However, the similarly short T1 relaxation constants allow substantial signal averaging, whereas the large chemical shift range (> 300 ppm) improves the spectral resolution of 17O NMR. Here it is shown that high-quality, natural abundance 17O NMR spectra can be obtained from rat brain in vivo at 11.74 T. The chemical shifts and line widths of more than 20 oxygen-containing metabolites are established and the sensitivity and potential for 17O-enriched NMR studies are estimated. PMID:18456525

  20. Hyperpolarized NMR of plant and cancer cell extracts at natural abundance.

    PubMed

    Dumez, Jean-Nicolas; Milani, Jonas; Vuichoud, Basile; Bornet, Aurélien; Lalande-Martin, Julie; Tea, Illa; Yon, Maxime; Maucourt, Mickaël; Deborde, Catherine; Moing, Annick; Frydman, Lucio; Bodenhausen, Geoffrey; Jannin, Sami; Giraudeau, Patrick

    2015-09-01

    Natural abundance (13)C NMR spectra of biological extracts are recorded in a single scan provided that the samples are hyperpolarized by dissolution dynamic nuclear polarization combined with cross polarization. Heteronuclear 2D correlation spectra of hyperpolarized breast cancer cell extracts can also be obtained in a single scan. Hyperpolarized NMR of extracts opens many perspectives for metabolomics. PMID:26215673

  1. δ15N as a Potential Paleoenvironmental Proxy for Nitrogen Loading in Chesapeake Bay

    NASA Astrophysics Data System (ADS)

    Black, H. D.; Andrus, C. F.; Rick, T.; Hines, A.

    2013-12-01

    Stable isotope analysis of Eastern Oyster (Crassostrea virginica) and other mollusk shells from archaeological sites is a useful means of acquiring paleoenvironmental data. Recently, nitrogen isotopes have been identified as a potential new proxy in these shells. δ15N content in mollusk shells is affected by numerous anthropogenic and natural influences and may be used as an environmental proxy for nitrogen loading conditions. Chesapeake Bay is well known for both historic and modern pollution problems from numerous anthropogenic sources, such as fertilizer runoff, sewage discharge, and densely populated land use and serves as an ideal study location for long-term nitrogen loading processes. Longer records of these processes may be recorded in abundant archaeological remains around the bay, however, little is known about the stability of δ15N and %N in shell material over recent geologic time. In this study, 90 archaeological C. virginica shells were collected by the Smithsonian Institution from the Rhode River Estuary within Chesapeake Bay and range in age from ~150 to 3200 years old. Twenty-two modern C. virginica shells were also collected from nearby beds in the bay. All shell samples were subsampled from the resilifer region of the calcitic shell using a hand-held micro drill and were analyzed using EA-IRMS analysis to determine the potential temporal variability of δ15N and %N as well as creating a baseline for ancient nitrogen conditions in the bay area. Modern POM water samples and C. virginica soft tissues were also analyzed in this study to determine the degree of seasonal variation of δ15N and %N in Chesapeake Bay.

  2. Global modeling of the 15N216O line positions within the framework of the polyad model of effective Hamiltonian and a room temperature 15N216O line list

    NASA Astrophysics Data System (ADS)

    Tashkun, S. A.; Perevalov, V. I.; Liu, A.-W.; Hu, S.-M.

    2016-05-01

    The global modeling of 15N216O line positions in the 4-12,516 cm-1 region has been performed using the polyad model of effective Hamiltonian. The effective Hamiltonian parameters were fitted to the line positions collected from an exhaustive review of the literature. The dimensionless weighted standard deviation of the fit is 1.31. The fitted set of 109 parameters allowed reproducing more than 18,000 measured line positions with an RMS value of 0.001 cm-1. A line list was calculated for a reference temperature 296 K, natural abundance (1.32×10-5), and an intensity cutoff 10-30 cm/molecule. The line list is based on the fitted set of the effective Hamiltonian parameters for 15N216O obtained in this work and the effective dipole moment parameters of the 15N216O and 14N216O isotopologues. Accurate values of the 15N216O total partition function are also given.

  3. Structure, dynamics and mapping of membrane-binding residues of micelle-bound antimicrobial peptides by natural abundance (13)C NMR spectroscopy.

    PubMed

    Wang, Guangshun

    2010-02-01

    Worldwide bacterial resistance to traditional antibiotics has drawn much research attention to naturally occurring antimicrobial peptides (AMPs) owing to their potential as alternative antimicrobials. Structural studies of AMPs are essential for an in-depth understanding of their activity, mechanism of action, and in guiding peptide design. Two-dimensional solution proton NMR spectroscopy has been the major tool. In this article, we describe the applications of natural abundance (13)C NMR spectroscopy that provides complementary information to 2D (1)H NMR. The correlation of (13)Calpha secondary shifts with both 3D structure and heteronuclear (15)N NOE values indicates that natural abundance carbon chemical shifts are useful probes for backbone structure and dynamics of membrane peptides. Using human LL-37-derived peptides (GF-17, KR-12, and RI-10), as well as amphibian antimicrobial and anticancer peptide aurein 1.2 and its analog LLAA, as models, we show that the cross peak intensity plots of 2D (1)H-(13)Calpha HSQC spectra versus residue number present a wave-like pattern (HSQC wave) where key hydrophobic residues of micelle-bound peptides are located in the troughs with weaker intensities, probably due to fast exchange between the free and bound forms. In all the cases, the identification of aromatic phenylalanines as a key membrane-binding residue is consistent with previous intermolecular Phe-lipid NOE observations. Furthermore, mutation of one of the key hydrophobic residues of KR-12 to Ala significantly reduced the antibacterial activity of the peptide mutants. These results illustrate that natural abundance heteronuclear-correlated NMR spectroscopy can be utilized to probe backbone structure and dynamics, and perhaps to map key membrane-binding residues of peptides in complex with micelles. (1)H-(13)Calpha HSQC wave, along with other NMR waves such as dipolar wave and chemical shift wave, offers novel insights into peptide-membrane interactions from different angles. PMID:19682427

  4. Natural abundance 17O DNP two-dimensional and surface-enhanced NMR spectroscopy

    DOE PAGESBeta

    Perras, Frédéric A.; Kobayashi, Takeshi; Pruski, Marek

    2015-06-22

    Due to its extremely low natural abundance and quadrupolar nature, the 17O nuclide is very rarely used for spectroscopic investigation of solids by NMR without isotope enrichment. Additionally, the applicability of dynamic nuclear polarization (DNP), which leads to sensitivity enhancements of 2 orders of magnitude, to 17O is wrought with challenges due to the lack of spin diffusion and low polarization transfer efficiency from 1H. Here, we demonstrate new DNP-based measurements that extend 17O solid-state NMR beyond its current capabilities. The use of the PRESTO technique instead of conventional 1H–17O cross-polarization greatly improves the sensitivity and enables the facile measurementmore » of undistorted line shapes and two-dimensional 1H–17O HETCOR NMR spectra as well as accurate internuclear distance measurements at natural abundance. This was applied for distinguishing hydrogen-bonded and lone 17O sites on the surface of silica gel; the one-dimensional spectrum of which could not be used to extract such detail. As a result, this greatly enhanced sensitivity has enabled, for the first time, the detection of surface hydroxyl sites on mesoporous silica at natural abundance, thereby extending the concept of DNP surface-enhanced NMR spectroscopy to the 17O nuclide.« less

  5. Natural Genetic Variation Influences Protein Abundances in C. elegans Developmental Signalling Pathways.

    PubMed

    Singh, Kapil Dev; Roschitzki, Bernd; Snoek, L Basten; Grossmann, Jonas; Zheng, Xue; Elvin, Mark; Kamkina, Polina; Schrimpf, Sabine P; Poulin, Gino B; Kammenga, Jan E; Hengartner, Michael O

    2016-01-01

    Complex traits, including common disease-related traits, are affected by many different genes that function in multiple pathways and networks. The apoptosis, MAPK, Notch, and Wnt signalling pathways play important roles in development and disease progression. At the moment we have a poor understanding of how allelic variation affects gene expression in these pathways at the level of translation. Here we report the effect of natural genetic variation on transcript and protein abundance involved in developmental signalling pathways in Caenorhabditis elegans. We used selected reaction monitoring to analyse proteins from the abovementioned four pathways in a set of recombinant inbred lines (RILs) generated from the wild-type strains N2 (Bristol) and CB4856 (Hawaii) to enable quantitative trait locus (QTL) mapping. About half of the cases from the 44 genes tested showed a statistically significant change in protein abundance between various strains, most of these were however very weak (below 1.3-fold change). We detected a distant QTL on the left arm of chromosome II that affected protein abundance of the phosphatidylserine receptor protein PSR-1, and two separate QTLs that influenced embryonic and ionizing radiation-induced apoptosis on chromosome IV. Our results demonstrate that natural variation in C. elegans is sufficient to cause significant changes in signalling pathways both at the gene expression (transcript and protein abundance) and phenotypic levels. PMID:26985669

  6. Natural Genetic Variation Influences Protein Abundances in C. elegans Developmental Signalling Pathways

    PubMed Central

    Singh, Kapil Dev; Roschitzki, Bernd; Snoek, L. Basten; Grossmann, Jonas; Zheng, Xue; Elvin, Mark; Kamkina, Polina; Schrimpf, Sabine P.; Poulin, Gino B.; Kammenga, Jan E.; Hengartner, Michael O.

    2016-01-01

    Complex traits, including common disease-related traits, are affected by many different genes that function in multiple pathways and networks. The apoptosis, MAPK, Notch, and Wnt signalling pathways play important roles in development and disease progression. At the moment we have a poor understanding of how allelic variation affects gene expression in these pathways at the level of translation. Here we report the effect of natural genetic variation on transcript and protein abundance involved in developmental signalling pathways in Caenorhabditis elegans. We used selected reaction monitoring to analyse proteins from the abovementioned four pathways in a set of recombinant inbred lines (RILs) generated from the wild-type strains N2 (Bristol) and CB4856 (Hawaii) to enable quantitative trait locus (QTL) mapping. About half of the cases from the 44 genes tested showed a statistically significant change in protein abundance between various strains, most of these were however very weak (below 1.3-fold change). We detected a distant QTL on the left arm of chromosome II that affected protein abundance of the phosphatidylserine receptor protein PSR-1, and two separate QTLs that influenced embryonic and ionizing radiation-induced apoptosis on chromosome IV. Our results demonstrate that natural variation in C. elegans is sufficient to cause significant changes in signalling pathways both at the gene expression (transcript and protein abundance) and phenotypic levels. PMID:26985669

  7. Indirect Measurement of {sup 15}N(p,{alpha}){sup 12}C and {sup 18}O(p,{alpha}){sup 15}N. Applications to the AGB Star Nucleosynthesis

    SciTech Connect

    La Cognata, M.; Spitaleri, C.; Cherubini, S.; Crucilla, V.; Gulino, M.; Lamia, L.; Pizzone, R. G.; Puglia, S. M. R.; Rapisarda, G. G.; Romano, S.; Sergi, M. L.; Tumino, A.; Tribble, R.; Al-Abdullah, T.; Banu, A.; Fu, C.; Goldberg, V.; Mukhamedzhanov, A.; Tabacaru, G.; Trache, L.

    2008-04-06

    The Trojan Horse Method has been recently applied to the study of reactions involved in fluorine nucleosynthesis inside AGB stars. Fluorine abundance is important since it allows to constrain mixing models from the comparison of the observed fluorine abundances with the ones predicted by models. Anyway direct measurements of the cross section do not extend down to the Gamow peak, which is the astrophysically relevant energy region. In particular the study focuses on the {sup 15}N(p,{alpha}){sup 12}C and the {sup 18}O(p,{alpha}){sup 15}N reactions which can influence fluorine yield as they are part of {sup 19}F production/destruction network.

  8. High-resolution NMR of hydrogen in organic solids by DNP enhanced natural abundance deuterium spectroscopy

    NASA Astrophysics Data System (ADS)

    Rossini, Aaron J.; Schlagnitweit, Judith; Lesage, Anne; Emsley, Lyndon

    2015-10-01

    We demonstrate that high field (9.4 T) dynamic nuclear polarization (DNP) at cryogenic (∼100 K) sample temperatures enables the rapid acquisition of natural abundance 1H-2H cross-polarization magic angle spinning (CPMAS) solid-state NMR spectra of organic solids. Spectra were obtained by impregnating substrates with a solution of the stable DNP polarizing agent TEKPol in tetrachloroethane. Tetrachloroethane is a non-solvent for the solids, and the unmodified substrates are then polarized through spin diffusion. High quality natural abundance 2H CPMAS spectra of histidine hydrochloride monohydrate, glycylglycine and theophylline were acquired in less than 2 h, providing direct access to hydrogen chemical shifts and quadrupolar couplings. The spectral resolution of the 2H solid-state NMR spectra is comparable to that of 1H spectra obtained with state of the art homonuclear decoupling techniques.

  9. Increased natural mortality at low abundance can generate an Allee effect in a marine fish

    PubMed Central

    Kuparinen, Anna; Hutchings, Jeffrey A.

    2014-01-01

    Negative density-dependent regulation of population dynamics promotes population growth at low abundance and is therefore vital for recovery following depletion. Inversely, any process that reduces the compensatory density-dependence of population growth can negatively affect recovery. Here, we show that increased adult mortality at low abundance can reverse compensatory population dynamics into its opposite—a demographic Allee effect. Northwest Atlantic cod (Gadus morhua) stocks collapsed dramatically in the early 1990s and have since shown little sign of recovery. Many experienced dramatic increases in natural mortality, ostensibly attributable in some populations to increased predation by seals. Our findings show that increased natural mortality of a magnitude observed for overfished cod stocks has been more than sufficient to fundamentally alter the dynamics of density-dependent population regulation. The demographic Allee effect generated by these changes can slow down or even impede the recovery of depleted populations even in the absence of fishing. PMID:26064531

  10. Increased natural mortality at low abundance can generate an Allee effect in a marine fish.

    PubMed

    Kuparinen, Anna; Hutchings, Jeffrey A

    2014-10-01

    Negative density-dependent regulation of population dynamics promotes population growth at low abundance and is therefore vital for recovery following depletion. Inversely, any process that reduces the compensatory density-dependence of population growth can negatively affect recovery. Here, we show that increased adult mortality at low abundance can reverse compensatory population dynamics into its opposite-a demographic Allee effect. Northwest Atlantic cod (Gadus morhua) stocks collapsed dramatically in the early 1990s and have since shown little sign of recovery. Many experienced dramatic increases in natural mortality, ostensibly attributable in some populations to increased predation by seals. Our findings show that increased natural mortality of a magnitude observed for overfished cod stocks has been more than sufficient to fundamentally alter the dynamics of density-dependent population regulation. The demographic Allee effect generated by these changes can slow down or even impede the recovery of depleted populations even in the absence of fishing. PMID:26064531

  11. Determination of lithium isotopes at natural abundance levels by atomic absorption spectrometry

    USGS Publications Warehouse

    Meier, A.L.

    1982-01-01

    The relationships of the absorption of 6Li and 7Li hollow cathode lamp emissions are used to determine lithium isotopic composition in the natural abundance range of geologic materials. Absorption was found to have a nonlinear dependence upon total lithium concentration and isotopic composition. A method using nonlinear equations to describe the relationship of the absorption of 6Li and 7Li lamp radiation is proposed as a means of calculating isotopic composition that is independent of total lithium concentration.

  12. [Utilization of 15N-labeled urea in laying hens. 8. 15N-incorporation in the amino acids of the oviduct].

    PubMed

    Gruhn, K; Graf, H

    1987-02-01

    3 colostomized laying hybrids received orally with a conventional ration 1% urea with 96.06 atom-% 15N excess (15N'). over a period of 6 days. In the period of the experiment every hen consumed 2.87 g 15N'. After another 2 days, on which they received conventional feed urea, the animals were butchered. 15N' was determined in the total N and in 15 amino acids of the oviduct. Of the 15 amino acids the labelling of glutamic acid, glycine and serine was highest and on average amounted to 0.80, 0.66 and 0.67 atom-% 15N'. In lysine and arginine only 0.10 and 0.11 atom-% 15N' could be detected. The amino acid N with natural isotopic frequency amounted to a quarter for the basic amino acids, a tenth for the branched chain ones and for the non-essential ones (glutamic acid, aspartic acid, serine, glycine, alanine, proline) a third of the total oviduct 14N, The average quota of 15N' is only 3.6%, that of the branched chain amino acids 4.5 and that of the non-essential ones 21.1%. Consequently, the 15N' of the urea is mainly used for the synthesis of the non-essential amino acids of the oviduct. PMID:3689132

  13. Effect of Environmental Factors on Cyanobacterial Abundance and Cyanotoxins Production in Natural and Drinking Water, Bangladesh.

    PubMed

    Affan, Abu; Khomavis, Hisham S; Al-Harbi, Salim Marzoog; Haque, Mahfuzul; Khan, Saleha

    2015-02-01

    Cyanobacterial blooms commonly appear during the summer months in ponds, lakes and reservoirs in Bangladesh. In these areas, fish mortality, odorous water and fish and human skin irritation and eye inflammation have been reported. The influence of physicochemical factors on the occurrence of cyanobacteria and its toxin levels were evaluated in natural and drinking water in Bangladesh. A highly sensitive immunosorbent assay was used to detect microcystins (MCs). Cyanobacteria were found in 22 of 23 samples and the dominant species were Microcystis aeruginosa, followed by Microcystisflosaquae, Anabeana crassa and Aphanizomenon flosaquae. Cyanobacterial abundance varied from 39 to 1315 x 10(3) cells mL(-1) in natural water and 31 to 49 x 10(3) cells mL(-1) in tap water. MC concentrations were 25-82300 pg mL(-1) with the highest value measured in the fish research pond, followed by Ishakha Lake. In tap water, MC concentrations ranged from 30-32 pg mL(-1). The correlation between nitrate-nitrogen (NO3-N) concentration and cyanobacterial cell abundance was R2 = 0.62 while that between cyanobacterial abundance and MC concentration was R2 = 0.98. The increased NO3-N from fish feed, organic manure, poultry and dairy farm waste and fertilizer from agricultural land eutrophicated the water bodies and triggered cyanobacterial bloom formation. The increased amount of cyanobacteria produced MCs, subsequently reducing the water quality. PMID:26364354

  14. Understanding the Nature of Stellar Chemical Abundance Distributions in Nearby Stellar Systems

    NASA Astrophysics Data System (ADS)

    Lee, Duane

    2013-01-01

    Since stars retain, in large part, a signature of their galactic origin in their chemical compositions, we can exploit the chemical abundance distributions that we observe in stellar systems to put constraints on the nature of their progenitors. In this talk, I will discuss the results from two projects aimed at understanding how high resolution spectroscopic observations of nearby stellar systems might be interpreted. The first project presents one possible scenario for the origin of peculiar abundance distributions observed in ultra-faint dwarf satellites of the Milky Way. The second project explores to what extent the distribution of chemical elements in the stellar halo might be used to trace Galactic accretion history from its birth to the present day.

  15. Understanding the Nature of Stellar Chemical Abundance Distributions in Nearby Stellar Systems

    NASA Astrophysics Data System (ADS)

    Lee, Duane Morris

    Since stars retain signatures of their galactic origins in their chemical compositions, we can exploit the chemical abundance distributions that we observe in stellar systems to put constraints on the nature of their progenitors. In this thesis, I present results from three projects aimed at understanding how high resolution spectroscopic observations of nearby stellar systems might be interpreted. The first project presents one possible explanation for the origin of peculiar abundance distributions observed in ultra-faint dwarf satellites of the Milky Way. The second project explores to what extent the distribution of chemical elements in the stellar halo can be used to trace Galactic accretion history from the birth of the Galaxy to the present day. Finally, a third project focuses on developing an input optimization algorithm for the second project to produce better estimates of halo accretion histories. In conclusion, I propose some other new ways to use statistical models and techniques along with chemical abundance distribution data to uncover galactic histories.

  16. Natural abundances of carbon isotopes in acetate from a coastal marine sediment

    NASA Technical Reports Server (NTRS)

    Blair, N. E.; Martens, C. S.; Des Marais, D. J.

    1987-01-01

    Measurements of the natural abundances of carbon isotopes were made in acetate samples isolated from the anoxic marine sediment of Cape Lookout Bight, North Carolina. The typical value of the total acetate carbon isotope ratio (delta 13C) was -16.1 +/- 0.2 per mil. The methyl and carboxyl groups were determined to be -26.4 +/- 0.3 and -6.0 +/- 0.3 per mil, respectively, for one sample. The isotopic composition of the acetate is thought to have resulted from isotopic discriminations that occurred during the cycling of that molecule. Measurements of this type, which have not been made previously in the natural environment, may provide information about the dominant microbial pathways in anoxic sediments as well as the processes that influence the carbon isotopic composition of biogenic methane from many sources.

  17. Changes in protein abundance are observed in bacterial isolates from a natural host

    PubMed Central

    Rees, Megan A.; Stinear, Timothy P.; Goode, Robert J. A.; Coppel, Ross L.; Smith, Alexander I.; Kleifeld, Oded

    2015-01-01

    Bacterial proteomic studies frequently use strains cultured in synthetic liquid media over many generations. It is uncertain whether bacterial proteins expressed under these conditions will be the same as the repertoire found in natural environments, or when bacteria are infecting a host organism. Thus, genomic and proteomic characterization of bacteria derived from the host environment in comparison to reference strains grown in the lab, should aid understanding of pathogenesis. Isolates of Corynebacterium pseudotuberculosis were obtained from the lymph nodes of three naturally infected sheep and compared to a laboratory reference strain using bottom-up proteomics, after whole genome sequencing of each of the field isolates. These comparisons were performed following growth in liquid media that allowed us to reach the required protein amount for proteomic analysis. Over 1350 proteins were identified in the isolated strains, from which unique proteome features were revealed. Several of the identified proteins demonstrated a significant abundance difference in the field isolates compared to the reference strain even though there were no obvious differences in the DNA sequence of the corresponding gene or in nearby non-coding DNA. Higher abundance in the field isolates was observed for proteins related to hypoxia and nutrient deficiency responses as well as to thiopeptide biosynthesis. PMID:26528441

  18. Changes in protein abundance are observed in bacterial isolates from a natural host.

    PubMed

    Rees, Megan A; Stinear, Timothy P; Goode, Robert J A; Coppel, Ross L; Smith, Alexander I; Kleifeld, Oded

    2015-01-01

    Bacterial proteomic studies frequently use strains cultured in synthetic liquid media over many generations. It is uncertain whether bacterial proteins expressed under these conditions will be the same as the repertoire found in natural environments, or when bacteria are infecting a host organism. Thus, genomic and proteomic characterization of bacteria derived from the host environment in comparison to reference strains grown in the lab, should aid understanding of pathogenesis. Isolates of Corynebacterium pseudotuberculosis were obtained from the lymph nodes of three naturally infected sheep and compared to a laboratory reference strain using bottom-up proteomics, after whole genome sequencing of each of the field isolates. These comparisons were performed following growth in liquid media that allowed us to reach the required protein amount for proteomic analysis. Over 1350 proteins were identified in the isolated strains, from which unique proteome features were revealed. Several of the identified proteins demonstrated a significant abundance difference in the field isolates compared to the reference strain even though there were no obvious differences in the DNA sequence of the corresponding gene or in nearby non-coding DNA. Higher abundance in the field isolates was observed for proteins related to hypoxia and nutrient deficiency responses as well as to thiopeptide biosynthesis. PMID:26528441

  19. Abundance and phenology patterns of two pond-breeding salamanders determine species interactions in natural populations.

    PubMed

    Anderson, Thomas L; Hocking, Daniel J; Conner, Christopher A; Earl, Julia E; Harper, Elizabeth B; Osbourn, Michael S; Peterman, William E; Rittenhouse, Tracy A G; Semlitsch, Raymond D

    2015-03-01

    Phenology often determines the outcome of interspecific interactions, where early-arriving species often dominate interactions over those arriving later. The effects of phenology on species interactions are especially pronounced in aquatic systems, but the evidence is largely derived from experimental studies. We examined whether differences in breeding phenology between two pond-breeding salamanders (Ambystoma annulatum and A. maculatum) affected metamorph recruitment and demographic traits within natural populations, with the expectation that the fall-breeding A. annulatum would negatively affect the spring-breeding A. maculatum. We monitored populations of each species at five ponds over 4 years using drift fences. Metamorph abundance and survival of A. annulatum were affected by intra- and interspecific processes, whereas metamorph size and date of emigration were primarily influenced by intraspecific effects. Metamorph abundance, snout-vent length, date of emigration and survival for A. maculatum were all predicted by combinations of intra- and interspecific effects, but often showed negative relationships with A. annulatum metamorph traits and abundance. Size and date of metamorphosis were strongly correlated within each species, but in opposite patterns (negative for A. annulatum and positive for A. maculatum), suggesting that the two species use alternative strategies to enhance terrestrial survival and that these factors may influence their interactions. Our results match predictions from experimental studies that suggest recruitment is influenced by intra- and interspecific processes which are determined by phenological differences between species. Incorporating spatiotemporal variability when modeling population dynamics is necessary to understand the importance of phenology in species interactions, especially as shifts in phenology occur under climate change. PMID:25413866

  20. Effect of protein restriction on (15)N transfer from dietary [(15)N]alanine and [(15)N]Spirulina platensis into urea.

    PubMed

    Hamadeh, M J; Hoffer, L J

    2001-08-01

    Six normal men consumed a mixed test meal while adapted to high (1.5 g. kg(-1) x day(-1)) and low (0.3 g. kg(-1) x day(-1)) protein intakes. They completed this protocol twice: when the test meals included 3 mg/kg of [(15)N]alanine ([(15)N]Ala) and when they included 30 mg/kg of intrinsically labeled [(15)N]Spirulina platensis ([(15)N]SPI). Six subjects with insulin-dependent diabetes mellitus (IDDM) receiving conventional insulin therapy consumed the test meal with added [(15)N]Ala while adapted to their customary high-protein diet. Protein restriction increased serum alanine, glycine, glutamine, and methionine concentrations and reduced those of leucine. Whether the previous diet was high or low in protein, there was a similar increase in serum alanine, methionine, and branched-chain amino acid concentrations after the test meal and a similar pattern of (15)N enrichment in serum amino acids for a given tracer. When [(15)N]Ala was included in the test meal, (15)N appeared rapidly in serum alanine and glutamine, to a minor degree in leucine and isoleucine, and not at all in other circulating amino acids. With [(15)N]SPI, there was a slow appearance of the label in all serum amino acids analyzed. Despite the different serum amino acid labeling, protein restriction reduced the postmeal transfer of dietary (15)N in [(15)N]Ala or [(15)N]SPI into [(15)N]urea by similar amounts (38 and 43%, respectively, not significant). The response of the subjects with IDDM was similar to that of the normal subjects. Information about adaptive reductions in dietary amino acid catabolism obtained by adding [(15)N]Ala to a test meal appears to be equivalent to that obtained using an intrinsically labeled protein tracer. PMID:11440912

  1. Measuring denitrification after grassland renewal and grassland conversion to cropland by using the 15N gas-flux method

    NASA Astrophysics Data System (ADS)

    Buchen, Caroline; Eschenbach, Wolfram; Flessa, Heinz; Giesemann, Anette; Lewicka-Szczebak, Dominika; Well, Reinhard

    2015-04-01

    Denitrification, the reduction of oxidized forms of inorganic N to N2O and N2 is an important pathway of gaseous nitrogen losses. Measuring denitrification, especially the reduction of N2O to N2, expressed in the product ratio (N2O/(N2O + N2)), is rather difficult and hence rarely performed under field conditions. But using the 15N gas-flux method allows determining N transformation processes in their natural environment. In order to develop effective climate mitigation strategies understanding the N2O source is essential. We used the 15N gas-flux method to determine N2O and N2 emissions following grassland renewal and conversion techniques. Therefore we selected three different treatments: control (C), mechanical grassland renovation (GR) (autumn 2013) and grassland conversion to maize (GM) (spring 2014) from field plot trials on two different sites (Histic Gleysoil and Plaggic Anthrosol) near Oldenburg, Lower Saxony, Germany. We applied 15N labeled KNO3- (60 atom. % 15N) at a rate equivalent to common farming practices (150 kg N*ha-1) using needle injection of fertilizer solution in three different depths (10 cm, 15 cm, 20 cm) for homogeneous soil labeling up to 30 cm in microplots. During the first 10 days after application (May 2014) gas flux measurements from closed chambers were performed every second day and then weekly following a period of 8 weeks. Gas samples were analyzed for δ15N of N2 and N2O by IRMS according to Lewicka-Szczebak et al. (2013). Concentration and 15N enrichment of NO3- in soil water was determined on weekly samples using the SPIN-MAS technique (Stange et al. 2007). Fluxes of N2 and N2O evolved from the 15N labeled soil nitrogen pool were calculated using the equations of Spott et al. (2006). Peak events of N2 and N2O emissions occurred during the first 10 days of measurement, showing differences in soil types, as well as treatment variations. N2 fluxes up to 178 g*ha-1*day-1 and N2O fluxes up to 280 g*ha-1*day-1 were measured on the Plaggic Anthrosol in the GR treatment, while on the Histic Gleysoil, the GM treatment showed highest fluxes with N2 fluxes up to 1260 g*ha-1*day-1 and N2O fluxes up to 747 g*ha-1*day-1. Alike the product ratio of initial fluxes was higher on the Plaggic Anthrosol and lower on the Histic Gleysoil. Data analysis is still in progress and further results will be provided. References: Lewicka-Szczebak, D., R. Well, A. Giesemann, L. Rohe and U. Wolf (2013). "An enhanced technique for automated determination of 15N signatures of N2, (N2+N2O) and N2O in gas samples." Rapid Communications in Mass Spectrometry 27(13): 1548-1558. Spott, O., R. Russow, B. Apelt and C. F. Stange (2006). "A 15N-aided artificial atmosphere gas flow technique for online determination of soil N2 release using the zeolite Köstrolith SX6®." Rapid Communications in Mass Spectrometry 20(22): 3267-3274. Stange, F., O. Spott, B. Apelt and R. W. Russow (2007). "Automated and rapid online determination of 15N abundance and concentration of ammonium, nitrite, or nitrate in aqueous samples by the SPINMAS technique." Isotopes in Environmental and Health Studies 43(3): 227-236.

  2. Natural abundance variations in stable isotopes and their potential uses in animal physiological ecology.

    PubMed

    Gannes, L Z; Martínez del Rio, C; Koch, P

    1998-03-01

    Chemical, biological, and physical processes lead to distinctive "isotopic signatures" in biological materials that allow tracing of the origins of organic substances. Isotopic variation has been extensively used by plant physiological ecologists and by paleontologists, and recently ecologists have adopted the use of stable isotopes to measure ecosystem patterns and processes. To date, animal physiological ecologists have made minimal use of naturally occurring stable isotopes as tracers. Here we provide a review of the current and potential uses of naturally occurring stable isotopes in animal physiological ecology. We outline the physical and biological processes that lead to variation in isotopic abundance in plants and animals. We summarize current uses in animal physiological ecology (diet reconstruction and animal movement patterns), and suggest areas of research where the use of stable isotopes can be fruitful (protein balance and turnover and the allocation of dietary nutrients). We argue that animal physiological ecologists can benefit from including the measurement of naturally occurring stable isotopes in their battery of techniques. We also argue that animal physiologists can make an important contribution to the emerging field of stable isotopes in biology by testing experimentally the plethora of assumptions upon which the techniques rely. PMID:9683412

  3. Investigating microbial carbon cycling using natural abundance isotope analysis of PLFA

    NASA Astrophysics Data System (ADS)

    Slater, G. G.; Brady, A.; Cowie, B.

    2008-12-01

    Understanding microbial carbon sources and cycling is fundamental to our conceptualization of microbial ecosystems and their role in biogeochemical cycling in natural systems. Achieving this understanding requires application of a wide range of approaches. Natural abundance isotope analysis of individual compounds, particularly cellular components such as Phospholipids Fatty Acids (PLFA) can provide insights into the carbon sources and metabolic activities of the in situ microbial community from environmental samples. This is primarily because specific PLFA can be well resolved by gas chromatography even from complex matrices where confounding biological/organic compound abound. These PLFA can then be attributed to the viable microbial community, in some cases to specific components of this community and due to characteristic biosynthetic fractionations of stable isotope ratios, δ13C analysis of PLFA can: differentiate isotopically distinct primary carbon sources of heterotrophic communities; identify isotopic patterns characteristic of autotrophic versus heterotrophic processes; and elucidate microbial biosynthetic pathways. In cases where there δ13C cannot provide resolution of carbon sources, new approaches in Δ14C of PLFA can be applied. The vast range in Δ14C of ancient and modern carbon provides an easily traceable signal that can differentiate uptake and utilization of these carbon sources. This is particularly useful in cases such as contaminated sites where petroleum based contamination has occurred, or in natural systems where microbial communities may be utilizing geologic versus recently photosynthetically fixed carbon. This talk will present several examples demonstrating the utility of this approach.

  4. Natural abundant (17) O NMR in a 1.5-T Halbach magnet.

    PubMed

    Sørensen, Morten K; Bakharev, Oleg N; Jensen, Ole; Nielsen, Niels Chr

    2016-06-01

    We present mobile, low-field (17) O NMR as a means for monitoring oxygen in liquids. Whereas oxygen is one of the most important elements, oxygen NMR is limited by a poor sensitivity related to low natural abundance and gyro-magnetic ratio of the NMR active (17) O isotope. Here, we demonstrate (17) O NMR detection at a Larmor frequency of 8.74 MHz in a 1.5-T Halbach neodymium magnet with a home-built digital NMR instrument suitable for large-scale production and in-line monitoring applications. The proposed (17) O NMR sensor may be applied for direct, noninvasive measurements of water content in, for example, oil, manure, or food in automated quality or process control. Copyright © 2015 John Wiley & Sons, Ltd. PMID:25641664

  5. In Vivo Natural-Abundance 13C Nuclear Magnetic Resonance Studies of Living Ectomycorrhizal Fungi 1

    PubMed Central

    Martin, Francis; Canet, Daniel; Marchal, Jean-Pierre; Brondeau, Jean

    1984-01-01

    Natural-abundance 13C nuclear magnetic resonance spectroscopy has been used to study intact mycelia of the ectomycorrhizal fungi Cenococcum graniforme (Ascomycetes) and Hebeloma crustuliniforme (Basidiomycetes). A number of sharp resonances are observed in living fungi. These signals primarily arise from fatty acyl chains and carbohydrate nuclei. The spectra are interpreted in terms of relative concentrations of the major fatty acids present in the fungal triglycerides. The small line width of fatty acids (mainly oleic, linoleic, and palmitic acids) resonances and spin-lattice relaxation time are indicative of fast rotational reorientations and are consequently thought to arise from fatty acyl chains in fat droplets. We were able to locate the site of lipids accumulation within mycelia using light microscopy and histological staining. Many lipid droplets were observed in mycelia of both species. These results suggest that fatty acids droplets could be involved in carbon storage and metabolism from ectomycorrhizal fungi. PMID:16663561

  6. Natural abundance carbon 13 nuclear magnetic resonance spectroscopy of antifreeze glycoproteins

    SciTech Connect

    Berman, E.; Allerhand, A.; DeVries, A.L.

    1980-05-25

    Natural abundance /sup 13/C NMR spectroscopy (at 67.9 MHz) is used to study an aqueous solution of the antifreeze glycoproteins 3 to 6 from the Antarctic cod. Assignments of resonances to specific carbons of the known Ala-Ala-(..beta..Gal-(1 ..-->.. 3)-..cap alpha..-N-acetylgalactosamine)Thr repeating unit are presented. Some of the carbons of the proline residues of glycoprotein 6 are also identified. Spin-lattice relaxation times (at 30/sup 0/C), linewidths (at 35/sup 0/C), nuclear Overhauser enhancements (at 35/sup 0/C), and the temperature dependence of chemical shifts (from 0/sup 0/ to 75/sup 0/C) all strongly suggest that these antifreeze glycoproteins are predominantly or entirely flexible random coil polymers.

  7. Solid-State 87Sr NMR Spectroscopy at Natural Abundance and High Magnetic Field Strength.

    PubMed

    Faucher, Alexandra; Terskikh, Victor V; Ye, Eric; Bernard, Guy M; Wasylishen, Roderick E

    2015-12-10

    Twenty-five strontium-containing solids were characterized via (87)Sr NMR spectroscopy at natural abundance and high magnetic field strength (B0 = 21.14 T). Strontium nuclear quadrupole coupling constants in these compounds are sensitive to the strontium site symmetry and range from 0 to 50.5 MHz. An experimental (87)Sr chemical shift scale is proposed, and available data indicate a chemical shift range of approximately 550 ppm, from -200 to +350 ppm relative to Sr(2+)(aq). In general, magnetic shielding increased with strontium coordination number. Experimentally measured chemical shift anisotropy is reported for stationary samples of solid powdered SrCl2·6H2O, SrBr2·6H2O, and SrCO3, with δaniso((87)Sr) values of +28, +26, and -65 ppm, respectively. NMR parameters were calculated using CASTEP, a gauge including projector augmented wave (GIPAW) DFT-based program, which addresses the periodic nature of solids using plane-wave basis sets. Calculated NMR parameters are in good agreement with those measured. PMID:26565918

  8. Assessing microbial uptake of petroleum hydrocarbons in groundwater systems using natural abundance radiocarbon.

    PubMed

    Ahad, Jason M E; Burns, Leanne; Mancini, Silvia; Slater, Greg F

    2010-07-01

    Carbon sources utilized by the active microbial communities in shallow groundwater systems underlying three petroleum service stations were characterized using natural abundance radiocarbon ((14)C). Total organic carbon (TOC) Delta(14)C values ranged from -314 to -972 per thousand and petroleum-extracted residues (EXT-RES) ranged from -293 to -971 per thousand. Phospholipid fatty acids (PLFAs)-biomarkers for active microbial populations-ranged from -405 to -885 per thousand and a comparison of these values with potential carbon sources pointed to significant microbial assimilation of (14)C-free fossil carbon. The most (14)C-depleted PLFAs were found in the samples with the highest concentrations of total petroleum hydrocarbons (TPHs). A radiocarbon mass balance indicated up to 43% of the carbon in microbial PLFAs was derived from TPHs, providing direct evidence for biodegradation at two of three sites. At lower levels of TPHs Delta(14)C values of PLFAs were generally similar to or more enriched than all other carbon in the system indicating microbial utilization of a more (14)C-enriched carbon source and no resolvable evidence for microbial incorporation of petroleum-derived carbon. Results from this study suggest that it is possible to delineate petroleum biodegradation in groundwater systems using these techniques even in complex situations where there exists a wide range in the ages of natural organic matter (i.e., EXT-RES). PMID:20527914

  9. Using natural abundance of δ13C to partition ecosystem soil respiration

    NASA Astrophysics Data System (ADS)

    Hunt, J.; Millard, P.; Midwood, A. J.; Whitehead, D.

    2010-12-01

    Soil respiration constitutes a major component of the global carbon and is the single most important flux of CO2 from the terrestrial ecosystem. Recently assimilated (autotrophic) carbon and CO2 respired during microbial decomposition of older SOM (heterotrophic carbon) may differ in their response to environmental conditions. We make use of a natural stable carbon isotope differences (typically 2-4 ‰) between the CO2 from autotrophic respiration of roots and heterotrophic respiration of soil organic matter to partition total soil respiration in situ. We used a new, open steady-state chamber technique to measure the respiration rate and collect samples of soil surface CO2 efflux with minimal disturbance to the diffusion of 12CO2 and 13CO2 from the soil surface. Gas samples were collected in Tedlar bags and analysed with a tunable diode laser. We installed rings and at each ring the roots, litter and root-free soil were incubated in bags to determine the δ13C of the end members. During incubation δ13C became more depleted with time (5.2 ‰ after 8 h), so the initial value was calculated from a time course. The total flux was partitioned into two major sources. Two examples are presented of partitioning soil respiration in young forests using natural abundance δ13C discrimination. In a 23 year-old Kanuka forest, SOM was typically 5.5 ‰ enriched compared with the roots (-27.5 ‰). All 50 rings were successfully partitioned, resulting in a mean of 22 % of the soil surface respiration originated from SOM. In the young 4-year old Pinus radiata forest, SOM was again enriched (3.6 ‰) compared to the roots (-25.8‰). Some 42 % of the soil respiration was from SOM turnover. We have shown the possibility of using natural abundance δ13C discrimination to quantify the proportion of total soil respired CO2 originating from SOM turnover in young C3 forests.

  10. FOLIAR NITROGEN CONCENTRATIONS AND NATURAL ABUNDANCE OF 15N SUGGEST NITROGEN ALLOCATION PATTERNS OF DOUGLAS-FIR AND MYCORRHIZAL FUNGI DURING DEVELOPMENT IN ELEVATED CARBON DIOXIDE CONCENTRATION AND TEMPERATURE

    EPA Science Inventory

    In an experiment using Pseudotsuga menziesii (Mirb.) Franco (Douglas-fir) seedlings and a 2x2 factorial design in enclosed mesocosms, temperatures were maintained at ambient or +3.5 degrees C above ambient, and CO2 levels were maintained at ambient or 179 ppm above ambient. Two ...

  11. Archaeal community diversity and abundance changes along a natural salinity gradient in estuarine sediments

    PubMed Central

    Webster, Gordon; O'Sullivan, Louise A.; Meng, Yiyu; Williams, Angharad S.; Sass, Andrea M.; Watkins, Andrew J.; Parkes, R. John; Weightman, Andrew J.

    2014-01-01

    Archaea are widespread in marine sediments, but their occurrence and relationship with natural salinity gradients in estuarine sediments is not well understood. This study investigated the abundance and diversity of Archaea in sediments at three sites [Brightlingsea (BR), Alresford (AR) and Hythe (HY)] along the Colne Estuary, using quantitative real-time PCR (qPCR) of 16S rRNA genes, DNA hybridization, Archaea 16S rRNA and mcrA gene phylogenetic analyses. Total archaeal 16S rRNA abundance in sediments were higher in the low-salinity brackish sediments from HY (2–8 × 107 16S rRNA gene copies cm−3) than the high-salinity marine sites from BR and AR (2 × 104–2 × 107 and 4 × 106–2 × 107 16S rRNA gene copies cm−3, respectively), although as a proportion of the total prokaryotes Archaea were higher at BR than at AR or HY. Phylogenetic analysis showed that members of the ‘Bathyarchaeota’ (MCG), Thaumarchaeota and methanogenic Euryarchaeota were the dominant groups of Archaea. The composition of Thaumarchaeota varied with salinity, as only ‘marine’ group I.1a was present in marine sediments (BR). Methanogen 16S rRNA genes from low-salinity sediments at HY were dominated by acetotrophic Methanosaeta and putatively hydrogentrophic Methanomicrobiales, whereas the marine site (BR) was dominated by mcrA genes belonging to methylotrophic Methanococcoides, versatile Methanosarcina and methanotrophic ANME-2a. Overall, the results indicate that salinity and associated factors play a role in controlling diversity and distribution of Archaea in estuarine sediments. PMID:25764553

  12. Archaeal community diversity and abundance changes along a natural salinity gradient in estuarine sediments.

    PubMed

    Webster, Gordon; O'Sullivan, Louise A; Meng, Yiyu; Williams, Angharad S; Sass, Andrea M; Watkins, Andrew J; Parkes, R John; Weightman, Andrew J

    2015-02-01

    Archaea are widespread in marine sediments, but their occurrence and relationship with natural salinity gradients in estuarine sediments is not well understood. This study investigated the abundance and diversity of Archaea in sediments at three sites [Brightlingsea (BR), Alresford (AR) and Hythe (HY)] along the Colne Estuary, using quantitative real-time PCR (qPCR) of 16S rRNA genes, DNA hybridization, Archaea 16S rRNA and mcrA gene phylogenetic analyses. Total archaeal 16S rRNA abundance in sediments were higher in the low-salinity brackish sediments from HY (2-8 × 10(7) 16S rRNA gene copies cm(-3)) than the high-salinity marine sites from BR and AR (2 × 10(4)-2 × 10(7) and 4 × 10(6)-2 × 10(7) 16S rRNA gene copies cm(-3), respectively), although as a proportion of the total prokaryotes Archaea were higher at BR than at AR or HY. Phylogenetic analysis showed that members of the 'Bathyarchaeota' (MCG), Thaumarchaeota and methanogenic Euryarchaeota were the dominant groups of Archaea. The composition of Thaumarchaeota varied with salinity, as only 'marine' group I.1a was present in marine sediments (BR). Methanogen 16S rRNA genes from low-salinity sediments at HY were dominated by acetotrophic Methanosaeta and putatively hydrogentrophic Methanomicrobiales, whereas the marine site (BR) was dominated by mcrA genes belonging to methylotrophic Methanococcoides, versatile Methanosarcina and methanotrophic ANME-2a. Overall, the results indicate that salinity and associated factors play a role in controlling diversity and distribution of Archaea in estuarine sediments. PMID:25764553

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

    NASA Astrophysics Data System (ADS)

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

    2011-09-01

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

  14. A novel method for determination of the (15) N isotopic composition of Rubisco in wheat plants exposed to elevated atmospheric carbon dioxide.

    PubMed

    Aranjuelo, Iker; Molero, Gemma; Avice, Jean Christophe; Bourguignon, Jacques

    2015-02-01

    Although ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is mostly known as a key enzyme involved in CO2 assimilation during the Calvin cycle, comparatively little is known about its role as a pool of nitrogen storage in leaves. For this purpose, we developed a protocol to purify Rubisco that enables later analysis of its (15) N isotope composition (δ(15) N) at the natural abundance and (15) N-labeled plants. In order to test the utility of this protocol, durum wheat (Triticum durum var. Sula) exposed to an elevated CO2 concentration (700 vs 400 µmol mol(-1) ) was labeled with K(15) NO3 (enriched at 2 atom %) during the ear development period. The developed protocol proves to be selective, simple, cost effective and reproducible. The study reveals that (15) N labeling was different in total organic matter, total soluble protein and the Rubisco fraction. The obtained data suggest that photosynthetic acclimation in wheat is caused by Rubisco depletion. This depletion may be linked to preferential nitrogen remobilization from Rubisco toward grain filling. PMID:25272325

  15. Nitrogen Isotopic Ratios in Cometary NH2: Implication for 15N-fractionation in Ammonia

    NASA Astrophysics Data System (ADS)

    Shinnaka, Yoshiharu; Kawakita, Hideyo; Jehin, Emmanuël; Decock, Alice; Hutsemékers, Damien; Manfroid, Jean; Arai, Akira

    2015-11-01

    Isotopic ratios in cometary molecules are diagnostic for the physico-chemical conditions where molecules formed and are processed, from the interstellar medium to the solar nebula. Usually temperatures at the molecular formation control the fractionation of the heavier element in molecular species, e.g., D-fractionation in water.In cometary volatiles, the 14N/15N ratios in CN have been well observed (Manfroid et al. 2009, A&A, 503, 613, and reference therein) and is consistent with the ratio in HCN (a most probable parent of CN) measured in few comets (Bockelée-Morvan et al. 2008, ApJ, 679, L49). Those ratios are enriched compared to the proto-solar value by a factor of ~3. In contrast to those Nitriles, there are only few reports on 14N/15N ratios in Ammonia (as Amine) (Rousselot et al. 2014, ApJ, 780, L17; Shinnaka et al. 2014, ApJ, 782, L16). Ammonia (NH3) is usually the most abundant and HCN is the second most abundant N-bearing volatiles in cometary ice. Especially, recent observations of 15NH2 revealed the 14N/15N ratios in NH3 are comparable to those of CN. However, from the viewpoint of theoretical work, the enrichment of 15N in cometary NH3 cannot be reproduced by current chemical network models. Information about the diversity of the 14N/15N ratios in NH3 of individual comets is needed to understand the formation mechanisms/environments of NH3 in the early solar system.To clarify the diversity of the 14N/15N ratios in cometary NH3, we determine the 14N/15N ratios in NH3 for more than ten comets individually which include not only Oort cloud comets but also short period comets by using the high-resolution optical spectra of NH2. These spectra were obtained with both the UVES mounted on the VLT in Chile and the HDS on the Subaru Telescope in Hawaii.The derived 14N/15N ratios in NH3 for more than ten comets show high 15N-enrichment compared with the elemental abundances of nitrogen in the Sun by about factor of ~3 and has no large diversity depending on these dynamical properties. We discuss about the origin of the formation conditions of cometary NH3 and its physico-chemical evolution in the solar nebula based on our and other results.This work was supported by JSPS, 15J10864 (Y. Shinnaka).

  16. A 115-year δ15N record of cumulative nitrogen pollution in California serpentine grasslands

    NASA Astrophysics Data System (ADS)

    Vallano, D.; Zavaleta, E. S.

    2010-12-01

    Until the 1980s, California’s biodiverse serpentine grasslands were threatened primarily by development and protected by reserve creation. However, nitrogen (N) fertilization due to increasing fossil fuel emissions in the expanding Bay Area is thought to be contributing to rapid, recent invasion of these ecosystems by exotic annual grasses that are displacing rare and endemic serpentine species. Documenting the cumulative effects of N deposition in this ecosystem can direct policy and management actions to mitigate the role of N deposition in its transformation. Natural abundance stable isotopes of N in vegetation have been increasingly used as bio-indicators of N deposition patterns and subsequent changes to plant N cycling and assimilation. However, the long-term record of atmospheric reactive N enrichment and the resulting changes in ecosystem N dynamics have yet to be adequately reconstructed in many ecosystems. Museum archives of vascular plant tissue are valuable sources of materials to reconstruct temporal and spatial isotopic patterns of N inputs to ecosystems. Here, we present N stable isotope data from archived and current specimens of an endemic California serpentine grassland species, leather oak (Quercus durata), since 1895 across the greater San Francisco Bay region. We measured spatial and temporal trends in stable isotope composition (δ15N and δ13C) and concentration (%N and %C) of historical and current samples of leather oak leaves from sites within the Bay Area, impacted by increasing development, and sites northeast of the Bay Area, with significantly lower rates of urbanization and industrialization. Specifically, we sampled dry museum and fresh leaf specimens from serpentine sites within Lake (n=27) and Santa Clara (n=30) counties dating from 1895 to 2010. Leaf δ15N values were stable from 1895 to the 1950s and then decreased strongly throughout the last 50 years as fossil fuel emissions rapidly increased in the Bay Area, indicating that N pollution is being retained in serpentine grassland ecosystems. Leaf δ15N values in the high-deposition region declined at a rate of -0.041‰ yr-1, while leaf δ15N values in the low-deposition region did not show a strong pattern. In both regions, leaf δ13C values declined through time as atmospheric CO2 concentrations increased in response to fossil fuel combustion (the Suess effect). Leaf %N and %C values did not present any clear patterns at sites within or outside of the Bay Area. We conclude that using natural abundance stable isotope values in leaves can indicate variation in N pollution inputs across wide spatial and temporal scales and that archived plant samples can provide valuable baselines against which to assess changes in regional N cycling and subsequent ecological impacts on vegetation.

  17. Characterization of Stratum Corneum Molecular Dynamics by Natural-Abundance 13C Solid-State NMR

    PubMed Central

    Bouwstra, Joke A.; Sparr, Emma; Topgaard, Daniel

    2013-01-01

    Despite the enormous potential for pharmaceutical applications, there is still a lack of understanding of the molecular details that can contribute to increased permeability of the stratum corneum (SC). To investigate the influence of hydration and heating on the SC, we record the natural-abundance 13C signal of SC using polarization transfer solid-state NMR methods. Resonance lines from all major SC components are assigned. Comparison of the signal intensities obtained with the INEPT and CP pulse sequences gives information on the molecular dynamics of SC components. The majority of the lipids are rigid at 32°C, and those lipids co-exist with a small pool of mobile lipids. The ratio between mobile and rigid lipids increases with hydration. An abrupt change of keratin filament dynamics occurs at RH = 80–85%, from completely rigid to a structure with rigid backbone and mobile protruding terminals. Heating has a strong effect on the lipid mobility, but only a weak influence on the keratin filaments. The results provide novel molecular insight into how the SC constituents are affected by hydration and heating, and improve the understanding of enhanced SC permeability, which is associated with elevated temperatures and SC hydration. PMID:23626744

  18. Probing surface hydrogen bonding and dynamics by natural abundance, multidimensional, 17O DNP-NMR spectroscopy

    DOE PAGESBeta

    Perras, Frederic A.; Chaudhary, Umesh; Slowing, Igor I.; Pruski, Marek

    2016-05-06

    Dynamic nuclear polarization (DNP)-enhanced solid-state nuclear magnetic resonance (SSNMR) spectroscopy is increasingly being used as a tool for the atomic-level characterization of surface sites. DNP surface-enhanced SSNMR spectroscopy of materials has, however, been limited to studying relatively receptive nuclei, and the particularly rare 17O nuclide, which is of great interest for materials science, has not been utilized. We demonstrate that advanced 17O SSNMR experiments can be performed on surface species at natural isotopic abundance using DNP. We use 17O DNP surface-enhanced 2D SSNMR to measure 17O{1H} HETCOR spectra as well as dipolar oscillations on a series of thermally treatedmore » mesoporous silica nanoparticle samples having different pore diameters. These experiments allow for a nonintrusive and unambiguous characterization of hydrogen bonding and dynamics at the surface of the material; no other single experiment can give such details about the interactions at the surface. Lastly, our data show that, upon drying, strongly hydrogen-bonded surface silanols, whose motions are greatly restricted by the interaction when compared to lone silanols, are selectively dehydroxylated.« less

  19. Natural Abundance 17O Nuclear Magnetic Resonance and Computational Modeling Studies of Lithium Based Liquid Electrolytes

    SciTech Connect

    Deng, Xuchu; Hu, Mary Y.; Wei, Xiaoliang; Wang, Wei; Chen, Zhong; Liu, Jun; Hu, Jian Z.

    2015-07-01

    Natural abundance 17O NMR measurements were conducted on electrolyte solutions consisting of Li[CF3SO2NSO2CF3] (LiTFSI) dissolved in the solvents of ethylene carbonate (EC), propylene carbonate (PC), ethyl methyl carbonate (EMC), and their mixtures at various concentrations. It was observed that 17O chemical shifts of solvent molecules change with the concentration of LiTFSI. The chemical shift displacements of carbonyl oxygen are evidently greater than those of ethereal oxygen, strongly indicating that Li+ ion is coordinated with carbonyl oxygen rather than ethereal oxygen. To understand the detailed molecular interaction, computational modeling of 17O chemical shifts was carried out on proposed solvation structures. By comparing the predicted chemical shifts with the experimental values, it is found that a Li+ ion is coordinated with four double bond oxygen atoms from EC, PC, EMC and TFSI- anion. In the case of excessive amount of solvents of EC, PC and EMC the Li+ coordinated solvent molecules are undergoing quick exchange with bulk solvent molecules, resulting in average 17O chemical shifts. Several kinds of solvation structures are identified, where the proportion of each structure in the liquid electrolytes investigated depends on the concentration of LiTFSI.

  20. Relation of desert pupfish abundance to selected environmental variables in natural and manmade habitats in the Salton Sea basin

    USGS Publications Warehouse

    Martin, B.A.; Saiki, M.K.

    2005-01-01

    We assessed the relation between abundance of desert pupfish, Cyprinodon macularius, and selected biological and physicochemical variables in natural and manmade habitats within the Salton Sea Basin. Field sampling in a natural tributary, Salt Creek, and three agricultural drains captured eight species including pupfish (1.1% of the total catch), the only native species encountered. According to Bray-Curtis resemblance functions, fish species assemblages differed mostly between Salt Creek and the drains (i.e., the three drains had relatively similar species assemblages). Pupfish numbers and environmental variables varied among sites and sample periods. Canonical correlation showed that pupfish abundance was positively correlated with abundance of western mosquitofish, Gambusia affinis, and negatively correlated with abundance of porthole livebearers, Poeciliopsis gracilis, tilapias (Sarotherodon mossambica and Tilapia zillii), longjaw mudsuckers, Gillichthys mirabilis, and mollies (Poecilia latipinnaandPoecilia mexicana). In addition, pupfish abundance was positively correlated with cover, pH, and salinity, and negatively correlated with sediment factor (a measure of sediment grain size) and dissolved oxygen. Pupfish abundance was generally highest in habitats where water quality extremes (especially high pH and salinity, and low dissolved oxygen) seemingly limited the occurrence of nonnative fishes. This study also documented evidence of predation by mudsuckers on pupfish. These findings support the contention of many resource managers that pupfish populations are adversely influenced by ecological interactions with nonnative fishes. ?? Springer 2005.

  1. Elevated Bacterial Abundance in Laboratory-Grown and Naturally Occurring Frost Flowers Under Late Winter Conditions

    NASA Astrophysics Data System (ADS)

    Bowman, J. S.; Deming, J. W.

    2009-12-01

    Sea ice has been identified as an important microbial habitat, with bacteria and other microbes concentrated in the brine inclusions between ice crystals. Frost flowers, thought to draw brine from underlying sea ice, have not been characterized from a microbial standpoint. To test whether frost flowers serve as an upward vector of bacteria contained within sea ice brines we grew frost flowers in a freezer laboratory (air temperature of -21°C) from saline water spiked with the mesophilic (and thus passive under experimental conditions) bacterium Halomonas pacifica. Salinity of melted samples was measured and bacterial abundance determined by epifluorescent microscopy. Bacterial counts scaled to ice-melt volume averaged 2.82 x 106 ml-1 for frost flowers, compared to 9.47 x 105 ml-1 for underlying ice (3 x higher). Bacterial counts also correlated significantly with salinity (maximum value of 62.5 psu) for frost flowers, brine skim, and ice (df = 17, r = 0.59, p < 0.0001). Segregation coefficients were calculated to describe the efficiency of transport of both cells and salt from the starting solution into frost flowers. From these coefficients an enrichment index was calculated to test for bacterial concentration into frost flowers at a different rate than salt. Analysis with a Student’s T-test (df = 24, t = 0.306, p = .76) indicated that cells and salt were not transported into frost flowers with a significantly different efficiency. To test these findings in the field we then collected frost flowers (and related samples) from new sea ice near Barrow, Alaska in April 2009. Bacterial counts were significantly elevated (again, a 3-fold increase) in natural frost flowers (mean = 2.73 x 105 ml-1) compared to underlying sea ice (mean = 8.46 x 104 cells ml-1). For all field samples collected (frost flowers, underlying brine skim and sea ice, as well as snow), bacterial abundance correlated significantly with salinity (maximum value 124 psu, df = 40, r = 0.60, p < 0.0001). The presence of elevated numbers of bacteria in frost flowers may have implications for the previously observed chemical reactions that take place in them, especially if microbial activity can be shown to occur in this unique low temperature, low water activity microbial habitat.

  2. The Determination of the Natural Abundance of the Isotopes of Chlorine: An Introductory Experiment in Mass Spectrometry.

    ERIC Educational Resources Information Center

    O'Malley, Rebecca M.

    1982-01-01

    Describes a laboratory experiment which introduces basic principles and experimental techniques of mass spectrometry for fourth year undergraduate (B.Sc.) students. Laboratory procedures, background information, and discussion of results are provided for the experiment in which the natural isotopic abundance of chlorine is determined. (Author/JN)

  3. Seasonal variation in natural abundance of 2H and 18O in urine samples from rural Nigeria.

    PubMed

    Harbison, Justin E; Dugas, Lara R; Brieger, William; Tayo, Bamidele O; Alabi, Tunrayo; Schoeller, Dale A; Luke, Amy

    2015-07-01

    The doubly labeled water (DLW) method is used to measure free-living energy expenditure in humans. Inherent to this technique is the assumption that natural abundances of stable isotopes (2)H and (18)O in body water remain constant over the course of the measurement period and after elimination of the loading dose of DLW will return to the same predose level. To determine variability in the natural abundances of (2)H and (18)O in humans living in a region with seasonal shifts in rain patterns and sources of drinking water, over the course of 12 mo we collected weekly urine samples from four individuals living in southwest Nigeria as well as samples of their drinking water. From ongoing regional studies of hypertension, obesity, and energy expenditure, we estimated average water turnover rate, urine volumes, and sodium and potassium excretion. Results suggest that (2)H and (18)O in urine, mean concentrations of urinary sodium and potassium, urine volume, and total body turnover differed significantly from dry to rainy season. Additionally, seasonal weather variables (mean monthly maximum temperatures, total monthly rainfall, and minimum relative humidity) were all significantly associated with natural abundances in urine. No seasonal difference was observed in drinking water samples. Findings suggest that natural abundances in urine may not remain constant as assumed, and studies incorporating DLW measurements across the transition of seasons should interpret results with caution unless appropriate doses of the tracers are used. PMID:25977450

  4. Mapping monoclonal antibody structure by 2D 13C NMR at natural abundance.

    PubMed

    Arbogast, Luke W; Brinson, Robert G; Marino, John P

    2015-04-01

    Monoclonal antibodies (mAbs) represent an important and rapidly growing class of biotherapeutics. Correct folding of a mAb is critical for drug efficacy, while misfolding can impact safety by eliciting unwanted immune or other off-target responses. Robust methods are therefore needed for the precise measurement of mAb structure for drug quality assessment and comparability. To date, the perception in the field has been that NMR could not be applied practically to mAbs due to the size (∼150 kDa) and complexity of these molecules, as well as the insensitivity of the method. The feasibility of applying NMR methods to stable isotope-labeled, protease-cleaved, mAb domains (Fab and Fc) has been demonstrated from both E. coli and Chinese hamster ovaries (CHO) cell expression platforms; however, isotopic labeling is not typically available when analyzing drug products. Here, we address the issue of feasibility of NMR-based mapping of mAb structure by demonstrating for the first time the application of a 2D (13)C NMR methyl fingerprint method for structural mapping of an intact mAb at natural isotopic abundance. Further, we show that 2D (13)C NMR spectra of protease-cleaved Fc and Fab fragments can provide accurate reporters on the domain structures that can be mapped directly to the intact mAb. Through combined use of rapid acquisition and nonuniform sampling techniques, we show that these Fab and Fc fingerprint spectra can be rapidly acquired in as short as approximately 30 min. PMID:25728213

  5. Estimation of natural mortality coefficient from fish abundance and catch data using Virtual Population Analysis (VPA)

    NASA Astrophysics Data System (ADS)

    Wang, Yingbin; Liu, Qun; Wang, Yanjun

    2007-01-01

    Natural mortality coefficient ( M) was estimated from fish abundance ( N) and catch ( C) data using a Virtual Population Analysis (VPA) model. Monte Carlo simulations were used to evaluate the impact of different error distributions for the simulated data on the estimates of M. Among the four error structures (normal, lognormal, Poisson and gamma), simulations of normally distributed errors produced the most viable estimates for M, with the lowest relative estimation errors (REEs) and median mean absolute deviations (MADs) for the ratio of the true to the estimated Ms. In contrast, the lognormal distribution had the largest REE value. Errors with different coefficients of variation (CV) were added to N and C. In general, when CVs in the data were less than 10%, reliable estimates of M were obtained. For normal and lognormal distributions, the estimates of M were more sensitive to the CVs in N than in C; when only C had error the estimates were close to the true. For Poisson and gamma distributions, opposite results were obtained. For instance, the estimates were more sensitive to the CVs in C than in N, with the largest REE from the scenario of error only in C. Two scenarios of high and low fishing mortality coefficient ( F) were generated, and the simulation results showed that the method performed better for the scenario with low F. This method was also applied to the published data for the anchovy ( Engraulis japonicus) of the Yellow Sea. Viable estimates of M were obtained for young groups, which may be explained by the fact that the great uncertainties in N and C observed for older Yellow Sea anchovy introduced large variation in the corresponding estimates of M.

  6. Natural abundance deuterium nuclear magnetic resonance spectroscopy: Study of the biosynthesis of monoterpenes

    SciTech Connect

    Leopold, M.F.

    1990-01-01

    Deuterium NMR spectroscopy at natural abundance (D NMR-na) is a new technique for exploring the biosynthesis of small molecules such as monoterpenes. The analysis of relative site-specific deuterium integration values is an effective means of measuring isotope effects, and examining the regio- and stereochemistry of biosynthetic reactions. The deuterium integration values of linalyl acetate and limonene isolated from the same source were consistent and showed that proton abstraction from the postulated {alpha}-terpinyl cation intermediate to form limonene is regioselective from the methyl derived from the Cs methyl of the precursor, geranyl diphosphate. This regiochemistry was observed in limonene samples from different sources and the measured primary kinetic isotope effect ranged from 0.25 to in excess of 100 (no deuterium was removed within experimental error). Various {alpha}- and {beta}-pinene samples were isolated and D NMR-na analysis showed evidence of isotopically sensitive partitioning of the pinylcation in the formation of these products. This spectral analysis supported published radiolabeling studies but did not require synthesis of substrates or enzyme purification. The formation of 3-carene occurs without isomerization of the double bond which was previously postulated. The olefinic deuterium of the bicyclic compound was traced to the depleted deuterium at C{sub 2} of isopentyl diphosphate by D NMR-na data and this supported unpublished radiolabeling studies. Study of irregular monoterpenes, chrysanthemyl acetate and lyratyl acetate, showed partitioning of dimethylallyl diphosphate (DMAPP) by chrysanthemyl cyclase. The {alpha}-secondary kinetic isotope effect of 1.06-1.12, obtained from relative deuterium integration values, suggested that S{sub N}1 ionization of one molecule of DMAPP is the first step in the condensation reaction.

  7. [(15)N(2)]arginine as a first potential inhaled diagnostic agent to characterize respiratory diseases.

    PubMed

    Krumbiegel, P; Denk, E; Russow, R; Rolle-Kampczyk, U; Metzner, G; Herbarth, O

    2002-01-01

    Conventional diagnosis of the pulmonary tract uses physical methods such as spirometry and oscillometry. However, the inhalation of a chemical diagnostic agent ought to provide novel ways of more specific diagnosis, for instance of inflammatory states of the bronchial and lung mucosa. The stable isotope technique using a (15)N-labeled substrate appears to be a suitable tool for this application. In a pilot study, defined amounts of the amino acid L-[guanidino-(15)N(2)]arginine monohydrochloride (aqueous solution, 20 atom % (15)N) were inhaled as a diagnostic agent by healthy volunteers and pulmonary patients suffering from asthma bronchiale. The amino acid is resorbed and partly metabolized to (15)NO. The exhaled air was collected under defined conditions in 10-L breath bags and analyzed for NO using chemiluminescence. Under standardized test conditions, healthy persons (n = 6) exhaled 0.97 +/- 0.08 micromol NO/m(3) and asthmatic patients (n = 7) 1.17 +/- 0.14 micromol NO/m(3). A better distinction was expected comparing the (15)NO exhalation. The (15)N abundance of NO was determined using a Cryotrap gas chromatography - mass spectrometry set-up. Between 30 and 80 minutes after inhaling 700 mg [(15)N]arginine, a maximum with a plateau of the (15)NO abundance was found in the exhaled air. At this time, healthy and asthmatic subjects exhibited clear differences in their exhaled (15)NO amounts. Under standardized test conditions, the healthy persons (n = 6) exhaled 102.3 +/- 6.7 nmol (15)NO/m(3), whereas asthmatic patients (n = 7) exhaled only 76.1 +/- 10.9 nmol (15)NO/m(3). It is concluded that (15)NO yielded after the inhalation of (15)N-labeled arginine could be a potential marker for demonstrating pathophysiological changes in the lung epithelium. This method could pave a new diagnostic principle of "inhalative breath test." PMID:12396247

  8. A new method for the identification of the origin of natural products. Quantitative /sup 2/H NMR at the natural abundance level applied to the characterization of anetholes

    SciTech Connect

    Martin, G.J.; Martin, M.L.; Mabon, F.; Bricont, J.

    1982-05-05

    We have shown by high-field /sup 2/H NMR spectrometry at the natural abundance level that very spectacular differences exist in the interal distribution of /sup 2/H in organic molecules. This phenomenon has been exemplified in particular by the case of ethyl and vinyl derivatives. We show in this study of various anethole samples the potential of this new method as a very powerful tool for the characterization and identification of natural products from different origins.

  9. Compound-specific δ15N and chlorin preservation in surface sediments of the Peru Margin with implications for ancient bulk δ15N records

    NASA Astrophysics Data System (ADS)

    Junium, Christopher K.; Arthur, Michael A.; Freeman, Katherine H.

    2015-07-01

    Understanding the processes that control the preservation of paleoceanographic proxies is of clear importance. Surface sediments from the Peru Margin oxygen-minimum zone are subject to lateral and downslope transport by bottom currents that decrease organic matter (OM) quality. Indicators of bulk OM quality (pyrolysis hydrogen index, pyrolysis S1 + S2 and C/N) demonstrate significant degradation between 150 and 400 m water depth, within the oxygen-minimum zone. Concentrations of the three most abundant chlorins (chlorophyllone, pheophytin and pyropheophytin) decrease from 750 to 150 nmol g TOC-1 from 150 to 400 m water depth though the relative abundances of the chlorins in an individual sample do not change. This suggests that the three chlorins have similar reactivity over the ambient conditions. Values for δ15N of bulk sediments (δ15Nbulk) decrease by 3‰ from the inner shelf to the upper slope (1000 m) but co-occurring compound-specific δ15N values (δ15Nchlorin) do not decrease downslope. The low variability of δ15Nchlorin values supports a single source for the chlorins, and demonstrates the recalcitrance of δ15Nchlorin values despite degradation. This set of observation raises questions about which type of OM fraction best records 'primary' signatures. We assess two possible models to guide our interpretation of these disparate datasets (1) that decreasing δ15Nbulk values are the result of degradation of a 15N-enriched fraction during downslope transport, and that δ15Nchlorin values reflect primary values; (2) that δ15Nbulk values are primary and that chlorins are derived from material transported from upslope. These data reaffirm that in active sedimentary environments such as the Eastern Tropical Pacific, transport of OM can significantly alter bulk geochemical parameters of OM integrity, but the impacts on the δ15N record of bulk sediments and chlorins are less clear, and require more study to be thoroughly understood.

  10. Quantitative proteomics using uniform (15)N-labeling, MASCOT, and the trans-proteomic pipeline.

    PubMed

    Palmblad, Magnus; Bindschedler, Laurence V; Cramer, Rainer

    2007-10-01

    Stable isotope labeling combined with MS is a powerful method for measuring relative protein abundances, for instance, by differential metabolic labeling of some or all amino acids with (14)N and (15)N in cell culture or hydroponic media. These and most other types of quantitative proteomics experiments using high-throughput technologies, such as LC-MS/MS, generate large amounts of raw MS data. This data needs to be processed efficiently and automatically, from the mass spectrometer to statistically evaluated protein identifications and abundance ratios. This paper describes in detail an approach to the automated analysis of uniformly (14)N/(15)N-labeled proteins using MASCOT peptide identification in conjunction with the trans-proteomic pipeline (TPP) and a few scripts to integrate the analysis workflow. Two large proteomic datasets from uniformly labeled Arabidopsis thaliana were used to illustrate the analysis pipeline. The pipeline can be fully automated and uses only common or freely available software. PMID:17726679

  11. The natural 13C abundance of plasma glucose is a useful biomarker of recent dietary caloric sweetener intake.

    PubMed

    Cook, Chad M; Alvig, Amy L; Liu, Yu Qiu David; Schoeller, Dale A

    2010-02-01

    There is a need for objective biomarkers of dietary intake, because self-reporting is often subject to bias. We tested the validity of a biomarker for the fraction of dietary carbohydrate (CHO) from cane sugar and high fructose corn syrup (C(4) sugars) using natural (13)C abundance of plasma glucose. In a randomized, single-blinded, crossover design, 5 participants consumed 3 weight-maintaining diets for 7 d, with a 2-wk washout between diet periods. Diets differed in the fraction of total CHO energy from C(4) sugars (5, 16, or 32%). During each diet period, blood samples were drawn at hours 0800 and 1600 on d 1, 3, and 5 and at 0800, 1000, 1200, 1400, and 1600 on d 7. The delta(13)C abundance of plasma glucose was analyzed via GC- isotope ratio MS. Within each diet period, delta(13)C abundance of the 0800 fasting glucose did not change from baseline with increasing time during a diet period; however, there was a strong positive correlation (R(2) = 0.89) between delta(13)C abundance of the glucose concentration at 1000 on d 7 and the percent of breakfast CHO from C(4) sugars. Also, delta(13)C abundance of the combined plasma glucose samples on d 7 demonstrated a strong positive correlation (R(2) = 0.90) with the percent of total daily CHO from C(4) sugars. The natural delta(13)C abundance of postprandial plasma glucose relative to dietary C(4) CHO content was a valid biomarker for contributions of C(4) caloric sweeteners from the previous meal. PMID:20018804

  12. Natural abundance 17O DNP two-dimensional and surface-enhanced NMR spectroscopy

    SciTech Connect

    Perras, Frédéric A.; Kobayashi, Takeshi; Pruski, Marek

    2015-06-22

    Due to its extremely low natural abundance and quadrupolar nature, the 17O nuclide is very rarely used for spectroscopic investigation of solids by NMR without isotope enrichment. Additionally, the applicability of dynamic nuclear polarization (DNP), which leads to sensitivity enhancements of 2 orders of magnitude, to 17O is wrought with challenges due to the lack of spin diffusion and low polarization transfer efficiency from 1H. Here, we demonstrate new DNP-based measurements that extend 17O solid-state NMR beyond its current capabilities. The use of the PRESTO technique instead of conventional 1H–17O cross-polarization greatly improves the sensitivity and enables the facile measurement of undistorted line shapes and two-dimensional 1H–17O HETCOR NMR spectra as well as accurate internuclear distance measurements at natural abundance. This was applied for distinguishing hydrogen-bonded and lone 17O sites on the surface of silica gel; the one-dimensional spectrum of which could not be used to extract such detail. As a result, this greatly enhanced sensitivity has enabled, for the first time, the detection of surface hydroxyl sites on mesoporous silica at natural abundance, thereby extending the concept of DNP surface-enhanced NMR spectroscopy to the 17O nuclide.

  13. Is the multicolored Asian ladybeetle, Harmonia axyridis, the most abundant natural enemy to aphids in agroecosystems?

    PubMed

    Vandereycken, Axel; Durieux, Delphine; Joie, Emilie; Sloggett, John J; Haubruge, Eric; Verheggen, François J

    2013-01-01

    The multicolored Asian ladybeetle, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), was introduced into Western Europe in the late 1990s. Since the late 2000s, this species has been commonly considered one of the most abundant aphid predators in most Western European countries. In spite of the large amount of research on H. axyridis, information concerning its relative abundance in agroecosystems is lacking. This study aims to evaluate the abundance of H. axyridis within the aphidophage community in four crops situated in southern Belgium: wheat, Triticum aestivum L. (Poales: Poaceae), corn, Zea mays, potato, Solanum tuberosum (Solanales: Solanaceae), and broad bean Vicia faba (Fabales: Fabaceae). In order to assess the species diversity, the collected data were analyzed by considering (1) the species richness and (2) the evenness according to the Shannon diversity index. Eleven aphidophages were observed in every inventoried agroecosystem, including five abundant species: three coccinellids, the seven-spotted ladybug, Coccinella septempunctata L. (Coleoptera: Coccinellidae), the 14-spotted Ladybird, Propylea quatuordecimpunctata, and H. axyridis; one hoverfly, the marmalade hoverfly, Episyrphus balteatus De Geer (Diptera: Syrphidae); and one lacewing, the common green lacewing, Chrysoperla carnea Stephens sensu lato (= s.l.) (Neuroptera: Chrysopidae). Harmonia axyridis has been observed to thrive, breed, and reproduce on the four studied crops. Harmonia axyridis is the most abundant predator of aphids in corn followed by C. septempunctata, which is the main aphid predator observed in the three other inventoried crops. In wheat and potato fields, H. axyridis occurs in low numbers compared to other aphidophage. These observations suggest that H. axyridis could be considered an invasive species of agrosystems, and that potato and wheat may intermittently act as refuges for other aphidophages vulnerable to intraguild predation by this invader. Harmonia axyridis is not the most abundant aphid predator in the main Belgian crops. PMID:24785375

  14. Is the Multicolored Asian Ladybeetle, Harmonia axyridis, the Most Abundant Natural Enemy to Aphids in Agroecosystems?

    PubMed Central

    Vandereycken, Axel; Durieux, Delphine; Joie, Emilie; Sloggett, John J.; Haubruge, Eric; Verheggen, François J.

    2013-01-01

    The multicolored Asian ladybeetle, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), was introduced into Western Europe in the late 1990s. Since the late 2000s, this species has been commonly considered one of the most abundant aphid predators in most Western European countries. In spite of the large amount of research on H. axyridis, information concerning its relative abundance in agroecosystems is lacking. This study aims to evaluate the abundance of H. axyridis within the aphidophage community in four crops situated in southern Belgium: wheat, Triticum aestivum L. (Poales: Poaceae), corn, Zea mays, potato, Solanum tuberosum (Solanales: Solanaceae), and broad bean Vicia faba (Fabales: Fabaceae). In order to assess the species diversity, the collected data were analyzed by considering (1) the species richness and (2) the evenness according to the Shannon diversity index. Eleven aphidophages were observed in every inventoried agroecosystem, including five abundant species: three coccinellids, the seven-spotted ladybug, Coccinella septempunctata L. (Coleoptera: Coccinellidae), the 14-spotted Ladybird, Propylea quatuordecimpunctata, and H. axyridis; one hoverfly, the marmalade hoverfly, Episyrphus balteatus De Geer (Diptera: Syrphidae); and one lacewing, the common green lacewing, Chrysoperla carnea Stephens sensu lato (= s.l.) (Neuroptera: Chrysopidae). Harmonia axyridis has been observed to thrive, breed, and reproduce on the four studied crops. Harmonia axyridis is the most abundant predator of aphids in corn followed by C. septempunctata, which is the main aphid predator observed in the three other inventoried crops. In wheat and potato fields, H. axyridis occurs in low numbers compared to other aphidophage. These observations suggest that H. axyridis could be considered an invasive species of agrosystems, and that potato and wheat may intermittently act as refuges for other aphidophages vulnerable to intraguild predation by this invader. Harmonia axyridis is not the most abundant aphid predator in the main Belgian crops. PMID:24785375

  15. Increased seawater temperature increases the abundance and alters the structure of natural Vibrio populations associated with the coral Pocillopora damicornis

    PubMed Central

    Tout, Jessica; Siboni, Nachshon; Messer, Lauren F.; Garren, Melissa; Stocker, Roman; Webster, Nicole S.; Ralph, Peter J.; Seymour, Justin R.

    2015-01-01

    Rising seawater temperature associated with global climate change is a significant threat to coral health and is linked to increasing coral disease and pathogen-related bleaching events. We performed heat stress experiments with the coral Pocillopora damicornis, where temperature was increased to 31°C, consistent with the 2–3°C predicted increase in summer sea surface maxima. 16S rRNA amplicon sequencing revealed a large shift in the composition of the bacterial community at 31°C, with a notable increase in Vibrio, including known coral pathogens. To investigate the dynamics of the naturally occurring Vibrio community, we performed quantitative PCR targeting (i) the whole Vibrio community and (ii) the coral pathogen Vibrio coralliilyticus. At 31°C, Vibrio abundance increased by 2–3 orders of magnitude and V. coralliilyticus abundance increased by four orders of magnitude. Using a Vibrio-specific amplicon sequencing assay, we further demonstrated that the community composition shifted dramatically as a consequence of heat stress, with significant increases in the relative abundance of known coral pathogens. Our findings provide quantitative evidence that the abundance of potential coral pathogens increases within natural communities of coral-associated microbes as a consequence of rising seawater temperature and highlight the potential negative impacts of anthropogenic climate change on coral reef ecosystems. PMID:26042096

  16. Increased seawater temperature increases the abundance and alters the structure of natural Vibrio populations associated with the coral Pocillopora damicornis.

    PubMed

    Tout, Jessica; Siboni, Nachshon; Messer, Lauren F; Garren, Melissa; Stocker, Roman; Webster, Nicole S; Ralph, Peter J; Seymour, Justin R

    2015-01-01

    Rising seawater temperature associated with global climate change is a significant threat to coral health and is linked to increasing coral disease and pathogen-related bleaching events. We performed heat stress experiments with the coral Pocillopora damicornis, where temperature was increased to 31°C, consistent with the 2-3°C predicted increase in summer sea surface maxima. 16S rRNA amplicon sequencing revealed a large shift in the composition of the bacterial community at 31°C, with a notable increase in Vibrio, including known coral pathogens. To investigate the dynamics of the naturally occurring Vibrio community, we performed quantitative PCR targeting (i) the whole Vibrio community and (ii) the coral pathogen Vibrio coralliilyticus. At 31°C, Vibrio abundance increased by 2-3 orders of magnitude and V. coralliilyticus abundance increased by four orders of magnitude. Using a Vibrio-specific amplicon sequencing assay, we further demonstrated that the community composition shifted dramatically as a consequence of heat stress, with significant increases in the relative abundance of known coral pathogens. Our findings provide quantitative evidence that the abundance of potential coral pathogens increases within natural communities of coral-associated microbes as a consequence of rising seawater temperature and highlight the potential negative impacts of anthropogenic climate change on coral reef ecosystems. PMID:26042096

  17. Assessment of the natural variation of low abundant metabolic proteins in soybean seeds using proteomics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry, we investigated the distribution of the low abundant proteins that are involved in soybean seed development in four wild and twelve cultivated soybean genotypes. We found proteomic variation of these proteins within and...

  18. A natural abundance 33S solid-state NMR study of layered transition metal disulfides at ultrahigh magnetic field.

    PubMed

    Sutrisno, Andre; Terskikh, Victor V; Huang, Yining

    2009-01-01

    Using a series of layered transition metal disulfides we demonstrate that the wide-line natural abundance solid-state NMR spectra of 33S in a less symmetric environment can readily be obtained at ultrahigh magnetic field of 21.1 T and that surprisingly these closely related materials display a wide range of 33S quadrupole coupling constant and chemical shift anisotropy values. PMID:19099063

  19. Partitioning Respiration Between Plant and Microbial Sources Using Natural Abundance Stable Carbon Isotopes

    NASA Astrophysics Data System (ADS)

    Dawson, T. E.; Tu, K. P.

    2009-12-01

    Partitioning plant and microbial respiration is important for understanding the mechanistic basis of ecosystem respiration, as each can respond to changes in environmental conditions in different ways and at different timescales. In theory, natural abundance stable C isotopes can allow source partitioning when the isotopic difference between the sources can be resolved. The longstanding notion is that such differences do not exist, yet field measurements to support this conclusion are rare. The question remains as to how much isotopic difference exists between the plant and microbial respiration and whether or not this difference is sufficient for partitioning. We addressed this question by measuring the C isotope ratios (13C/12C) of plant, microbial, and whole ecosystem respiration from three contrasting ecosystems in California. We found significant variation in the 13C/12C ratios between plant and microbial sources. However, isotopic mass balance was not observed in more than half the cases. When isotopic mass balance was observed, the largest isotopic differences were always between CO2 evolved during leaf respiration and SOC decomposition, with leaf respiration more depleted by 1-8 per mil. Within plants, the leaf respiration was more depleted than rhizosphere respiration by 2-6 per mil. Among microbial sources, litter decomposition was more depleted than SOC decomposition by 1-5 per mil. The 13C/12C ratio of bulk C and respired CO2 exhibited similar trends, but bulk C values were clearly not a good surrogate for the 13C/12C ratios of respired CO2. Based on the 13C/12C ratios of respired CO2, belowground respiration accounted for 25% in the redwood forest, 37% in the grassland and 84% in the pine forest. Belowground respiration was further partitioned between rhizosphere, litter and SOC decomposition. Each contributed nearly equal amounts in the redwood forest (30/40/30) whereas litter respiration dominated in the grassland (20/70/10) and pine forest (15/65/20). Given that there were three sources and only one isotope, these estimates for belowground partitioning were only potential ranges. We also found large temporal variation in the 13C/12C ratios of plant respiration. At the pine forest, differences between night and day were as large as 4 per mil for leaf respiration and 2.5 per mil for rhizosphere respiration. Belowground respiration changed by less than 1 per mil, and this variation appeared to be driven entirely by rhizosphere respiration (r2= 94%). The fact that isotopic mass balance was not observed on many days may have been due to this temporal variability. For example, isotopic mass balance was observed in the redwood forest when all respiration samples were collected at the same time of day (pre-dawn), whereas isotopic mass balance was not observed when respiration samples were collected from different times of the day. Partitioning is therefore possible because of relatively large isotopic differences between leaf and SOC decomposition, but care must be taken to determine all source signatures at the same time due to large temporal variability.

  20. An Evolutionary Perspective on Global Vegetation \\delta15N Co-variance With Average Annual Precipitation

    NASA Astrophysics Data System (ADS)

    Sechrest, W.; Billmark, K. A.; Shields, L. G.; Swap, R. J.; Macko, S. A.

    2001-12-01

    Recent regional transect studies have shown a significant negative relationship between the \\delta15N value of vegetation and the mean annual precipitation. \\delta15N values provide an indication of the source nitrogen utilized by the plant owing to differential fractionations from biological and chemical processes. The fact that such a relationship exists across a variety of systems is surprising given the abundance of global plant species and the complexity of the nitrogen cycle. This observed pattern suggests that there may be some biological controls on plant utilization of nitrogen that are linked to available water. After observing this relationship in samples collected as a part of the Southern African Regional Science Initiative (SAFARI 2000), we have found that this relationship holds on a global scale for published values of \\delta15N. The critical question remains; is this relationship a result of shared characteristics of related species, or more broadly is there an evolutionary reason for this relationship? Since the \\delta15N values varied with rainfall, we hypothesize that the photosynthetic pathway utilized by plants play a role in this pattern. For example, plants that have evolved C4 metabolism, in other words, plants that have evolved mechanisms to allow for utilization of nutrients in a way more independent of available water than their C3 metabolizing counterparts, may exhibit different \\delta15N with respect to our original relationship. The data collected in conjunction with SAFARI 2000 confirms this hypothesis and as a result we have used modern phylogenetic techniques (using taxonomic information as a surrogate for true phylogenetic relationships between the included plant species) to test whether evolutionary history has played a role in the \\delta15N pattern. We anticipate that by using multidisciplinary tools this study will greatly advance ongoing research in the areas of nitrogen dynamics and vegetative biogeochemical cycling.

  1. Evaluating the source of streamwater nitrate using δ15N and δ18O in nitrate in two watersheds in New Hampshire, USA

    USGS Publications Warehouse

    Pardo, Linda H.; Kendall, Carol; Pett-Ridge, Jennifer; Chang, Cecily C.Y.

    2004-01-01

    The natural abundance of nitrogen and oxygen isotopes in nitrate can be a powerful tool for identifying the source of nitrate in streamwater in forested watersheds, because the two main sources of nitrate, atmospheric deposition and microbial nitrification, have distinct δ18O values. Using a simple mixing model, we estimated the relative fractions in streamwater derived from these sources for two forested watersheds with markedly different streamwater nitrate outputs. In this study, we monitored δ15N and δ18O of nitrate biweekly in atmospheric deposition and in streamwater for 20 months at the Hubbard Brook Experimental Forest, New Hampshire, USA (moderate nitrogen export), and monthly in streamwater at the Bowl Research Natural Area, New Hampshire, USA (high nitrogen export). For rain, δ18O values ranged from +47 to +77‰ (mean: +58‰) and δ15N from −5 to +1‰ (mean: −3‰); for snow, δ18O values ranged from +52 to +75‰ (mean: +67‰) and δ15N from −3 to +2‰ (mean: −1‰). Streamwater nitrate, in contrast to deposition, had δ18O values between +12 and +33‰ (mean: +18‰) and δ15N between −3 and +6‰ (mean: 0‰). Since nitrate produced by nitrification typically has δ18O values ranging from −5 to +15‰, our field data suggest that most of the nitrate lost from the watersheds in streamflow was nitrified within the catchment. Our results confirm the importance of microbial nitrogen transformations in regulating nitrogen losses from forested ecosystems and suggest that hydrologic storage may be a factor in controlling catchment nitrate losses.

  2. Unveiling the Nature of the "Green Pea" Galaxies: Oxygen and Nitrogen Chemical Abundances

    NASA Astrophysics Data System (ADS)

    Amorín, R. O.; Pérez-Montero, E.; Vílchez, J. M.

    2011-07-01

    We present recent results on the oxygen and nitrogen chemical abundances in the extremely compact, low-mass starburst galaxies at redshifts 0.1-0.3 usually referred to as "green pea" galaxies. We show that they are metal-poor galaxies (~1/5 solar) with lower oxygen abundances than star-forming galaxies of similar mass and N/O ratios unusually high for galaxies of the same metallicity. Recent, rapid, and massive inflows of cold gas, possibly coupled with enriched outflows from supernova winds, are used to explain the results. This is consistent with the known "pea" galaxy properties and suggest that these rare objects are experiencing a short and extreme phase in their evolution.

  3. Using natural isotopic abundances to determine the source of nitrous oxide (N2O) emissions

    NASA Astrophysics Data System (ADS)

    Mothet, A.; Sebilo, M.; Laverman, A. M.; Vaury, V.; Mariotti, A.

    2012-04-01

    Numerous greenhouse gas studies have focused on carbon dioxide (CO2), whereas nitrous oxide (N2O) also plays a major role in global warming. Indeed, while nitrous oxide is 1000 times less concentrated than CO2 in the atmosphere, it is 300 times more efficient in terms of global warming potential. In addition, its atmospheric concentration increases with 0,3 % per year. According to the literature, nitrous oxide is produced, in soils and sediments, by two major processes: (1) Nitrification, mediated by autotrophic nitrifying bacteria under oxic conditions; (2) Denitrification, mediated by heterotrophic denitrifying bacteria under anoxic conditions. Denitrification induces intensive, localized and instantaneous fluxes. N2O emissions can be easily measured and modeled. In contrast, nitrification induces weak emissions, but spatially and temporally extended. Therefore, this process could represent a large potential of N2O emissions from soils and sediments. The study of isotopomer's isotopic composition of N2O, i.e. the intramolecular distribution or site preference (SP) determined by 15N measurement allows the determination of the origin of N2O emissions (nitrification vs. denitrification). Recent studies on pure cultures have showed that SP associated with nitrification is 35 ‰ while SP associated with denitrification is 0 ‰. The aim of this study was to determine SP associated with denitrification in soils and sediments, taking into account the environmental denitrifying bacterial communities, and under different environmental variables. To this end, flow-through reactors were used to determine denitrification rates at different temperatures and varying substrate (nitrate) concentrations. Site preference was measured for the different experiments. Different experiments of denitrification were realized in sediment flow through reactors under denitrifying conditions (anoxia, presence of organic matter and nitrate). We used acetylene (25°C) to block the enzyme nitrous oxide reductase, resulting in accumulation of N2O originating only from denitrification. Despite the fact that the isotopic composition of the produced N2O (15N and 18O) varies, the SP did not change significantly (SP = 6 ‰). These results compared to those of chemical denitrification show that despite very different isotopic compositions, the SP value is independent of the type of denitrification. Different nitrate concentrations (5 mM, 3 mM, 1,5 mM and 1 mM) at ambient temperature (25°C). The results of N2O production kinetics were not related to nitrate concentrations. SP of N2O are currently being analyzed. Different temperatures (35°C, 25°C and 12°C) and a nitrate concentration of 5 mM. The results of N2O production kinetics at different temperatures show an increase in N2O emissions with increasing temperature. SP of N2O are currently being analyzed. The goal for future work is to study the SP in these systems relative to salinity, pH and carbon organic concentration in denitrifying conditions but also in nitrifying conditions.

  4. Nature's Starships. I. Observed Abundances and Relative Frequencies of Amino Acids in Meteorites

    NASA Astrophysics Data System (ADS)

    Cobb, Alyssa K.; Pudritz, Ralph E.

    2014-03-01

    The class of meteorites called carbonaceous chondrites are examples of material from the solar system which have been relatively unchanged from the time of their initial formation. These meteorites have been classified according to the temperatures and physical conditions of their parent planetesimals. We collate available data on amino acid abundance in these meteorites and plot the concentrations of different amino acids for each meteorite within various meteorite subclasses. We plot average concentrations for various amino acids across meteorites separated by subclass and petrologic type. We see a predominance in the abundance and variety of amino acids in CM2 and CR2 meteorites. The range in temperature corresponding to these subclasses indicates high degrees of aqueous alteration, suggesting aqueous synthesis of amino acids. Within the CM2 and CR2 subclasses, we identify trends in relative frequencies of amino acids to investigate how common amino acids are as a function of their chemical complexity. These two trends (total abundance and relative frequencies) can be used to constrain formation parameters of amino acids within planetesimals. Our organization of the data supports an onion shell model for the temperature structure of planetesimals. The least altered meteorites (type 3) and their amino acids originated near cooler surface regions. The most active amino acid synthesis likely took place at intermediate depths (type 2). The most altered materials (type 1) originated furthest toward parent body cores. This region is likely too hot to either favor amino acid synthesis or for amino acids to be retained after synthesis.

  5. Nature's starships. I. Observed abundances and relative frequencies of amino acids in meteorites

    SciTech Connect

    Cobb, Alyssa K.; Pudritz, Ralph E. E-mail: pudritz@physics.mcmaster.ca

    2014-03-10

    The class of meteorites called carbonaceous chondrites are examples of material from the solar system which have been relatively unchanged from the time of their initial formation. These meteorites have been classified according to the temperatures and physical conditions of their parent planetesimals. We collate available data on amino acid abundance in these meteorites and plot the concentrations of different amino acids for each meteorite within various meteorite subclasses. We plot average concentrations for various amino acids across meteorites separated by subclass and petrologic type. We see a predominance in the abundance and variety of amino acids in CM2 and CR2 meteorites. The range in temperature corresponding to these subclasses indicates high degrees of aqueous alteration, suggesting aqueous synthesis of amino acids. Within the CM2 and CR2 subclasses, we identify trends in relative frequencies of amino acids to investigate how common amino acids are as a function of their chemical complexity. These two trends (total abundance and relative frequencies) can be used to constrain formation parameters of amino acids within planetesimals. Our organization of the data supports an onion shell model for the temperature structure of planetesimals. The least altered meteorites (type 3) and their amino acids originated near cooler surface regions. The most active amino acid synthesis likely took place at intermediate depths (type 2). The most altered materials (type 1) originated furthest toward parent body cores. This region is likely too hot to either favor amino acid synthesis or for amino acids to be retained after synthesis.

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

    PubMed

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

    2010-12-30

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

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

    PubMed Central

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

    2013-01-01

    Distinguishing organic and conventional products is a major issue of food security and authenticity. Previous studies successfully used stable isotopes to separate organic and conventional products, but up to now, this approach was not tested for organic grassland hay and soil. Moreover, isotopic abundances could be a powerful tool to elucidate differences in ecosystem functioning and driving mechanisms of element cycling in organic and conventional management systems. Here, we studied the δ15N and δ13C isotopic composition of soil and hay samples of 21 organic and 34 conventional grasslands in two German regions. We also used Δδ15N15N plant - δ15N soil) to characterize nitrogen dynamics. In order to detect temporal trends, isotopic abundances in organic grasslands were related to the time since certification. Furthermore, discriminant analysis was used to test whether the respective management type can be deduced from observed isotopic abundances. Isotopic analyses revealed no significant differences in δ13C in hay and δ15N in both soil and hay between management types, but showed that δ13C abundances were significantly lower in soil of organic compared to conventional grasslands. Δδ15N values implied that management types did not substantially differ in nitrogen cycling. Only δ13C in soil and hay showed significant negative relationships with the time since certification. Thus, our result suggest that organic grasslands suffered less from drought stress compared to conventional grasslands most likely due to a benefit of higher plant species richness, as previously shown by manipulative biodiversity experiments. Finally, it was possible to correctly classify about two third of the samples according to their management using isotopic abundances in soil and hay. However, as more than half of the organic samples were incorrectly classified, we infer that more research is needed to improve this approach before it can be efficiently used in practice. PMID:24205126

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

    PubMed

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

    2013-01-01

    Distinguishing organic and conventional products is a major issue of food security and authenticity. Previous studies successfully used stable isotopes to separate organic and conventional products, but up to now, this approach was not tested for organic grassland hay and soil. Moreover, isotopic abundances could be a powerful tool to elucidate differences in ecosystem functioning and driving mechanisms of element cycling in organic and conventional management systems. Here, we studied the ?(15)N and ?(13)C isotopic composition of soil and hay samples of 21 organic and 34 conventional grasslands in two German regions. We also used ??(15)N (?(15)N plant - ?(15)N soil) to characterize nitrogen dynamics. In order to detect temporal trends, isotopic abundances in organic grasslands were related to the time since certification. Furthermore, discriminant analysis was used to test whether the respective management type can be deduced from observed isotopic abundances. Isotopic analyses revealed no significant differences in ?(13)C in hay and ?(15)N in both soil and hay between management types, but showed that ?(13)C abundances were significantly lower in soil of organic compared to conventional grasslands. ??(15)N values implied that management types did not substantially differ in nitrogen cycling. Only ?(13)C in soil and hay showed significant negative relationships with the time since certification. Thus, our result suggest that organic grasslands suffered less from drought stress compared to conventional grasslands most likely due to a benefit of higher plant species richness, as previously shown by manipulative biodiversity experiments. Finally, it was possible to correctly classify about two third of the samples according to their management using isotopic abundances in soil and hay. However, as more than half of the organic samples were incorrectly classified, we infer that more research is needed to improve this approach before it can be efficiently used in practice. PMID:24205126

  9. In vivo 13C and 15N NMR studies of methylamine metabolism in Pseudomonas species MA.

    PubMed

    Jones, J G; Bellion, E

    1991-06-25

    Pseudomonas species MA was grown with methylamine as a sole source of carbon and nitrogen enabling the total flow of carbon and nitrogen into this organism to be simultaneously monitored in vivo using 13C and 15N NMR. [13C]Methylamine was rapidly and extensively incorporated into the methyl group of N-methylglutamate during high oxygenation of the cell suspension, but when the oxygenation rate was lower, a significant portion was also found in the methyl group of gamma-glutamylmethylamide. At later times the carbon label was found in intermediates of the serine assimilation pathway, with glutamate derived from the tricarboxylic acid cycle being the most abundant product. Incorporation of [15N]methylamine was only detected as N-methyl[15N]glutamate, but when protein synthesis was inhibited, the label was also detected in the amino nitrogen of glutamate. When oxygenation rates were lower, the 15N-labeled methylamine was found in the methylamide group of gamma-glutamylmethylamide in addition to being incorporated into N-methylglutamate. gamma-Glutamylmethylamide formation was linked to the overall energy state of the cell and was not affected by inhibition of the carbon assimilation pathway. Neither 5-hydroxy-N-methylpyroglutamate nor N-methyl-alpha-ketoglutaramate were detected to any significant extent. A mechanism was proposed for the role of gamma-glutamylmethylamide in the regulation of endogenous nitrogen supplies in this organism. PMID:2050672

  10. Natural decay process affects the abundance and community structure of Bacteria and Archaea in Picea abies logs.

    PubMed

    Rinta-Kanto, J M; Sinkko, H; Rajala, T; Al-Soud, W A; Sørensen, S J; Tamminen, M V; Timonen, S

    2016-07-01

    Prokaryotes colonize decaying wood and contribute to the degradation process, but the dynamics of prokaryotic communities during wood decay is still poorly understood. We studied the abundance and community composition of Bacteria and Archaea inhabiting naturally decaying Picea abies logs and tested the hypothesis that the variations in archaeal and bacterial abundances and community composition are coupled with environmental parameters related to the decay process. The data set comprises >500 logs at different decay stages from five geographical locations in south and central Finland. The results show that Bacteria and Archaea are an integral and dynamic component of decaying wood biota. The abundances of bacterial and archaeal 16S rRNA genes increase as wood decay progresses. Changes in bacterial community composition are clearly linked to the loss of density of wood, while specific fungal-bacterial interactions may also affect the distribution of bacterial taxa in decaying wood. Thaumarchaeota were prominent members of the archaeal populations colonizing decaying wood, providing further evidence of the versatility and cosmopolitan nature of this phylum in the environment. The composition and dynamics of the prokaryotic community suggest that they are an active component of biota that are involved in processing substrates in decaying wood material. PMID:27127195

  11. δ15N patterns of Douglas-fir and red alder riparian forests in the Oregon Coast Range

    USGS Publications Warehouse

    Scott, E.E.; Perakis, S.S.; Hibbs, D.E.

    2008-01-01

    We used naturally occurring stable isotopes of N to compare N dynamics in near-stream and upslope environments along riparian catenas in N-fixing red alder (Alnus rubra) and Douglas-fir (Pseudotsuga menziesii) forests in the Coast Range of western Oregon. Based on the existing literature, we expected soil δ15N to be enriched closer to streams owing to inputs of isotopically heavy, marine-derived N by spawning salmon, higher rates of denitrification near the stream, or both. However, it has been unclear what effect red alder might have on soil δ15N patterns near streams. We found a consistent −1‰ δ15N signature in red alder foliage, and δ15N of total N in soils under red alder averaged 2.2‰ along sampling transects extending 20 m upslope from the stream. Surprisingly, δ15N of total N in soil under Douglas-fir was progressively depleted nearer to streams, opposite from the pattern expected from N losses by denitrification or N inputs from anadromous salmon. Instead, δ15N of total N in soil under Douglas-fir converged toward soil δ15N values typical of red alder sites. We consider that the historic presence of red alder may have contributed a legacy of lower soil δ15N nearer to streams on sites that are currently dominated by young Douglas-fir forest.

  12. NMR resonance assignments for sparsely 15N labeled proteins.

    PubMed

    Feng, Lianmei; Lee, Han-Seung; Prestegard, James H

    2007-07-01

    For larger proteins, and proteins not amenable to expression in bacterial hosts, it is difficult to deduce structures using NMR methods based on uniform (13)C, (15)N isotopic labeling and observation of just nuclear Overhauser effects (NOEs). In these cases, sparse labeling with selected (15)N enriched amino acids and extraction of a wider variety of backbone-centered structural constraints is providing an alternate approach. A limitation, however, is the absence of resonance assignment strategies that work without uniform (15)N, (13)C labeling or preparation of numerous samples labeled with pairs of isotopically labeled amino acids. In this paper an approach applicable to a single sample prepared with sparse (15)N labeling in selected amino acids is presented. It relies on correlation of amide proton exchange rates, measured from data on the intact protein and on digested and sequenced peptides. Application is illustrated using the carbohydrate binding protein, Galectin-3. Limitations and future applications are discussed. PMID:17487550

  13. Does human activity impact the natural antibiotic resistance background? Abundance of antibiotic resistance genes in 21 Swiss lakes.

    PubMed

    Czekalski, Nadine; Sigdel, Radhika; Birtel, Julia; Matthews, Blake; Bürgmann, Helmut

    2015-08-01

    Antibiotic resistance genes (ARGs) are emerging environmental contaminants, known to be continuously discharged into the aquatic environment via human and animal waste. Freshwater aquatic environments represent potential reservoirs for ARG and potentially allow sewage-derived ARG to persist and spread in the environment. This may create increased opportunities for an eventual contact with, and gene transfer to, human and animal pathogens via the food chain or drinking water. However, assessment of this risk requires a better understanding of the level and variability of the natural resistance background and the extent of the human impact. We have analyzed water samples from 21 Swiss lakes, taken at sampling points that were not under the direct influence of local contamination sources and analyzed the relative abundance of ARG using quantitative real-time PCR. Copy numbers of genes mediating resistance to three different broad-spectrum antibiotic classes (sulfonamides: sul1, sul2, tetracyclines: tet(B), tet(M), tet(W) and fluoroquinolones: qnrA) were normalized to copy numbers of bacterial 16S rRNA genes. We used multiple linear regression to assess if ARG abundance is related to human activities in the catchment, microbial community composition and the eutrophication status of the lakes. Sul genes were detected in all sampled lakes, whereas only four lakes contained quantifiable numbers of tet genes, and qnrA remained below detection in all lakes. Our data indicate higher abundance of sul1 in lakes with increasing number and capacity of wastewater treatment plants (WWTPs) in the catchment. sul2 abundance was rather related to long water residence times and eutrophication status. Our study demonstrates the potential of freshwater lakes to preserve antibiotic resistance genes, and provides a reference for ARG abundance from lake systems with low human impact as a baseline for assessing ARG contamination in lake water. PMID:25913323

  14. Proton-Detected 15N NMR-Spectroscopy and Imaging

    NASA Astrophysics Data System (ADS)

    Freeman, D.; Sailasuta, N.; Sukumar, S.; Hurd, R. E.

    1993-10-01

    Proton detection of nitrogen-15, using gradients for coherence selection, was determined to be an effective method for obtaining spectra of 15N-labeled metabolites from extracts and biopsies of tissue infused with [15N] ammonium chloride. The advantage of gradient selection of coherence was best demonstrated by the almost complete single-shot elimination of solvent water in extracts and tissue water in biopsies. As a single-acquisition editing method in which only protons attached to 15N are detected, the potential limitations of dynamic range and motion are also reduced. Gradient-enhanced heteronuclear multiple-quantum coherence (1H[15N] HMQC) was compared with conventional HMQC, and despite selection of only one of the two heteronuclear pathways, GE-HMQC was found to be more effective for resolving the desired signal for dilute solutions; and with a single scan. In addition, effective water elimination made it possible to use the resolution advantage of a frequency-encoding dimension in proton-detected 15N imaging experiments. The limit of detection of the method at 500 MHz was 0.7 mM in 16 scans from a total volume of 400 μl. Signals from tissue extracts were observable in less than one minute for kidney, heart, brain, and muscle. Proton-detected 15N GE-HMQC images with a voxel size of 39 × 78 × 625 μm were obtained at 600 MHz from a 4 mM (1.6 μmol) 15N urea sample in less than four hours. Distribution of [15N] urea in the kidney was observed in a 600 MHz GE-HMQC image of the papilla and some cortical structures.

  15. Plant community change mediates the response of foliar δ(15)N to CO 2 enrichment in mesic grasslands.

    PubMed

    Polley, H Wayne; Derner, Justin D; Jackson, Robert B; Gill, Richard A; Procter, Andrew C; Fay, Philip A

    2015-06-01

    Rising atmospheric CO2 concentration may change the isotopic signature of plant N by altering plant and microbial processes involved in the N cycle. CO2 may increase leaf δ(15)N by increasing plant community productivity, C input to soil, and, ultimately, microbial mineralization of old, (15)N-enriched organic matter. We predicted that CO2 would increase aboveground productivity (ANPP; g biomass m(-2)) and foliar δ(15)N values of two grassland communities in Texas, USA: (1) a pasture dominated by a C4 exotic grass, and (2) assemblages of tallgrass prairie species, the latter grown on clay, sandy loam, and silty clay soils. Grasslands were exposed in separate experiments to a pre-industrial to elevated CO2 gradient for 4 years. CO2 stimulated ANPP of pasture and of prairie assemblages on each of the three soils, but increased leaf δ(15)N only for prairie plants on a silty clay. δ(15)N increased linearly as mineral-associated soil C declined on the silty clay. Mineral-associated C declined as ANPP increased. Structural equation modeling indicted that CO2 increased ANPP partly by favoring a tallgrass (Sorghastrum nutans) over a mid-grass species (Bouteloua curtipendula). CO2 may have increased foliar δ(15)N on the silty clay by reducing fractionation during N uptake and assimilation. However, we interpret the soil-specific, δ(15)N-CO2 response as resulting from increased ANPP that stimulated mineralization from recalcitrant organic matter. By contrast, CO2 favored a forb species (Solanum dimidiatum) with higher δ(15)N than the dominant grass (Bothriochloa ischaemum) in pasture. CO2 enrichment changed grassland δ(15)N by shifting species relative abundances. PMID:25604918

  16. δ15N Value Does Not Reflect Fasting in Mysticetes

    PubMed Central

    Aguilar, Alex; Giménez, Joan; Gómez–Campos, Encarna; Cardona, Luís; Borrell, Asunción

    2014-01-01

    The finding that tissue δ15N values increase with protein catabolism has led researchers to apply this value to gauge nutritive condition in vertebrates. However, its application to marine mammals has in most occasions failed. We investigated the relationship between δ15N values and the fattening/fasting cycle in a model species, the fin whale, a migratory capital breeder that experiences severe seasonal variation in body condition. We analyzed two tissues providing complementary insights: one with isotopic turnover (muscle) and one that keeps a permanent record of variations in isotopic values (baleen plates). In both tissues δ15N values increased with intensive feeding but decreased with fasting, thus contradicting the pattern previously anticipated. The apparent inconsistency during fasting is explained by the fact that a) individuals migrate between different isotopic isoscapes, b) starvation may not trigger significant negative nitrogen balance, and c) excretion drops and elimination of 15N-depleted urine is minimized. Conversely, when intensive feeding is resumed in the northern grounds, protein anabolism and excretion start again, triggering 15N enrichment. It can be concluded that in whales and other mammals that accrue massive depots of lipids as energetic reserves and which have limited access to drinking water, the δ15N value is not affected by fasting and therefore cannot be used as an indicatior of nutritive condition. PMID:24651388

  17. Characterization of α-helix structures in polypeptides, revealed by 13CO⋯H- 15N hydrogen bond lengths determined by 13C REDOR NMR

    NASA Astrophysics Data System (ADS)

    Kimura, S.; Naito, A.; Saitô, H.; Ogawa, K.; Shoji, A.

    2001-05-01

    We have determined accurate 13C- 15N interatomic distances of mono-dispersed sequential oligopeptides, [1- 13C]Leu 8-, [ 15N]Ala 12-labeled (Phe-Leu-Ala) 6, and a variety of its derivatives in which Leu, at the residue11, was replaced by Ala, Phe, Gly or D-Ala, by means of rotational echo double resonance (REDOR) experiments. In order to minimize a plausible contribution from the peak intensities of natural abundant signals from amino-acid residues other than labeled, 13C NMR signals from such contributions were subtracted from the full echo amplitude of the doubly labeled spectra as reference. We have determined the interatomic distance as 4.5±0.1 Å for five kinds of sample examined without any significant deviation among them. Because [1- 13C]Leu 8 and [ 15N]Ala 12 form CO⋯HN hydrogen bond in the case of α-helical structure, this finding evidently showed the presence of α-helix and is consistent with the data from the conformation-dependent 13C NMR chemical shifts of corresponding 13C NMR signals. It turned out, therefore, that characterization of the α-helix structure based on REDOR experiment is straightforward and practical, if careful setting of experimental conditions is achieved to arrive at the meaningful data. It is also proved that no correction from the REDOR effect from the neighboring chains is necessary as far as α-helix structures are concerned, because fully and 30% labeled samples gave rise to identical REDOR results. Further, it is concluded that unusual displacement of 15N chemical shifts present in the sequential oligopeptide should be ascribed to a contribution other than any modified α-helical structures.

  18. Changes of natural isotopic abundances in the KOSI comet simulation experiments

    NASA Astrophysics Data System (ADS)

    Roessler, K.; Eich, G.; Klinger, J.; Trimborn, P.

    1992-04-01

    Results are presented of the KOSI-7 comet simulation experiment in which samples were taken from the analog body at 90-110 K in a liquid nitrogen-cooled glove box. The H2O phase was analyzed for its D and O-18 abundances. An enrichment of both isotopes (from delta(D) of -47.7 per mil in the unirradiated sample to delta(D) of -35.7 per mil in the insolated and delta(O-18) of -6.40 per mil to delta(O-18) of -4.27 per mil) was observed for the crustal region formed via sintering near the surface. The interior did not exhibit isotopic anomalies, not even at the CO2 zone of recrystallization.

  19. Nitrate dynamics in natural plants: insights based on the concentration and natural isotope abundances of tissue nitrate.

    PubMed

    Liu, Xue-Yan; Koba, Keisuke; Makabe, Akiko; Liu, Cong-Qiang

    2014-01-01

    The dynamics of nitrate (NO(-) 3), a major nitrogen (N) source for natural plants, has been studied mostly through experimental N addition, enzymatic assay, isotope labeling, and genetic expression. However, artificial N supply may not reasonably reflect the N strategies in natural plants because NO(-) 3 uptake and reduction may vary with external N availability. Due to abrupt application and short operation time, field N addition, and isotopic labeling hinder the elucidation of in situ NO(-) 3-use mechanisms. The concentration and natural isotopes of tissue NO(-) 3 can offer insights into the plant NO(-) 3 sources and dynamics in a natural context. Furthermore, they facilitate the exploration of plant NO(-) 3 utilization and its interaction with N pollution and ecosystem N cycles without disturbing the N pools. The present study was conducted to review the application of the denitrifier method for concentration and isotope analyses of NO(-) 3 in plants. Moreover, this study highlights the utility and advantages of these parameters in interpreting NO(-) 3 sources and dynamics in natural plants. We summarize the major sources and reduction processes of NO(-) 3 in plants, and discuss the implications of NO(-) 3 concentration in plant tissues based on existing data. Particular emphasis was laid on the regulation of soil NO(-) 3 and plant ecophysiological functions in interspecific and intra-plant NO(-) 3 variations. We introduce N and O isotope systematics of NO(-) 3 in plants and discuss the principles and feasibilities of using isotopic enrichment and fractionation factors; the correlation between concentration and isotopes (N and O isotopes: δ(18)O and Δ(17)O); and isotope mass-balance calculations to constrain sources and reduction of NO(-) 3 in possible scenarios for natural plants are deliberated. Finally, we offer a preliminary framework of intraplant δ(18)O-NO(-) 3 variation, and summarize the uncertainties in using tissue NO(-) 3 parameters to interpret plant NO(-) 3 utilization. PMID:25101106

  20. Nitrate dynamics in natural plants: insights based on the concentration and natural isotope abundances of tissue nitrate

    PubMed Central

    Liu, Xue-Yan; Koba, Keisuke; Makabe, Akiko; Liu, Cong-Qiang

    2014-01-01

    The dynamics of nitrate (NO−3), a major nitrogen (N) source for natural plants, has been studied mostly through experimental N addition, enzymatic assay, isotope labeling, and genetic expression. However, artificial N supply may not reasonably reflect the N strategies in natural plants because NO−3 uptake and reduction may vary with external N availability. Due to abrupt application and short operation time, field N addition, and isotopic labeling hinder the elucidation of in situ NO−3-use mechanisms. The concentration and natural isotopes of tissue NO−3 can offer insights into the plant NO−3 sources and dynamics in a natural context. Furthermore, they facilitate the exploration of plant NO−3 utilization and its interaction with N pollution and ecosystem N cycles without disturbing the N pools. The present study was conducted to review the application of the denitrifier method for concentration and isotope analyses of NO−3 in plants. Moreover, this study highlights the utility and advantages of these parameters in interpreting NO−3 sources and dynamics in natural plants. We summarize the major sources and reduction processes of NO−3 in plants, and discuss the implications of NO−3 concentration in plant tissues based on existing data. Particular emphasis was laid on the regulation of soil NO−3 and plant ecophysiological functions in interspecific and intra-plant NO−3 variations. We introduce N and O isotope systematics of NO−3 in plants and discuss the principles and feasibilities of using isotopic enrichment and fractionation factors; the correlation between concentration and isotopes (N and O isotopes: δ18O and Δ17O); and isotope mass-balance calculations to constrain sources and reduction of NO−3 in possible scenarios for natural plants are deliberated. Finally, we offer a preliminary framework of intraplant δ18O-NO−3 variation, and summarize the uncertainties in using tissue NO−3 parameters to interpret plant NO−3 utilization. PMID:25101106

  1. Natural expansion and experimental manipulation of seagrass ( Zostera spp.) abundance and the response of infaunal invertebrates

    NASA Astrophysics Data System (ADS)

    Harrison, Paul G.

    1987-06-01

    A study of an expanding seagrass bed on the south-west coast of British Columbia, Canada involved documentation and explanation of the pattern of expansion of the vegetation as well as documentation and experimental investigation of the accompanying changes in the distribution of infaunal invertebrates. Expansion followed a major environmental change, improved water clarity initiated in 1969 when a causeway blocked access to the site for silty Fraser River water. The original eelgrass ( Zostera marina) bed expanded landward over 30 m year -1, probably causing more and more water to be retained in the bed at low tide and thus improving its own habitat. More rapid expansion occurred from 1979-1983 after Z.japonica colonized at the landward edge of the eelgrass bed and in a separate intertidal area. Expansion ceased around 1983 coincident with, but probably independent of, further construction. Burrowing shrimp ( Callianassa californiensis) decreased in abundance between 1977 and 1984 in areas where the two seagrasses colonized. Short-term experiment over one summer and fall showed that removal of all shoots allowed adult shrimp and tube worms to colonize the sediment while addition of shoots of Z. japonica by transplanting caused temporary decreases in shrimp abundance. After a few weeks an established shrimp population destroyed the transplants. Among reasonable alternate hypotheses for the decline in Callianassa, the effect of sediment texture can be eliminated, but either direct negative effects of seagrass, i.e. inhibition of the burrowing of adult shrimp or of settlement of juveniles, or indirect effects, i.e. the harboring of more predators of shrimp in seagrass beds, deserve further study.

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

    USGS Publications Warehouse

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

    2003-01-01

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

  3. 15N investigation into the effect of a pollutant on the nitrogen metabolism of Tetrahymena pyriformis as a model for environmental medical research.

    PubMed Central

    Arndt, K; Hofmann, D; Gehre, M; Krumbiegel, P

    1998-01-01

    A pilot study was performed to examine the potential of stable isotope techniques for monitoring the impact of a harmful substance on the cellular nitrogen metabolism in the ciliate species Tetrahymena pyriformis. After identical cultivation periods of control cells and toluene-exposed cells in a defined culture medium enriched with [guanidino-15N2]l-arginine, a number of nitrogen-containing pools were analyzed: 1) quantity and 15N abundance of ammonia as the end product of nitrogen metabolism in the system; 2) pattern and 15N abundances of the protein-bound amino acids in the cells; 3) pattern and 15N abundances of free amino acids in the cells; and 4) pattern and 15N abundances of the amino acids in the culture medium. In addition to 15N emission spectrometry, a new gas chromatography/combustion interface-isotope ratio mass spectrometry/mass spectrometry analytical system was used. The production and 15N content of ammonia were higher in the toluene-exposed system by 30% and 43%, respectively, indicating higher deamination rates and greater arginine consumption. The toluene-exposed cells exhibited increased 15N abundances of protein-bound amino acids in alanine, aspartic acid, glutamic acid, and tyrosine. Furthermore, structural analyses revealed the presence of N[Omega]-acetylarginine and pyrrolidonecarboxylic acid--compounds that had not previously been detected in Tetrahymena pyriformis. Differences in the 15N-enrichment of free amino acids were also evident. This new effect-monitoring system designed to investigate the impact of a pollutant on protein metabolism by using a stable isotope-labeled cell culture is a powerful tool for environmental medical research. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:9681977

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

    NASA Astrophysics Data System (ADS)

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

    2012-12-01

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

  5. 15N/14N variations in Cretaceous Atlantic sedimentary sequences: implication for past changes in marine nitrogen biogeochemistry

    USGS Publications Warehouse

    Rau, G.H.; Arthur, M.A.; Dean, W.E.

    1987-01-01

    At two locations in the Atlantic Ocean (DSDP Sites 367 and 530) early to middle Cretaceous organic-carbon-rich beds ("black shales") were found to have significantly lower ??15N values (lower 15N/14N ratios) than adjacent organic-carbon-poor beds (white limestones or green claystones). While these lithologies are of marine origin, the black strata in particular have ??15N values that are significantly lower than those previously found in the marine sediment record and most contemporary marine nitrogen pools. In contrast, black, organic-carbon-rich beds at a third site (DSDP Site 603) contain predominantly terrestrial organic matter and have C- and N-isotopic compositions similar to organic matter of modern terrestrial origin. The recurring 15N depletion in the marine-derived Cretaceous sequences prove that the nitrogen they contain is the end result of an episodic and atypical biogeochemistry. Existing isotopic and other data indicate that the low 15N relative abundance is the consequence of pelagic rather than post-depositional processes. Reduced ocean circulation, increased denitrification, and, hence, reduced euphotic zone nitrate availability may have led to Cretaceous phytoplankton assemblages that were periodically dominated by N2-fixing blue-green algae, a possible source of this sediment 15N-depletion. Lack of parallel isotopic shifts in Cretaceous terrestrially-derived nitrogen (Site 603) argues that the above change in nitrogen cycling during this period did not extend beyond the marine environment. ?? 1987.

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

    PubMed Central

    Truhlar, Stephanie M.E.; Cervantes, Carla F.; Torpey, Justin W.; Kjaergaard, Magnus; Komives, Elizabeth A.

    2008-01-01

    Advances in NMR spectroscopy have enabled the study of larger proteins that typically have significant overlap in their spectra. Specific 15N-amino acid incorporation is a powerful tool for reducing spectral overlap and attaining reliable sequential assignments. However, scrambling of the label during protein expression is a common problem. We describe a rapid method to evaluate the fidelity of specific 15N-amino acid incorporation. The selectively labeled protein is proteolyzed, and the resulting peptides are analyzed using MALDI mass spectrometry. The 15N incorporation is determined by analyzing the isotopic abundance of the peptides in the mass spectra using the program DEX. This analysis determined that expression with a 10-fold excess of unlabeled amino acids relative to the 15N-amino acid prevents the scrambling of the 15N label that is observed when equimolar amounts are used. MALDI TOF-TOF MS/MS data provide additional information that shows where the “extra” 15N labels are incorporated, which can be useful in confirming ambiguous assignments. The described procedure provides a rapid technique to monitor the fidelity of selective labeling that does not require a lot of protein. These advantages make it an ideal way of determining optimal expression conditions for selectively labeled NMR samples. PMID:18567787

  7. New method for estimating bacterial cell abundances in natural samples by use of sublimation

    NASA Technical Reports Server (NTRS)

    Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

    2004-01-01

    We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500 degrees C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approximately 10(5) to 10(9) E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.

  8. A thalium-doped sodium iodide well counter for radioactive tracer applications with naturally-abundant 40K

    NASA Astrophysics Data System (ADS)

    Parker, Andrew J.; Boxall, Colin; Joyce, Malcolm J.; Schotanus, Paul

    2013-09-01

    The use of a thallium-doped sodium-iodide well-type scintillation detector for the assay of the low-activity radioisotope 40K, in open-source potassium chloride aqueous solutions, is described. The hazards, safety concerns and radiowaste generation associated with using open-source radioactive isotopes can present significant difficulties, the use of hot cells and escalated costs in radioanalytical laboratory research. A solution to this is the use of low-hazard alternatives that mimic the migration and dispersion characteristics of notable fission products (in this case 137Cs). The use of NaI(Tl) as a detection medium for naturally-abundant levels of 40K in a range of media is widespread, but the use of 40K as a radioactive tracer has not been reported. The use of such low-activity sources is often complicated by the ability to detect them efficiently. In this paper a scintillator detector designed to detect the naturally-abundant 40K present in potassium chloride in tracer applications is described. Examples of the use of potassium chloride as a tracer are given in the context of ion exchange and electrochemical migration studies, and comparisons in performance are drawn from literature with hyper pure germanium semiconductor detectors, which are more commonly utilised detectors in high-resolution counting applications.

  9. Inferring the nature of anthropogenic threats from long-term abundance records.

    PubMed

    Shoemaker, Kevin T; Akçakaya, H Resit

    2015-02-01

    Diagnosing the processes that threaten species persistence is critical for recovery planning and risk forecasting. Dominant threats are typically inferred by experts on the basis of a patchwork of informal methods. Transparent, quantitative diagnostic tools would contribute much-needed consistency, objectivity, and rigor to the process of diagnosing anthropogenic threats. Long-term census records, available for an increasingly large and diverse set of taxa, may exhibit characteristic signatures of specific threatening processes and thereby provide information for threat diagnosis. We developed a flexible Bayesian framework for diagnosing threats on the basis of long-term census records and diverse ancillary sources of information. We tested this framework with simulated data from artificial populations subjected to varying degrees of exploitation and habitat loss and several real-world abundance time series for which threatening processes are relatively well understood: bluefin tuna (Thunnus maccoyii) and Atlantic cod (Gadus morhua) (exploitation) and Red Grouse (Lagopus lagopus scotica) and Eurasian Skylark (Alauda arvensis) (habitat loss). Our method correctly identified the process driving population decline for over 90% of time series simulated under moderate to severe threat scenarios. Successful identification of threats approached 100% for severe exploitation and habitat loss scenarios. Our method identified threats less successfully when threatening processes were weak and when populations were simultaneously affected by multiple threats. Our method selected the presumed true threat model for all real-world case studies, although results were somewhat ambiguous in the case of the Eurasian Skylark. In the latter case, incorporation of an ancillary source of information (records of land-use change) increased the weight assigned to the presumed true model from 70% to 92%, illustrating the value of the proposed framework in bringing diverse sources of information into a common rigorous framework. Ultimately, our framework may greatly assist conservation organizations in documenting threatening processes and planning species recovery. PMID:25065712

  10. A New Method for Estimating Bacterial Abundances in Natural Samples using Sublimation

    NASA Technical Reports Server (NTRS)

    Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

    2004-01-01

    We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert were heated to a temperature of 500 C for several seconds under reduced pressure. The sublimate was collected on a cold finger and the amount of adenine released from the samples then determined by high performance liquid chromatography (HPLC) with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approx. l0(exp 5) to l0(exp 9) E. coli cell equivalents per gram. For most of these samples, the sublimation based cell counts were in agreement with total bacterial counts obtained by traditional DAPI staining. The simplicity and robustness of the sublimation technique compared to the DAPI staining method makes this approach particularly attractive for use by spacecraft instrumentation. NASA is currently planning to send a lander to Mars in 2009 in order to assess whether or not organic compounds, especially those that might be associated with life, are present in Martian surface samples. Based on our analyses of the Atacama Desert soil samples, several million bacterial cells per gam of Martian soil should be detectable using this sublimation technique.

  11. Refining cotton-wick method for 15N plant labelling.

    NASA Astrophysics Data System (ADS)

    Fustec, Jolle; Mahieu, Stphanie

    2010-05-01

    The symbiosis Fabaceae/Rhizobiaceae plays a critical role in the nitrogen cycle. It gives the plant the ability to fix high amounts of atmospheric N. A part of this N can be transferred to the soil via rhizodeposition. The contribution of Fabaceae to the soil N pool is difficult to measure, since it is necessary for assessing N benefits for other crops, for soil biological activity, and for reducing water pollution in sustainable agriculture (Fustec, 2009). The aim of this study was to test and improve the reliability of the 15N cotton-wick method for measuring the soil N derived from plant rhizodeposition (Mahieu et al., 2007). The effects of the concentration of the 15N-urea labelling solution and of the feeding frequency (continuous or pulses) on the assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.) and the non-nodulating isoline P2. The plant parts and the soil were prepared for 15N:14N measurements for assessing N rhizodeposition (Mahieu et al., 2009). The fraction of plants' belowground nitrogen allocated to rhizodeposition in both Frisson pea and P2 was 20 to more than 50% higher when plants were labelled continuously than when they were labelled using fortnightly pulses. Our results suggested that when 15N root enrichment was high, nitrogen rhizodeposition was underestimated only for plants that were 15N-fed by fortnightly pulses, and not in plants 15N-fed continuously. This phenomenon was especially observed for plants relying on symbiotic N fixation for N acquisition; it may be linked to the concentration of the labelling solution. In conclusion, N rhizodeposition assessment was strongly influenced by the 15N-feeding frequency and the concentration of the labelling solution. The estimation of N rhizodeposition was more reliable when plants were labelled continuously with a dilute solution of 15N urea. Fustec et al. 2009. Agron. Sustain. Dev., DOI 10.1051/agro/2009003, in press. Mahieu et al. 2007. Plant Soil 295, 193-205. Mahieu et al. 2009. Soil Biol. Biochem. 41, 2236-2243.

  12. The 15N NMR powder spectra of semicrystalline nylon 6

    NASA Astrophysics Data System (ADS)

    Powell, Douglas G.; Mathias, Lon J.

    1989-05-01

    The solid state 15N NMR chemical shift anisotropy (CSA) spectra of nylon 6 is reported. Nylon 6 (20 percent 15N enriched) was prepared by anionic polymerization of isotopically enriched caprolactam. The samples observed were prepared by extraction with methanol to remove unreacted caprolactam, then either annealed to promote crystallization or artificially plasticized by immersion in molten caprolactam. The anisotropic linewidth was approximately 200 ppm, intermediate between the range reported for sp2 and sp3 hybridized nitrogens and consistent with the partial double-bond character of the carbon-nitrogen bond in amides.

  13. Peptides and proteins in a confined environment: NMR spectra at natural isotopic abundance.

    PubMed

    Pastore, Annalisa; Salvadori, Severo; Temussi, Piero Andrea

    2007-05-01

    Confinement of proteins and peptides in a small inert space mimics the natural environment of the cell, allowing structural studies in conditions that stabilize folded conformations. We have previously shown that confinement in polyacrylamide gels (PAGs) is sufficient to induce a change in the viscosity of the aqueous solution without changing the composition and temperature of the solvent. The main limitation of a PAG to run NMR experiments in a confined environment is the need for labelling the peptides. Here we report the use of the agarose gel to run the NMR spectra of proteins and peptides. We show that agarose gels are completely transparent in NMR experiments, relieving the need for labelling. Although it is necessary to expose biomolecules to fairly high temperatures during sample preparation, we believe that this is not generally an obstacle to the study of peptides, and found that the method is also compatible with temperature-resistant proteins. The mesh of agarose gels is too wide for direct effects of confinement on the stability of proteins but confinement can be easily exploited to interact the proteins with other reagents, including crowding macromolecules that can eventually lead to fold stabilization. The use of these gels is ideally suited for low-temperature studies; we show that a very flexible peptide at subzero temperatures is stabilized into a well-folded conformation. PMID:17436341

  14. Coffinite, USiO4, Is Abundant in Nature: So Why Is It So Difficult To Synthesize?

    PubMed

    Mesbah, Adel; Szenknect, Stephanie; Clavier, Nicolas; Lozano-Rodriguez, Janeth; Poinssot, Christophe; Den Auwer, Christophe; Ewing, Rodney C; Dacheux, Nicolas

    2015-07-20

    Coffinite, USiO4, is the second most abundant U(4+) mineral on Earth, and its formation by the alteration of the UO2 in spent nuclear fuel in a geologic repository may control the release of radionuclides to the environment. Despite its abundance in nature, the synthesis and characterization of coffinite have eluded researchers for decades. On the basis of the recent synthesis of USiO4, we can now define the experimental conditions under which coffinite is most efficiently formed. Optimal formation conditions are defined for four parameters: pH, T, heating time, and U/Si molar ratio. The adjustment of pH between 10 and 12 leads probably to the formation of a uranium(IV) hydroxo-silicate complex that acts as a precursor of uranium(IV) silicate colloids and then of coffinite. Moreover, in this pH range, the largest yield of coffinite formation (as compared with those of the two competing byproduct phases, nanometer-scale UO2 and amorphous SiO2) is obtained for 250 °C, 7 days, and 100% excess silica. PMID:26145720

  15. Determining the isotopic abundance of a labeled compound by mass spectrometry and how correcting for natural abundance distribution using analogous data from the unlabeled compound leads to a systematic error.

    PubMed

    Schenk, David J; Lockley, William J S; Elmore, Charles S; Hesk, Dave; Roberts, Drew

    2016-04-01

    When the isotopic abundance or specific activity of a labeled compound is determined by mass spectrometry (MS), it is necessary to correct the raw MS data to eliminate ion intensity contributions, which arise from the presence of heavy isotopes at natural abundance (e.g., a typical carbon compound contains ~1.1% (13) C per carbon atom). The most common approach is to employ a correction in which the mass-to-charge distribution of the corresponding unlabeled compound is used to subtract the natural abundance contributions from the raw mass-to-charge distribution pattern of the labeled compound. Following this correction, the residual intensities should be due to the presence of the newly introduced labeled atoms only. However, this will only be the case when the natural abundance mass isotopomer distribution of the unlabeled compound is the same as that of the labeled species. Although this may be a good approximation, it cannot be accurate in all cases. The implications of this approximation for the determination of isotopic abundance and specific activity have been examined in practice. Isotopically mixed stable-atom labeled valine batches were produced, and both these and [(14) C6 ]carbamazepine were analyzed by MS to determine the extent of the error introduced by the approach. Our studies revealed that significant errors are possible for small highly-labeled compounds, such as valine, under some circumstances. In the case with [(14) C6 ]carbamazepine, the errors introduced were minor but could be significant for (14) C-labeled compounds with particular isotopic distributions. This source of systematic error can be minimized, although not eliminated, by the selection of an appropriate isotopic correction pattern or by the use of a program that varies the natural abundance distribution throughout the correction. PMID:26916110

  16. The contamination of commercial 15N2 gas stocks with 15N-labeled nitrate and ammonium and consequences for nitrogen fixation measurements.

    PubMed

    Dabundo, Richard; Lehmann, Moritz F; Treibergs, Lija; Tobias, Craig R; Altabet, Mark A; Moisander, Pia H; Granger, Julie

    2014-01-01

    We report on the contamination of commercial 15-nitrogen (15N) N2 gas stocks with 15N-enriched ammonium, nitrate and/or nitrite, and nitrous oxide. 15N2 gas is used to estimate N2 fixation rates from incubations of environmental samples by monitoring the incorporation of isotopically labeled 15N2 into organic matter. However, the microbial assimilation of bioavailable 15N-labeled N2 gas contaminants, nitrate, nitrite, and ammonium, is liable to lead to the inflation or false detection of N2 fixation rates. 15N2 gas procured from three major suppliers was analyzed for the presence of these 15N-contaminants. Substantial concentrations of 15N-contaminants were detected in four Sigma-Aldrich 15N2 lecture bottles from two discrete batch syntheses. Per mole of 15N2 gas, 34 to 1900 µmoles of 15N-ammonium, 1.8 to 420 µmoles of 15N-nitrate/nitrite, and ≥21 µmoles of 15N-nitrous oxide were detected. One 15N2 lecture bottle from Campro Scientific contained ≥11 µmoles of 15N-nitrous oxide per mole of 15N2 gas, and no detected 15N-nitrate/nitrite at the given experimental 15N2 tracer dilutions. Two Cambridge Isotopes lecture bottles from discrete batch syntheses contained ≥0.81 µmoles 15N-nitrous oxide per mole 15N2, and trace concentrations of 15N-ammonium and 15N-nitrate/nitrite. 15N2 gas equilibrated cultures of the green algae Dunaliella tertiolecta confirmed that the 15N-contaminants are assimilable. A finite-differencing model parameterized using oceanic field conditions typical of N2 fixation assays suggests that the degree of detected 15N-ammonium contamination could yield inferred N2 fixation rates ranging from undetectable, <0.01 nmoles N L(-1) d(-1), to 530 nmoles N L(-1) d(-1), contingent on experimental conditions. These rates are comparable to, or greater than, N2 fixation rates commonly detected in field assays. These results indicate that past reports of N2 fixation should be interpreted with caution, and demonstrate that the purity of commercial 15N2 gas must be ensured prior to use in future N2 fixation rate determinations. PMID:25329300

  17. Quantitative structural constraints for organic powders at natural isotopic abundance using dynamic nuclear polarization solid-state NMR spectroscopy.

    PubMed

    Mollica, Giulia; Dekhil, Myriam; Ziarelli, Fabio; Thureau, Pierre; Viel, Stéphane

    2015-05-11

    A straightforward method is reported to quantitatively relate structural constraints based on (13)C-(13)C double-quantum build-up curves obtained by dynamic nuclear polarization (DNP) solid-state NMR to the crystal structure of organic powders at natural isotopic abundance. This method relies on the significant gain in NMR sensitivity provided by DNP (approximately 50-fold, lowering the experimental time from a few years to a few days), and is sensitive to the molecular conformation and crystal packing of the studied powder sample (in this case theophylline). This method allows trial crystal structures to be rapidly and effectively discriminated, and paves the way to three-dimensional structure elucidation of powders through combination with powder X-ray diffraction, crystal-structure prediction, and density functional theory computation of NMR chemical shifts. PMID:25809550

  18. The natural abundance of 13C with different agricultural management by NIRS with fibre optic probe technology.

    PubMed

    Fuentes, Mariela; González-Martín, Inmaculada; Hernández-Hierro, Jose Miguel; Hidalgo, Claudia; Govaerts, Bram; Etchevers, Jorge; Sayre, Ken D; Dendooven, Luc

    2009-06-30

    In the present study the natural abundance of (13)C is quantified in agricultural soils in Mexico which have been submitted to different agronomic practices, zero and conventional tillage, retention of crop residues (with and without) and rotation of crops (wheat and maize) for 17 years, which have influenced the physical, chemical and biological characteristics of the soil. The natural abundance of C13 is quantified by near infrared spectra (NIRS) with a remote reflectance fibre optic probe, applying the probe directly to the soil samples. Discriminate partial least squares analysis of the near infrared spectra allowed to classify soils with and without residues, regardless of the type of tillage or rotation systems used with a prediction rate of 90% in the internal validation and 94% in the external validation. The NIRS calibration model using a modified partial least squares regression allowed to determine the delta(13)C in soils with or without residues, with multiple correlation coefficients 0.81 and standard error prediction 0.5 per thousand in soils with residues and 0.92 and 0.2 per thousand in soils without residues. The ratio performance deviation for the quantification of delta(13)C in soil was 2.5 in soil with residues and 3.8 without residues. This indicated that the model was adequate to determine the delta(13)C of unknown soils in the -16.2 per thousand to -20.4 per thousand range. The development of the NIR calibration permits analytic determinations of the values of delta(13)C in unknown agricultural soils in less time, employing a non-destructive method, by the application of the fibre optic probe of remote reflectance to the soil sample. PMID:19376340

  19. Influence of niche differentiation on the abundance of methanogenic archaea and methane production potential in natural wetland ecosystems across China

    NASA Astrophysics Data System (ADS)

    Liu, D.; Ding, W.; Jia, Z.; Cai, Z.

    2010-10-01

    Methane (CH4) emissions from natural wetland ecosystems exhibit large spatial variability. To understand the underlying factors that induce differences in CH4 emissions from natural wetlands around China, we measured the CH4 production potential and the abundance of methanogenic archaea in vertical profile soils sampled from the Poyang wetland in the subtropical zone, the Hongze wetland in the warm temperate zone, the Sanjiang marsh in the cold temperate zone, and the Ruoergai peatland in the Qinghai-Tibetan Plateau. The top soil layer had the highest population of methanogens (1.07-8.29109 cells g-1 soil) in all wetlands except the Ruoergai peatland and exhibited the maximum CH4 production potential measured at the mean in situ summer temperature. There is a significant logarithmic correlation between the abundance of methanogenic archaea and the soil organic carbon (R2=0.718, P<0.001, n=13) and between the abundance of methanogenic archaea and the total nitrogen concentrations (R2=0.758, P<0.001, n=13) in wetland soils. This indicates that the amount of soil organic carbon may affect the population of methanogens in wetland ecosystems. While the CH4 production potential is not significantly related to methanogen population (R2=0.011, P>0.05, n=13), it is related to the dissolved organic carbon concentration (R2=0.305, P=0.05, n=13). This suggests that the methanogen population is not an effective index for predicting the CH4 production in wetland ecosystems. The CH4 production rate of the top soil layer increases with increasing latitude, from 274 ?g CH4 kg-1 soil d-1 in the Poyang wetland to 665 ?g CH4 kg-1 soil d-1 in the Carex lasiocarpa marsh of the Sanjiang Plain. The CH4 production potential in the freshwater wetlands of Eastern China is affected by the supply of methanogenic substrates rather than by temperature, whereas the supply of substrates was mainly affected by the position and stability of the wetland water table. In contrast, low summer temperatures at high elevations in the Ruoergai peatland of the Qinghai-Tibetan Plateau result in the presence of dominant species of methanogens with low CH4 production potential rather than the reduction of the supply of methanogenic substrates, which in turn suppresses CH4 production.

  20. Stepwise enrichment of 15N along food chains: Further evidence and the relation between δ 15N and animal age

    NASA Astrophysics Data System (ADS)

    Minagawa, Masao; Wada, Eitaro

    1984-05-01

    The isotopic composition of nitrogen was measured in marine and fresh-water animals from the East China Sea, The Bering Sea, Lake Ashinoko and Usujiri intertidal zone. Primary producers, showed average δ15Nversus atmospheric nitrogen of +5.0%. (+3.4 to +7.5) in the Bering Sea and Lake Ashinoko, and +6.8%. (+6.0 to +7.6) in Usujiri intertidal zone. Blue green algae from the East China Sea show an average -0.55%. (-0.8 to +1.2). All consumers, Zooplankton, fish and bird exhibited Stepwise enrichment of 15N with increasing trophic level. The 15N enrichment at a single feeding process ranged from +1.3 to +5.3 averaging +3.4 ± 1.1%.. This isotopic fractionation seems to be independent of habitat. The effect of age in animals was obtained by analyzing two marine mussels. The soft tissue nitrogen showed +2.0%. enrichment relative to that of primary producers, and the magnitude was almost constant with shell ages ranging from 0 to 8 years. A similar 15N enrichment occurs in all Molluscs, Crustaceans, Insecta, Amphibia, Fish, Ave and Mammal species regardless of the difference in the form of excreted nitrogen and in laboratory cultured fish, brine shrimp and mice (+2.9 to +4.9%.). The excreted ammonia from guppy was sufficiently light to balance the concentration of 15N to animal body.

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

    PubMed

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

    2000-12-01

    Backbone dynamics of uniformly (15)N-labeled barstar have been studied at 32 degrees C, pH 6.7, by using (15)N relaxation data obtained from proton-detected 2D (1)H-(15)N NMR spectroscopy. (15)N spin-lattice relaxation rate constants (R(1)), spin-spin relaxation rate constants (R(2)), and steady-state heteronuclear (1)H-(15)N NOEs have been determined for 69 of the 86 (excluding two prolines and the N-terminal residue) backbone amide (15)N at a magnetic field strength of 14.1 Tesla. The primary relaxation data have been analyzed by using the model-free formalism of molecular dynamics, using both isotropic and axially symmetric diffusion of the molecule, to determine the overall rotational correlation time (tau(m)), the generalized order parameter (S(2)), the effective correlation time for internal motions (tau(e)), and NH exchange broadening contributions (R(ex)) for each residue. As per the axially symmetric diffusion, the ratio of diffusion rates about the unique and perpendicular axes (D( parallel)/D( perpendicular)) is 0.82 +/- 0.03. The two results have only marginal differences. The relaxation data have also been used to map reduced spectral densities for the NH vectors of these residues at three frequencies: 0, omega(H), and omega(N), where omega(H),(N) are proton and nitrogen Larmor frequencies. The value of tau(m) obtained from model-free analysis of the relaxation data is 5.2 ns. The reduced spectral density analysis, however, yields a value of 5.7 ns. The tau(m) determined here is different from that calculated previously from time-resolved fluorescence data (4.1 ns). The order parameter ranges from 0.68 to 0.98, with an average value of 0.85 +/- 0.02. A comparison of the order parameters with the X-ray B-factors for the backbone nitrogens of wild-type barstar does not show any considerable correlation. Model-free analysis of the relaxation data for seven residues required the inclusion of an exchange broadening term, the magnitude of which ranges from 2 to 9.1 s(-1), indicating the presence of conformational averaging motions only for a small subset of residues. PMID:11056034

  2. Female offspring desertion and male-only care increase with natural and experimental increase in food abundance

    PubMed Central

    Eldegard, Katrine; Sonerud, Geir A.

    2009-01-01

    In species with biparental care, one parent may escape the costs of parental care by deserting and leaving the partner to care for the offspring alone. A number of theoretical papers have suggested a link between uniparental offspring desertion and ecological factors, but empirical evidence is scarce. We investigated the relationship between uniparental desertion and food abundance in a natural population of Tengmalm's owl Aegolius funereus, both by means of a 5-year observational study and a 1-year experimental study. Parents and offspring were fitted with radio-transmitters in order to reveal the parental care strategy (i.e. care or desert) of individual parents, and to keep track of the broods post-fledging. We found that 70 per cent of the females from non-experimental nests deserted, while their partner continued to care for their joint offspring alone. Desertion rate was positively related to natural prey population densities and body reserves of the male partner. In response to food supplementation, a larger proportion of the females deserted, and females deserted the offspring at an earlier age. Offspring survival during the post-fledging period tended to be lower in deserted than in non-deserted broods. We argue that the most important benefit of deserting may be remating (sequential polyandry). PMID:19324835

  3. Poly-ethers from Winogradskyella poriferorum: Antifouling potential, time-course study of production and natural abundance.

    PubMed

    Dash, Swagatika; Nogata, Yasuyuki; Zhou, Xiaojian; Zhang, Yifan; Xu, Ying; Guo, Xianrong; Zhang, Xixiang; Qian, Pei-Yuan

    2011-08-01

    A sponge-associated bacterium, Winogradskyella poriferorum strain UST030701-295T was cultured up to 100l for extraction of antifouling bioactive compounds. Five poly-ethers were isolated and partially characterized based on nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS); two of them showed inhibitory effects on biofilm formation of marine bacteria and larval settlement of macro-foulers but did not produce any adverse effects on the phenotypes of zebra fish embryos at a concentration of 5μg ml(-1). The effect of culture duration on the production of the poly-ethers and the bioactivity of the relevant extracts was monitored over a period of 12 days. The total crude poly-ether production increased from day 2 to day 5 and the highest bioactivity was observed on day 3. The poly-ethers were found to be localized in the cellular fraction of the extracts, implying their natural occurrence. The potent bioactivity of these poly-ethers together with their high natural abundance in bacteria makes them promising candidates as ingredients in antifouling applications. PMID:21664125

  4. Depth profiling of nitrogen using 429 keV and 897 keV resonances in the 15N(p, ??) 12C reaction

    NASA Astrophysics Data System (ADS)

    Kumar, Sanjiv; Vikram Kumar, S.; Reddy, G. L. N.; Kain, Vivekanand; Ramana, J. V.; Raju, V. S.

    2005-11-01

    Resonances at 429 keV and 897 keV in the 15N(p, ??)12C reaction were investigated for depth profiling nitrogen in materials containing nitrogen isotopes in natural abundances. Both resonances exhibit identical sensitivity, however the resonance at 897 keV is prone to interferences from light elements, F and Al in particular. These resonances were employed to depth profile nitrogen in binary and ternary nitride films and in stainless steel wires that had fractured while in use in an ammonia converter vessel of a heavy water plant. The studies indicated the ingress of nitrogen into the interiors of the wires under operating conditions of the plant that lead to nitriding causing embrittlement of the components.

  5. Marking Drosophila suzukii (Diptera: Drosophilidae) With Rubidium or 15N.

    PubMed

    Klick, J; Yang, W Q; Bruck, D J

    2015-06-01

    Drosophila suzukii Matsumura (Diptera: Drosophilidae) has caused significant economic damage to berry and stone fruit production regions. Markers that are systemic in plants and easily transferred to target organisms are needed to track D. suzukii exploitation of host resources and trophic interactions. High and low concentrations of the trace element, rubidium (Rb), and the stable isotope, 15N, were tested to mark D. suzukii larvae feeding on fruits of enriched strawberry plants grown in containers under greenhouse conditions. Fly marker content and proportion of flies marked 1, 7, and 14 d after emergence from enriched fruits and fly dry mass were analyzed. Nearly 100% of the flies analyzed 14 d after emerging from 15N-enriched plants were marked, whereas only 30-75% and 0-3% were marked 14 d after emerging from high and low Rb concentration plants, respectively. Rapid Rb decay, strong 15N persistence, and the economics of using these markers in the field to elucidate D. suzukii pest ecology are discussed. PMID:26470275

  6. Factors Driving the Abundance of Ixodes ricinus Ticks and the Prevalence of Zoonotic I. ricinus-Borne Pathogens in Natural Foci

    PubMed Central

    Fernández-de-Mera, Isabel G.; Acevedo, Pelayo; Gortázar, Christian; de la Fuente, José

    2012-01-01

    Environmental factors may drive tick ecology and therefore tick-borne pathogen (TBP) epidemiology, which determines the risk to animals and humans of becoming infected by TBPs. For this reason, the aim of this study was to analyze the influence of environmental factors on the abundance of immature-stage Ixodes ricinus ticks and on the prevalence of two zoonotic I. ricinus-borne pathogens in natural foci of endemicity. I. ricinus abundance was measured at nine sites in the northern Iberian Peninsula by dragging the vegetation with a cotton flannelette, and ungulate abundance was measured by means of dung counts. In addition to ungulate abundance, data on variables related to spatial location, climate, and soil were gathered from the study sites. I. ricinus adults, nymphs, and larvae were collected from the vegetation, and a representative subsample of I. ricinus nymphs from each study site was analyzed by PCR for the detection of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum DNA. Mean prevalences of these pathogens were 4.0% ± 1.8% and 20.5% ± 3.7%, respectively. Statistical analyses confirmed the influence of spatial factors, climate, and ungulate abundance on I. ricinus larva abundance, while nymph abundance was related only to climate. Interestingly, cattle abundance rather than deer abundance was the main driver of B. burgdorferi sensu lato and A. phagocytophilum prevalence in I. ricinus nymphs in the study sites, where both domestic and wild ungulates coexist. The increasing abundance of cattle seems to increase the risk of other hosts becoming infected by A. phagocytophilum, while reducing the risk of being infected by B. burgdorferi sensu lato. Controlling ticks in cattle in areas where they coexist with wild ungulates would be more effective for TBP control than reducing ungulate abundance. PMID:22286986

  7. [Isotopic signature (15N/14N and 13C/12C) confirms similarity of trophic niches of millipedes (Myriapoda, Diplopoda) in a temperate deciduous forest].

    PubMed

    Semeniuk, I I; Tiunov, A V

    2011-01-01

    The species composition, abundance, and isotopic signature of millipedes (Myriapoda, Diplopoda) were investigated in seven biotopes of Kaluzhskie Zaseki State Nature Reserve. Nine Diplopoda species were found in total, and the local species diversity (within a sampling plot) reached seven species. The Diplopoda tissues were similar to the plant litter in the isotopic composition of nitrogen (delta15N was by 0.4% per hundred higher, on average), but were more strongly enriched in heavy carbon (delta13C was by 4% per hundred higher, on average). Removal of mineral carbon from the cuticle reduced delta13C of Diplopoda by about 1.4% per hundred on average. Differences in the delta15N and delta13C values between the species did not exceed 2.5 per hundred. Differences in the isotopic compositions of the considered species are insignificant, and thus, it is impossible to distinguish particular trophic guilds in the Diplopoda community. Analysis of the published data confirmed that isotopic differentiation of millipedes was much less pronounced than in other investigated groups of soil animals. Hence, millipedes of the deciduous forest form a uniform trophic group. PMID:21789998

  8. Ureide assay for measuring nitrogen fixation by nodulated soybean calibrated by sup 15 N methods. [Glycine max

    SciTech Connect

    Herridge, D.F. ); Peoples, M.B. )

    1990-06-01

    We report experiments to quantify the relationships between the relative abundance of ureide-N in root-bleeding sap, vacuum-extracted sap, and hot water extracts of stems and petioles of nodulated soybean (Glycine max (L.) Merrill cv Bragg) and the proportion of plant N derived from nitrogen fixation. Additional experiments examined the effects of plant genotype and strain of rhizobia on these relationships. In each of the five experiments reported, plants of cv Bragg (experiment 1), cv Lincoln (experiments 3, 4, 5), or six cultivars/genotypes (experiment 2) were grown in a sand:vermiculite mixture in large pots in a naturally lit, temperature-controlled glasshouse during summer. Pots were inoculated at sowing with effective Bradyrhizobium japonicum CB 1809 (USDA 136) or with one of 21 different strains of rhizobia. The proportions of plant N derived from nitrogen fixation were determined using {sup 15}N dilution. Results show that assessment of nitrogen fixation by soybean using the ureide technique should now be possible with the standard curves presented, irrespective of genotype or strain of rhizobia occupying the nodules.

  9. Two-Dimensional Infrared Study of (13)C-Natural Abundant Vibrational Transition Reveals Intramolecular Vibrational Redistribution Rather than Fluxional Exchange in Mn(CO)5Br.

    PubMed

    Yang, Fan; Zhao, Juan; Wang, Jianping

    2016-02-25

    In this work, molecular-symmetry enhanced (13)CO natural abundant isotopic infrared transition was identified in Mn(CO)5Br dissolved in CCl4 by FTIR spectroscopy. Diagonal and associated off-diagonal two-dimensional IR (2D IR) peaks of the (13)CO-species were found to be spectrally separated from the all-(12)CO species, allowing a direct probe of the (13)C natural abundant ensemble. Temperature-dependent FTIR experiment showed no evidence of ligand exchange in the metal carbonyl complex. Intramolecular vibrational redistribution dynamics among the CO stretching vibrational states were extracted using population-time dependent 2D IR diagonal and off-diagonal peaks for both radial mono-(13)CO and all-(12)CO isotopomers. This work demonstrates the potential use of natural abundant isotopic molecular species as a probe for revealing equilibrium and nonequilibrium structural dynamics in condensed-phase molecular systems. PMID:26836759

  10. Abundance of non-native crabs in intertidal habitats of New England with natural and artificial structure.

    PubMed

    Lovely, Christina M; O'Connor, Nancy J; Judge, Michael L

    2015-01-01

    Marine habitats containing complex physical structure (e.g., crevices) can provide shelter from predation for benthic invertebrates. To examine effects of natural and artificial structure on the abundance of intertidal juvenile crabs, 2 experiments were conducted in Kingston Bay, Massachusetts, USA, from July to September, 2012. In the first experiment, structure was manipulated in a two-factor design that was placed in the high intertidal for 3 one-week periods to test for both substrate type (sand vs. rock) and the presence or absence of artificial structure (mesh grow-out bags used in aquaculture, ∼0.5 m(2) with 62 mm(2) mesh openings). The Asian shore crab, Hemigrapsus sanguineus, and small individuals of the green crab, Carcinus maenas, were observed only in the treatments of rocks and mesh bag plus rocks. Most green crabs were small (<6 mm in carapace width) whereas H. sanguineus occurred in a wide range of sizes. In the second experiment, 3 levels of oyster-shell treatments were established using grow-out bags placed on a muddy sand substrate in the low intertidal zone: mesh grow-out bags without shells, grow-out bags with oyster shells, and grow-out bags containing live oysters. Replicate bags were deployed weekly for 7 weeks in a randomized complete block design. All crabs collected in the bags were juvenile C. maenas (1-15 mm carapace width), and numbers of crabs differed 6-fold among treatments, with most crabs present in bags with live oysters (29.5 ± 10.6 m(-2) [mean ± S.D.]) and fewest in bags without shells (4.9 ± 3.7 m(-2)). Both C. maenas and H. sanguineus occurred in habitats with natural structure (cobble rocks). The attraction of juvenile C. maenas to artificial structure consisting of plastic mesh bags containing both oyster shells and living oysters could potentially impact oyster aquaculture operations. PMID:26401456

  11. Abundance of non-native crabs in intertidal habitats of New England with natural and artificial structure

    PubMed Central

    Lovely, Christina M.; Judge, Michael L.

    2015-01-01

    Marine habitats containing complex physical structure (e.g., crevices) can provide shelter from predation for benthic invertebrates. To examine effects of natural and artificial structure on the abundance of intertidal juvenile crabs, 2 experiments were conducted in Kingston Bay, Massachusetts, USA, from July to September, 2012. In the first experiment, structure was manipulated in a two-factor design that was placed in the high intertidal for 3 one-week periods to test for both substrate type (sand vs. rock) and the presence or absence of artificial structure (mesh grow-out bags used in aquaculture, ∼0.5 m2 with 62 mm2 mesh openings). The Asian shore crab, Hemigrapsus sanguineus, and small individuals of the green crab, Carcinus maenas, were observed only in the treatments of rocks and mesh bag plus rocks. Most green crabs were small (<6 mm in carapace width) whereas H. sanguineus occurred in a wide range of sizes. In the second experiment, 3 levels of oyster-shell treatments were established using grow-out bags placed on a muddy sand substrate in the low intertidal zone: mesh grow-out bags without shells, grow-out bags with oyster shells, and grow-out bags containing live oysters. Replicate bags were deployed weekly for 7 weeks in a randomized complete block design. All crabs collected in the bags were juvenile C. maenas (1–15 mm carapace width), and numbers of crabs differed 6-fold among treatments, with most crabs present in bags with live oysters (29.5 ± 10.6 m−2 [mean ± S.D.]) and fewest in bags without shells (4.9 ± 3.7 m−2). Both C. maenas and H. sanguineus occurred in habitats with natural structure (cobble rocks). The attraction of juvenile C. maenas to artificial structure consisting of plastic mesh bags containing both oyster shells and living oysters could potentially impact oyster aquaculture operations. PMID:26401456

  12. In vivo quantitation of cerebral metabolite concentrations using natural abundance 13C MRS at 1.5 T.

    PubMed

    Blüml, S

    1999-02-01

    A method for the quantitation of cerebral metabolites on a clinical MR scanner by natural abundance 13C MRS in vivo is described. Proton-decoupled spectra were acquired with a power deposition within FDA guidelines using a novel coil design. myo-Inositol, quantified by a separate proton MRS, and readily detectable in 13C MRS, was used as an internal reference. Normal concentrations, measured in four control subjects, age 7 months to 12 years, were glutamate 9.9 +/- 0.7, glutamine 5.6 +/- 1.0, and NAA 8.8 +/- 2.8 mmol/kg. In a patient diagnosed with Canavan disease, examined four times, glutamate was reduced to 46% of normal, 4.6 +/- 0.5 mmol/kg. NAA was increased by 50% to 13.2 +/- 1.6 mmol/kg in 13C MRS, consistent with the 41% increase to 12.3 +/- 1.1 from control 8.7 +/- 1.1 mmol/kg assayed by 1H MRS. Limited concentration of glutamate may impact on glutamatergic neurons and excitatory neurotransmission in Canavan disease. Quantitation of cerebral glutamate in human brain may have clinical value in human neuropathologies in which glutamate is believed to play a central role. PMID:9986765

  13. Molecular characterization of dissolved organic matter in glacial ice: coupling natural abundance 1H NMR and fluorescence spectroscopy.

    PubMed

    Pautler, Brent G; Woods, Gwen C; Dubnick, Ashley; Simpson, André J; Sharp, Martin J; Fitzsimons, Sean J; Simpson, Myrna J

    2012-04-01

    Glaciers and ice sheets are the second largest freshwater reservoir in the global hydrologic cycle, and the onset of global climate warming has necessitated an assessment of their contributions to sea-level rise and the potential release of nutrients to nearby aquatic environments. In particular, the release of dissolved organic matter (DOM) from glacier melt could stimulate microbial activity in both glacial ecosystems and adjacent watersheds, but this would largely depend on the composition of the material released. Using fluorescence and (1)H NMR spectroscopy, we characterize DOM at its natural abundance in unaltered samples from a number of glaciers that differ in geographic location, thermal regime, and sample depth. Parallel factor analysis (PARAFAC) modeling of DOM fluorophores identifies components in the ice that are predominantly proteinaceous in character, while (1)H NMR spectroscopy reveals a mixture of small molecules that likely originate from native microbes. Spectrofluorescence also reveals a terrestrial contribution that was below the detection limits of NMR; however, (1)H nuclei from levoglucosan was identified in Arctic glacier ice samples. This study suggests that the bulk of the DOM from these glaciers is a mixture of biologically labile molecules derived from microbes. PMID:22385100

  14. Natural Abundance 43Ca NMR as a Tool for Exploring Calcium Biomineralization: Renal Stone Formation and Growth

    SciTech Connect

    Bowers, Geoffrey M.; Kirkpatrick, Robert J.

    2011-12-07

    Renal stone diseases are a global health issue with little effective therapeutic recourse aside from surgery and shock-wave lithotripsy, primarily because the fundamental chemical mechanisms behind calcium biomineralization are poorly understood. In this work, we show that natural abundance 43Ca NMR at 21.1 T is an effective means to probe the molecular-level Ca2+ structure in oxalate-based kidney stones. We find that the 43Ca NMR resonance of an authentic oxalate-based kidney stone cannot be explained by a single pure phase of any common Ca2+-bearing stone mineral. Combined with XRD results, our findings suggest an altered calcium oxalate monohydrate-like Ca2+ coordination environment for some fraction of Ca2+ in our sample. The evidence is consistent with existing literature hypothesizing that nonoxalate organic material interacts directly with Ca2+ at stone surfaces and is the primary driver of renal stone aggregation and growth. Our findings show that 43Ca NMR spectroscopy may provide unique and crucial insight into the fundamental chemistry of kidney stone formation, growth, and the role organic molecules play in these processes.

  15. Studies of minute quantities of natural abundance molecules using 2D heteronuclear correlation spectroscopy under 100kHz MAS

    SciTech Connect

    Nishiyama, Y.; Kobayashi, T.; Malon, M.; Singappuli-Arachchige, D.; Slowing, I. I.; Pruski, M.

    2015-02-16

    Two-dimensional 1H{13C} heteronuclear correlation solid-state NMR spectra of naturally abundant solid materials are presented, acquired using the 0.75-mm magic angle spinning (MAS) probe at spinning rates up to 100 kHz. In spite of the miniscule sample volume (290 nL), high-quality HSQC-type spectra of bulk samples as well as surface-bound molecules can be obtained within hours of experimental time. The experiments are compared with those carried out at 40 kHz MAS using a 1.6-mm probe, which offered higher overall sensitivity due to a larger rotor volume. The benefits of ultrafast MAS in such experiments include superior resolution in 1H dimension without resorting to 1H–1H homonuclear RF decoupling, easy optimization, and applicability to mass-limited samples. As a result, the HMQC spectra of surface-bound species can be also acquired under 100 kHz MAS, although the dephasing of transverse magnetization has significant effect on the efficiency transfer under MAS alone.

  16. Studies of minute quantities of natural abundance molecules using 2D heteronuclear correlation spectroscopy under 100kHz MAS

    DOE PAGESBeta

    Nishiyama, Y.; Kobayashi, T.; Malon, M.; Singappuli-Arachchige, D.; Slowing, I. I.; Pruski, M.

    2015-02-16

    Two-dimensional 1H{13C} heteronuclear correlation solid-state NMR spectra of naturally abundant solid materials are presented, acquired using the 0.75-mm magic angle spinning (MAS) probe at spinning rates up to 100 kHz. In spite of the miniscule sample volume (290 nL), high-quality HSQC-type spectra of bulk samples as well as surface-bound molecules can be obtained within hours of experimental time. The experiments are compared with those carried out at 40 kHz MAS using a 1.6-mm probe, which offered higher overall sensitivity due to a larger rotor volume. The benefits of ultrafast MAS in such experiments include superior resolution in 1H dimensionmore » without resorting to 1H–1H homonuclear RF decoupling, easy optimization, and applicability to mass-limited samples. As a result, the HMQC spectra of surface-bound species can be also acquired under 100 kHz MAS, although the dephasing of transverse magnetization has significant effect on the efficiency transfer under MAS alone.« less

  17. Implications of scaled δ15N fractionation for community predator-prey body mass ratio estimates in size-structured food webs.

    PubMed

    Reum, Jonathan C P; Jennings, Simon; Hunsicker, Mary E

    2015-11-01

    Nitrogen stable isotope ratios (δ(15) N) may be used to estimate community-level relationships between trophic level (TL) and body size in size-structured food webs and hence the mean predator to prey body mass ratio (PPMR). In turn, PPMR is used to estimate mean food chain length, trophic transfer efficiency and rates of change in abundance with body mass (usually reported as slopes of size spectra) and to calibrate and validate food web models. When estimating TL, researchers had assumed that fractionation of δ(15) N (Δδ(15) N) did not change with TL. However, a recent meta-analysis indicated that this assumption was not as well supported by data as the assumption that Δδ(15) N scales negatively with the δ(15) N of prey. We collated existing fish community δ(15) N-body size data for the Northeast Atlantic and tropical Western Arabian Sea with new data from the Northeast Pacific. These data were used to estimate TL-body mass relationships and PPMR under constant and scaled Δδ(15) N assumptions, and to assess how the scaled Δδ(15) N assumption affects our understanding of the structure of these food webs. Adoption of the scaled Δδ(15) N approach markedly reduces the previously reported differences in TL at body mass among fish communities from different regions. With scaled Δδ(15) N, TL-body mass relationships became more positive and PPMR fell. Results implied that realized prey size in these size-structured fish communities are less variable than previously assumed and food chains potentially longer. The adoption of generic PPMR estimates for calibration and validation of size-based fish community models is better supported than hitherto assumed, but predicted slopes of community size spectra are more sensitive to a given change or error in realized PPMR when PPMR is small. PMID:26046788

  18. Identifying diazotrophs by incorporation of nitrogen from (15)N(2) into RNA.

    PubMed

    Addison, Sarah L; McDonald, Ian R; Lloyd-Jones, Gareth

    2010-08-01

    The diversity and abundance of active diazotrophs was investigated in a New Zealand pulp and paper wastewater by enrichment with (15)N(2). Purified (15)N-RNA was analysed by reverse transcription, molecular cloning and sequence analysis of 16S rRNA to reveal a diverse community of bacteria as indicated by a Shannon Weaver Index value of > 2.8. The major class represented in the enriched culture were the gamma-Proteobacteria at 85% with a secondary group of the phylum Firmicutes present at 8.2%, the remaining sequences were affiliated with the alpha- and beta-Proteobacterial classes (1.4% and 4.3%, respectively). Three dominant genera, Aeromonas, Pseudomonas and Bacillus, were identified by comparison with published sequences and phylogenetic analysis. To confirm that representatives of the taxonomic groups identified from the active enriched nitrogen-fixing community were capable of fixing nitrogen Aeromonas and Pseudomonas species were cultivated and shown to possess nifH genes. In wastewater, fluorescence in situ hybridisation probing revealed that the dominant nitrogen-fixing population identified in this study were present in the population, but at lower levels. The population is, therefore, reliant on a small sub-population of diazotrophs to supply the community's nitrogen needs above that already present in the wastewater. PMID:20582411

  19. Acceleration of natural-abundance solid-state MAS NMR measurements on bone by paramagnetic relaxation from gadolinium-DTPA

    NASA Astrophysics Data System (ADS)

    Mroue, Kamal H.; Zhang, Rongchun; Zhu, Peizhi; McNerny, Erin; Kohn, David H.; Morris, Michael D.; Ramamoorthy, Ayyalusamy

    2014-07-01

    Reducing the data collection time without affecting the signal intensity and spectral resolution is one of the major challenges for the widespread application of multidimensional nuclear magnetic resonance (NMR) spectroscopy, especially in experiments conducted on complex heterogeneous biological systems such as bone. In most of these experiments, the NMR data collection time is ultimately governed by the proton spin-lattice relaxation times (T1). For over two decades, gadolinium(III)-DTPA (Gd-DTPA, DTPA = Diethylene triamine pentaacetic acid) has been one of the most widely used contrast-enhancement agents in magnetic resonance imaging (MRI). In this study, we demonstrate that Gd-DTPA can also be effectively used to enhance the longitudinal relaxation rates of protons in solid-state NMR experiments conducted on bone without significant line-broadening and chemical-shift-perturbation side effects. Using bovine cortical bone samples incubated in different concentrations of Gd-DTPA complex, the 1H T1 values were calculated from data collected by 1H spin-inversion recovery method detected in natural-abundance 13C cross-polarization magic angle spinning (CPMAS) NMR experiments. Our results reveal that the 1H T1 values can be successfully reduced by a factor of 3.5 using as low as 10 mM Gd-DTPA without reducing the spectral resolution and thus enabling faster data acquisition of the 13C CPMAS spectra. These results obtained from 13C-detected CPMAS experiments were further confirmed using 1H-detected ultrafast MAS experiments on Gd-DTPA doped bone samples. This approach considerably improves the signal-to-noise ratio per unit time of NMR experiments applied to bone samples by reducing the experimental time required to acquire the same number of scans.

  20. Cumulative fission yields of short-lived isotopes under natural-abundance-boron-carbide-moderated neutron spectrum

    SciTech Connect

    Finn, Erin C.; Metz, Lori A.; Greenwood, Lawrence R.; Pierson, Bruce; Wittman, Richard S.; Friese, Judah I.; Kephart, Rosara F.

    2015-04-09

    The availability of gamma spectroscopy data on samples containing mixed fission products at short times after irradiation is limited. Due to this limitation, data interpretation methods for gamma spectra of mixed fission product samples, where the individual fission products have not been chemically isolated from interferences, are not well-developed. The limitation is particularly pronounced for fast pooled neutron spectra because of the lack of available fast reactors in the United States. Samples containing the actinide isotopes 233, 235, 238U, 237Np, and 239Pu individually were subjected to a 2$ pulse in the Washington State University 1 MW TRIGA reactor. To achieve a fission-energy neutron spectrum, the spectrum was tailored using a natural abundance boron carbide capsule to absorb neutrons in the thermal and epithermal region of the spectrum. Our tailored neutron spectrum is unique to the WSU reactor facility, consisting of a soft fission spectrum that contains some measurable flux in the resonance region. This results in a neutron spectrum at greater than 0.1 keV with an average energy of 70 keV, similar to fast reactor spectra and approaching that of 235U fission. Unique fission product gamma spectra were collected from 4 minutes to 1 week after fission using single-crystal high purity germanium detectors. Cumulative fission product yields measured in the current work generally agree with published fast pooled fission product yield values from ENDF/B-VII, though a bias was noted for 239Pu. The present work contributes to the compilation of energy-resolved fission product yield nuclear data for nuclear forensic purposes.

  1. Using natural abundance radiocarbon to trace the flux of petrocarbon to the seafloor following the Deepwater Horizon oil spill.

    PubMed

    Chanton, Jeffrey; Zhao, Tingting; Rosenheim, Brad E; Joye, Samantha; Bosman, Samantha; Brunner, Charlotte; Yeager, Kevin M; Diercks, Arne R; Hollander, David

    2015-01-20

    In 2010, the Deepwater Horizon accident released 4.6–6.0 × 10(11) grams or 4.1 to 4.6 million barrels of fossil petroleum derived carbon (petrocarbon) as oil into the Gulf of Mexico. Natural abundance radiocarbon measurements on surface sediment organic matter in a 2.4 × 10(10) m(2) deep-water region surrounding the spill site indicate the deposition of a fossil-carbon containing layer that included 1.6 to 2.6 × 10(10) grams of oil-derived carbon. This quantity represents between 0.5 to 9.1% of the released petrocarbon, with a best estimate of 3.0–4.9%. These values may be lower limit estimates of the fraction of the oil that was deposited on the seafloor because they focus on a limited mostly deep-water area of the Gulf, include a conservative estimate of thickness of the depositional layer, and use an average background or prespill radiocarbon value for sedimentary organic carbon that produces a conservative value. A similar approach using hopane tracer estimated that 4–31% of 2 million barrels of oil that stayed in the deep sea settled on the bottom. Converting that to a percentage of the total oil that entered into the environment (to which we normalized our estimate) converts this range to 1.8 to 14.4%. Although extrapolated over a larger area, our independent estimate produced similar values. PMID:25494527

  2. An sup 15 N NMR method for the characterization of organic sulfur in coal and coal products via iminosulfurane formation

    SciTech Connect

    Franz, J.A.; Lamb, C.N.; Linehan, J.C.

    1991-09-01

    The indirect of organic sulfur by {sup 15}N NMR spectroscopy in the solid state is feasible by facile reactions providing the iminosulfurane structures. Unfortunately, nitrogen chemical shifts appear to be insufficiently sensitive to the nature of the sulfur substituent to be useful for structural studies. Further work is underway to determine the {sup 15}N chemical shifts of iminosulfuranes formed from dibenzothiophene, 4-4{prime}-dimethoxydiphenyl sulfide, and a sulfur-containing, methylated asphaltene to determine the sensitivity of {sup 15}N shifts to a broader variation of aromatic structure. Although double cross-polarization experiments or rotational echo experiments could make use of iminosulfurane formation for detection of carbon in proximity to sulfur, the difficulties in quantitation using these methods are not encouraging for coal product mixtures. 6 refs., 1 fig., 1 tab.

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

    NASA Astrophysics Data System (ADS)

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

    2011-11-01

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

  4. Toward hyperpolarized molecular imaging of HIV: synthesis and longitudinal relaxation properties of (15) N-Azidothymidine.

    PubMed

    Shchepin, Roman V; Chekmenev, Eduard Y

    2014-08-01

    Previously unreported (15) N labeled Azidothymidine (AZT) was prepared as an equimolar mixture of two isotopomers: 1-(15) N-AZT and 3-(15) N-AZT. Polarization decay of (15) N NMR signal was studied in high (9.4?T) and low (~50?mT) magnetic fields. (15) N T1 values were 45??5?s (1-(15) N-AZT) and 37??2?s (3-(15) N-AZT) at 9.4?T, and 140??16?s (3-(15) N-AZT) at 50?mT. (15) N-AZT can be potentially (15) N hyperpolarized by several methods. These sufficiently long (15) N-AZT T1 values potentially enable hyperpolarized in vivo imaging of (15) N-AZT, because of the known favorable efficient (i.e., of the time scale shorter than the longest reported here (15) N T1 ) kinetics of uptake of injected AZT. Therefore, 3-(15) N-AZT can be potentially used for HIV molecular imaging using hyperpolarized magnetic resonance imaging. PMID:25156931

  5. (15)N in tree rings as a bio-indicator of changing nitrogen cycling in tropical forests: an evaluation at three sites using two sampling methods.

    PubMed

    van der Sleen, Peter; Vlam, Mart; Groenendijk, Peter; Anten, Niels P R; Bongers, Frans; Bunyavejchewin, Sarayudh; Hietz, Peter; Pons, Thijs L; Zuidema, Pieter A

    2015-01-01

    Anthropogenic nitrogen deposition is currently causing a more than twofold increase of reactive nitrogen input over large areas in the tropics. Elevated (15)N abundance (δ(15)N) in the growth rings of some tropical trees has been hypothesized to reflect an increased leaching of (15)N-depleted nitrate from the soil, following anthropogenic nitrogen deposition over the last decades. To find further evidence for altered nitrogen cycling in tropical forests, we measured long-term δ(15)N values in trees from Bolivia, Cameroon, and Thailand. We used two different sampling methods. In the first, wood samples were taken in a conventional way: from the pith to the bark across the stem of 28 large trees (the "radial" method). In the second, δ(15)N values were compared across a fixed diameter (the "fixed-diameter" method). We sampled 400 trees that differed widely in size, but measured δ(15)N in the stem around the same diameter (20 cm dbh) in all trees. As a result, the growth rings formed around this diameter differed in age and allowed a comparison of δ(15)N values over time with an explicit control for potential size-effects on δ(15)N values. We found a significant increase of tree-ring δ(15)N across the stem radius of large trees from Bolivia and Cameroon, but no change in tree-ring δ(15)N values over time was found in any of the study sites when controlling for tree size. This suggests that radial trends of δ(15)N values within trees reflect tree ontogeny (size development). However, for the trees from Cameroon and Thailand, a low statistical power in the fixed-diameter method prevents to conclude this with high certainty. For the trees from Bolivia, statistical power in the fixed-diameter method was high, showing that the temporal trend in tree-ring δ(15)N values in the radial method is primarily caused by tree ontogeny and unlikely by a change in nitrogen cycling. We therefore stress to account for tree size before tree-ring δ(15)N values can be properly interpreted. PMID:25914707

  6. 15N in tree rings as a bio-indicator of changing nitrogen cycling in tropical forests: an evaluation at three sites using two sampling methods

    PubMed Central

    van der Sleen, Peter; Vlam, Mart; Groenendijk, Peter; Anten, Niels P. R.; Bongers, Frans; Bunyavejchewin, Sarayudh; Hietz, Peter; Pons, Thijs L.; Zuidema, Pieter A.

    2015-01-01

    Anthropogenic nitrogen deposition is currently causing a more than twofold increase of reactive nitrogen input over large areas in the tropics. Elevated 15N abundance15N) in the growth rings of some tropical trees has been hypothesized to reflect an increased leaching of 15N-depleted nitrate from the soil, following anthropogenic nitrogen deposition over the last decades. To find further evidence for altered nitrogen cycling in tropical forests, we measured long-term δ15N values in trees from Bolivia, Cameroon, and Thailand. We used two different sampling methods. In the first, wood samples were taken in a conventional way: from the pith to the bark across the stem of 28 large trees (the “radial” method). In the second, δ15N values were compared across a fixed diameter (the “fixed-diameter” method). We sampled 400 trees that differed widely in size, but measured δ15N in the stem around the same diameter (20 cm dbh) in all trees. As a result, the growth rings formed around this diameter differed in age and allowed a comparison of δ15N values over time with an explicit control for potential size-effects on δ15N values. We found a significant increase of tree-ring δ15N across the stem radius of large trees from Bolivia and Cameroon, but no change in tree-ring δ15N values over time was found in any of the study sites when controlling for tree size. This suggests that radial trends of δ15N values within trees reflect tree ontogeny (size development). However, for the trees from Cameroon and Thailand, a low statistical power in the fixed-diameter method prevents to conclude this with high certainty. For the trees from Bolivia, statistical power in the fixed-diameter method was high, showing that the temporal trend in tree-ring δ15N values in the radial method is primarily caused by tree ontogeny and unlikely by a change in nitrogen cycling. We therefore stress to account for tree size before tree-ring δ15N values can be properly interpreted. PMID:25914707

  7. N-15 NMR spectra of naturally abundant nitrogen in soil and aquatic natural organic matter samples of the International Humic Substances Society

    SciTech Connect

    Thorn, Kevin A.; Cox, Larry G.

    2009-02-28

    The naturally abundant nitrogen in soil and aquatic NOM samples from the International Humic Substances Society has been characterized by solid state CP/MAS ¹⁵N NMR. Soil samples include humic and fulvic acids from the Elliot soil, Minnesota Waskish peat and Florida Pahokee peat, as well as the Summit Hill soil humic acid and the Leonardite humic acid. Aquatic samples include Suwannee River humic, fulvic and reverse osmosis isolates, Nordic humic and fulvic acids and Pony Lake fulvic acid. Additionally, Nordic and Suwannee River XAD-4 acids and Suwannee River hydrophobic neutral fractions were analyzed. Similar to literature reports, amide/aminoquinone nitrogens comprised the major peaks in the solid state spectra of the soil humic and fulvic acids, along with heterocyclic and amino sugar/terminal amino acid nitrogens. Spectra of aquatic samples, including the XAD-4 acids, contain resolved heterocyclic nitrogen peaks in addition to the amide nitrogens. The spectrum of the nitrogen enriched, microbially derived Pony Lake, Antarctica fulvic acid, appeared to contain resonances in the region of pyrazine, imine and/or pyridine nitrogens, which have not been observed previously in soil or aquatic humic substances by ¹⁵N NMR. Liquid state ¹⁵N NMR experiments were also recorded on the Elliot soil humic acid and Pony Lake fulvic acid, both to examine the feasibility of the techniques, and to determine whether improvements in resolution over the solid state could be realized. For both samples, polarization transfer (DEPT) and indirect detection (¹H–¹⁵N gHSQC) spectra revealed greater resolution among nitrogens directly bonded to protons. The amide/aminoquinone nitrogens could also be observed by direct detection experiments.

  8. Natural abundance (δ¹⁵N) indicates shifts in nitrogen relations of woody taxa along a savanna-woodland continental rainfall gradient.

    PubMed

    Soper, Fiona M; Richards, Anna E; Siddique, Ilyas; Aidar, Marcos P M; Cook, Garry D; Hutley, Lindsay B; Robinson, Nicole; Schmidt, Susanne

    2015-05-01

    Water and nitrogen (N) interact to influence soil N cycling and plant N acquisition. We studied indices of soil N availability and acquisition by woody plant taxa with distinct nutritional specialisations along a north Australian rainfall gradient from monsoonal savanna (1,600-1,300 mm annual rainfall) to semi-arid woodland (600-250 mm). Aridity resulted in increased 'openness' of N cycling, indicated by increasing δ(15)N(soil) and nitrate:ammonium ratios, as plant communities transitioned from N to water limitation. In this context, we tested the hypothesis that δ(15)N(root) xylem sap provides a more direct measure of plant N acquisition than δ(15)N(foliage). We found highly variable offsets between δ(15)N(foliage) and δ(15)N(root) xylem sap, both between taxa at a single site (1.3-3.4 ‰) and within taxa across sites (0.8-3.4 ‰). As a result, δ(15)N(foliage) overlapped between N-fixing Acacia and non-fixing Eucalyptus/Corymbia and could not be used to reliably identify biological N fixation (BNF). However, Acacia δ(15)N(root) xylem sap indicated a decline in BNF with aridity corroborated by absence of root nodules and increasing xylem sap nitrate concentrations and consistent with shifting resource limitation. Acacia dominance at arid sites may be attributed to flexibility in N acquisition rather than BNF capacity. δ(15)N(root) xylem sap showed no evidence of shifting N acquisition in non-mycorrhizal Hakea/Grevillea and indicated only minor shifts in Eucalyptus/Corymbia consistent with enrichment of δ(15)N(soil) and/or decreasing mycorrhizal colonisation with aridity. We propose that δ(15)N(root) xylem sap is a more direct indicator of N source than δ(15)N(foliage), with calibration required before it could be applied to quantify BNF. PMID:25502440

  9. Coronal Abundances

    NASA Astrophysics Data System (ADS)

    Fludra, A.; Saba, J. L. R.; Henoux, J.-C.; Murphy, R. J.; Reames, D. V.; Lemen, J. R.; Strong, K. T.; Sylwester, J.; Widing, K. G.

    Flare X-Ray Measurements from BCS Calcium Abundance Fe/H and Fe/Ca Abundance Relative Abundances of Ar. Ca. and Fe in Flares Factors Affecting Abundance Determinations from X-Ray Spectra FCS Abundances FCS Active-Region Abundances Abundance Variability in Active Regions Impact of Resonance Scattering Assessment of FCS Active-Region Abundance Results FCS Flare Abundance Studies Coronal CI/S and Ar/S Measurements Dem Studies of Flare Abuncances Determination of Solar Abundances by Solar Flare γ-Ray Spectrometry γ-Ray Spectral Analysis γ-Ray Results Solar Energetic Particles Major Proton Events CIR Events from Coronal Holes Impulsive Flare Events Theory of Abundance Fractionation Gravitational Settling Pressure Gradient and Stationary Diffusion Ion-Neutral Separation Due to Currents Ion-Neutral Separation Due to Electromagnetic Forces Discussion Summary

  10. Estimating modal abundances from the spectra of natural and laboratory pyroxene mixtures using the modified Gaussian model

    NASA Technical Reports Server (NTRS)

    Sunshine, Jessica M.; Pieters, Carle M.

    1993-01-01

    The modified Gaussian model (MGM) is used to explore spectra of samples containing multiple pyroxene components as a function of modal abundance. The MGM allows spectra to be analyzed directly, without the use of actual or assumed end-member spectra and therefore holds great promise for remote applications. A series of mass fraction mixtures created from several different particle size fractions are analyzed with the MGM to quantify the properties of pyroxene mixtures as a function of both modal abundance and grain size. Band centers, band widths, and relative band strengths of absorptions from individual pyroxenes in mixture spectra are found to be largely independent of particle size. Spectral properties of both zoned and exsolved pyroxene components are resolved in exsolved samples using the MGM, and modal abundances are accurately estimated to within 5-10 percent without predetermined knowledge of the end-member spectra.

  11. Bacterial abundance and aerobic microbial activity across natural and oyster aquaculture habitats during summer conditions in a northeastern Pacific estuary.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We measured sediment properties and the abundance and functional diversity of microbes in Willapa Bay, Washington, USA, to test the response of sediment microbes to oyster aquaculture. Sites spanned the estuary gradient (salinity 24-30) and six different habitat types: eelgrass (Zostera marina), uns...

  12. The metabolic effect of resistant starch and yoghurt on the renal and faecal nitrogen and ammonia excretion in humans as measured by lactose-[(15)N2]ureide.

    PubMed

    Wutzke, Klaus D; Scholübbers, Debora

    2013-01-01

    Resistant starch (RS) and Lactobacillus acidophilus yoghurt (LC1) were supplemented simultaneously in healthy adults to evaluate the effect on the urinary and faecal nitrogen and ammonia excretion by means of lactose-[(15)N2]ureide ((15)N-LU) degradation. Nineteen subjects received a regular daily diet either without or with supplementation of an RS-LC1-mixture composed of fibre of potatoes (RS type 1), wrinkle pea starch (RS type 2), and LC1 over a 20-day period in randomised order. Thereafter, (15)N-LU was administered together with breakfast. Urine and faeces were collected over a period of 48 and 72 h, respectively. The (15)N abundances were measured by isotope ratio mass spectrometry. The intake of the pre- and probiotic mixture composed of RS of type 1, type 2 and of LC1 significantly lowered the colonic generation and the renal excretion of toxic (15)NH3 and functioned as an ammonia shift from urinary to faecal (15)N excretion when using (15)N-LU as a xenobiotic marker. PMID:23937067

  13. Advancements in natural abundance solid-state 33S MAS NMR: characterization of transition-metal M=S bonds in ammonium tetrathiometallates.

    PubMed

    Jakobsen, Hans J; Hove, Anders R; Bildsøe, Henrik; Skibsted, Jørgen; Brorson, Michael

    2007-04-28

    We report the first (33)S chemical shift anisotropy (CSA) data as obtained from a combined determination of (33)S CSA and quadrupole coupling parameters utilizing the observation of both the (33)S (I = 3/2) central and satellite transitions in a natural abundance (33)S MAS NMR study aimed at characterizing the two important tetrathiometallates (NH4)(2)MoS(4) and (NH4)(2)WS(4). PMID:17530082

  14. Using PLFA Biomarkers and Natural Abundance Stable and Radiocarbon Isotopes to Characterize the Microbial Ecology and Metabolism of Methane Cycling

    NASA Astrophysics Data System (ADS)

    Mills, C. T.; Mandernack, K. W.; Slater, G. F.; Dias, R. F.

    2008-12-01

    Methane generated in the subsurface is a major source of atmospheric CH4, but its release is mitigated by CH4-oxidizing bacteria (methanotrophs). Therefore, it is important to understand the ecology of methanotroph communities in various environments. Phospholipid fatty acid (PLFA) analyses are a particularly useful method for characterizing these communities for two reasons: (1) Many type I and II methanotrophs produce specific PLFA biomarkers that can be used to estimate their populations, and (2) because CH4 is often very depleted in 13C and sometimes 14C, natural abundance δ13CPLFA and Δ14CPLFA values can be used to trace the flow of CH4- derived carbon through microbial ecosystems. We used these tools to evaluate the role of methanotrophs in carbon flow in three different environments: (1) a soil column overlying a coal bed methane (CBM) seep in southwest CO, and pristine, oligotrophic groundwaters within (2) sedimentary and (3) granitic host rocks in Japan. In the soil column impacted by CBM seepage, concentrations of the biomarker PLFAs for type I (16:1ω8cis) and type II (18:1ω8cis) methanotrophs were as high as 13 and 18 nmoles (g dry soil)-1, respectively. Depth profiles of methanotroph PLFA concentrations varied over different sampling dates indicating dynamic populations. δ13CPLFA values of the CBM soils (-25.1 to - 66.9‰) were substantially more negative than those for the control soil (-14.5 to -32.5‰) indicating that CBM is an important carbon source for the CBM-impacted soil microbial community. Δ14CPLFA values (-351 to -936‰) indicate the importance of 14C-dead CBM as a carbon source to the microbial communities, contributing 32 to 66% of total carbon in PLFA structures isolated from shallow soils and 67 to 97% for those isolated from deeper soils. The biomarker for type II methanotrophs, comprised 3 and 18% of total PLFAs in sedimentary and granitic groundwaters, respectively. The Δ14C values determined for type II methanotroph PLFAs in the sedimentary (- 861‰) and granite (-867‰) waters were very similar to the Δ14C values of dissolved inorganic carbon (DIC) in each water (ca -850‰) suggesting that type II methanotrophs ultimately derive all of their carbon from DIC. In contrast, δ13C values of type II PLFAs in the sedimentary (- 93‰) and granite (-60‰) waters indicate that these organisms use different carbon assimilation schemes in each environment. These studies show the utility of PLFA biomarkers and δ13CPLFA and Δ14CPLFA values to characterize the in situ metabolisms of methanotrophic bacteria and overall CH4 recycling in diverse environments.

  15. Natural Carbon Isotope Abundance of Plasma Metabolites and Liver Tissue Differs between Diabetic and Non-Diabetic Zucker Diabetic Fatty Rats

    PubMed Central

    Godin, Jean-Philippe; Ross, Alastair B.; Cléroux, Marilyn; Pouteau, Etienne; Montoliu, Ivan; Moser, Mireille; Kochhar, Sunil

    2013-01-01

    Background ‘You are what you eat’ is an accurate summary for humans and animals when it comes to carbon isotope abundance. In biological material, natural13C/12C ratio is subject to minute variations due to diet composition (mainly from ingestion of C3 and C4 metabolism plants) and to the discrimination between ‘light’ and ‘heavy’ isotopes during biochemical reactions (isotope effects and isotopic fractionation). Methodology/Principal Findings Carbon isotopic abundance was measured in ZDF (fa/+) and ZDF (fa/fa), (lean and obese-diabetic rats respectively) fed the same diet. By analysing plasma metabolites (glucose and non-esterified fatty acids), breath and liver tissue by high-precision isotope ratio mass spectrometry, we demonstrate for the first time statistically distinguishable metabolic carbon isotope abundance between ZDF (fa/+) and ZDF (fa/fa) rats based on plasma glucose, palmitic, oleic, linoleic, arachidonic acids and bulk analysis of liver tissue (P<0.005) resulting into clear isotopic fingerprints using principal component analysis. We studied the variation of isotopic abundance between both groups for each metabolite and through the metabolic pathways using the precursor/product approach. We confirmed that lipids were depleted in 13C compared to glucose in both genotypes. We found that isotopic abundance of linoleic acid (C18: 2n-6), even though both groups had the same feed, differed significantly between both groups. The likely reason for these changes between ZDF (fa/+) and ZDF (fa/fa) are metabolic dysregulation associated with various routing and fluxes of metabolites. Conclusion/Significance This work provides evidence that measurement of natural abundance isotope ratio of both bulk tissue and individual metabolites can provide meaningful information about metabolic changes either associated to phenotype or to genetic effects; irrespective of concentration. In the future measuring the natural abundance δ13C of key metabolites could be used as endpoints for studying in vivo metabolism, especially with regards to metabolic dysregulation, and development and progression of metabolic diseases. PMID:24086387

  16. Compound specific amino acid δ15N in marine sediments: A new approach for studies of the marine nitrogen cycle

    NASA Astrophysics Data System (ADS)

    Batista, Fabian C.; Ravelo, A. Christina; Crusius, John; Casso, Michael A.; McCarthy, Matthew D.

    2014-10-01

    The nitrogen (N) isotopic composition (δ15N) of bulk sedimentary N (δ15Nbulk) is a common tool for studying past biogeochemical cycling in the paleoceanographic record. Empirical evidence suggests that natural fluctuations in the δ15N of surface nutrient N are reflected in the δ15N of exported planktonic biomass and in sedimentary δ15Nbulk. However, δ15Nbulk is an analysis of total combustible sedimentary N, and therefore also includes mixtures of N sources and/or selective removal or preservation of N-containing compounds. Compound-specific nitrogen isotope analyses of individual amino acids (δ15NAA) are novel measurements with the potential to decouple δ15N changes in nutrient N from trophic effects, two main processes that can influence δ15Nbulk records. As a proof of concept study to examine how δ15NAA can be applied in marine sedimentary systems, we compare the δ15NAA signatures of surface and sinking POM sources with shallow surface sediments from the Santa Barbara Basin, a sub-oxic depositional environmental that exhibits excellent preservation of sedimentary organic matter. Our results demonstrate that δ15NAA signatures of both planktonic biomass and sinking POM are well preserved in such surface sediments. However, we also observed an unexpected inverse correlation between δ15N value of phenylalanine (δ15NPhe; the best AA proxy for N isotopic value at the base of the food web) and calculated trophic position. We used a simple N isotope mass balance model to confirm that over long time scales, δ15NPhe values should in fact be directly dependent on shifts in ecosystem trophic position. While this result may appear incongruent with current applications of δ15NAA in food webs, it is consistent with expectations that paleoarchives will integrate N dynamics over much longer timescales. We therefore propose that for paleoceanographic applications, key δ15NAA parameters are ecosystem trophic position, which determines relative partitioning of 15N into source AA versus trophic AA pools, and the integrated δ15NAA of all common protein AA (δ15NTHAA), which serves as a proxy for the δ15N of nutrient N. Together, we suggest that these can provide a coupled picture of regime shifts in planktonic ecosystem structure, δ15N at the base of food webs, and possibly additional information about nutrient dynamics.

  17. Using δ15N of Chironomidae as an index of nitrogen sources and processing within watersheds as part of EPA's National Aquatic Resource Surveys

    NASA Astrophysics Data System (ADS)

    Brooks, J. R.; Compton, J.; Herlihy, A.; Sobota, D. J.; Stoddard, J.; Weber, M.

    2014-12-01

    Nitrogen (N) removal in watersheds is an important regulating ecosystem service that can help reduce N pollution in the nation's waterways. However, processes that remove N such as denitrification are generally determined at point locations. Measures that integrate N processing within watersheds and over time would be particularly useful for assessing the degree of this vital service. Because most N removal processes isotopically enrich the N remaining, δ15N from basal food-chain organisms in aquatic ecosystems can provide information on watershed N processing. As part of EPA's National Aquatic Resource Surveys (NARS), we measured δ15N of Chironomidae in lakes, rivers and streams because these larval aquatic insects were found in abundance in almost every lake and stream in the U.S. Using information on nitrogen loading to the watershed, and total N concentrations within the water, we assessed when elevated chironomid δ15N would indicate N removal rather than possible enriched sources of N. Chironomid δ15N values ranged from -4 to +20 ‰, and were higher in rivers and streams than in lakes (median = 7.6 ‰ vs. 4.8 ‰, respectively), indicating that N was processed to a greater degree in lotic chironomids than in lentic ones. For both, δ15N increased with watershed-level agricultural land cover and N loading, and decreased as precipitation increased. In rivers and streams with high synthetic N loading, we found lower N concentrations in streams with higher chironomid δ15N values, suggesting greater N removal. At low levels of synthetic N loading, the pattern reversed, and streams with enriched chironomid δ15N had higher N concentrations, suggesting enriched sources such as manure or sewage. Our results indicate that chironomid δ15N values can provide valuable information about watershed-level N inputs and processing for national water quality monitoring efforts.

  18. 15N2 formation and fast oxygen isotope exchange during pulsed 15N18O exposure of MnOx/CeO2

    SciTech Connect

    Kwak, Ja Hun; Szanyi, Janos

    2014-12-23

    Pulsing 15N18O onto an annealed 1% Mn16Ox/Ce16O2 catalyst resulted in very fast oxygen isotope exchange and 15N2 formation at 295 K. In the 1st 15N18O pulse, due to the presence of large number of surface oxygen defects, extensive 15N218O and 15N2 formations were observed. In subsequent pulses oxygen isotope exchange dominated as a result of highly labile oxygen in the oxide. We gratefully acknowledge the US Department of Energy (DOE), Office of Energy Efficiency and Renewable Energy/Vehicle Technologies Program for the support of this work. The research described in this paper was performed at the Environmental Molecular Sciences Laboratory (EMSL), a national scientific user facility sponsored by the DOE’s Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory (PNNL). PNNL is operated for the US DOE by Battelle.

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

    NASA Astrophysics Data System (ADS)

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

    1997-11-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2011-11-01

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

  1. Trophic guilds of generalist feeders in soil animal communities as indicated by stable isotope analysis (15N/14N).

    PubMed

    Oelbermann, K; Scheu, S

    2010-10-01

    We investigated if the commonly used aggregation of organisms into trophic guilds, such as detritivores and predators, in fact represent distinct trophic levels. Soil arthropods of a forest-meadow transect were ascribed a priori to trophic guilds (herbivores, detritivores, predators and necrovores), which are often used as an equivalent to trophic levels. We analysed natural variations in 15N/14N ratios of the animals in order to investigate the trophic similarity of organisms within (a priori defined) trophic guilds. Using trophic guilds as an equivalent to trophic level, the assumed stepwise enrichment of 15N by 3.4 per thousand per trophic level did not apply to detritivores; they were only enriched in 15N by on average 1.5 per thousand compared to litter materials. Predators on average were enriched in 15N by 3.5 per thousand compared to detritivores. Within detritvores and predators delta15N signatures varied markedly, indicating that these trophic guilds are dominated by generalist feeders which form a gradient of organisms feeding on different resources. The results indicate that commonly used trophic guilds, in particular detritivores and predators, do not represent trophic levels but consist of subguilds, i.e. subsets of organisms differing in resource utilization. In particular, in soil and litter food webs where trophic level omnivory is common, the use of distinct trophic levels may be inappropriate. Guilds of species delineated by natural variations of stable isotope ratios are assumed to more adequately represent the structure of litter and soil food webs allowing a more detailed understanding of their functioning. PMID:20109270

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

    NASA Astrophysics Data System (ADS)

    Welti, N.; Bondar-Kunze, E.; Mair, M.; Bonin, P.; Wanek, W.; Pinay, G.; Hein, T.

    2012-11-01

    Floodplain restoration changes the nitrate delivery pattern and dissolved organic matter pool in backwaters, though the effects these changes have are not yet well known. We performed two mesocosm experiments on floodplain sediments to quantify the nitrate metabolism in two types of floodplains. Rates of denitrification, dissimilatory nitrate reduction to ammonium (DNRA) and anammox were measured using 15N-NO3 tracer additions in mesocosms of undisturbed floodplain sediments originating from (1) restored and (2) disconnected sites in the Alluvial Zone National Park on the Danube River downstream of Vienna, Austria. DNRA rates were an order of magnitude lower than denitrification and neither rate was affected by changes in nitrate delivery pattern or organic matter quality. Anammox was not detected at any of the sites. Denitrification was out-competed by assimilation, which was estimated to use up to 70% of the available nitrate. Overall, denitrification was higher in the restored sites, with mean rates of 5.7 ± 2.8 mmol N m-2 h-1 compared to the disconnected site (0.6 ± 0.5 mmol N m-2 h-1). In addition, ratios of N2O : N2 were lower in the restored site indicating a more complete denitrification. Nitrate addition had neither an effect on denitrification, nor on the N2O : N2 ratio. However, DOM (dissolved organic matter) quality significantly changed the N2O : N2 ratio in both sites. Addition of riverine-derived organic matter lowered the N2O : N2 ratio in the disconnected site, whereas addition of floodplain-derived organic matter increased the N2O : N2 ratio in the restored site. These results demonstrate that increasing floodplains hydrological connection to the main river channel increases nitrogen retention and decreases nitrous oxide emissions.

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

    PubMed

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

    2012-06-01

    Alternative metabolic pathways inside a cell can be deduced using stable isotopically labeled substrates. One prerequisite is accurate measurement of the labeling pattern of targeted metabolites. Experiments are generally limited to the use of single-element isotopes, mainly (13)C. Here, we demonstrate the application of direct infusion nanospray, ultrahigh-resolution Fourier transform ion cyclotron resonance-mass spectrometry (FTICR-MS) for metabolic studies using differently labeled elemental isotopes simultaneously--i.e., (13)C and (15)N--in amino acids of a total protein hydrolysate. The optimized strategy for the analysis of metabolism by a hybrid linear ion trap-FTICR-MS comprises the collection of multiple adjacent selected ion monitoring scans. By limiting both the width of the mass range and the number of ions entering the ICR cell with automated gain control, sensitive measurements of isotopologue distribution were possible without compromising mass accuracy and isotope intensity mapping. The required mass-resolving power of more than 60,000 is only achievable on a routine basis by FTICR and Orbitrap mass spectrometers. Evaluation of the method was carried out by comparison of the experimental data to the natural isotope abundances of selected amino acids and by comparison to GC/MS results obtained from a labeling experiment with (13)C-labeled glucose. The developed method was used to shed light on the complexity of the yeast Saccharomyces cerevisiae carbon-nitrogen co-metabolism by administering both (13)C-labeled glucose and (15)N-labeled alanine. The results indicate that not only glutamate but also alanine acts as an amino donor during alanine and valine synthesis. Metabolic studies using FTICR-MS can exploit new possibilities by the use of multiple-labeled elemental isotopes. PMID:22543713

  4. 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. PMID:24596018

  5. Patterns of diversity and abundance of carrion insect assemblages in the Natural Park "Hoces del Río Riaza" (central Spain).

    PubMed

    Baz, Arturo; Cifrián, Blanca; Martín-Vega, Daniel

    2014-01-01

    The patterns of diversity and abundance of the carrion insect species in the different habitats of the Natural Park "Hoces del Río Riaza" (central Spain) were studied with the use of carrion-baited traps. Representativeness of the inventories was assessed with the calculation of randomized species richness curves and nonparametric estimators. Coleoptera families, Silphidae and Dermestidae, and Diptera families, Calliphoridae and Muscidae, were dominant in every sampling habitat, but differences in the patterns of diversity and abundance were found. Lusitanian oakwood and riparian forest were the most diverse habitats with high abundance of saprophagous species, whereas more open (i.e., exposed to continuous sunlight during the day) habitats showed lower diversity values and a different species composition and distribution of species abundance, favoring thermophilous species and necrophagous species with high tolerance to different environmental conditions. Differences in the bioclimatical features of the sampled habitats are suggested to explain the composition and diversity of the carrion insect assemblages in different environments. PMID:25368080

  6. Nutrient regime shift in the western North Atlantic indicated by compound-specific δ15N of deep-sea gorgonian corals

    PubMed Central

    Sherwood, Owen A.; Lehmann, Moritz F.; Schubert, Carsten J.; Scott, David B.; McCarthy, Matthew D.

    2011-01-01

    Despite the importance of the nitrogen (N) cycle on marine productivity, little is known about variability in N sources and cycling in the ocean in relation to natural and anthropogenic climate change. Beyond the last few decades of scientific observation, knowledge depends largely on proxy records derived from nitrogen stable isotopes (δ15N) preserved in sediments and other bioarchives. Traditional bulk δ15N measurements, however, represent the combined influence of N source and subsequent trophic transfers, often confounding environmental interpretation. Recently, compound-specific analysis of individual amino acids (δ15N-AA) has been shown as a means to deconvolve trophic level versus N source effects on the δ15N variability of bulk organic matter. Here, we demonstrate the first use of δ15N-AA in a paleoceanographic study, through analysis of annually secreted growth rings preserved in the organic endoskeletons of deep-sea gorgonian corals. In the Northwest Atlantic off Nova Scotia, coral δ15N is correlated with increasing presence of subtropical versus subpolar slope waters over the twentieth century. By using the new δ15N-AA approach to control for variable trophic processing, we are able to interpret coral bulk δ15N values as a proxy for nitrate source and, hence, slope water source partitioning. We conclude that the persistence of the warm, nutrient-rich regime since the early 1970s is largely unique in the context of the last approximately 1,800 yr. This evidence suggests that nutrient variability in this region is coordinated with recent changes in global climate and underscores the broad potential of δ15N-AA for paleoceanographic studies of the marine N cycle. PMID:21199952

  7. 15N tracer studies of protein metabolism in low birth weight preterm infants: a comparison of 15N glycine and 15N yeast protein hydrolysate and of human milk- and formula-fed babies.

    PubMed

    Stack, T; Reeds, P J; Preston, T; Hay, S; Lloyd, D J; Aggett, P J

    1989-02-01

    Nitrogen flux and protein synthesis and degradation were estimated using a single oral bolus of 15N glycine or 15N yeast protein hydrolysate and measuring the 15N enrichment of urinary ammonia in five low birth wt infants fed a low birth wt formula and in six who were receiving their own mother's breast milk. Results derived from using 15N-glycine and 15N-yeast hydrolysate tracers in a randomized crossover study in 10 studies on seven infants showed, with one exception, higher turnover rates and more interindividual variation with the 15N yeast. Both tracers showed good reproducibility in two infants who had repeated studies. Although wt gain was similar in both groups, nitrogen intake and retention were greater (p less than 0.01) in the formula-fed group. Mean nitrogen turnover was similar in both groups, but there was a greater variance in the human milk-fed group which also had a greater nitrogen turnover/U absorbed nitrogen (p less than 0.025) and a lower excretion of nitrogen/U flux. PMID:2919131

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

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

    Soil organic matter (SOM) contains three times more carbon than in the atmosphere or terrestrial vegetation. This major pool of organic carbon is sensitive to climate change, but the mechanisms for carbon stabilization in soils are still not well understood and the ultimate potential for carbon stabilization is unknown. For predicting SOM dynamics, it is necessary to gain information on the turnover rates or stability of different soil organic carbon pools. The common method to determine stability and age of SOM is the 14C radio carbon technique, which is very expensive and therefore limited in use. Conen et al. (2008) developed a model to estimate the SOM stability based on the isotopic discrimination of 15N natural abundance by soil micro-organisms, and the decreasing C:N ratio during organic matter decomposition. This model has been developed for permanent grasslands in the Swiss Alps under steady-state conditions. The objective of our study was to validate whether this model could be used or adapted, in combination with 13C isotope signatures of SOM, to predict the relative age and stability of SOM fractions in more disturbed agricultural ecosystems. The present study was carried out on soils collected from six long-term experimental trials (from 12 to 50 years) under different agricultural management practices (e.g. no tillage vs conventional tillage, and mulch, fertilizer, green or animal manure application), located in Austria, Belgium, Kenya and China. Top and subsoil were sampled until 80-100 cm depth. Particulate organic matter (POM) fraction was obtained by wet sieving (> 63μm) after sonification and density separation (<1.8 g cm-3). Carbon and nitrogen contents and their stable isotopic ratios (i.e. 15N and 13C) were measured in POM and bulk soils. The mineral associated matter fraction (mOM), as the protected carbon, was calculated by difference to the bulk soil organic carbon. The relative age of the SOM was calculated using the Conen model and preliminary validated by 14C dating. At all sites, the POM has a higher C:N ratio and a lower δ15N signature compared to the mOM fraction. The POM in top soil layers (<30 cm) has a lower C:N ratio than in deep soil. The C:N ratio and δ15N of POM was influenced by agricultural management. The mOM fraction has 53 to 2063 times longer turnover rate than POM, the relative age of the SOM raised with increasing soil depth. The combination of the above results with δ13C data lead to a more comprehensive understanding of the processes underlying SOM dynamics. Tillage practices increased the bulk δ13C signature of the SOM in the deeper subsoil, suggesting the presence of more stable decomposed materials. The results of this research seem to indicate that the model, developed for grasslands, can be used to determine the stability of SOM in agricultural ecosystems. The C:N ratio and δ15N signature of the POM and mOM fraction follow the expected model pattern. The isotopic δ13C signature can further enhance the understanding of the processes driving SOM stability.

  9. Use of 15N Label in Organic Synthesis and Spectroscopy. Part I: Preparation of 15N-Labeled tert-Butylamine

    NASA Astrophysics Data System (ADS)

    Talaty, Erach R.; Boese, Christopher A.; Adewale, Sanni M.; Ismail, Mohammed S.; Provenzano, Frank A.; Utz, Melissa J.

    2002-02-01

    The preparation of 15N-labeled tert-butylamine involves the conversion of the correspondingly labeled potassium cyanide into the 15N-labeled tert-butylformamide via the Ritter reaction in 85% yield, followed by hydrolysis with either aqueous sodium hydroxide or hydrochloric acid. The NMR spectra of the compounds provide a valuable opportunity for discussing several important topics in NMR spectroscopy, such as cis-trans isomerism due to restricted rotation and 15N coupling. Comparison of the IR spectra of the labeled and unlabeled compounds permits a forum for discussing the theory of vibrational frequencies.

  10. Abundance and seasonal activity of questing Ixodes ricinus ticks in their natural habitats in southern Germany in 2011.

    PubMed

    Schulz, Michaela; Mahling, Monia; Pfister, Kurt

    2014-06-01

    Questing ticks were sampled monthly over a period of 11 months from February, 2011 to December, 2011 at 13 sites in southern Germany using the flagging method. The ticks were identified to species, gender, and stadium. Although both I. ricinus and D. reticulatus were sampled, this study concentrated on I. ricinus, since it was the most abundant tick to be found. Additional weather data (air and soil temperature, relative air humidity, precipitation, sunshine duration) were recorded on each sampling site and the local vegetation described. A total of 14, 394 ticks was collected (7,862 larvae, 5,568 nymphs, 964 adults) and their activity was recorded in order to determine the seasonal activity pattern over different periods of the year. In contrast to the widely accepted pattern of a bimodal seasonal activity in moderate areas with a dominant peak in spring and a minor peak in autumn, a unimodal activity pattern was found for all development stages on six of the 12 sampling sites. Tick abundance was compared to weather variables. Tick host-seeking activity was found to be significantly dependent on the temperature at ground level, precipitation, and sunshine duration as well as relative air humidity. Adult ticks showed a positive correlation with the duration of sunshine, whereas nymphs were mostly unaffected by this phenomenon. PMID:24820556

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

    USGS Publications Warehouse

    Johnson, C.A.

    1996-01-01

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

  12. Abiotic and biotic factors associated with the presence of Anopheles arabiensis immatures and their abundance in naturally occurring and man-made aquatic habitats

    PubMed Central

    2012-01-01

    Background Anopheles arabiensis (Diptera: Culicidae) is a potential malaria vector commonly present at low altitudes in remote areas in Reunion Island. Little attention has been paid to the environmental conditions driving larval development and abundance patterns in potential habitats. Two field surveys were designed to determine whether factors that discriminate between aquatic habitats with and without An. arabiensis larvae also drive larval abundance, comparatively in man-made and naturally occurring habitats. Methods In an initial preliminary survey, a representative sample of aquatic habitats that would be amenable to an intensive long-term study were selected and divided into positive and negative sites based on the presence or absence of Anopheles arabiensis larvae. Subsequently, a second survey was prompted to gain a better understanding of biotic and abiotic drivers of larval abundance, comparatively in man-made and naturally occurring habitats in the two studied locations. In both surveys, weekly sampling was performed to record mosquito species composition and larval density within individual habitats, as well as in situ biological characteristics and physico-chemical properties. Results Whilst virtually any stagnant water body could be a potential breeding ground for An. arabiensis, habitats occupied by their immatures had different structural and biological characteristics when compared to those where larvae were absent. Larval occurrence seemed to be influenced by flow velocity, macrofauna diversity and predation pressure. Interestingly, the relative abundance of larvae in man-made habitats (average: 0.55 larvae per dip, 95%CI [0.3–0.7]) was significantly lower than that recorded in naturally occurring ones (0.74, 95%CI [0.5–0.8]). Such differences may be accounted for in part by varying pressures that could be linked to a specific habitat. Conclusions If the larval ecology of An. arabiensis is in general very complex and factors affecting breeding site productivity sometimes not easy to highlight, our results, however, highlight lower populations of An. arabiensis immatures compared to those reported in comparable studies conducted in the African continent. Overall, this low larval abundance, resulting from both abiotic and biotic factors, suggests that vector control measures targeting larval habitats are likely to be successful in Reunion, but these could be better implemented by taking environmental variability into account. PMID:22608179

  13. A novel experiment for the quantitative measurement of CSA(1H(N))/CSA(15N) cross-correlated relaxation in 15N-labeled proteins.

    PubMed

    Tessari, M; Vuister, G W

    2000-02-01

    An experiment is presented which allows for the quantitative measurement of the relaxation interference between the 1H(N) CSA and 15N CSA interactions in 15N labeled proteins. A constant-time buildup scheme is used to measure the differential relaxation rate, eta, between double-quantum (DQ) and zero-quantum (ZQ) 1H(N)-15N coherences. The CSA/CSA experiment was recorded at three different Bo field strengths. The CSA(1H(N))/CSA(15N) cross-correlation rate was obtained from the linear fit of the measured rate, eta, versus Bo2 for 77 residues of the EH2 domain from mouse Eps15. PMID:10723996

  14. Constraints on abundance, composition, and nature of X-ray amorphous components of soils and rocks at Gale crater, Mars

    NASA Astrophysics Data System (ADS)

    Dehouck, Erwin; McLennan, Scott M.; Meslin, Pierre-Yves; Cousin, Agnès.

    2014-12-01

    X-ray diffraction patterns of the three samples analyzed by Curiosity's Chemistry and Mineralogy (CheMin) instrument during the first year of the Mars Science Laboratory mission—the Rocknest sand, and the John Klein and Cumberland drill fines, both extracted from the Sheepbed mudstone—show evidence for a significant amorphous component of unclear origin. We developed a mass balance calculation program that determines the range of possible chemical compositions of the crystalline and amorphous components of these samples within the uncertainties of mineral abundances derived from CheMin data. In turn, the chemistry constrains the minimum abundance of amorphous component required to have realistic compositions (all oxides ≥ 0 wt %): 21-22 wt % for Rocknest and 15-20 wt % for Cumberland, in good agreement with estimates derived from the diffraction patterns (~27 and ~31 wt %, respectively). Despite obvious differences between the Rocknest sand and the Sheepbed mudstone, the amorphous components of the two sites are chemically very similar, having comparable concentrations of SiO2, TiO2, Al2O3, Cr2O3, FeOT, CaO, Na2O, K2O, and P2O5. MgO tends to be lower in Rocknest, although it may also be comparable between the two samples depending on the exact composition of the smectite in Sheepbed. The only unambiguous difference is the SO3 content, which is always higher in Rocknest. The observed similarity suggests that the two amorphous components share a common origin or formation process. The individual phases possibly present within the amorphous components include: volcanic (or impact) glass, hisingerite (or silica + ferrihydrite), amorphous sulfates (or adsorbed SO42-), and nanophase ferric oxides.

  15. Screening enoxaparin tetrasaccharide SEC fractions for 3-O-sulfo-N-sulfoglucosamine residues using [(1)H,(15)N] HSQC NMR.

    PubMed

    Beecher, Consuelo N; Manighalam, Matthew S; Nwachuku, Adanma F; Larive, Cynthia K

    2016-02-01

    Heparin and heparan sulfate (HS) are important in mediating a variety of biological processes through binding to myriad different proteins. Specific structural elements along the polysaccharide chains are essential for high affinity protein binding, such as the 3-O-sulfated N-sulfoglucosamine (GlcNS3S) residue, a relatively rare modification essential for heparin's anticoagulant activity. The isolation of 3-O-sulfated oligosaccharides from complex mixtures is challenging because of their low abundance. Although methods such as affinity chromatography are useful in isolating oligosaccharides that bind specific proteins with high affinity, other important 3-O-sulfated oligosaccharides may easily be overlooked. Screening preparative-scale size-exclusion chromatography (SEC) fractions of heparin or HS digests using [(1)H,(15)N] HSQC NMR allows the identification of fractions containing 3-O-sulfated oligosaccharides through the unique (1)H and (15)N chemical shifts of the GlcNS3S residue. Those SEC fractions containing 3-O-sulfated oligosaccharides can then be isolated using strong anion-exchange (SAX)-HPLC. Compared with the results obtained by pooling the fractions comprising a given SEC peak, SAX-HPLC analysis of individual SEC fractions produces a less complicated chromatogram in which the 3-O-sulfated oligosaccharides are enriched relative to more abundant components. The utility of this approach is demonstrated for tetrasaccharide SEC fractions of the low molecular weight heparin drug enoxaparin facilitating the isolation and characterization of an unsaturated 3-O-sulfated tetrasaccharide containing a portion of the antithrombin-III binding sequence. Graphical Abstract [(1)H, (15)N] HSQC NMR spectrum of a mixture of two tetrasaccharides, one of which contains the relatively rare 3-O-sulfated N-sulfoglucosamine residue. PMID:26758598

  16. TEDOR with adiabatic inversion pulses: Resonance assignments of 13C/15N labelled RNAs.

    PubMed

    Riedel, Kerstin; Leppert, Jörg; Ohlenschläger, Oliver; Görlach, Matthias; Ramachandran, Ramadurai

    2005-01-01

    We have examined via numerical simulations the performance characteristics of different 15N RF pulse schemes employed in the transferred echo double resonance (TEDOR) experimental protocol for generating 13C-15N dipolar chemical shift correlation spectra of isotopically labelled biological systems at moderate MAS frequencies (omega(r) approximately 10 kHz). With an 15N field strength of approximately 30-35 kHz that is typically available in 5 mm triple resonance MAS NMR probes, it is shown that a robust TEDOR sequence with significant tolerance to experimental imperfections sa as H1 inhomogeneity and resonance offsets can be effectively implemented using adiabatic heteronuclear dipolar recoupling pulse schemes. TEDOR-based 15N-13C and 15N-13C-13C chemical shift correlation experiments were carried out for obtaining 13C and 15N resonance assignments of an RNA composed of 97 (CUG) repeats which has been implicated in the neuromuscular disease myotonic dystrophy. PMID:15692738

  17. Resolving the bulk δ 15N values of ancient human and animal bone collagen via compound-specific nitrogen isotope analysis of constituent amino acids

    NASA Astrophysics Data System (ADS)

    Styring, Amy K.; Sealy, Judith C.; Evershed, Richard P.

    2010-01-01

    Stable nitrogen isotope analysis is a fundamental tool in assessing dietary preferences and trophic positions within contemporary and ancient ecosystems. In order to assess more fully the dietary contributions to human tissue isotope values, a greater understanding of the complex biochemical and physiological factors which underpin bulk collagen δ 15N values is necessary. Determinations of δ 15N values of the individual amino acids which constitute bone collagen are necessary to unravel these relationships, since different amino acids display different δ 15N values according to their biosynthetic origins. A range of collagen isolates from archaeological faunal and human bone ( n = 12 and 11, respectively), representing a spectrum of terrestrial and marine protein origins and diets, were selected from coastal and near-coastal sites at the south-western tip of Africa. The collagens were hydrolysed and δ 15N values of their constituent amino acids determined as N-acetylmethyl esters (NACME) via gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The analytical approach employed accounts for 56% of bone collagen nitrogen. Reconstruction of bulk bone collagen δ 15N values reveals a 2‰ offset from bulk collagen δ 15N values which is attributable to the δ 15N value of the amino acids which cannot currently be determined by GC-C-IRMS, notably arginine which comprises 53% of the nitrogen unaccounted for (23% of the total nitrogen). The δ 15N values of individual amino acids provide insights into both the contributions of various amino acids to the bulk δ 15N value of collagen and the factors influencing trophic position and the nitrogen source at the base of the food web. The similarity in the δ 15N values of alanine, glutamate, proline and hydroxyproline reflects the common origin of their amino groups from glutamate. The depletion in the δ 15N value of threonine with increasing trophic level indicates a fundamental difference between the biosynthetic pathway of threonine and the other amino acids. The δ 15N value of phenylalanine does not change significantly with trophic level, reflecting its conservative nature as an essential amino acid, and thus represents the isotopic composition of the nitrogen at the base of the food web. Δ 15N Glu-Phe values in particular are shown to reflect trophic level nitrogen sources within a food web. In relation to the reconstruction of ancient human diet the contribution of marine and terrestrial protein are strongly reflected in Δ 15N Glu-Phe values. Differences in nitrogen metabolism are also shown to have an influence upon individual amino acid δ 15N values with Δ 15N Glu-Phe values emphasising differences between the different physiological adaptations. The latter is demonstrated in tortoises, which can excrete nitrogen in the form of uric acid and urea and display negative Δ 15N Glu-Phe values whereas those for marine and terrestrial mammals are positive. The findings amplify the potential advantages of compound-specific nitrogen isotope analysis in the study of nitrogen flow within food webs and in the reconstruction of past human diets.

  18. Soil processes drive seasonal variation in retention of 15N tracers in a deciduous forest catchment.

    PubMed

    Goodale, Christine L; Fredriksen, Guinevere; Weiss, Marissa S; McCalley, K; Sparks, Jed P; Thomas, Steven A

    2015-10-01

    Seasonal patterns of stream nitrate concentration have long been interpreted as demonstrating the central role of plant uptake in regulating stream nitrogen loss from forested catchments. Soil processes are rarely considered as important drivers of these patterns. We examined seasonal variation in N retention in a deciduous forest using three whole-ecosystem 15N tracer additions: in late April (post-snowmelt, pre-leaf-out), late July (mid-growing- season), and late October (end of leaf-fall). We expected that plant 15N uptake would peak in late spring and midsummer, that immobilization in surface litter and soil would peak the following autumn leaf-fall, and that leaching losses would vary inversely with 15N retention. Similar to most other 15N tracer studies, we found that litter and soils dominated ecosystem retention of added 15N. However, 15N recovery in detrital pools varied tremendously by season, with > 90% retention in spring and autumn and sharply reduced 15N retention in late summer. During spring, over half of the 15N retained in soil occurred within one day in the heavy (mineral-associated) soil fraction. During summer, a large decrease in 15N retention one week after addition coincided with increased losses of 15NO3- to soil leachate and seasonal increases in soil and stream NO3- concentrations, although leaching accounted for only a small fraction of the lost 15N (< 0.2%). Uptake of 15N into roots did not vary by season and accounted for < 4% of each tracer addition. Denitrification or other processes that lead to N gas loss may have consumed the rest. These measurements of 15N movement provide strong evidence for the dominant role of soil processes in regulating seasonal N retention and losses in this catchment and perhaps others with similar soils. PMID:26649387

  19. Seasonal variation in species composition and abundance of demersal fish and invertebrates in a Seagrass Natural Reserve on the eastern coast of the Shandong Peninsula, China

    NASA Astrophysics Data System (ADS)

    Xu, Qiang; Guo, Dong; Zhang, Peidong; Zhang, Xiumei; Li, Wentao; Wu, Zhongxin

    2015-09-01

    Seagrass habitats are structurally complex ecosystems, which support high productivity and biodiversity. In temperate systems the density of seagrass may change seasonally, and this may influence the associated fish and invertebrate community. Little is known about the role of seagrass beds as possible nursery areas for fish and invertebrates in China. To study the functioning of a seagrass habitat in northern China, demersal fish and invertebrates were collected monthly using traps, from February 2009 to January 2010. The density, leaf length and biomass of the dominant seagrass Zostera marina and water temperature were also measured. The study was conducted in a Seagrass Natural Reserve (SNR) on the eastern coast of the Shandong Peninsula, China. A total of 22 fish species and five invertebrate species were recorded over the year. The dominant fish species were Synechogobius ommaturus, Sebastes schlegelii, Pholis fangi, Pagrus major and Hexagrammos otakii and these species accounted for 87% of the total number of fish. The dominant invertebrate species were Charybdis japonica and Octopus variabilis and these accounted for 98% of the total abundance of invertebrates. There was high temporal variation in species composition and abundance. The peak number of fish species occurred in August-October 2009, while the number of individual fish and biomass was highest during November 2009. Invertebrate numbers and biomass was highest in March, April, July and September 2009. Temporal changes in species abundance of fishes and invertebrates corresponded with changes in the shoot density and leaf length of the seagrass, Zostera marina.

  20. Seasonal variation in species composition and abundance of demersal fish and invertebrates in a Seagrass Natural Reserve on the eastern coast of the Shandong Peninsula, China

    NASA Astrophysics Data System (ADS)

    Xu, Qiang; Guo, Dong; Zhang, Peidong; Zhang, Xiumei; Li, Wentao; Wu, Zhongxin

    2016-03-01

    Seagrass habitats are structurally complex ecosystems, which support high productivity and biodiversity. In temperate systems the density of seagrass may change seasonally, and this may influence the associated fish and invertebrate community. Little is known about the role of seagrass beds as possible nursery areas for fish and invertebrates in China. To study the functioning of a seagrass habitat in northern China, demersal fish and invertebrates were collected monthly using traps, from February 2009 to January 2010. The density, leaf length and biomass of the dominant seagrass Zostera marina and water temperature were also measured. The study was conducted in a Seagrass Natural Reserve (SNR) on the eastern coast of the Shandong Peninsula, China. A total of 22 fish species and five invertebrate species were recorded over the year. The dominant fish species were Synechogobius ommaturus, Sebastes schlegelii, Pholis fangi, Pagrus major and Hexagrammos otakii and these species accounted for 87% of the total number of fish. The dominant invertebrate species were Charybdis japonica and Octopus variabilis and these accounted for 98% of the total abundance of invertebrates. There was high temporal variation in species composition and abundance. The peak number of fish species occurred in August-October 2009, while the number of individual fish and biomass was highest during November 2009. Invertebrate numbers and biomass was highest in March, April, July and September 2009. Temporal changes in species abundance of fishes and invertebrates corresponded with changes in the shoot density and leaf length of the seagrass, Zostera marina.

  1. Formation and abundance of doubly-substituted methane isotopologues ( 13CH 3D) in natural gas systems

    NASA Astrophysics Data System (ADS)

    Ma, Qisheng; Wu, Sheng; Tang, Yongchun

    2008-11-01

    Formation of the Carbon-13 ( 13C) and deuterium (D) doubly-substituted methane isotopologues ( 13CH 3D) in natural gases is studied utilizing both first-principle quantum mechanism molecular calculation and direct FTIR laboratorial measurements of specifically synthesized high isotope concentration methane gas. For 13CH 3D, the symmetrically breathing mode A0 emerges as IR-detectable attributed to the molecular symmetry lowering to C3v from Td of the non-isotopic methane (CH 4), along with a large vibrational frequency shift from ˜3000 to ˜2250 cm -1. Our studies also indicate that the concentration of 13CH 3D is dependent on the environmental temperature through isotope exchanges among methane isotopologues; and the Gibbs' Free Energy difference due to Quantum Mechanics Zero-Point vibrational motions has the major contribution to this temperature dependency. Potential geologic applications of the 13CH 3D measurement to natural gas exploration and assessments are also discussed. In order to detect the 13CH 3D concentration change of each 50 °C in the natural gas system, a 10 -9 resolution is desirable. Such a measurement could provide important add-on information to distinguish natural gas origin and distribution.

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

    NASA Astrophysics Data System (ADS)

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

    2011-12-01

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

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

    SciTech Connect

    Modi, S.; Behere, D.V.; Mitra, S. )

    1989-05-30

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

  4. Determination of organic milk authenticity using carbon and nitrogen natural isotopes.

    PubMed

    Chung, Ill-Min; Park, Inmyoung; Yoon, Jae-Yeon; Yang, Ye-Seul; Kim, Seung-Hyun

    2014-10-01

    Natural stable isotopes of carbon and nitrogen ((12)C, (13)C, (14)N, (15)N) have abundances unique to each living creature. Therefore, measurement of the stable isotope ratio of carbon and nitrogen (δ(13)C=(13)C/(12)C, δ(15)N=(15)N/(14)N) in milk provides a reliable method to determine organic milk (OM) authenticity. In the present study, the mean δ(13)C value of OM was higher than that of conventional milk (CM), whereas the mean δ(15)N value of OM was lower than that of CM; nonetheless both δ(13)C and δ(15)N values were statistically different for the OM and CM (P<0.05). Furthermore, the values of δ(13)C and δ(15)N were found to differ statistically with the collection date and the milk brand (P<0.05). The combination of δ(13)C and δ(15)N values was more effective than either value alone in distinguishing between OM and CM. The results of the present study, which is based on preliminary data from a limited sample size and sampling period, could be highly valuable and helpful for consumers, the food industry, and/or government regulatory agencies as it can prevent fraudulent labelling of organic food. Further studies include additional analyses of other milk brands and analyses over longer time periods in order to accurately determine OM authenticity using stable isotopes of carbon and nitrogen. PMID:24799230

  5. Variations in the natural ¹⁵N abundance of Brassica chinensis grown in uncultivated soil affected by different nitrogen fertilizers.

    PubMed

    Yuan, Yuwei; Hu, Guixian; Zhao, Ming; Chen, Tianjin; Zhang, Yongzhi; Zhu, Jiahong; Wang, Qiang

    2014-11-26

    To further investigate the method of using δ(15)N as a marker for organic vegetable discrimination, the effects of different fertilizers on the δ(15)N in different growing stages of Brassica chinensis (B. chinensis) grown in uncultivated soil were investigated with a pot experiment. B. chinensis was planted with uncultivated soil and different fertilizer treatments and then harvested three times in three seasons consecutively. For the spring experiments in the years of 2011 and 2012, the δ(15)N value of B. chinensis, which increased due to organic manure application and decreased due to chemical fertilizer application, was significantly different (p < 0.05) with manure treatment and chemical treatment. The δ(15)N value of vegetables varied among three growing stages and ranged from +8.6‰ to +11.5‰ for the control, from +8.6‰ to +12.8‰ for the compost chicken manure treatment, from +2.8‰ to +7.7‰ for the chemical fertilizer urea treatment, and from +7.7‰ to +10.9‰ for the compost-chemical fertilizer treatment. However, the δ(15)N values observed in the autumn experiment of 2011 without any fertilizer application increased ranging from +13.4‰ to +15.4‰, + 11.2‰ to +17.7‰, +10.7‰ to +17.1‰, and +10.6‰ to +19.1‰, respectively, for the same treatments mentioned above. This result was not significantly different between manure treatment and chemical treatment. The δ(15)N values of soil obtained in the spring of 2011 during three growing stages were slightly affected by fertilizers and varied in the range of +1.6‰ to +2.5‰ for CK, +4.7‰ to +6.5‰ for compost treatment, +2.1‰ to +2.4‰ for chemical treatment, and +2.7‰ to +4.6‰ for chemical-compost treatment, respectively. High δ(15)N values of B. chinensis were observed in these experiments, which would be useful to supplement a δ(15)N database for discriminating organic vegetables. Although there was a significant difference between manure treatment and chemical treatment, it was still difficult to discriminate whether a labeled organic vegetable was really grown without chemical fertilizer just with a fixed high δ(15)N value, especially for the vegetables planted simultaneously with chemical and compost fertilizer. PMID:25369912

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

    USGS Publications Warehouse

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

    2011-01-01

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

  7. Sensitivity enhancement in natural-abundance solid-state 33S MAS NMR spectroscopy employing adiabatic inversion pulses to the satellite transitions

    NASA Astrophysics Data System (ADS)

    Hansen, Michael Ryan; Brorson, Michael; Bildsøe, Henrik; Skibsted, Jørgen; Jakobsen, Hans J.

    2008-02-01

    The WURST (wideband uniform rate smooth truncation) and hyperbolic secant (HS) pulse elements have each been employed as pairs of inversion pulses to induce population transfer (PT) between the four energy levels in natural abundance solid-state 33S (spin I = 3/2) MAS NMR, thereby leading to a significant gain in intensity for the central transition (CT). The pair of inversion pulses are applied to the satellite transitions for a series of inorganic sulfates, the sulfate ions in the two cementitious materials ettringite and thaumasite, and the two tetrathiometallates (NH 4) 2WS 4 and (NH 4) 2MoS 4. These materials all exhibit 33S quadrupole coupling constants ( CQ) in the range 0.1-1.0 MHz, with precise CQ values being determined from analysis of the PT enhanced 33S MAS NMR spectra. The enhancement factors for the WURST and HS elements are quite similar and are all in the range 1.74-2.25 for the studied samples, in excellent agreement with earlier reports on HS enhancement factors (1.6-2.4) observed for other spin I = 3/2 nuclei with similar CQ values (0.3-1.2 MHz). Thus, a time saving in instrument time by a factor up to five has been achieved in natural abundance 33S MAS NMR, a time saving which is extremely welcome for this important low-γ nucleus.

  8. Sensitivity enhancement in natural-abundance solid-state 33S MAS NMR spectroscopy employing adiabatic inversion pulses to the satellite transitions.

    PubMed

    Hansen, Michael Ryan; Brorson, Michael; Bildsøe, Henrik; Skibsted, Jørgen; Jakobsen, Hans J

    2008-02-01

    The WURST (wideband uniform rate smooth truncation) and hyperbolic secant (HS) pulse elements have each been employed as pairs of inversion pulses to induce population transfer (PT) between the four energy levels in natural abundance solid-state (33)S (spin I=3/2) MAS NMR, thereby leading to a significant gain in intensity for the central transition (CT). The pair of inversion pulses are applied to the satellite transitions for a series of inorganic sulfates, the sulfate ions in the two cementitious materials ettringite and thaumasite, and the two tetrathiometallates (NH(4))(2)WS(4) and (NH(4))(2)MoS(4). These materials all exhibit (33)S quadrupole coupling constants (C(Q)) in the range 0.1-1.0 MHz, with precise C(Q) values being determined from analysis of the PT enhanced (33)S MAS NMR spectra. The enhancement factors for the WURST and HS elements are quite similar and are all in the range 1.74-2.25 for the studied samples, in excellent agreement with earlier reports on HS enhancement factors (1.6-2.4) observed for other spin I=3/2 nuclei with similar C(Q) values (0.3-1.2 MHz). Thus, a time saving in instrument time by a factor up to five has been achieved in natural abundance (33)S MAS NMR, a time saving which is extremely welcome for this important low-gamma nucleus. PMID:18082436

  9. 15N CSA tensors and 15N-1H dipolar couplings of protein hydrophobic core residues investigated by static solid-state NMR

    NASA Astrophysics Data System (ADS)

    Vugmeyster, Liliya; Ostrovsky, Dmitry; Fu, Riqiang

    2015-10-01

    In this work, we assess the usefulness of static 15N NMR techniques for the determination of the 15N chemical shift anisotropy (CSA) tensor parameters and 15N-1H dipolar splittings in powder protein samples. By using five single labeled samples of the villin headpiece subdomain protein in a hydrated lyophilized powder state, we determine the backbone 15N CSA tensors at two temperatures, 22 and -35 °C, in order to get a snapshot of the variability across the residues and as a function of temperature. All sites probed belonged to the hydrophobic core and most of them were part of α-helical regions. The values of the anisotropy (which include the effect of the dynamics) varied between 130 and 156 ppm at 22 °C, while the values of the asymmetry were in the 0.32-0.082 range. The Leu-75 and Leu-61 backbone sites exhibited high mobility based on the values of their temperature-dependent anisotropy parameters. Under the assumption that most differences stem from dynamics, we obtained the values of the motional order parameters for the 15N backbone sites. While a simple one-dimensional line shape experiment was used for the determination of the 15N CSA parameters, a more advanced approach based on the "magic sandwich" SAMMY pulse sequence (Nevzorov and Opella, 2003) was employed for the determination of the 15N-1H dipolar patterns, which yielded estimates of the dipolar couplings. Accordingly, the motional order parameters for the dipolar interaction were obtained. It was found that the order parameters from the CSA and dipolar measurements are highly correlated, validating that the variability between the residues is governed by the differences in dynamics. The values of the parameters obtained in this work can serve as reference values for developing more advanced magic-angle spinning recoupling techniques for multiple labeled samples.

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

    PubMed Central

    Svensson, Elisabeth; Schouten, Stefan; Hopmans, Ellen C.; Middelburg, Jack J.; Sinninghe Damsté, Jaap S.

    2016-01-01

    Lipid extraction of biomass prior to stable isotope analysis is known to cause variable changes in the stable nitrogen isotopic composition (δ15N) of residual biomass. However, the underlying factors causing these changes are not yet clear. Here we address this issue by comparing the δ15N of bulk and residual biomass of several marine animal tissues (fish, crab, cockle, oyster, and polychaete), as well as the δ15N of the extracted lipids. As observed previously, lipid extraction led to a variable offset in δ15N of biomass (differences ranging from -2.3 to +1.8 ‰). Importantly, the total lipid extract (TLE) was highly depleted in 15N compared to bulk biomass, and also highly variable (differences ranging from -14 to +0.7 ‰). The TLE consisted mainly of phosphatidylcholines, a group of lipids with one nitrogen atom in the headgroup. To elucidate the cause for the 15N-depletion in the TLE, the δ15N of amino acids was determined, including serine because it is one of the main sources of nitrogen to N-containing lipids. Serine δ15N values differed by -7 to +2 ‰ from bulk biomass δ15N, and correlated well with the 15N depletion in TLEs. On average, serine was less depleted (-3‰) than the TLE (-7 ‰), possibly due to fractionation during biosynthesis of N-containing headgroups, or that other nitrogen-containing compounds, such as urea and choline, or recycled nitrogen contribute to the nitrogen isotopic composition of the TLE. The depletion in 15N of the TLE relative to biomass increased with the trophic level of the organisms. PMID:26731720

  11. Seasonal Abundance and Natural Inoculativity of Insect Vectors of Xylella fastidiosa in Oklahoma Tree Nurseries and Vineyards.

    PubMed

    Overall, Lisa M; Rebek, Eric J

    2015-12-01

    Xylella fastidiosa is the causative agent of diseases of perennial plants including peach, plum, elm, oak, pecan, and grape. This bacterial pathogen is transmitted by xylem-feeding insects. In recent years, Pierce's disease of grape has been detected in 10 counties in central and northeastern Oklahoma, prompting further investigation of the disease epidemiology in this state. We surveyed vineyards and tree nurseries in Oklahoma for potential insect vectors to determine species composition, infectivity, and natural inoculativity of commonly captured insect vectors. Yellow sticky cards were used to sample insect fauna at each location. Insects were removed from sticky cards and screened for X. fastidiosa using immunocapture-PCR to determine their infectivity. A second objective was to test the natural inoculativity of insect vectors that are found in vineyards. Graphocephala versuta (Say), Graphocephala coccinea (Forster), Paraulacizes irrorata (F.), Oncometopia orbona (F.), Cuerna costalis (F.), and Entylia carinata Germar were collected from vineyards and taken back to the lab to determine their natural inoculativity. Immunocapture-PCR was used to test plant and insect samples for presence of X. fastidiosa. The three most frequently captured species from vineyards and tree nurseries were G. versuta, Clastoptera xanthocephala Germar, and O. orbona. Of those insects screened for X. fastidiosa, 2.4% tested positive for the bacterium. Field-collected G. versuta were inoculative to both ragweed and alfalfa. Following a 7-d inoculation access period, a higher percentage of alfalfa became infected than ragweed. Results from this study provide insight into the epidemiology of X. fastidiosa in Oklahoma. PMID:26331482

  12. What's on the menu? Assessing microbial carbon sources and cycling in soils using natural abundance radiocarbon analysis

    NASA Astrophysics Data System (ADS)

    Mahmoudi, N.; Burns, L.; Mancini, S.; Fulthorpe, R.; Slater, G. F.

    2011-12-01

    Organic matter in soils is composed of diverse materials in various stages of decomposition. Soil organic matter is not in a single pool but rather in multiple carbon pools with different intrinsic turnover times that can be on annual to decadal and even millennial timescales. Microorganisms can influence the total amount of carbon stored in soils and the turnover rates of these different pools. However, the links between microbes and their ability to utilize these various carbon pools are not well understood. Moreover, microbes have been shown to co-utilize a number of available carbon sources rather than a single carbon source under soil conditions which creates difficulties in identifying microbial carbon sources in the natural environment. Compound-specific radiocarbon analysis of microbial phospholipid fatty acids (PLFA) has become a useful tool in elucidating microbial carbon sources in complex environments with multiple carbon sources. We investigated microbial carbon cycling at an industrial site in Ontario which included a variety of carbon sources including vegetation, PAHs and natural organic matter (NOM). Using this approach, the 14C content of microbial membrane lipids (which reflects their carbon source) can be compared to surrounding carbon sources in order to assess which carbon source they are metabolizing and incorporating into their lipids. In addition, we assessed microbial community structure and diversity by analyzing amplified bacterial, eukaryotic and archaeal rDNA fragments with denaturing gel gradient electrophoresis (DGGE). The Δ14C value for PLFAs ranged from +54 to -697% which indicates that microbial carbon sources across soils differ. The Δ14CPLFA for some soils is consistent with modern carbon sources while Δ14CPLFA for other soils is consistent with natural organic matter including older pools of carbon. The microbial communities at this site are not metabolizing PAHs but rather they are utilizing various pools of natural organic matter as a carbon source. Moreover, our radiocarbon analysis of total organic carbon (TOC) and non-solvent extracted residue (representative of NOM) support the notion that soil organic matter is not homogenous and is comprised of multiple carbon fractions with different intrinsic turnover times. DNA fingerprinting revealed consistent levels of diversity across soils with the greatest diversity observed in the bacteria, followed by eukarya and finally archaea which had the lowest observed diversity. It was previously thought that there is a pool of refractory organic compounds that are biologically inert in soil however our findings indicate that microbes are able to utilize almost all available carbon sources in soil. These finding brings forth questions regarding the influence of microbes on soil organic matter stabilization and the bioavailability of different carbon pools.

  13. Determination of Natural 14C Abundances in Dissolved Organic Carbon in Organic-Rich Marine Sediment Porewaters by Thermal Sulfate Reduction

    NASA Astrophysics Data System (ADS)

    Johnson, L.; Komada, T.

    2010-12-01

    The abundances of natural 14C in dissolved organic carbon (DOC) in the marine environment hold clues regarding the processes that influence the biogeochemical cycling of this large carbon reservoir. At present, UV irradiation is the widely accepted method for oxidizing seawater DOC for determination of their 14C abundances. This technique yields precise and accurate values with low blanks, but it requires a dedicated vacuum line, and hence can be difficult to implement. As an alternative technique that can be conducted on a standard preparatory vacuum line, we modified and tested a thermal sulfate reduction method that was previously developed to determine δ13C values of marine DOC (Fry B. et al., 1996. Analysis of marine DOC using a dry combustion method. Mar. Chem., 54: 191-201.) to determine the 14C abundances of DOC in marine sediment porewaters. In this method, the sample is dried in a 100 ml round-bottom Pyrex flask in the presence of excess oxidant (K2SO4) and acid (H3PO4), and combusted at 550 deg.C. The combustion products are cryogenically processed to collect and quantify CO2 using standard procedures. Materials we have oxidized to date range from 6-24 ml in volume, and 95-1500 μgC in size. The oxidation efficiency of this method was tested by processing known amounts of reagent-grade dextrose and sucrose (as examples of labile organic matter), tannic acid and humic acid (as examples of complex natural organic matter), and porewater DOC extracted from organic-rich nearshore sediments. The carbon yields for all of these materials averaged 99±4% (n=18). The 14C abundances of standard materials IAEA C-6 and IAEA C-5 processed by this method using >1mgC aliquots were within error of certified values. The size and the isotopic value of the blank were determined by a standard dilution technique using IAEA C-6 and IAEA C-5 that ranged in size from 150 to 1500 μgC (n=4 and 2, respectively). This yielded a blank size of 6.7±0.7 μgC, and a blank isotopic value of 0.54±0.05 fMC. The size of the blank agreed well with that determined directly by processing variable volumes of UV-irradiated deionized water (5.6±0.7 μgC, n=9). The size of the blank amounts to <~5% of the size of porewater DOC samples that are typically recovered from organic-rich sediment cores (~100-500 μgC). The fMC value of the blank suggests that there may be multiple sources of extraneous carbon that range in 14C abundance. In order to assess the fidelity of 14C abundances in natural porewater DOC oxidized by thermal sulfate reduction, we oxidized porewater DOC samples collected from the central floor of the Santa Monica Basin, California Borderland, using both this method and UV irradiation (the latter carried out at the Druffel laboratory, University of California Irvine). The fMC values obtained by the two methods agreed within error. Carbon yields from the two methods also agreed closely. These findings show that thermal sulfate reduction may be a promising method to oxidize small, concentrated marine DOC samples for 14C analysis.

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

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

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

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

    PubMed

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

    2013-09-01

    Dietary analysis using δ(15)N values of human remains such as bone and hair is usually based on general principles and limited data sets. Even for modern humans, the direct ascertainment of dietary δ(15)N is difficult and laborious, due to the complexity of metabolism and nitrogen fractionation, differing dietary habits and variation of δ(15)N values of food items. The objective of this study was to summarize contemporary regional experimental and global literature data to ascertain mean representative δ(15)N values for distinct food categories. A comprehensive data set of more than 12,000 analyzed food samples was summarized from the literature. Data originated from studies dealing with (1) authenticity tracing or origin control of food items, and (2) effects of fertilization or nutrition on δ(15)N values of plants or animals. Regional German food δ(15)N values revealed no major differences compared with the mean global values derived from the literature. We found that, in contrast to other food categories, historical faunal remains of pig and poultry are significantly enriched in (15)N compared to modern samples. This difference may be due to modern industrialized breeding practices. In some food categories variations in agricultural and feeding regimens cause significant differences in δ(15)N values that may lead to misinterpretations when only limited information is available. PMID:23921599

  16. Increased Plant Uptake of Nitrogen from 15N Depleted Fertilizer Using Plant Growth-Promoting Rhizobacteria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The techniques of 15N isotope have been very useful for determining the behavior and fate of N in soil, including the use efficiency of applied N fertilizers by plants. Our objective in this study was to use 15N isotope techniques to demonstrate that a model plant growth-promoting rhizobacteria (PGP...

  17. Tracking wind-dispersed seeds using (15)N-isotope enrichment.

    PubMed

    Forster, C; Herrmann, J D

    2014-11-01

    Seed dispersal influences a wide range of ecological processes. However, measuring dispersal patterns, particularly long-distance dispersal, has been a difficult task. Marking bird-dispersed seeds with stable (15)N isotopes has been shown to be a user-friendly method to trace seed dispersal. In this study, we determined whether (15)N urea solution could be used to enrich seeds of two common wind-dispersed plants, Eupatorium glaucescens (Asteraceae) and Sericocarpus tortifolius (Asteraceae). We further tested if the water type (distilled versus tap) in (15)N urea solutions influences the level and variability of enrichment of plant seeds, and if increasing spraying frequency per se increases enrichment. Because droughts may lower seed set or kill plants, we wanted to investigate if the additional use of an externally applied anti-transpirant affects the intake of externally applied (15)N into seeds. The results demonstrate that (15)N enrichment of seeds can facilitate dispersal experiments with wind-dispersed plants. The use of distilled water in (15)N urea solutions did not increase (15)N enrichment compared to tap water. Further, enrichment was more efficient at lower spray frequencies. Both the use of tap water and low frequencies could lower time, effort and project costs. The results suggest that species can be protected from drought using an anti-transpirant without decreasing the incorporation of (15)N into seeds. PMID:25174806

  18. Disturbance and topography shape nitrogen availability and ?15N over long-term forest succession

    EPA Science Inventory

    Forest disturbance and long-term succession can promote open N cycling that increases N loss and soil ?15N values. We examined soil and foliar patterns in N and ?15N, and soil N mineralization, across a topographically complex montane forest landscape influenced by human logging ...

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

    EPA Science Inventory

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

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

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

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

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

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

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

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

    ERIC Educational Resources Information Center

    Thompson, Laura E.; Rovnyak, David

    2007-01-01

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

  3. Variability assessment of (15)N metabolic labeling-based proteomics workflow in mouse plasma and brain.

    PubMed

    Filiou, Michaela D; Soukupova, Magdalena; Rewerts, Christiane; Webhofer, Christian; Turck, Chris W; Maccarrone, Giuseppina

    2015-06-01

    (15)N metabolic labeling-based quantitative proteomics is used for the identification of disease- and phenotype-related alterations in live organisms. The variability of (15)N metabolic labeling proteomics workflows has been assessed in plants and bacteria. However, no study has addressed this topic in mice. We have investigated the repeatability of a quantitative in vivo(15)N metabolic labeling proteomics workflow in mice by assessing LC variability, peptide and protein profiling characteristics and overall (15)N/(14)N protein quantification accuracy in technical replicates of plasma and brain specimens. We furthermore examined how sample preparation affects these parameters in plasma and brain. We found that specimen type (i.e. plasma or brain) influences the variability of the (15)N metabolic labeling workflow in an LC-independent manner. PMID:25782008

  4. 15N and13C NMR investigation of hydroxylamine-derivatized humic substances

    USGS Publications Warehouse

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

    1992-01-01

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

  5. The effect of manuring on cereal and pulse amino acid δ(15)N values.

    PubMed

    Styring, Amy K; Fraser, Rebecca A; Bogaard, Amy; Evershed, Richard P

    2014-06-01

    Amino acid δ(15)N values of barley (Hordeum vulgare) and bread wheat (Triticum aestivum) grains and rachis and broad bean (Vicia faba) and pea (Pisum sativum) seeds, grown in manured and unmanured soil at the experimental farm stations of Rothamsted, UK and Bad Lauchstädt, Germany, were determined by GC-C-IRMS. Manuring was found to result in a consistent (15)N-enrichment of cereal grain amino acid δ(15)N values, indicating that manuring did not affect the metabolic routing of nitrogen (N) into cereal grain amino acids. The increase in cereal grain δ(15)N values with manuring is therefore due to a (15)N-enrichment in the δ(15)N value of assimilated inorganic-N. Greater variation was observed in the (15)N-enrichment of rachis amino acids with manuring, possibly due to enhanced sensitivity to changes in growing conditions and higher turnover of N in rachis cells compared to cereal grains. Total amino acid δ(15)N values of manured and unmanured broad beans and peas were very similar, indicating that the legumes assimilated N2 from the atmosphere rather than N from the soil, since there was no evidence for routing of (15)N-enriched manure N into any of the pulse amino acids. Crop amino acid δ(15)N values thus provide insights into the sources of N assimilated by non N2-fixing and N2-fixing crops grown on manured and unmanured soils, and reveal an effect of manure on N metabolism in different crop species and plant parts. PMID:24631496

  6. Disturbance and topography shape nitrogen availability and δ15 N over long-term forest succession

    USGS Publications Warehouse

    Perakis, Steven; Tepley, Alan J.; Compton, Jana

    2015-01-01

    Forest disturbance and long-term succession towards old-growth are thought to increase nitrogen (N) availability and N loss, which should increase soil δ15N values. We examined soil and foliar patterns in N and δ15N, and soil N mineralization, across 800 years of forest succession in a topographically complex montane landscape influenced by human logging and wildfire. In contrast to expectations, we found that disturbance caused declines in surface mineral soil δ15N values, both in logged forests measured 40–50 years after disturbance, and in unlogged forests disturbed by severe wildfire within the last 200 years. Both symbiotic N fixation and N transfers from disturbed vegetation and detritus could lower soil δ15N values after disturbance. A more important role for symbiotic N fixation is suggested by lower soil δ15N values in slow-successional sites with slow canopy closure, which favors early-successional N fixers. Soil δ15N values increased only marginally throughout 800 years of succession, reflecting soil N uptake by vegetation and strong overall N retention. Although post-disturbance N inputs lowered surface soil δ15N values, steady-state mass balance calculations suggest that wildfire combustion of vegetation and detritus can dominate long-term N loss and increase whole-ecosystem δ15N. On steeper topography, declining soil δ15N values highlight erosion and accelerated soil turnover as an additional abiotic control on N balances. We conclude for N-limited montane forests that soil δ15N and N availability are less influenced by nitrate leaching and denitrification loss than by interactions between disturbance, N fixation, and erosion.

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

    NASA Astrophysics Data System (ADS)

    Charteris, Alice; Michaelides, Katerina; Evershed, Richard

    2015-04-01

    Organic N concentrations far exceed those of inorganic N in most soils and despite much investigation, the composition and cycling of this complex pool of SOM remains poorly understood. A particular problem has been separating more recalcitrant soil organic N from that actively cycling through the soil system; an important consideration in N cycling studies and for the soil's nutrient supplying capacity. The use of 15N-labelled substrates as stable isotope tracers has contributed much to our understanding of the soil system, but the complexity and heterogeneity of soil organic N prevents thorough compound-specific 15N analyses of organic N compounds and makes it difficult to examine any 15N-labelled organic products in any detail. As a result, a significant proportion of previous work has either simply assumed that since the majority of soil N is organic, all of the 15N retained in the soil is organic N (e.g. Sebilo et al., 2013) or subtracted 15N-labelled inorganic compounds from bulk values (e.g. Pilbeam et al., 1997). While the latter approach is more accurate, these methods only provide an estimate of the bulk 15N value of an extremely complex and non-uniformly labelled organic pool. A more detailed approach has been to use microbial biomass extraction (Brookes et al., 1985) and subsequent N isotopic analysis to determine the 15N value of biomass-N, representing the fraction of 15N assimilated by microbes or the 15N cycling through the 'living' or 'active' portion of soil organic N. However, this extraction method can only generate estimates and some lack of confidence in its validity and reliability remains. Here, we present an alternative technique to obtain a measure of the assimilation of an applied 15N substrate by the soil microbial biomass and an estimate of the newly synthesized soil protein, which is representative of the magnitude of the active soil microbial biomass. The technique uses a stable isotope tracer and compound-specific 15N analysis, but unlike previous works analyses for amino acids (representing organic products) rather than ammonium (NH4+) and nitrate (NO3-). Amino acids are commonly referred to as 'the building blocks of life' as they form the proteins which regulate life's essential biochemical reactions. Proteinaceous matter generally comprises 20-40% of total soil N and is ubiquitous in living organisms, so is a likely 'organic product' of microbial activity/assimilation. Hence, we consider it likely that amino acids represent the major organic nitrogenous products and a reasonable 'proxy' for/measure of the assimilation of an applied 15N substrate by the soil microbial biomass and an estimate of the newly synthesized soil protein. Brookes, P. C. et al. Soil Biol Biochem. 1985, 17, 837-842. Jenkinson, D. S. et al. Soil Biol Biochem. 2004, 36, 5-7. Nannipieri, P. et al. Plant Soil. 1999, 208, 43-56. Pilbeam, C. J. et al. J Agr Sci. 1997, 128, 415-424. Sebilo, M. et al. PNAS. 2013, 110, 18185-18189.

  8. Cadmium burden of men and women who report regular consumption of confectionery sunflower kernels containing a natural abundance of cadmium.

    PubMed Central

    Reeves, P G; Vanderpool, R A

    1997-01-01

    Because of inherent genetic and physiological characteristics, the natural concentration of cadmium in the kernels of sunflowers grown in uncontaminated soils of the northern Great Plains region of the United States is higher than in most other grains. We tested the hypothesis that a habitual consumption of sunflower kernels will increase the body burden and health effects of cadmium in humans. Sixty-six men and women who reported consuming various amounts of sunflower kernels were recruited and divided by sex and kernel consumption: those who consumed less than or equal to 1 ounce(oz)/week and those who consumed more than 1 oz/week. Cadmium intake was assessed by calculation from 7-day food diaries, cadmium burden by whole blood cadmium, red blood cell (RBC) cadmium and urine cadmium concentrations, and health effects by urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) activity and beta2-microglobulin (beta2MG). The results showed that high intakes of sunflower kernels (>1 oz/day) significantly increased the intake of cadmium (p<0.004). However, the amount of cadmium in whole blood or RBCs was not affected by cadmium intake. Urinary excretion of cadmium also was not affected by cadmium intake. Urine NAG activity and the amount of urinary beta2MG were significantly elevated in the urine of high sunflower kernel consumers when the values were expressed on a urine volume basis (p<0.03), but not when expressed on a creatinine basis (p>0.05). Because normal ranges for the excretion of these protein markers have not been established, it was not possible to determine if these elevated values were meaningful. However, given the knowledge that habitual consumption of sunflower kernels with natural cadmium concentrations higher than most other food products will increase the average intake of dietary cadmium, the potential exists for an increased body burden of cadmium. Controlled feeding studies in humans should be pursued in order to determine if the body burden does indeed increase and, if so, is it a cause for concern. PMID:9349833

  9. EFFECT OF HIGH-ENERGY RESONANCES ON THE {sup 18}O(p, {alpha}){sup 15}N REACTION RATE AT AGB AND POST-AGB RELEVANT TEMPERATURES

    SciTech Connect

    La Cognata, M.; Spitaleri, C.; Mukhamedzhanov, A. M.

    2010-11-10

    The {sup 18}O(p, {alpha}){sup 15}N reaction is of great importance in several astrophysical scenarios, as it influences the production of key isotopes such as {sup 19}F, {sup 18}O, and {sup 15}N. Fluorine is synthesized in the intershell region of asymptotic giant branch (AGB) stars, together with s-elements, by {alpha} radiative capture on {sup 15}N, which in turn is produced in the {sup 18}O proton-induced destruction. Peculiar {sup 18}O abundances are observed in R-Coronae Borealis stars, having {sup 16}O/{sup 18}O {approx}< 1, hundreds of times smaller than the galactic value. Finally, there is no definite explanation of the {sup 14}N/{sup 15}N ratio in pre-solar grains formed in the outer layers of AGB stars. Again, such an isotopic ratio is influenced by the {sup 18}O(p, {alpha}){sup 15}N reaction. In this work, a high accuracy {sup 18}O(p, {alpha}){sup 15}N reaction rate is proposed, based on the simultaneous fit of direct measurements and of the results of a new Trojan Horse experiment. Indeed, current determinations are uncertain because of the poor knowledge of the resonance parameters of key levels of {sup 19}F. In particular, we have focused on the study of the broad 660 keV 1/2{sup +} resonance corresponding to the 8.65 MeV level of {sup 19}F. Since {Gamma} {approx} 100-300 keV, it determines the low-energy tail of the resonant contribution to the cross section and dominates the cross section at higher energies. Here, we provide a reaction rate that is a factor of two larger above T {approx} 0.5 10{sup 9} K based on our new improved determination of its resonance parameters, which could strongly influence present-day astrophysical model predictions.

  10. The perils and promises of microbial abundance: novel natures and model ecosystems, from artisanal cheese to alien seas.

    PubMed

    Paxson, Heather; Helmreich, Stefan

    2014-04-01

    Microbial life has been much in the news. From outbreaks of Escherichia coli to discussions of the benefits of raw and fermented foods to recent reports of life forms capable of living in extreme environments, the modest microbe has become a figure for thinking through the presents and possible futures of nature, writ large as well as small. Noting that dominant representations of microbial life have shifted from an idiom of peril to one of promise, we argue that microbes--especially when thriving as microbial communities--are being upheld as model ecosystems in a prescriptive sense, as tokens of how organisms and human ecological relations with them could, should, or might be. We do so in reference to two case studies: the regulatory politics of artisanal cheese and the speculative research of astrobiology. To think of and with microbial communities as model ecosystems offers a corrective to the scientific determinisms we detect in some recent calls to attend to the materiality of scientific objects. PMID:24941610

  11. Climate-Dependence of Plant-Soil 15N/14N Interactions Across Tropical Rainforests

    NASA Astrophysics Data System (ADS)

    Houlton, B. Z.; Sigman, D. M.; Hedin, L. O.

    2005-12-01

    In most areas of the world, the 15N/14N of bulk soils is higher than that of plant leaves, and the isotopic signatures of these two ecosystem N pools progressively diverge with increasing rainfall. However, both the cause for this isotopic trend and its implications for understanding interactions between climate and N cycles are largely unknown. We report 15N/14N measurements of nitrate, ammonium, and total dissolved N in soil extracts from a highly constrained rainfall sequence in Hawaii, across which this trend in ecosystem 15N/14N is captured, to examine the competing explanations for plant-soil 15N/14N uncouplings. While the isotopic influences of microbial transfers of N between nitrate and ammonium pools and plant-mycorrhizae interactions have been posited in plant-soil 15N/14N relationships, our data did not support an important role for either of these mechanisms. Instead, preferential regeneration of 14N during the breakdown of DON to ammonium explains why the 15N/14N of plants is lower than that of bulk soils. Fractionation at this step leads to two isotopically distinct N subcycles in each forest, a lower-15N/14N subcycle composed of ammonium, nitrate, and bulk plant biomass N that `spins' rapidly and a higher-15N/14N subcycle composed of bulk soil N and DON that is much less dynamic. The increased difference between soil and plant 15N/14N is due to changes in the impacts of nitrification and denitrification on the 15N/14N of ammonium and nitrate, coupled with a switch from nitrate to ammonium uptake by plants under the wettest conditions. For instance, the particularly large (~6 per mil) 15N/14N difference between plants and soils in the wettest sites is due to the lack of 15N-enrichment of ammonium by nitrification coupled with plant dependence on ammonium uptake only. Our results highlight the importance of interactions between DON breakdown, ecosystem N recycling, and gaseous N losses in the explaining the interactions between the 15N signatures of soils and plants across these tropical rainforests. We also show that these interactions are climate-dependent.

  12. 15N Content Reflects Development of Mycorrhizae and Nitrogen Dynamics During Primary Succession

    NASA Astrophysics Data System (ADS)

    Hobbie, E. A.; Jumpponen, A.

    2004-05-01

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

  13. Linking nitrogen dynamics to climate variability off central California: a 51 year record based on 15N/ 14N in CalCOFI zooplankton

    NASA Astrophysics Data System (ADS)

    Rau, Greg H.; Ohman, Mark D.; Pierrot-Bults, Annelies

    2003-08-01

    Long-term variability in zooplankton 15N/ 14N was investigated in two species of calanoid copepods ( Calanus pacificus and Eucalanus californicus) and two chaetognaths ( Sagitta bierii and Sagitta euneritica) sampled in the spring of selected years from 1951 to 2001 off the central California coast. No statistically significant trend in 15N/ 14N was detected for any of the four species, with isotopic ratios in 2001 resembling those in copepods and chaetognaths sampled five decades earlier. Zooplankton body lengths also showed no long-term trends. With respect to proposed regime shifts in this region, heterogeneity in 15N/ 14N was detected only for S. bierii when comparing the periods 1951-1975, 1978-1998, and 1999-2001. In this species the 15N/ 14N in the most recent, brief period (1999-2001) averaged slightly lower than in the previous period. Three of the four species ( C. pacificus, S. bierii, and S. euneritica) showed significant increases in 15N/ 14N during major El Niños. El Niño-related enrichment in 15N could arise as a consequence of increased nitrate demand:supply at the base of the food web or advection of 15N-enriched nitrate from more southerly waters. While a range of physical and climate indices were evaluated, anomalies of 15N/ 14N from the long-term mean were found to be significantly related only to: (i) the Southern Oscillation Index in the case of both chaetognath species, (ii) a regional surface water temperature record ( S. bierii only), (iii) an index of wind-driven coastal upwelling for the surface-dwelling C. pacificus, and (iv) variability in the Pacific Decadal Oscillation for the somewhat deeper-dwelling E. californicus. The relationships among each species' 15N/ 14N averaged over the total sampling period was: E. californicus≈C. pacificus≪S. euneritica < S. bierii, consistent with trophic 15N biomagnification and the predatory nature of Sagitta.

  14. Protonation studies of modified adenine and adenine nucleotides by theoretical calculations and (15)N NMR.

    PubMed

    Major, Dan T; Laxer, Avital; Fischer, Bilha

    2002-02-01

    The acid/base character of nucleobases affects phenomena such as self-association, interaction with metal ions, molecular recognition by proteins, and nucleic acid base-pairing. Therefore, the investigation of proton-transfer equilibria of natural and synthetic nucleos(t)ides is of great importance to obtain a deeper understanding of these phenomena. For this purpose, a set of ATP prototypes was investigated using (15)N NMR spectroscopy, and the corresponding adenine bases were investigated by theoretical calculations. (15)N NMR measurements provided not only acidity constants but also information on the protonation site(s) on the adenine ring and regarding the ratio of the singly protonated species in equilibrium. Substituents of different nature and position on the adenine ring did not change the preferred protonation site, which remained N1. However, for 2-thioether-ATP derivatives a mixed population of N1 and N7 singly protonated species was observed. Reduction of basicity of 0.4-1 pK(a) units relative to ATP was also observed for all evaluated ATP derivatives, except for 2-Cl-ATP, for which K(a) was ca. 10,000-fold lower. To explain the substitution-dependent variations in the experimental pK(a) values of the ATP analogues, gas-phase proton affinities (PA), Delta Delta G(hyd), and pK(a) values of the corresponding adenine bases were calculated using quantum mechanical methods. The computed PA and Delta Delta G(hyd) values successfully explained the experimental pK(a) values. A computational procedure for the prediction of accurate pK(a) values was developed using density functional theory and polarizable continuum model calculations. In this procedure, we developed a set of parameters for the polarizable continuum model that was fitted to reproduce experimental pK(a) values of nitrogen heterocycles. This method is proposed for the prediction of pK(a) values and protonation site(s) of purine analogues that have not been synthesized or analyzed. PMID:11856021

  15. Gas chromatography/combustion/isotope-ratio-monitoring mass spectrometric analysis of methylboronic derivatives of monosaccharides: a new method for determining natural 13C abundances of carbohydrates.

    PubMed

    van Dongen, B E; Schouten, S; Damsté, J S

    2001-01-01

    Monosaccharides were derivatized using methylboronic acid and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), and the delta13C values of these derivatives measured by gas chromatography/combustion/isotope-ratio-monitoring mass spectrometry to determine the original 13C-content of the monosaccharides. Comparison with the measured off-line delta13 values of the monosaccharides shows that no fractionation in 13C takes place during derivatization. The methylboronic derivatization method has proven to be a new method for natural abundance isotopic analysis of intact monosaccharides (arabinose, xylose, fucose, fructose and glucose). The method is rapid, does not involve isotopic fractionation during derivatization, and gives more precise delta13C values than other methods reported. The method was successfully applied to determine the delta13C value of glucose of the freshwater alga Scenedesmus communis. PMID:11268134

  16. Nitrogen stable isotope composition (δ15N) of vehicle-emitted NOx.

    PubMed

    Walters, Wendell W; Goodwin, Stanford R; Michalski, Greg

    2015-02-17

    The nitrogen stable isotope ratio of NOx (δ(15)N-NOx) has been proposed as a regional indicator for NOx source partitioning; however, knowledge of δ(15)N values from various NOx emission sources is limited. This study presents a detailed analysis of δ(15)N-NOx emitted from vehicle exhaust, the largest source of anthropogenic NOx. To accomplish this, NOx was collected from 26 different vehicles, including gasoline and diesel-powered engines, using a modification of a NOx collection method used by the United States Environmental Protection Agency, and δ(15)N-NOx was analyzed. The vehicles sampled in this study emitted δ(15)N-NOx values ranging from -19.1 to 9.8‰ that negatively correlated with the emitted NOx concentrations (8.5 to 286 ppm) and vehicle run time because of kinetic isotope fractionation effects associated with the catalytic reduction of NOx. A model for determining the mass-weighted δ(15)N-NOx from vehicle exhaust was constructed on the basis of average commute times, and the model estimates an average value of -2.5 ± 1.5‰, with slight regional variations. As technology improvements in catalytic converters reduce cold-start emissions in the future, it is likely to increase current δ(15)N-NOx values emitted from vehicles. PMID:25621737

  17. Steroselective synthesis and application of L-( sup 15 N) amino acids

    SciTech Connect

    Unkefer, C.J. ); Lodwig, S.N. . Div. of Science)

    1991-01-01

    We have developed two general approaches to the stereoselective synthesis of {sup 15}N- and {sup 13}C-labeled amino acids. First, labeled serine, biosynthesized using the methylotrophic bacterium M. extorquens AM1, serves as a chiral precursor for the synthesis of other amino acids. For example, pyridoxal phosphate enzymes can be used for the conversion of L-({alpha}-{sup 15}N)serine to L-({alpha}-{sup 15}N)tyrosine, L-({alpha}-{sup 15}N)tryptophan, and L-({alpha}-{sup 15}N)cysteine. In the second approach, developed by Oppolzer and Tamura, an electrophilic amination'' reagent, 1-chloro-1-nitrosocyclohexane, was used to convert chiral enolates into L-{alpha}-amino acids. We prepared 1-chloro-1-({sup 15}N) nitrosocyclohexane and used it to aminate chiral enolates to produce L-({alpha}-{sup 15}N)amino acids. The stereoselectivity of this scheme using the Oppolzer sultam chiral auxiliary is remarkable, producing enantiomer ratios of 200 to 1. 22 refs., 4 figs.

  18. Changes in Natural Abundance Carbon Stable isotopes of Human Blood and Saliva After 24 Days of Controlled Carbohydrate Supplementation

    NASA Astrophysics Data System (ADS)

    Kraft, R. A.; Jahren, A. H.; Baer, D. J.; Caballero, B.

    2008-12-01

    With the advent of corporate agriculture, large-scale economic decisions have given rise to unique global environmental effects. Emphasis on corn production results in dramatic changes in nitrogen and water cycling via the intensive cultivation practices necessary to support Zea mays (Tilman, 1998). In particular, consumption of corn derived food additive high-fructose corn syrup (HFCS) has increased more than 1000% since 1970 and may be associated with the epidemics of obesity and diabetes (Bray et al., 2004). Plausible mechanisms for an adverse effect of fructose load on glucose homeostasis have been proposed (Havel, 2005). The unusually heavy 13C signature of corn, as compared to other plants, offers the opportunity to develop a biomarker for sugar consumption. Among the many experiments that are needed to establish such a technique, the demonstration of change in 13C signature of human tissues with known change in carbohydrate consumption is foremost. Here we report on a controlled feeding study performed in cooperation with the United States Department of Agriculture (USDA), to test the effect of supplementation of human diet with carbohydrate of known δ13C value. During this study, 13 individuals were fed a typical American diet (32% calories from fat, 15% calories from protein, 53% carbohydrate) for ~six months. Each participant was fed a random sequence of carbohydrate supplements (50 grams of supplement per day): 1. resistant maltodextrin (δ13C = -10.59‰); 2. maltodextrin (δ13C = -23.95‰); 3. a 50-50 mixture of the two (δ13C = -15.94‰). After 24 days of feeding, subjects showed enrichment in blood serum that was significantly correlated (p = 0.0038) with the δ13C value of the supplement. However, blood clot and saliva showed no such correlation, suggesting that the half-lives of these substrates may render them unsuitable for carbohydrate dietary reconstruction over day-to-month timescales. All subjects of the study showed a net enrichment in the δ13C value of their blood and saliva relative to baseline: blood clot was enriched by 0.27‰; blood serum by 0.50‰ and saliva by 1.12‰. We believe this overall enrichment resulted from a 13C-enriched bulk diet (δ13C = - 20.42‰) relative to the subjects free-living diet. Evidence for this derives from inspection of foods within the bulk diet provided, compared to published profiles of the typical American diet. We will discuss possible complicating factors, such as differential absorption and metabolism of the supplements according to solubility and caloric value. These results are encouraging for the development of a δ13C blood serum biomarker that, in the company of other tests, could be used to indicate a change in carbohydrate intake. Bray, G.A., Nielsen, S.J. and Popkin, B.M., 2004. Consumption of high-fructose corn syrup in beverages may play a role in the epidemic of obesity. American Journal of Clinical Nutrition, 79: 537-543. Havel, P.J., 2005. Dietary fructose: Implications for dysregulation of energy homeostasis and lipid/carbohydrate metabolism. Nutrition Reviews, 63(5): 133-157. Tilman D., 1998. The greening of the green revolution. Nature, 396:211-212.

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

    PubMed

    Inamura, O; Zhang, J; Minagawa, M

    2012-01-15

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

  20. Preservation of proteinaceous material during the degradation of the green alga Botryococcus braunii: A solid-state 2D 15N 13C NMR spectroscopy study

    NASA Astrophysics Data System (ADS)

    Zang, Xu; Nguyen, Reno T.; Harvey, H. Rodger; Knicker, Heike; Hatcher, Patrick G.

    2001-10-01

    Using solid-state cross-polarization-magic-angle-spinning (CPMAS) 13C and 15N nuclear magnetic resonance (NMR) and 2-D double cross polarization (DCP) MAS 15N 13C NMR techniques, microbially degraded Botryococcus braunii was analyzed to study the chemical nature of organic nitrogen in the algal residue. The amide linkage, as found in protein, was observed as the major nitrogen component in 201-day-old degraded algae. No significant amount of heterocyclic nitrogen, or evidence for melanoidin products, was found. The results strongly suggest that proteinaceous material can survive early diagenesis and be preserved via its encapsulation by refractory, macromolecular, organic matter.

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

    NASA Astrophysics Data System (ADS)

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

    2012-05-01

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

  2. Application of capillary gas chromatography-reaction interface/mass spectrometry to the selective detection of sup 13 C-, sup 15 N-, sup 2 H-, and sup 14 C-labeled drugs and their metabolites

    SciTech Connect

    Chace, D.H.

    1989-01-01

    A novel reaction interface/mass spectrometer (RIMS) technique has been applied to the selective detection of {sup 13}C-, {sup 15}N-, {sup 2}H-, and {sup 14}C-labeled phenytoin and its metabolites in urine following separation by capillary gas chromatography. The microwave-powered reaction interface converts materials from their original forms into small molecules whose mass spectra serve to identify and quantify the nuclides. The presence of each element is followed by monitoring the isotopic variants of CO{sub 2}, NO, H{sub 2}, or CH{sub 4} that are produced by the reaction interface. Chromatograms showing only enriched {sup 13}C and {sup 15}N were produced using the net {sup 13}CO{sub 2} or {sup 15}NO signal derived by subtracting the abundance of naturally occurring isotopes from the observed M + 1 signal. When hydrogen was used as a reactant gas, a selective chromatogram of {sup 2}H (D) was obtained by measuring HD at m/Z 3.0219, and a chromatogram showing {sup 14}C was obtained by measuring {sup 14}CH{sub 4} at m/Z 18.034 with a high resolution. For a stable isotope detection, metabolites representing less than 1.5% of the total labeled compounds could be detected in the chromatogram. Detection limits of 170 pCi/mL (34 pCi on column that is equivalent to 187 pg) of a {sup 14}C- labeled metabolite was detected. To identify many of these labeled peaks (metabolites), the chromatographic analysis was repeated with the reaction interface turned off and mass spectra obtained at the retention times found in the RIMS experiment. In addition to the ability of GC-RIMS to detect the presence of {sup 13}C-, {sup 15}N-, and {sup 2}H- (D), it can also quantify the level of enrichment. Enrichment of {sup 13}C and {sup 15}N is quantified by measuring the ratio of excess {sup 13}CO{sub 2} to total {sup 12}CO{sub 2} or excess {sup 15}NO to total {sup 14}NO.

  3. Nitrogen dynamics in a Western Boundary Upwelling System (Cabo Frio, Brazil) based on δ15N-nitrate and δ15N of sinking particle signals

    NASA Astrophysics Data System (ADS)

    Fontana, L.; Belem, A. L.; Venancio, I.; Duarte, C.; Chiara, S. D.; Albuquerque, A. L.

    2014-12-01

    To improve the efficiency of upwelling to control nitrogen dynamic in the ocean, better understanding of the occurring processes is necessary. This research explores δ15N of nitrate and sinking particles on a western boundary upwelling System (Cabo Frio, Brazil). The Continental Shelf of southeastern Brazil is dominated by the oligotrophic Brazil Current, whose instabilities promote the coastal upwelling of South Atlantic Central Water (SACW), and consequently increases of primary productivity. The coastal upwelling system plays an important role in the nitrogen dynamics on the Cabo Frio Upwelling System (CFUS). However, the interactions between biological induced processes, including biological N-fixation and nitrate inputs from upwelled waters in CFUS still have not been well explored. Then, this study aims clarify N-dynamics on CFUS based on a cross-shelf approach. δ15N-nitrate was characterized for each water masses present on the shelf (South Atlantic Central Water, Tropical Water and Coastal Water) and associated with physicochemical parameters (T/S, nutrients), as well as the δ15N of sinking particles at different depths (from surface to the bottom water). Samples were collected in a time interval of 1 month during ~4 years (2011 to 2014). Cross-shelf gradients of nitrogen species concentration (ammonium + nitrite + nitrate) and stable isotopes were observed. The δ15N of nitrate and sinking particles were interpreted according to the prevailing processes of the N-transformations. Considering the region as N-limited (N:P < 16), processes as biological N-fixation seems to be dominant on oligotrophic Tropical Waters. Coastal upwelled SACW showed δ15N-nitrate signature within the global average of deep ocean (5-6‰) characterizing the inner and mid-shelf conditions, where the input of new nitrate from upwelling is rapidly used by organisms in the euphotic zone without any fractionation. On the other hands, the dominance of N-limited Tropical Waters on the outer shelf provide a δ15N-nitrate and δ15N-sinking particles signals (-2.0 to 3.0‰) lower than the global average of deep ocean range indicating the importance of N-biological fixation as a dominant source of nitrogen.

  4. Continuous field measurement of N2O isotopologues using FTIR spectroscopy following 15N addition

    NASA Astrophysics Data System (ADS)

    Phillips, R. L.; Griffith, D. W.; Dijkstra, F. A.; Lugg, G.; Lawrie, R.; Macdonald, B.

    2012-12-01

    Anthropogenic additions of fertilizer nitrogen (N) have significantly increased the mole fraction of nitrous oxide (N2O) in the troposphere. Tracking the fate of fertilizer N and its transformation to N2O is important to advance knowledge of greenhouse gas emissions from soils. Transport and transformations are frequently studied using 15N labeling experiments, but instruments capable of continuous measurements of 15N-N2O at the surface of soil have only recently come to the fore. Our primary aim was to quantify emissions of N2O and the fraction of 15N emitted as N2O from an agricultural soil following 15N addition using a mobile Fourier Transform Infrared (FTIR) spectrometer. We set up a short-term field experiment on a coastal floodplain site near Nowra, New South Wales. We deployed an automated chamber system connected to a multi-pass cell (optical pathlength 24 m) and low resolution FTIR spectrometer to measure fluxes of all N2O isotopologues collected from five 0.25 m2 chambers every three hours. We measured N2O fluxes pre and post-application of 15N-labeled substrate as potassium nitrate (KNO3) or urea [CO(NH2)2] to the soil surface. Root mean square uncertainties for all isotopologue measurements were less than 0.3 nmol mol-1 for 1 minute average concentration measurements, and minimum detectable fluxes for each isotopologue were <0.1 ng N m-2 s-1. Emissions of all N2O isotopologues were evident immediately following 15N addition. Emissions of 14N15NO, 15N14NO and 15N15NO isotopologues subsided within 10 d, but 14N14NO fluxes were evident over the entire experiment. The figure provides an overview of the emissions. Cumulative 15N-N2O fluxes (sum of the three 15N isotopologues) per chamber for the 14 days following 15N addition ranged from 1.5 to 10.3 mg 15N-N2O m-2. The chambers were destructively sampled after 2 weeks and 15N analyzed in soil and plant material using isotope ratio mass spectrometry. Approximately 1% (range 0.7 - 1.9%) of the total amount of 15N applied was emitted as N2O. Average fractions of 15N recovered in soil, root, shoot, and microbial biomass pools varied between trials but were approximately 0.4, 0.08, 0.1 and 0.03, respectively. The results indicate that the portable FTIR spectroscopic technique can effectively trace transfer of 15N to the atmosphere as N2O after 15N addition, allowing for powerful quantification of N2O emissions under field conditions.

  5. Covalent binding of reduced metabolites of [{sup 15}N{sub 3}]TNT to soil organic matter during a bioremediation process analyzed by {sup 15}N NMR spectroscopy

    SciTech Connect

    Achtnich, C.; Fernandes, E.; Bollag, J.M.; Knackmuss, H.J.; Lenke, H.

    1999-12-15

    Evidence is presented for the covalent binding of biologically reduced metabolites of 2,4,6-{sup 15}N{sub 3}-trinitrotoluene (TNT) to different soil fractions, using liquid {sup 15}N NMR spectroscopy. A silylation procedure was used to release soil organic matter from humin and whole soil for spectroscopic measurements. TNT-contaminated soil was spiked with 2,4,6-{sup 15}N{sub 3}-trinitrotoluene and {sup 14}C-ring labeled TNT, before treatment in a soil slurry reactor. During the anaerobic/aerobic incubation the amount of radioactivity detected in the fulvic and humic acid fractions did not change significantly whereas the radioactivity bound to humin increased to 71%. The {sup 15}N NMR spectra of the fulvic acid samples were dominated by a large peak that corresponded to aliphatic amines or ammonia. In the early stages of incubation, {sup 15}N NMR analysis of the humic acids indicated bound azoxy compounds. The signals arising from nitro and azoxy groups disappeared with further anaerobic treatment. At the end of incubation, the NMR shifts showed that nitrogen was covalently bound to humic acid as substituted amines and amides. The NMR spectra of the silylated humin suggest formation of azoxy compounds and imine linkages. Bound metabolites possessing nitro groups were also detected. Primary amines formed during the anaerobic incubation disappeared during the aerobic treatment. Simultaneously, the amount of amides and tertiary amines increased. Nitro and azoxy groups of bound molecules were still present in humin at the end of the incubation period. Formation of azoxy compounds from partially reduced TNT followed by binding and further reduction appears to be an important mechanism for the immobilization of metabolites of TNT to soil.

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

    NASA Astrophysics Data System (ADS)

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

    2015-08-01

    Carbon-rich grains with isotopic anomalies compared to the Sun are found in primitive meteorites. They were made by stars, and carry the original stellar nucleosynthesis signature. Silicon carbide grains of Type X and C and low-density (LD) graphites condensed in the ejecta of core-collapse supernovae. We present a new set of models for the explosive He shell and compare them with the grains showing 12C/13C and 14N/15N ratios lower than solar. In the stellar progenitor H was ingested into the He shell and not fully destroyed before the explosion. Different explosion energies and H concentrations are considered. If the supernova shock hits the He-shell region with some H still present, the models can reproduce the C and N isotopic signatures in C-rich grains. Hot-CNO cycle isotopic signatures are obtained, including a large production of 13C and 15N. The short-lived radionuclides 22Na and 26Al are increased by orders of magnitude. The production of radiogenic 22Ne from the decay of 22Na in the He shell might solve the puzzle of the Ne-E(L) component in LD graphite grains. This scenario is attractive for the SiC grains of type AB with 14N/15N ratios lower than solar, and provides an alternative solution for SiC grains originally classified as nova grains. Finally, this process may contribute to the production of 14N and 15N in the Galaxy, helping to produce the 14N/15N ratio in the solar system.

  7. On the measurement of 15N-{1H} nuclear Overhauser effects.

    PubMed

    Ferrage, Fabien; Piserchio, Andrea; Cowburn, David; Ghose, Ranajeet

    2008-06-01

    Accurate quantification of the 15N-{1H} steady-state NOE is central to current methods for the elucidation of protein backbone dynamics on the fast, sub-nanosecond time scale. This experiment is highly susceptible to systematic errors arising from multiple sources. The nature of these errors and their effects on the determined NOE ratio is evaluated by a detailed analysis of the spin dynamics during the pair of experiments used to measure this ratio and possible improvements suggested. The experiment that includes 1H irradiation, is analyzed in the framework of Average Liouvillian Theory and a modified saturation scheme that generates a stable steady-state and eliminates the need to completely saturate 1H nuclei is presented. The largest source of error, however, in 1H-dilute systems at ultra-high fields is found to be an overestimation of the steady-state NOE value as a consequence of the incomplete equilibration of the magnetization in the so-called "reference experiment". The use of very long relaxation delays is usually an effective, but time consuming, solution. Here, we introduce an alternative reference experiment, designed for larger, deuterated systems, that uses the fastest relaxing component of the longitudinal magnetization as a closer approximation to the equilibrium state for shorter relaxation delays. The utility of the modified approach is illustrated through simulations on realistic spin systems over a wide range of time scales and experimentally verified using a perdeuterated sample of human ubiquitin. PMID:18417394

  8. Chlorophyll a-specific Δ14C, δ13C and δ15N values in stream periphyton: implications for aquatic food web studies

    NASA Astrophysics Data System (ADS)

    Ishikawa, N. F.; Yamane, M.; Suga, H.; Ogawa, N. O.; Yokoyama, Y.; Ohkouchi, N.

    2015-11-01

    Periphytic algae attached to a streambed substrate (periphyton) are an important primary producer in stream ecosystems. We determined the isotopic composition of chlorophyll a in periphyton collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, a pure aquatic primary producer (Cladophora sp.) and a terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Epeorus latifolium). Periphyton Δ14Cchl values (-258 ‰ in April and -190 ‰ in October) were slightly lower than Δ14Cbulk values (-228 ‰ in April and -179 ‰ in October) but were close to the Δ14C value for dissolved inorganic carbon (DIC; -217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = -1000 ‰), CO2 derived from aquatic and terrestrial organic matters (variable Δ14C) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (-215 ‰ in April and -199 ‰ in October) and Cladophora sp. (-210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

  9. Chlorophyll a specific Δ14C, δ13C and δ15N values in stream periphyton: implications for aquatic food web studies

    NASA Astrophysics Data System (ADS)

    Ishikawa, N. F.; Yamane, M.; Suga, H.; Ogawa, N. O.; Yokoyama, Y.; Ohkouchi, N.

    2015-07-01

    We determined the isotopic composition of chlorophyll a in periphytic algae attached to a streambed substrate (periphyton). The samples were collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, pure aquatic primary producer (Cladophora sp.) and terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Mayfly larva, Epeorus latifolium), suggesting that periphyton δ13C values do not faithfully trace carbon transfer between primary producers and primary consumers. Periphyton Δ14Cchl values (-258 ‰ in April and -190 ‰ in October) were slightly lower than Δ14Cbulk values (-228 ‰ in April and -179 ‰ in October), but were close to the Δ14C value for dissolved inorganic carbon (DIC) (-217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = -1000 ‰) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (-215 ‰ in April and -199 ‰ in October) and Cladophora sp. (-210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

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

    DOEpatents

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

    2001-01-01

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

  11. Eastern oyster (Crassostrea virginica) delta(15)N as a bioindicator of nitrogen sources: Observations and modeling.

    PubMed

    Fertig, B; Carruthers, T J B; Dennison, W C; Fertig, E J; Altabet, M A

    2010-08-01

    Stable nitrogen isotopes (delta(15)N) in bioindicators are increasingly employed to identify nitrogen sources in many ecosystems and biological characteristics of the eastern oyster (Crassostrea virginica) make it an appropriate species for this purpose. To assess nitrogen isotopic fractionation associated with assimilation and baseline variations in oyster mantle, gill, and muscle tissue delta(15)N, manipulative fieldwork in Chesapeake Bay and corresponding modeling exercises were conducted. This study (1) determined that five individuals represented an optimal sample size; (2) verified that delta(15)N in oysters from two locations converged after shared deployment to a new location reflecting a change in nitrogen sources; (3) identified required exposure time and temporal integration (four months for muscle, two to three months for gill and mantle); and (4) demonstrated seasonal delta(15)N increases in seston (summer) and oysters (winter). As bioindicators, oysters can be deployed for spatial interpolation of nitrogen sources, even in areas lacking extant populations. PMID:20381097

  12. Secondary sup 15 N isotope effects on the reactions catalyzed by alcohol and formate dehydrogenases

    SciTech Connect

    Rotberg, N.S.; Cleland, W.W. )

    1991-04-23

    Secondary {sup 15}N isotope effects at the N-1 position of 3-acetylpyridine adenine dinucleotide have been determined, by using the internal competition technique, for horse liver alcohol dehydrogenase (LADH) with cyclohexanol as a substrate and yeast formate dehydrogenase (FDH) with formate as a substrate. On the basis of less precise previous measurements of these {sup 15}N isotope effects, the nicotinamide ring of NAD has been suggested to adopt a boat conformation with carbonium ion character at C-4 during hydride transfer. If this mechanism were valid, as N-1 becomes pyramidal an {sup 15}N isotope effect for the reaction catalyzed by LADH was measured. These values suggest that a significant {sup 15}N kinetic isotope effect is not associated with hydride transfer for LADH and FDH. Thus, in contrast with the deformation mechanism previously postulated, the pyridine ring of the nucleotide apparently remains planar during these dehydrogenase reactions.

  13. Eastern oyster (Crassostrea virginica) δ15N as a bioindicator of nitrogen sources: Observations and modeling

    PubMed Central

    Fertig, B.; Carruthers, T.J.B.; Dennison, W.C.; Fertig, E.J.; Altabet, M.A.

    2013-01-01

    Stable nitrogen isotopes (δ15N) in bioindicators are increasingly employed to identify nitrogen sources in many ecosystems and biological characteristics of the eastern oyster (Crassostrea virginica) make it an appropriate species for this purpose. To assess nitrogen isotopic fractionation associated with assimilation and baseline variations in oyster mantle, gill, and muscle tissue δ15N, manipulative fieldwork in Chesapeake Bay and corresponding modeling exercises were conducted. This study (1) determined that five individuals represented an optimal sample size; (2) verified that δ15N in oysters from two locations converged after shared deployment to a new location reflecting a change in nitrogen sources; (3) identified required exposure time and temporal integration (four months for muscle, two to three months for gill and mantle); and (4) demonstrated seasonal δ15N increases in seston (summer) and oysters (winter). As bioindicators, oysters can be deployed for spatial interpolation of nitrogen sources, even in areas lacking extant populations. PMID:20381097

  14. Position-Specific Isotope Analysis of Xanthines: A (13)C Nuclear Magnetic Resonance Method to Determine the (13)C Intramolecular Composition at Natural Abundance.

    PubMed

    Diomande, Didier G; Martineau, Estelle; Gilbert, Alexis; Nun, Pierrick; Murata, Ariaki; Yamada, Keita; Watanabe, Naoharu; Tea, Illa; Robins, Richard J; Yoshida, Naohiro; Remaud, Gérald S

    2015-07-01

    The natural xanthines caffeine, theobromine, and theophylline are of major commercial importance as flavor constituents in coffee, cocoa, tea, and a number of other beverages. However, their exploitation for authenticity, a requirement in these commodities that have a large origin-based price-range, by the standard method of isotope ratio monitoring by mass spectrometry (irm-MS) is limited. We have now developed a methodology that overcomes this deficit that exploits the power of isotopic quantitative (13)C nuclear magnetic resonance (NMR) spectrometry combined with chemical modification of the xanthines to enable the determination of positional intramolecular (13)C/(12)C ratios (δ(13)Ci) with high precision. However, only caffeine is amenable to analysis: theobromine and theophylline present substantial difficulties due to their poor solubility. However, their N-methylation to caffeine makes spectral acquisition feasible. The method is confirmed as robust, with good repeatability of the δ(13)Ci values in caffeine appropriate for isotope fractionation measurements at natural abundance. It is shown that there is negligible isotope fractionation during the chemical N-methylation procedure. Thus, the method preserves the original positional δ(13)Ci values. The method has been applied to measure the position-specific variation of the (13)C/(12)C distribution in caffeine. Not only is a clear difference between caffeine isolated from different sources observed, but theobromine from cocoa is found to show a (13)C pattern distinct from that of caffeine. PMID:26067163

  15. 15N fractionation in star-forming regions and Solar System objects

    NASA Astrophysics Data System (ADS)

    Wirström, Eva; Milam, Stefanie; Adande, Gilles; Charnley, Steven B.; Cordiner, Martin A.

    2015-08-01

    A central issue for understanding the formation and evolution of matter in the early Solar System is the relationship between the chemical composition of star-forming interstellar clouds and that of primitive Solar System materials. The pristine molecular content of comets, interplanetary dust particles and carbonaceous chondrites show significant bulk nitrogen isotopic fractionation relative to the solar value, 14N/15N ~ 440. In addition, high spatial resolution measurements in primitive materials locally show even more extreme enhancements of 14N/15N < 100.The coherent 15N enrichment in comets from different formation zones suggests that these isotopic enhancements are remnants of the interstellar chemistry in the natal molecular cloud core and the outer protosolar nebula. Indeed, early chemical models of gas-phase ion-molecule nitrogen fractionation showed that HCN and HNC (nitriles) can hold significant 15N enrichments in cold dark clouds where CO is depleted onto dust grains. In addition, 15N fractionation in nitriles and amines (NH2, NH3) follow different chemical pathways. More recently we have shown that once the spin-state dependence in rates of reactions with H2 is included in the models, amines can either be enhanced or depleted in 15N, depending on the core’s evolutionary stage. Observed 15N fractionation in amines and nitriles therefore cannot be expected to be the same, instead their ratio is a potential chemical clock.Observations of molecular isotope ratios in dark cores are challenging. Limited published results in general show higher 15N/14N ratios in HCN and HNC than ammonia, but more measurements are necessary to confirm these trends. We will present recent results from our ongoing observing campaign of 14N/15N isotopic ratios in HCN, HNC and NH3 in dense cores and protostars which seem consistent with significant fractionation in nitriles as compared to other molecules in each object. The few 14N/15N ratios observed in N2H+ are similar to those in NH3, contrary to our model results which predict a significant 15N enhancement in N2 and N2H+. Model upgrades which may address this discrepancy will be presented and discussed.

  16. 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.

  17. Water proton spin saturation affects measured protein backbone 15 N spin relaxation rates

    NASA Astrophysics Data System (ADS)

    Chen, Kang; Tjandra, Nico

    2011-12-01

    Protein backbone 15N NMR spin relaxation rates are useful in characterizing the protein dynamics and structures. To observe the protein nuclear-spin resonances a pulse sequence has to include a water suppression scheme. There are two commonly employed methods, saturating or dephasing the water spins with pulse field gradients and keeping them unperturbed with flip-back pulses. Here different water suppression methods were incorporated into pulse sequences to measure 15N longitudinal T1 and transversal rotating-frame T1ρ spin relaxation. Unexpectedly the 15N T1 relaxation time constants varied significantly with the choice of water suppression method. For a 25-kDa Escherichiacoli. glutamine binding protein (GlnBP) the T1 values acquired with the pulse sequence containing a water dephasing gradient are on average 20% longer than the ones obtained using a pulse sequence containing the water flip-back pulse. In contrast the two T1ρ data sets are correlated without an apparent offset. The average T1 difference was reduced to 12% when the experimental recycle delay was doubled, while the average T1 values from the flip-back measurements were nearly unchanged. Analysis of spectral signal to noise ratios ( s/ n) showed the apparent slower 15N relaxation obtained with the water dephasing experiment originated from the differences in 1H N recovery for each relaxation time point. This in turn offset signal reduction from 15N relaxation decay. The artifact becomes noticeable when the measured 15N relaxation time constant is comparable to recycle delay, e.g., the 15N T1 of medium to large proteins. The 15N relaxation rates measured with either water suppression schemes yield reasonable fits to the structure. However, data from the saturated scheme results in significantly lower Model-Free order parameters (< S2> = 0.81) than the non-saturated ones (< S2> = 0.88), indicating such order parameters may be previously underestimated.

  18. 15N techniques and analytical procedures. Indo/U. S. science and technology initiative. Research report

    SciTech Connect

    Porter, L.K.; Mosier, A.R.

    1992-05-01

    (15)N technology is used to explore many agricultural research topics, including the movement of nitrates to groundwater, use of fertilizer nitrogen by plants, ways to increase nitrogen fixation, and effects of management practices on denitrification. The publication reviews (15)N procedures and methods for handling and collecting samples, introducing isotopes into plants and soils, and for performing Kjeldahl analyses, isotope dilutions, Rittenberg oxidation conversions for isotope-ration analyses, and automated Dumas isotope-ratio analyses.

  19. Efficient Synthesis of Nicotinamide-1-(15)N for Ultrafast NMR Hyperpolarization Using Parahydrogen.

    PubMed

    Shchepin, Roman V; Barskiy, Danila A; Mikhaylov, Dmitry M; Chekmenev, Eduard Y

    2016-04-20

    Nicotinamide (a vitamin B3 amide) is one of the key vitamins as well as a drug for treatment of M. tuberculosis, HIV, cancer, and other diseases. Here, an improved Zincke reaction methodology is presented allowing for straightforward and scalable synthesis of nicotinamide-1-(15)N with an excellent isotopic purity (98%) and good yield (55%). (15)N nuclear spin label in nicotinamide-1-(15)N can be NMR hyperpolarized in seconds using parahydrogen gas. NMR hyperpolarization using the process of temporary conjugation between parahydrogen and to-be-hyperpolarized biomolecule on hexacoordinate iridium complex via the Signal Amplification By Reversible Exchange (SABRE) method significantly increases detection sensitivity (e.g., >20 000-fold for nicotinamide-1-(15)N at 9.4 T) as has been shown by Theis T. et al. (J. Am. Chem. Soc. 2015, 137, 1404), and hyperpolarized in this fashion, nicotinamide-1-(15)N can be potentially used to probe metabolic processes in vivo in future studies. Moreover, the presented synthetic methodology utilizes mild reaction conditions, and therefore can also be potentially applied to synthesis of a wide range of (15)N-enriched N-heterocycles that can be used as hyperpolarized contrast agents for future in vivo molecular imaging studies. PMID:26999571

  20. Elastic and inelastic scattering of 15N ions by 9Be at 84 MeV

    NASA Astrophysics Data System (ADS)

    Rudchik, A. T.; Chercas, K. A.; Kemper, K. W.; Rusek, K.; Rudchik, A. A.; Herashchenko, O. V.; Koshchy, E. I.; Pirnak, Val. M.; Piasecki, E.; Trzcińska, A.; Sakuta, S. B.; Siudak, R.; Strojek, I.; Stolarz, A.; Ilyin, A. P.; Ponkratenko, O. A.; Stepanenko, Yu. M.; Shyrma, Yu. O.; Szczurek, A.; Uleshchenko, V. V.

    2016-03-01

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

  1. Molecular Investigation of the Short-term Sequestration of Natural Abundance 13C -labelled Cow Dung in the Surface Horizons of a Temperate Grassland Soil

    NASA Astrophysics Data System (ADS)

    Dungait, J.; Bol, R.; Evershed, R. P.

    2004-12-01

    An adequate understanding of the carbon (C) sequestration potential of grasslands requires that the quantity and residence times of C inputs be measured. Herbivore dung is largely comprised of plant cell wall material, a significant source of stable C in intensively grazed temperate grassland ecosystems that contributes to the soil carbon budget. Our work uses compound-specific isotope analysis to identify the pattern of input of dung-derived compounds from natural abundance 13C/-labelled cow dung into the surface horizons of a temperate grassland soil over one year. C4 dung (δ 13C \\-12.6 ‰ ) from maize fed cows was applied to a temperate grassland surface (δ 13C \\-29.95 ‰ ) at IGER-North Wyke (Devon, UK), and dung remains and soil cores beneath the treatments collected at ŧ = 7, 14, 28, 56, 112, 224 and 372 days. Bulk dung carbon present in the 0\\-1 cm and 1\\-5 cm surface horizons of a grassland soil over one year was estimated using Δ 13C between C4 dung and C3 dung, after Bol {\\et al.} (2000). The major biochemical components of dung were quantified using proximate forage fibre analyses, after Goering and Van Soest (1970) and identified using `wet' chemical and GC-MS methods. Plant cell wall polysaccharides and lignin were found to account for up to 67 {%} of dung dry matter. Hydrolysed polysaccharides were prepared as alditol acetates for analyses (after Docherty {\\et al.}, 2001), and a novel application of an off-line pyrolysis method applied to measure lignin-derived phenolic compounds (after Poole & van Bergen, 2002). This paper focuses on major events in the incorporation of dung carbon, estimated using natural abundance 13C&-slash;labelling technique. This revealed a major bulk input of dung carbon after a period of significant rainfall with a consequent decline in bulk soil δ 13C values until the end of the experiment (Dungait {\\et al.}, submitted). Findings will be presented revealing contribution of plant cell wall polysaccharides and lignin to these bulk δ 13C values, and their potential for sequestration considered. References: Bol, R., Amelung, W., Friedrich, C. Ostle, N. (2000). Tracing dung-derived carbon in temperate grassland using 13C natural abundance measurements. Soil Biology and Biochemistry, 32, 1337-1343. Goering and Van Soest (1970). Forage fibre analysis (apparatus, reagents, procedures and some applications). In: USDA-ARS Agricultural Handbook, 379. U. S. Government Printing Office, Washington D.C. Docherty, G., Jones, V. and Evershed, R.P. (2001). Practical and theoretical considerations in the gas chromatography/combustion/isotope ratio mass spectrometry δ 13C analysis of small polyfunctional compounds. Rapid Communications in Mass Spectrometry, 15, 730-738. Poole, I. & van Bergen, P. F. (2002). Carbon isotope ratio analysis of organic moieties from fossil mummified wood: establishing optimum conditions for off-line pyrolysis extraction using gas chromatography/mass spectrometry. Rapid Communications in Mass Spectrometry, 16, 1976-1981. Dungait, J. A. J., Bol, R. and Evershed, R.P. (submitted). The Fate of Dung Carbon in Temperate Grassland Soil: 1. Preliminary Findings Based on Bulk Stable Carbon Isotope Determinations. Isotopes in Health and Environmental Studies

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

    PubMed

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

    2015-10-01

    A simple triple resonance NMR experiment that leads to the correlation of the backbone amide resonances of each amino acid residue 'i' with that of residues 'i-1' and 'i+1' in ((13)C, (15)N) labelled intrinsically disordered proteins (IDPs) is presented. The experimental scheme, {HN-NCA heteronuclear TOCSY-NH}, exploits the favourable relaxation properties of IDPs and the presence of (1) J CαN and (2) J CαN couplings to transfer the (15)N x magnetisation from amino acid residue 'i' to adjacent residues via the application of a band-selective (15)N-(13)C(α) heteronuclear cross-polarisation sequence of ~100 ms duration. Employing non-uniform sampling in the indirect dimensions, the efficacy of the approach has been demonstrated by the acquisition of 3D HNN chemical shift correlation spectra of α-synuclein. The experimental performance of the RF pulse sequence has been compared with that of the conventional INEPT-based HN(CA)NH pulse scheme. As the availability of data from both the HCCNH and HNN experiments will make it possible to use the information extracted from one experiment to simplify the analysis of the data of the other and lead to a robust approach for unambiguous backbone and side-chain resonance assignments, a time-saving strategy for the simultaneous collection of HCCNH and HNN data is also described. PMID:26282620

  3. Sewage derive [sup 15]N in the Baltic traced in fucus

    SciTech Connect

    Hobbie, J.E.; Fry, B. ); Larsson, U.; Elmgren, R. )

    1990-01-09

    Himmerfjarden, a fjord-like bay on the eastern shore of the Baltic, receives treated sewage from 250,000 inhabitants. Because the inorganic N in the effluent is enriched in [sup 15]N through denitrification, nitrification, and ammonia volatilization, an analysis of the distribution of [sup 15]N in the Bay tells how far from the source the sewage nitrogen moves. The attached macroalga Fucus vesiculosus was collected in early May from rocky shore at 0-0.5 m depth and the [sup 15]N content of the tips of the fronds analyzed. This N represents uptake and storage during the previous six months and growth during March and April. The [delta][sup 15]N was uniformly high (11-13[per thousand]) in the main body of the Bay within 15 km from the sewage source. Beyond 15 km values decreased with distance to a low of 4.6[per thousand] at 35 km, where the Bay ends and the coastal waters begin. Using the 11-13 and 4.6[per thousand] as endmembers, the percentage of sewage N making up the Fucus at any point may be calculated. The [delta][sub 15]N of particulate organic matter in the offshore Baltic waters was around 0[per thousand] and Fucus had an [delta][sup 15]N about 1.5[per thousand] higher than the POM. From this and other evidence we conclude that there is a belt of coastal water with an elevated [delta][sup 15]N lying along the east coast of the Baltic. This presumably derives from sewage and perhaps from agriculture and is potentially of use as a tracer of coastal zone/pelagic zone interactions.

  4. Natural abundance 17O, 6Li NMR and molecular modeling studies of the solvation structures of lithium bis(fluorosulfonyl)imide/1,2-dimethoxyethane liquid electrolytes

    NASA Astrophysics Data System (ADS)

    Wan, Chuan; Hu, Mary Y.; Borodin, Oleg; Qian, Jiangfeng; Qin, Zhaohai; Zhang, Ji-Guang; Hu, Jian Zhi

    2016-03-01

    Natural abundance 17O and 6Li NMR experiments, quantum chemistry and molecular dynamics studies were employed to investigate the solvation structures of Li+ at various concentrations of LiFSI in DME electrolytes. It was found that the chemical shifts of both 17O and 6Li changed with the concentration of LiFSI, indicating the changes of solvation structures with concentration. For the quantum chemistry calculations, the coordinated cluster LiFSI(DME)2 forms at first, and its relative ratio increases with increasing LiFSI concentration to 1 M. Then the solvation structure LiFSI(DME) become the dominant component. As a result, the coordination of forming contact ion pairs between Li+ and FSI- ion increases, but the association between Li+ and DME molecule decreases. Furthermore, at LiFSI concentration of 4 M the solvation structures associated with Li+(FSI-)2(DME), Li+2(FSI-)(DME)4 and (LiFSI)2(DME)3 become the dominant components. For the molecular dynamics simulation, with increasing concentration, the association between DME and Li+ decreases, and the coordinated number of FSI- increases, which is in perfect accord with the DFT results.

  5. Studies of individual carbon sites of proteins in solution by natural abundance carbon 13 nuclear magnetic resonance spectroscopy. Strategies for assignments.

    PubMed

    Oldfield, E; Norton, R S; Allerhand, A

    1975-08-25

    Natural abundance 13C Fourier transform NMR spectra (at 15.18 MHz, in 20-mm sample tubes) of aqueous native proteins yield numerous narrow single carbon resonances of nonprotonated aromatic carbons. Techniques for the assignment of these resonances are presented. Each technique is applied to one or more of the following proteins: ferricytochrome c from horse heart and Candida krusei, ferrocytochrome c and cyanoferricytochrome c from horse heart, lysozyme from hen egg white, cyanoferrimyoglobins from horse and sperm whale skeletal muscle, and carbon monoxide myoglobin from horse. In all of the protein spectra we have examined, methine aromatic carbons give rise to broad bands. Studies of the narrow resonances of nonprotonated aromatic carbons of proteins are facilitated by removal of these broad bands by means of the convolution-difference method, preferably from spectra recorded under conditions of noise-modulated off-resonance proton decoupling. We present a summary of the chemical shift ranges for the various types of nonprotonated aromatic carbons of amino acid residues and hemes of diamagnetic proteins, based on our results for hen egg white lysozyme, horse heart ferrocytochrome c, horse carbon monoxide myoglobin, and carbon monoxide hemoglobins from various species... PMID:169240

  6. 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. PMID:21306158

  7. The physical state of osmoregulatory solutes in unicellular algae. A natural-abundance carbon-13 nuclear-magnetic-resonance relaxation study.

    PubMed Central

    Norton, R S; MacKay, M A; Borowitzka, L J

    1982-01-01

    Natural-abundance 13C n.m.r. spin-lattice relaxation-time measurements have been carried out on intact cells of the unicellular blue--green alga Synechococcus sp. and the unicellular green alga Dunaliella salina, with the aim of characterizing the environments of the organic osmoregulatory solutes in these salt-tolerant organisms. In Synechococcus sp., all of the major organic osmoregulatory solute, 2-O-alpha-D-glucopyranosylglycerol, is visible in spectra of intact cells. Its rotational motion in the cell is slower by a factor of approx. 2.4 than in aqueous solution, but the molecule is still freely mobile and therefore able to contribute to the osmotic balance. In D. salina, only about 60% of the osmoregulatory solute glycerol is visible in spectra of intact cells. The rotational mobility of this observable fraction is approximately half that found in aqueous solution, but the data also indicate that there is a significant concentration of some paramagnetic species in D. salina which contributes to the overall spin-lattice relaxation of the glycerol carbon atoms. The non-observable fraction, which must correspond to glycerol molecules that have very broad 13C resonances and that are in slow exchange with bulk glycerol, has not been properly characterized as yet, but may represent glycerol in the chloroplast. The implications of these findings in relation to the physical state of the cytoplasm and the mechanism of osmoregulation in these cells are discussed. PMID:6807296

  8. 3D15N/15N/1H Chemical Shift Correlation Experiment Utilizing an RFDR-based 1H/1H Mixing Period at 100 kHz MAS

    PubMed Central

    Nishiyama, Yusuke; Malon, Michal; Ishii, Yuji; Ramamoorthy, Ayyalusamy

    2014-01-01

    Homonuclear correlation NMR experiments are commonly used in the high-resolution structural studies of proteins. While 13C/13C chemical shift correlation experiments utilizing dipolar recoupling techniques are fully utilized under MAS, correlation of the chemical shifts of 15N nuclei in proteins has been a challenge. Previous studies have shown that the negligible 15N-15N dipolar coupling in peptides or proteins necessitates the use of a very long mixing time (typically several seconds) for effective spin diffusion to occur and considerably slows down a 15N/15N correlation experiment. In this study, we show that the use of mixing proton magnetization, instead of 15N, via the recoupled 1H-1H dipolar couplings enable faster 15N/15N correlation. In addition, the use of proton-detection under ultrafast MAS overcomes the sensitivity loss due to multiple magnetization transfer (between 1H and 15N nuclei) steps. In fact, less than 300 nL (~1.1 micromole quantity) sample is sufficient to acquire the 3D spectrum within 5 hours. Our results also demonstrate that a 3D 15N/15N/1H experiment can render higher resolution spectra that will be useful in the structural studies of proteins at ultrafast MAS frequencies. 3D 15N/15N/1H and 2D radio frequency-driven dipolar recoupling (RFDR)-based 1H/1H experimental results obtained from a powder sample of N-acetyla-L-15N-valyl-L-15N-leucine at 70 and 100 kHz MAS frequencies are presented. PMID:24801998

  9. 3D 15N/15N/1H chemical shift correlation experiment utilizing an RFDR-based 1H/1H mixing period at 100 kHz MAS

    NASA Astrophysics Data System (ADS)

    Nishiyama, Yusuke; Malon, Michal; Ishii, Yuji; Ramamoorthy, Ayyalusamy

    2014-07-01

    Homonuclear correlation NMR experiments are commonly used in the high-resolution structural studies of proteins. While 13C/13C chemical shift correlation experiments utilizing dipolar recoupling techniques are fully utilized under MAS, correlation of the chemical shifts of 15N nuclei in proteins has been a challenge. Previous studies have shown that the negligible 15N-15N dipolar coupling in peptides or proteins necessitates the use of a very long mixing time (typically several seconds) for effective spin diffusion to occur and considerably slows down a 15N/15N correlation experiment. In this study, we show that the use of mixing proton magnetization, instead of 15N, via the recoupled 1H-1H dipolar couplings enable faster 15N/15N correlation. In addition, the use of proton-detection under ultrafast MAS overcomes the sensitivity loss due to multiple magnetization transfer (between 1H and 15N nuclei) steps. In fact, less than 300 nL (?1.1 micromole quantity) sample is sufficient to acquire the 3D spectrum within 5 h. Our results also demonstrate that a 3D 15N/15N/1H experiment can render higher resolution spectra that will be useful in the structural studies of proteins at ultrafast MAS frequencies. 3D 15N/15N/1H and 2D radio frequency-driven dipolar recoupling (RFDR)-based 1H/1H experimental results obtained from a powder sample of N-acetyla-L-15N-valyl-L-15N-leucine at 70 and 100 kHz MAS frequencies are presented.

  10. Rivermouth Alteration of Agricultural Impacts on Consumer Tissue δ15N

    PubMed Central

    Larson, James H.; Richardson, William B.; Vallazza, Jon M.; Nelson, John C.

    2013-01-01

    Terrestrial agricultural activities strongly influence riverine nitrogen (N) dynamics, which is reflected in the δ15N of riverine consumer tissues. However, processes within aquatic ecosystems also influence consumer tissue δ15N. As aquatic processes become more important terrestrial inputs may become a weaker predictor of consumer tissue δ15N. In a previous study, this terrestrial-consumer tissue δ15N connection was very strong at river sites, but was disrupted by processes occurring in rivermouths (the ‘rivermouth effect’). This suggested that watershed indicators of N loading might be accurate in riverine settings, but could be inaccurate when considering N loading to the nearshore of large lakes and oceans. In this study, the rivermouth effect was examined on twenty-five sites spread across the Laurentian Great Lakes. Relationships between agriculture and consumer tissue δ15N occurred in both upstream rivers and at the outlets where rivermouths connect to the nearshore zone, but agriculture explained less variation and had a weaker effect at the outlet. These results suggest that rivermouths may sometimes be significant sources or sinks of N, which would cause N loading estimates to the nearshore zone that are typically made at discharge gages further upstream to be inaccurate. Identifying definitively the controls over the rivermouth effect on N loading (and other nutrients) will require integration of biogeochemical and hydrologic models. PMID:23935980

  11. Rivermouth alteration of agricultural impacts on consumer tissue δ15N

    USGS Publications Warehouse

    Larson, James H.; Richardson, William B.; Vallazza, Jonathan M.; Nelson, J. C.

    2013-01-01

    Terrestrial agricultural activities strongly influence riverine nitrogen (N) dynamics, which is reflected in the δ15N of riverine consumer tissues. However, processes within aquatic ecosystems also influence consumer tissue δ15N. As aquatic processes become more important terrestrial inputs may become a weaker predictor of consumer tissue δ15N. In a previous study, this terrestrial-consumer tissue δ15N connection was very strong at river sites, but was disrupted by processes occurring in rivermouths (the ‘rivermouth effect’). This suggested that watershed indicators of N loading might be accurate in riverine settings, but could be inaccurate when considering N loading to the nearshore of large lakes and oceans. In this study, the rivermouth effect was examined on twenty-five sites spread across the Laurentian Great Lakes. Relationships between agriculture and consumer tissue δ15N occurred in both upstream rivers and at the outlets where rivermouths connect to the nearshore zone, but agriculture explained less variation and had a weaker effect at the outlet. These results suggest that rivermouths may sometimes be significant sources or sinks of N, which would cause N loading estimates to the nearshore zone that are typically made at discharge gages further upstream to be inaccurate. Identifying definitively the controls over the rivermouth effect on N loading (and other nutrients) will require integration of biogeochemical and hydrologic models.

  12. 13C and 15N fractionation of CH4/N2 mixtures during photochemical aerosol formation: Relevance to Titan

    NASA Astrophysics Data System (ADS)

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

    2016-05-01

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

  13. Nitrogen Retention in Three Streams in the Piedmont Region of North Carolina Using a 15N Tracer Addition

    NASA Astrophysics Data System (ADS)

    Rushforth, H. M.; Hershey, A. E.

    2005-05-01

    Stream restoration is often done as a compensatory mitigation project, but it is unresolved as to what extent restoration helps restore natural nutrient cycling within streams. During the summer of 2004 we ran a stable isotope drip of 15 NH 4Cl for 4 weeks in a forested pristine stream, an unrestored urban stream (but with a restored riparian zone), and a restored urban stream (restored channel and riparian zone). All three streams were located in the piedmont region of North Carolina. By collecting and analyzing samples along a 500m reach in each stream we were able to trace 15N uptake into the food web components. Preliminary results indicate that the restored stream was less retentive of 15N in periphyton and other food web components than the unrestored stream. Differences between these two urban streams likely reflect continued nitrogen loading rather than the instream conditions. The unrestored urban stream had consistently higher mean algal biomass than the other two streams. Grazers were consistently more enriched than periphyton in all streams. Hydropsychid caddisflies were isotopically similar to grazers in the forested and unrestored stream, but tracked seston in the restored stream indicating considerable variability in their trophic status.

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

    NASA Astrophysics Data System (ADS)

    Radabaugh, Kara R.; Peebles, Ernst B.

    2014-08-01

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

  15. Nitrate removal in two relict oxbow urban wetlands: a 15N mass-balance approach

    NASA Astrophysics Data System (ADS)

    Harrison, M. D.; Groffman, P. M.; Mayer, P. M.; Kaushal, S.

    2012-12-01

    A mass-balance approach was used to directly determine the flow of 15NO3- to plants, algae, and sediments,with unaccounted for 15N assumed to be denitrified. During the summer, plant and algal uptake accounted for 42%, of the added 15NO3 - in oxbow 1, less than 1% remained in the water column and 57% was unaccounted for. In oxbow 2 during the summer, plant and algal uptake accounted for 63% of the added 15NO3 -, with 1% remaining in the water column and 38% unaccounted for. During the early spring, plant and algal uptake were much lower in both oxbows, ranging from 0.05 to 13.3% of the 15N added, with 97 and 87% was unaccounted for in oxbow 1 and 2, respectively. The amount of unaccounted for 15N was equivalent to estimated areal denitrification rates of 12 and 6 mg N m-2 d-1 in the summer and 78 and 15 mg N m-2 d-1 in the spring, in oxbow 1 and oxbow 2, respectively. However, the uncertainty of these estimates is high as it was difficult to detect accumulation of 15N in the sediments which could have accounted for a very large percentage of the added 15N. Our results suggest that the two relict oxbow wetlands are sinks for NO3 - during both summer and spring. Plane view of Ox1 (A) and Ox2 (B) wetlands with closed contour intervals (color scale) and surrounding stream and upland elevations (labeled in black) located at Minebank Run, near Glen Arms, MD. 15N enrichment (atom %) of measured N pools prior to (Day 0) and after (Day 5) the end of the experiment in July 2009 and April 2010 for Ox1 and Ox2. Values are mean atom % (n = 2 algae, macrophytes and sediment; n = 6 for water samples).

  16. Determination of γ -ray widths in 15N using nuclear resonance fluorescence

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

    Background: The stable nucleus 15N is the mirror of 15O, the bottleneck in the hydrogen burning CNO cycle. Most of the 15N level widths below the proton emission threshold are known from just one nuclear resonance fluorescence (NRF) measurement, with limited precision in some cases. A recent experiment with the AGATA demonstrator array determined level lifetimes using the Doppler shift attenuation method in 15O. As a reference and for testing the method, level lifetimes in 15N have also been determined in the same experiment. Purpose: The latest compilation of 15N level properties dates back to 1991. The limited precision in some cases in the compilation calls for a new measurement to enable a comparison to the AGATA demonstrator data. The widths of several 15N levels have been studied with the NRF method. Method: The solid nitrogen compounds enriched in 15N have been irradiated with bremsstrahlung. The γ rays following the deexcitation of the excited nuclear levels were detected with four high-purity germanium detectors. Results: Integrated photon-scattering cross sections of 10 levels below the proton emission threshold have been measured. Partial γ -ray widths of ground-state transitions were deduced and compared to the literature. The photon-scattering cross sections of two levels above the proton emission threshold, but still below other particle emission energies have also been measured, and proton resonance strengths and proton widths were deduced. Conclusions: Gamma and proton widths consistent with the literature values were obtained, but with greatly improved precision.

  17. Determination of the mutual orientation of the 15N and 13C NMR chemical shift tensors of 13- 15N double labeled model peptides for silk fibroin from the dipolar-coupled powder patterns

    NASA Astrophysics Data System (ADS)

    Asakura, Tetsuo; Yamazaki, Yasunobu; Seng, Koo Wey; Demura, Makoto

    1998-05-01

    The 15N and 13C chemical shift tensors, and the orientation of the principal axis system relative to the molecular symmetry axes were determined for 15N and 13C carbonyl carbon sites of 13C 15N double labeled model peptides for Bombyx mori silk fibroin, that is, Boc-[1- 13C]Ala[ 15N]Gly-OMe, Boc-[1- 13C]Ala[ 15N]GlyAlaGly-OPac, Boc-AlaGly[1- 13C]Ala[ 15N]GlyAlaGly-OPac, Boc-[1- 13C]Gly[ 15N]AlaGlyAla-OPac, Boc-GlyAla[1- 13C]Gly[ 15N]AlaGlyAla-OPac and Boc-[1- 13C]Gly[ 15N]ValGlyAla-OPac, where Boc is t-butoxycarbonyl, OMe is methyl ester, OPac is phenacyl ester, Ala is alanine, Gly is glycine and Val is valine. From the comparisons of the 15N chemical shift tensors and the orientations of the principal axis system relative to the molecular symmetry axes among three compounds having [1- 13C]Ala[ 15N]Gly units, it is concluded that the intermolecular interactions such as hydrogen bonding are different between Boc-[1- 13C]Ala[ 15N]Gly-OMe and two compounds, Boc-[1- 13C]Ala[ 15N]GlyAlaGly-OPac and Boc-AlaGly[1- 13C]Ala[ 15N]GlyAlaGly-OPac although the latter two compounds have similar structures. A similar conclusion has also been obtained from the 13C chemical shift tensors of these compounds.

  18. 15N Fractionation in Star-Forming Regions and Solar System Objects

    NASA Technical Reports Server (NTRS)

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

    2015-01-01

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

  19. The effect of drought and interspecific interactions on depth of water uptake in deep- and shallow-rooting grassland species as determined by δ18O natural abundance

    NASA Astrophysics Data System (ADS)

    Hoekstra, N. J.; Finn, J. A.; Hofer, D.; Lüscher, A.

    2014-08-01

    Increased incidence of drought, as predicted under climate change, has the potential to negatively affect grassland production. Compared to monocultures, vertical belowground niche complementarity between shallow- and deep-rooting species may be an important mechanism resulting in higher yields and higher resistance to drought in grassland mixtures. However, very little is known about the belowground responses in grassland systems and increased insight into these processes may yield important information both to predict the effect of future climate change and better design agricultural systems to cope with this. This study assessed the effect of a 9-week experimental summer drought on the depth of water uptake of two shallow-rooting species (Lolium perenne L. and Trifolium repens L.) and two deep-rooting species (Cichorium intybus L. and Trifolium pratense L.) in grassland monocultures and four-species mixtures by using the natural abundance δ18O isotope method. We tested the following three hypotheses: (1) drought results in a shift of water uptake to deeper soil layers, (2) deep-rooting species take up a higher proportion of water from deeper soil layers relative to shallow-rooting species, and (3) as a result of interspecific interactions in mixtures, the water uptake of shallow-rooting species becomes shallower when grown together with deep-rooting species and vice versa, resulting in reduced niche overlap. The natural abundance δ18O technique provided novel insights into the depth of water uptake of deep- and shallow- rooting grassland species and revealed large shifts in depth of water uptake in response to drought and interspecific interactions. Compared to control conditions, drought reduced the proportional water uptake from 0-10 cm soil depth (PCWU0-10) of L. perenne, T. repens and C. intybus in monocultures by on average 54%. In contrast, the PCWU0-10 of T. pratense in monoculture increased by 44%, and only when grown in mixture did the PCWU0-10 of T. pratense decrease under drought conditions. In line with hypothesis (2), in monoculture, the PCWU0-10 of shallow-rooting species L. perenne and T. repens was 0.53 averaged over the two drought treatments, compared to 0.16 for the deep-rooting C. intybus. Surprisingly, in monoculture, water uptake by T. pratense was shallower than for the shallow-rooting species (PCWU0-10 = 0.68). Interspecific interactions in mixtures resulted in a shift in the depth of water uptake by the different species. As hypothesised, the shallow-rooting species L. perenne and T. repens tended to become shallower, and the deep-rooting T. pratense made a dramatic shift to deeper soil layers (reduction in PCWU0-10 of 58% on average) in mixture compared to monoculture. However, these shifts did not result in a reduction in the proportional similarity of the proportional water uptake from different soil depth intervals (niche overlap) in mixtures compared to monocultures. There was no clear link between interspecific differences in depth of water uptake and the reduction of biomass production under drought compared to control conditions (drought resistance). Cichorium intybus, the species with water uptake from the deepest soil layers was one of the species most affected by drought. Interestingly, T. pratense, which was least affected by drought, also had the greatest plasticity in depth of water uptake. This suggests that there may be an indirect effect of rooting depth on drought resistance, as it determines the potential plasticity in the depth of water uptake.

  20. First results on the incorporation and excretion of 15N from orally administered urea in lactating pony mares.

    PubMed

    Schubert, R; Zander, R; Gruhn, K; Hennig, A

    1991-05-01

    Two lactating pony mares were given oral offers of 20 g 15N urea [95 atom-% 15N-excess (15N')] on 6 subsequent days. About 80% of the consumed 15N' were excreted via urine and faeces, but only about 2% via milk. The 15N' secreted via milk-lysine only amounted to 0.04% of the 15N' intake. The recovery was about 90% in each case. Tissues with active metabolism had an unexpectedly high labelling (greater than 0.3 atom-% 15N'). The low extent of the conversion of oral urea N into milk-lysine speaks against an essential participation of the enteral synthesis in meeting the amino acid requirement of lactating mares. It was already concluded from this results that the determination of the amino acid requirement will be necessary for this group of performance. PMID:1888274

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

    PubMed

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

    2014-06-30

    Many studies show that seagrass ?(15)N ratios increase with the amount of urbanization in coastal watersheds. However, there is little information on the relationship between urbanization and seagrass ?(15)N ratios on a global scale. We performed a meta-analysis on seagrass samples from 79 independent locations to test if seagrass ?(15)N ratios correlate with patterns of population density and fertilizer use within a radius of 10-200 km around the sample locations. Our results show that seagrass ?(15)N ratios are more influenced by intergeneric and latitudinal differences than the degree of urbanization or the amount of fertilizer used in nearby watersheds. The positive correlation between seagrass ?(15)N ratios and latitude hints at an underlying pattern in discrimination or a latitudinal gradient in the (15)N isotopic signature of nitrogen assimilated by the plants. The actual mechanisms responsible for the correlation between ?(15)N and latitude remain unknown. PMID:23866922

  2. δ15N measurement of organic and inorganic substances by EA-IRMS: a speciation-dependent procedure.

    PubMed

    Gentile, Natacha; Rossi, Michel J; Delémont, Olivier; Siegwolf, Rolf T W

    2013-01-01

    Little attention has been paid so far to the influence of the chemical nature of the substance when measuring δ(15)N by elemental analysis (EA)-isotope ratio mass spectrometry (IRMS). Although the bulk nitrogen isotope analysis of organic material is not to be questioned, literature from different disciplines using IRMS provides hints that the quantitative conversion of nitrate into nitrogen presents difficulties. We observed abnormal series of δ(15)N values of laboratory standards and nitrates. These unexpected results were shown to be related to the tailing of the nitrogen peak of nitrate-containing compounds. A series of experiments were set up to investigate the cause of this phenomenon, using ammonium nitrate (NH(4)NO(3)) and potassium nitrate (KNO(3)) samples, two organic laboratory standards as well as the international secondary reference materials IAEA-N1, IAEA-N2-two ammonium sulphates [(NH(4))(2)SO(4)]-and IAEA-NO-3, a potassium nitrate. In experiment 1, we used graphite and vanadium pentoxide (V(2)O(5)) as additives to observe if they could enhance the decomposition (combustion) of nitrates. In experiment 2, we tested another elemental analyser configuration including an additional section of reduced copper in order to see whether or not the tailing could originate from an incomplete reduction process. Finally, we modified several parameters of the method and observed their influence on the peak shape, δ(15)N value and nitrogen content in weight percent of nitrogen of the target substances. We found the best results using mere thermal decomposition in helium, under exclusion of any oxygen. We show that the analytical procedure used for organic samples should not be used for nitrates because of their different chemical nature. We present the best performance given one set of sample introduction parameters for the analysis of nitrates, as well as for the ammonium sulphate IAEA-N1 and IAEA-N2 reference materials. We discuss these results considering the thermochemistry of the substances and the analytical technique itself. The results emphasise the difference in chemical nature of inorganic and organic samples, which necessarily involves distinct thermochemistry when analysed by EA-IRMS. Therefore, they should not be processed using the same analytical procedure. This clearly impacts on the way international secondary reference materials should be used for the calibration of organic laboratory standards. PMID:23099528

  3. Core-shell molecularly imprinted polymer nanoparticles with assistant recognition polymer chains for effective recognition and enrichment of natural low-abundance protein.

    PubMed

    Liu, Dejing; Yang, Qian; Jin, Susu; Song, Yingying; Gao, Junfei; Wang, Ying; Mi, Huaifeng

    2014-02-01

    Core-shell molecular imprinting of nanomaterials overcomes difficulties with template transfer and achieves higher binding capacities for macromolecular imprinting, which are more important to the imprinting of natural low-abundance proteins from cell extracts. In the present study, a novel strategy of preparing core-shell nanostructured molecularly imprinted polymers (MIPs) was developed that combined the core-shell approach with assistant recognition polymer chains (ARPCs). Vinyl-modified silica nanoparticles were used as support and ARPCs were used as additional functional monomers. Immunoglobulin heavy chain binding protein (BiP) from the endoplasmic reticulum (ER) was chosen as the model protein. The cloned template protein BiP was selectively assembled with ARPCs from their library, which contained numerous limited-length polymer chains with randomly distributed recognition and immobilization sites. The resulting complex was copolymerized onto the surface of vinyl-modified silica nanoparticles under low concentrations of the monomers. After template removal, core-shell-structured nanoparticles with a thin imprinted polymer layer were produced. The particles demonstrated considerably high adsorption capacity, fast adsorption kinetics and selective binding affinities toward the template BiP. Furthermore, the synthesized MIP nanoparticles successfully isolated cloned protein BiP from protein mixtures and highly enriched BiP from an ER extract containing thousands of kinds of proteins. The enrichment reached 115-fold and the binding capacity was 5.4 μg g(-1), which were higher than those achieved by using traditional MIP microspheres. The advantageous properties of MIP nanoparticles hold promise for further practical applications in biology, such as protein analysis and purification. PMID:24140608

  4. A critical size and period hypothesis to explain natural regulation of salmon abundance and the linkage to climate and climate change

    NASA Astrophysics Data System (ADS)

    Beamish, R. J.; Mahnken, Conrad

    We hypothesise that salmon year class strength is determined in two stages during the first year in the ocean. There is an early natural mortality that is mostly related to predation, which is followed by a physiologically-based mortality. Juvenile salmon that fail to reach a critical size by the end of their first marine summer do not survive the following winter. In this study we describe our initial tests of this critical size and critical period hypothesis using data from ocean surveys of juvenile salmon and from experimental feeding studies on coho. Conservative swept volume abundance estimates for juvenile coho, and possibly chinook, indicate that there is high mortality in fall and winter during their first year in the sea. Studies of otolith weight show that the length and otolith-weight relationship for young coho changes in the early fall of their first ocean year. Studies of growth and associated hormone levels in feeding studies show that slow growing juvenile coho are stunted and deficient in an insulin-like growth factor-I (IGF-I). Juvenile coho sampled in September had low IGF-I values, indicative of poor growth. The results of these studies provide evidence for the general hypothesis that growth-related mortality occurs late in the first marine year and may be important in determining the strength of the year class (brood year). The link between total mortality and climate could be operating via the availability of nutrients regulating the food supply and hence competition for food (i.e. bottom-up regulation).

  5. Rapid and automated processing of MALDI-FTICR/MS data for 15N-metabolic labeling in a shotgun proteomics analysis

    NASA Astrophysics Data System (ADS)

    Jing, Li; Amster, I. Jonathan

    2009-10-01

    Offline high performance liquid chromatography combined with matrix assisted laser desorption and Fourier transform ion cyclotron resonance mass spectrometry (HPLC-MALDI-FTICR/MS) provides the means to rapidly analyze complex mixtures of peptides, such as those produced by proteolytic digestion of a proteome. This method is particularly useful for making quantitative measurements of changes in protein expression by using 15N-metabolic labeling. Proteolytic digestion of combined labeled and unlabeled proteomes produces complex mixtures with many mass overlaps when analyzed by HPLC-MALDI-FTICR/MS. A significant challenge to data analysis is the matching of pairs of peaks which represent an unlabeled peptide and its labeled counterpart. We have developed an algorithm and incorporated it into a computer program which significantly accelerates the interpretation of 15N-metabolic labeling data by automating the process of identifying unlabeled/labeled peak pairs. The algorithm takes advantage of the high resolution and mass accuracy of FTICR mass spectrometry. The algorithm is shown to be able to successfully identify the 15N/14N peptide pairs and calculate peptide relative abundance ratios in highly complex mixtures from the proteolytic digest of a whole organism protein extract.

  6. [Investigations of the metabolism of colostomised laying hens with 15N-labelled wheat. 5. Incorporation and distribution of 15N in the contents of the intestinal tract, the tissue of stomachs and the segments of the intestines].

    PubMed

    Gruhn, K

    1980-06-01

    In an experiement with 3 colostomised laying hybrids each animal received 80 g pelleted mixed feed and 40 g 15N-labelled wheat with 20.13 atom-% 15N-excess (15N') over a period of four days. On the following four days the hens received rations composed in the same way with uulabelled wheat, however. Of the individual tissues and organs extracted from the slaughtered hens 15N' was determined in the total N and the amino acids lysine, histidine and arginine in the individual segments both of the gasto-intestinal tract and in its contents. On a average, the amount of 15N' in the stomachs, the small intestines and the colon was 43.7%, 27.2% and 29.1%. The tissue of the small intestines contained, on an average, the highest 15N' in lysine of all the basic amino acids. It amounts to 0.82 atom-% 15N' for lysine, O.55% for histidine and 0.63 atom-% 15N' for arginine. On an average, the percentage of the 15N' of the basic amino acids from the corresponding total 15N'-amount of the charges is in the contents of the gasto-intestinal tract 20.5%, in the stomach tissue 28.0% and in the tissue of the small intestines 24.4%, of the caecum 21.5% and of the rectum 25.7%. PMID:7416967

  7. Compound-Specific δ15N Amino Acid Measurements in Littoral Mussels in the California Upwelling Ecosystem: A New Approach to Generating Baseline δ15N Isoscapes for Coastal Ecosystems

    PubMed Central

    Vokhshoori, Natasha L.; McCarthy, Matthew D.

    2014-01-01

    We explored δ15N compound-specific amino acid isotope data (CSI-AA) in filter-feeding intertidal mussels (Mytilus californianus) as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ15N gradients in the California Upwelling Ecosystem (CUE), determining bulk δ15N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ15N values showed a strong linear trend with latitude, increasing from North to South (from ∼7‰ to ∼12‰, R2 = 0.759). In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ15N trend is therefore most consistent with a baseline δ15N gradient, likely due to the mixing of two source waters: low δ15N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC), with15N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ15N values of phenylalanine (δ15NPhe), the best AA proxy for baseline δ15N values. We hypothesize δ15NPhe values in intertidal mussels can approximate annual integrated δ15N values of coastal phytoplankton primary production. We therefore used δ15NPhe values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ15N values. We propose that δ15NPhe isoscapes derived from filter feeders can directly characterize baseline δ15N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives. PMID:24887109

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

    EPA Science Inventory

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

  9. Transfer of (15)N from oral lactose-ureide to lysine in normal adults.

    PubMed

    Jackson, Alan A; Gibson, Neil R; Bundy, Rafe; Hounslow, Angela; Millward, D Joe; Wootton, Stephen A

    2004-09-01

    The metabolic fate of salvaged urea-nitrogen was explored in normal adults who had consumed a diet that provided 36 g protein/day for 7 days. We hypothesised that the colonic microflora utilise nitrogen derived from urea salvage to synthesise lysine in functionally significant amounts for the host. Oral lactose-[(15)N(15)N]ureide is resistant to digestion but is fermented by the colonic microflora to release (15)NH3, which can be used for amino acid synthesis. Prime and intermittent oral doses of lactose-[(15)N(15)N]ureide were ingested for 18 h, urine was collected every 3 h and stools were collected for a further 2 days. Amino acids were isolated from urine and from faecal bacterial protein and the enrichment measured. Compared with baseline values, there was significant enrichment (atoms per cent excess) in faecal bacterial glycine (0.0526), alanine (0.117), lysine (0.0875) and histidine (0.0487), and in urinary glycine (0.016), alanine (0.0144) and lysine (0.0098), but not hisitidine. These data show that the gastrointestinal bacteria can utilise urea-nitrogen in the formation of essential and non-essential amino acids that are available to the host. We estimate that on this low protein diet the amount of lysine from bacterial synthesis and available to the host may be 30 mg/kg/day. These data have important implications for our current perceptions for the dietary requirements for essential amino acids. PMID:15762310

  10. The use of delta(15)N in assessing sewage stress on coral reefs.

    PubMed

    Risk, Michael J; Lapointe, Brian E; Sherwood, Owen A; Bedford, Bradley J

    2009-06-01

    While coral reefs decline, scientists argue, and effective strategies to manage land-based pollution lag behind the extent of the problem. There is need for objective, cost-effective, assessment methods. The measurement of stable nitrogen isotope ratios, delta(15)N, in tissues of reef organisms shows promise as an indicator of sewage stress. The choice of target organism will depend upon study purpose, availability, and other considerations such as conservation. Algae are usually plentiful and have been shown faithfully to track sewage input. The organic matrix of bivalve shells can provide time series spanning, perhaps, decades. Gorgonians have been shown to track sewage, and can provide records potentially centuries-long. In areas where baseline data are lacking, which is almost everywhere, delta(15)N in gorgonians can provide information on status and trends. In coral tissue, delta(15)N combined with insoluble residue determination can provide information on both sewage and sediment stress in areas lacking baseline data. In the developed world, delta(15)N provides objective assessment in a field complicated by conflicting opinions. Sample handling and processing are simple and analysis costs are low. This is a method deserving widespread application. PMID:19286230

  11. Modelling postsilking nitrogen fluxes in maize (Zea mays) using 15N-labelling field experiments.

    PubMed

    Gallais, André; Coque, Marie; Quilléré, Isabelle; Prioul, Jean-Louis; Hirel, Bertrand

    2006-01-01

    In maize (Zea mays), nitrogen (N) remobilization and postflowering N uptake are two processes that provide amino acids for grain protein synthesis. To study the way in which N is allocated to the grain and to the stover, two different 15N-labelling techniques were developed. 15NO(3-) was provided to the soil either at the beginning of stem elongation or after silking. The distribution of 15N in the stover and in the grain was monitored by calculating relative 15N-specific allocation (RSA). A nearly linear relationship between the RSA of the kernels and the RSA of the stover was found as a result of two simultaneous N fluxes: N remobilization from the stover to the grain, and N allocation to the stover and to the grain originating from N uptake. By modelling the 15N fluxes, it was possible to demonstrate that, as a consequence of protein turnover, a large proportion of the amino acids synthesized from the N taken up after silking were integrated into the proteins of the stover, and these proteins were further hydrolysed to provide N to the grain. PMID:17096795

  12. 15N Uptake from Manure and Fertilizer Sousrces by Three Consecutive Crops Under Controlled Conditions.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The North Central “La Comarca Lagunera” region (25° 31’ N, 103° 14’ W), is one of the most important dairy production areas of Mexico. We conducted the first isotopic nitrogen (15N) labeled manure study in Mexico to assess the potential to supply nitrogen (N) to three consecutive forage crops with a...

  13. Using a Macroalgal δ15N Bioassay to Detect Cruise Ship Waste Water Effluent Inputs

    EPA Science Inventory

    Nitrogen stable isotopes are a powerful tool for tracking sources of N to marine ecosystems. I used green macroalgae as a bioassay organism to evaluate if the δ15N signature of cruise ship waste water effluent (CSWWE) could be detected in Skagway Harbor, AK. Opportunistic green...

  14. Tracing Nitrogen through Landscapes to Coastal Wetlands using d15N of Larval Fish

    EPA Science Inventory

    Our objective was to evaluate the use of the nitrogen stable isotope value (d15N) of larval fish as an indicator of incipient anthropogenic nitrogen loading to coastal wetlands in the Great Lakes. We sampled coastal wetlands in five Lake Superior south shore tributaries that had ...

  15. {sup 15}N(p,{alpha}{sub 0}){sup 12}C S factor

    SciTech Connect

    Barker, F. C.

    2008-10-15

    Experimental values of the astrophysical S factor for the {sup 15}N(p,{alpha}{sub 0}){sup 12}C reaction are available both from direct measurements and from the Trojan horse method. We here use R-matrix formulas to fit these values and to extrapolate to zero energy to obtain values of S(0)

  16. Benchmark Theoretical and Experimental Study on (15)N NMR Shifts of Oxidatively Damaged Guanine.

    PubMed

    Dra?nsk, Martin; la, Michal; Klepet?ov, Blanka; ebera, Jakub; Fukal, Ji?; Hole?kov, Veronika; Tanaka, Yoshiyuki; Nencka, Radim; Sychrovsk, Vladimr

    2016-02-11

    The (15)N NMR shifts of 9-ethyl-8-oxoguanine (OG) were calculated and measured in liquid DMSO and in crystal. The OG molecule is a model for oxidatively damaged 2'-deoxyguanosine that occurs owing to oxidative stress in cell. The DNA lesion is repaired with human 8-oxoguanine glycosylase 1 (hOGG1) base-excision repair enzyme, however, the exact mechanism of excision of damaged nucleobase with hOGG1 is currently unknown. This benchmark study on (15)N NMR shifts of OG aims their accurate structural interpretation and calibration of the calculation protocol utilizable in future studies on mechanism of hOGG1 enzyme. The effects of NMR reference, DFT functional, basis set, solvent, structure, and dynamics on calculated (15)N NMR shifts were first evaluated for OG in crystal to calibrate the best performing calculation method. The effect of large-amplitude motions on (15)N NMR shifts of OG in liquid was calculated employing molecular dynamics. The B3LYP method with Iglo-III basis used for B3LYP optimized geometry with 6-311++G(d,p) basis and including effects of solvent and molecular dynamic was the calculation protocol used for calculation of (15)N NMR shifts of OG. The NMR shift of N9 nitrogen of OG was particularly studied because the atom is involved in an N-glycosidic bond that is cleaved with hOGG1. The change of N9 NMR shift owing to oxidation of 9-ethylguanine (G) measured in liquid was -27.1 ppm. The calculated N9 NMR shift of OG deviated from experiment in crystal and in liquid by 0.45 and 0.65 ppm, respectively. The calculated change of N9 NMR shift owing to notable N9-pyramidalization of OG in one previously found polymorph was 20.53 ppm. We therefore assume that the pyramidal geometry of N9 nitrogen that could occur for damaged DNA within hOGG1 catalytic site might be detectable with (15)N NMR spectroscopy. The calculation protocol can be used for accurate structural interpretation of (15)N NMR shifts of oxidatively damaged guanine DNA residue. PMID:26727398

  17. Variable δ15N Diet-Tissue Discrimination Factors among Sharks: Implications for Trophic Position, Diet and Food Web Models

    PubMed Central

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

    2013-01-01

    The application of stable isotopes to characterize the complexities of a species foraging behavior and trophic relationships is dependent on assumptions of δ15N diet-tissue discrimination factors (∆15N). As ∆15N values have been experimentally shown to vary amongst consumers, tissues and diet composition, resolving appropriate species-specific ∆15N values can be complex. Given the logistical and ethical challenges of controlled feeding experiments for determining ∆15N values for large and/or endangered species, our objective was to conduct an assessment of a range of reported ∆15N values that can hypothetically serve as surrogates for describing the predator-prey relationships of four shark species that feed on prey from different trophic levels (i.e., different mean δ15N dietary values). Overall, the most suitable species-specific ∆15N values decreased with increasing dietary-δ15N values based on stable isotope Bayesian ellipse overlap estimates of shark and the principal prey functional groups contributing to the diet determined from stomach content analyses. Thus, a single ∆15N value was not supported for this speciose group of marine predatory fishes. For example, the ∆15N value of 3.7‰ provided the highest percent overlap between prey and predator isotope ellipses for the bonnethead shark (mean diet δ15N = 9‰) whereas a ∆15N value < 2.3‰ provided the highest percent overlap between prey and predator isotope ellipses for the white shark (mean diet δ15N = 15‰). These data corroborate the previously reported inverse ∆15N-dietary δ15N relationship when both isotope ellipses of principal prey functional groups and the broader identified diet of each species were considered supporting the adoption of different ∆15N values that reflect the predators’ δ15N-dietary value. These findings are critical for refining the application of stable isotope modeling approaches as inferences regarding a species’ ecological role in their community will be influenced with consequences for conservation and management actions. PMID:24147026

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

    PubMed

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

    2013-01-01

    The application of stable isotopes to characterize the complexities of a species foraging behavior and trophic relationships is dependent on assumptions of δ(15)N diet-tissue discrimination factors (∆(15)N). As ∆(15)N values have been experimentally shown to vary amongst consumers, tissues and diet composition, resolving appropriate species-specific ∆(15)N values can be complex. Given the logistical and ethical challenges of controlled feeding experiments for determining ∆(15)N values for large and/or endangered species, our objective was to conduct an assessment of a range of reported ∆(15)N values that can hypothetically serve as surrogates for describing the predator-prey relationships of four shark species that feed on prey from different trophic levels (i.e., different mean δ(15)N dietary values). Overall, the most suitable species-specific ∆(15)N values decreased with increasing dietary-δ(15)N values based on stable isotope Bayesian ellipse overlap estimates of shark and the principal prey functional groups contributing to the diet determined from stomach content analyses. Thus, a single ∆(15)N value was not supported for this speciose group of marine predatory fishes. For example, the ∆(15)N value of 3.7‰ provided the highest percent overlap between prey and predator isotope ellipses for the bonnethead shark (mean diet δ(15)N = 9‰) whereas a ∆(15)N value < 2.3‰ provided the highest percent overlap between prey and predator isotope ellipses for the white shark (mean diet δ(15)N = 15‰). These data corroborate the previously reported inverse ∆(15)N-dietary δ(15)N relationship when both isotope ellipses of principal prey functional groups and the broader identified diet of each species were considered supporting the adoption of different ∆(15)N values that reflect the predators' δ(15)N-dietary value. These findings are critical for refining the application of stable isotope modeling approaches as inferences regarding a species' ecological role in their community will be influenced with consequences for conservation and management actions. PMID:24147026

  19. A pilot study for the intrinsic labeling of egg proteins with 15N and 13C.

    PubMed

    Fromentin, Claire; Sanders, Pascal; Nau, Françoise; Anton, Marc; Fromentin, Gilles; Tomé, Daniel; Thibault, Jean-Noël; Gaudichon, Claire

    2012-01-15

    The aim of this study was to produce intrinsically and uniformly doubly (15)N-(13)C-labeled proteins. These proteins can be used as intrinsic tracers of dietary amino acids, both α-amino groups and carbon skeletons, during postprandial metabolic utilization. Two (Rhodes) laying hens were fed for 16 days with a standard poultry diet supplemented with 0, 0.2% or 0.4% of a mixture of 20 doubly (15)N-(13)C-labeled AAs. A third hen was given a non-enriched diet, as the control. The eggs laid were collected over 24 days, from 3 days before to 4 days after supplementation. The (15)N and (13)C enrichments in proteins from white and yolk were measured by EA-IRMS and GC-C-IRMS for enrichment in individual amino acids. After 10 days of supplementation, the (15)N enrichment reached an isotopic plateau at 1500 to 3000 ‰, depending on the supplementation level, in both white and yolk while the (13)C enrichment was 220 to 650 ‰ in white and was 100 to 250 ‰ in yolk. The (15)N enrichment was similar among the amino acids, except for the aromatic ones in which the enrichment was lower. The δ(13)C values were variable among amino acids in both white and yolk, ranging from 77 ‰ for tyrosine to 555 ‰ for proline with the 0.2 % supplementation level. In conclusion, the incorporation of 0.2 % labeled amino acids in the hen diet allowed us to achieve sufficient enrichment for metabolic studies. However, due to the non-homogeneity of the (13)C labeling, adequate (13)C enrichment of individual amino acids must be considered depending on the investigated metabolic pathway. PMID:22215577

  20. **1**5N-NMR INVESTIGATION OF HYDROXYLAMINE DERIVATIZED HUMIC SUBSTANCES.

    USGS Publications Warehouse

    Thorn, Kevin A.; Arterburn, Jeffrey B.; Mikita, Michael A.

    1986-01-01

    Humic substances are the most abundant naturally occurring refactory organic compounds in soils and water. They have a broad range of physical, chemical and physiological properties. In soils, humic substances contribute to the cation exchange capacity, help maintain the physical structure, and play a role in plant growth and nutrition. In aquatic systems, humic substances serve to regulate the levels of inorganic constituents, yield trihalomethanes upon chlorination, and transport or concentrate organic and inorganic pollutants. The oxygen containing functional groups of humic and fulvic acids are believed to play a key role in the chemical properties of humic substances. This study was undertaken to gain additional information on the specific types of oxygen functionalities in humic substances. Since the analysis of hydroxyl moieties had been earlier established, we focused our attention on the analysis of ketone and aldehyde functional groups in humic substances.

  1. A Survey of \\delta18O and \\delta15N Ratios in Ground Water from an Agricultural Community in the San Joaquin Valley, California

    NASA Astrophysics Data System (ADS)

    Glowacki, S. D.; Suen, C. J.

    2004-12-01

    We studied ground water samples from domestic and monitoring wells in an agricultural community in the eastern side of the San Joaquin Valley, California. The study area is rich in alluvial soils creating an extremely fertile farmland. Livestock farms and agricultural fields are abundant in the area. Fifty-four ground water samples were analyzed for \\delta18O and \\delta15N in dissolved nitrate, in addition to nutrients and major minerals. Nitrate concentration levels in groundwater are elevated and affected by agricultural and other activities. Possible sources of nutrients include: a municipal waste-water treatment facility, a raisin processing plant, a meat processing plant, a turkey farm, diary operations, and agricultural fields. However, except for the turkey farm and a diary, we found no statistical significant contribution of nitrate from the other facilities as compared to the rest of the area. The \\delta18O versus \\delta15N ratios plot of dissolved ground water nitrate shows most samples clustered around an area consistent with soil organic nitrogen. In addition, the rest of the samples show a trend that is indicative of denitrification process. Generally, high \\delta15N values are associated with low nitrate concentrations. The isotopic signal of denitrification is particularly pronounced in samples in the vicinity of the waste water treatment facility, where the highest values of \\delta15N and the lowest nitrate concentrations are observed. However, these samples also have elevated chloride concentrations indicating a waste-water source. These data suggest that the denitrification in the subsurface may have been enhanced by bacteria species introduced by the effluence of the plant. [This study was performed with the collaboration of Steven R Silva of USGS, Menlo Park, and Iris Yamagata and Holly Jo Ferrin of California Department of Water Resources.

  2. Leaf δ15N as a physiological indicator of the responsiveness of N2-fixing alfalfa plants to elevated [CO2], temperature and low water availability

    PubMed Central

    Ariz, Idoia; Cruz, Cristina; Neves, Tomé; Irigoyen, Juan J.; Garcia-Olaverri, Carmen; Nogués, Salvador; Aparicio-Tejo, Pedro M.; Aranjuelo, Iker

    2015-01-01

    The natural 15N/14N isotope composition (δ15N) of a tissue is a consequence of its N source and N physiological mechanisms in response to the environment. It could potentially be used as a tracer of N metabolism in plants under changing environmental conditions, where primary N metabolism may be complex, and losses and gains of N fluctuate over time. In order to test the utility of δ15N as an indicator of plant N status in N2-fixing plants grown under various environmental conditions, alfalfa (Medicago sativa L.) plants were subjected to distinct conditions of [CO2] (400 vs. 700 μmol mol−1), temperature (ambient vs. ambient +4°C) and water availability (fully watered vs. water deficiency—WD). As expected, increased [CO2] and temperature stimulated photosynthetic rates and plant growth, whereas these parameters were negatively affected by WD. The determination of δ15N in leaves, stems, roots, and nodules showed that leaves were the most representative organs of the plant response to increased [CO2] and WD. Depletion of heavier N isotopes in plants grown under higher [CO2] and WD conditions reflected decreased transpiration rates, but could also be related to a higher N demand in leaves, as suggested by the decreased leaf N and total soluble protein (TSP) contents detected at 700 μmol mol−1 [CO2] and WD conditions. In summary, leaf δ15N provides relevant information integrating parameters which condition plant responsiveness (e.g., photosynthesis, TSP, N demand, and water transpiration) to environmental conditions. PMID:26322051

  3. Biogenic N2 and δ15 N-N2 As Proxies for N-Loss in the Eastern Tropical North Pacific: A Lagrangian Float Experiment.

    NASA Astrophysics Data System (ADS)

    Bourbonnais, A.; Altabet, M. A.; McNeil, C. L.; Larkum, J.; Reed, A. C.; D'Asaro, E. A.

    2014-12-01

    A large portion of the ocean's bioavailable N, a macronutrient limiting primary productivity, is lost in oxygen minimum zones (OMZs). Mesoscale processes (e.g. eddies, meandering currents), can transport highly productive waters from the coasts, increasing the downward flux of organic material, a substrate for N-loss, and thus can act as N-loss hotspots in OMZs. However, due to their transient nature, these mesoscale events are difficult to monitor using traditional shipboard observations. We deployed biogeochemical Lagrangian floats in the eastern tropical North Pacific during a research cruise in May/June 2014, where transport of high chlorophyll waters from the coast were inferred from satellite imagery. These Lagrangian floats are automonous platforms with the ability to follow isopycnals and were equipped with a suite of gas tension devices and other sensors to measure N2(g), O2, NO3- and NO2- concentrations. We concurrently collected discrete samples to calibrate and complement float measurements. We present here biogenic N2, i.e. N2 produced by local N-loss processes and derived from measured N2/Ar and δ15N-N2 anomaly, i.e. the difference between δ15N-N2 observed and at equilibrium for in-situ temperature and salinity, during a ~4 weeks Lagrangian experiment. During N-loss, the product (N2) is depleted in 15N because of kinetic isotope fractionation. While biogenic N2 only reached up to ~10 µmol/kg, the δ15N-N2 anomalies were relatively low (down to ~-0.4‰). The δ15N-N2 anomalies are low compared to values always >-0.1‰) for equivalent biogenic N2 in the OMZ of the eastern tropical South Pacific. We will discuss the implication of these results for the global oceanic N budget.

  4. Leaf δ(15)N as a physiological indicator of the responsiveness of N2-fixing alfalfa plants to elevated [CO2], temperature and low water availability.

    PubMed

    Ariz, Idoia; Cruz, Cristina; Neves, Tomé; Irigoyen, Juan J; Garcia-Olaverri, Carmen; Nogués, Salvador; Aparicio-Tejo, Pedro M; Aranjuelo, Iker

    2015-01-01

    The natural (15)N/(14)N isotope composition (δ(15)N) of a tissue is a consequence of its N source and N physiological mechanisms in response to the environment. It could potentially be used as a tracer of N metabolism in plants under changing environmental conditions, where primary N metabolism may be complex, and losses and gains of N fluctuate over time. In order to test the utility of δ(15)N as an indicator of plant N status in N2-fixing plants grown under various environmental conditions, alfalfa (Medicago sativa L.) plants were subjected to distinct conditions of [CO2] (400 vs. 700 μmol mol(-1)), temperature (ambient vs. ambient +4°C) and water availability (fully watered vs. water deficiency-WD). As expected, increased [CO2] and temperature stimulated photosynthetic rates and plant growth, whereas these parameters were negatively affected by WD. The determination of δ(15)N in leaves, stems, roots, and nodules showed that leaves were the most representative organs of the plant response to increased [CO2] and WD. Depletion of heavier N isotopes in plants grown under higher [CO2] and WD conditions reflected decreased transpiration rates, but could also be related to a higher N demand in leaves, as suggested by the decreased leaf N and total soluble protein (TSP) contents detected at 700 μmol mol(-1) [CO2] and WD conditions. In summary, leaf δ(15)N provides relevant information integrating parameters which condition plant responsiveness (e.g., photosynthesis, TSP, N demand, and water transpiration) to environmental conditions. PMID:26322051

  5. Influence of open ocean nitrogen supply on the skeletal δ15N of modern shallow-water scleractinian corals

    NASA Astrophysics Data System (ADS)

    Wang, Xingchen T.; Sigman, Daniel M.; Cohen, Anne L.; Sinclair, Daniel J.; Sherrell, Robert M.; Cobb, Kim M.; Erler, Dirk V.; Stolarski, Jarosław; Kitahara, Marcelo V.; Ren, Haojia

    2016-05-01

    The isotopic composition of skeleton-bound organic nitrogen in shallow-water scleractinian corals (hereafter, CS-δ15N) is an emerging tool for studying the marine nitrogen cycle in the past. The CS-δ15N has been shown to reflect the δ15N of nitrogen (N) sources to corals, with most applications to date focusing on the anthropogenic/terrestrial N inputs to reef environments. However, many coral reefs receive their primary N sources from the open ocean, and the CS-δ15N of these corals may provide information on past changes in the open ocean regional and global N cycle. Using a recently developed persulfate/denitrifier-based method, we measured CS-δ15N in modern shallow-water scleractinian corals from 8 sites proximal to the open ocean. At sites with low open ocean surface nitrate concentrations typical of the subtropics and tropics, measured CS-δ15N variation on seasonal and annual timescales is most often less than 2‰. In contrast, a broad range in CS-δ15N (of ∼10‰) is measured across these sites, with a strong correlation between CS-δ15N and the δ15N of the deep nitrate supply to the surface waters near the reefs. While CS-δ15N can be affected by other N sources as well and can vary in response to local reef conditions as well as coral/symbiont physiological changes, this survey indicates that, when considering corals proximal to the open ocean, the δ15N of the subsurface nitrate supply to surface waters drives most of the CS-δ15N variation across the global ocean. Thus, CS-δ15N is a promising proxy for reconstructing the open ocean N cycle in the past.

  6. Reevaluation of siderophile element abundances and ratios across the Cretaceous-Paleogene (K-Pg) boundary: Implications for the nature of the projectile

    NASA Astrophysics Data System (ADS)

    Goderis, S.; Tagle, R.; Belza, J.; Smit, J.; Montanari, A.; Vanhaecke, F.; Erzinger, J.; Claeys, Ph.

    2013-11-01

    The discovery over 30 years ago at Gubbio (Italy) and Caravaca (Spain) of an enrichment in the concentrations of iridium (Ir) and the other platinum group elements (PGE) by up to four orders of magnitude (Irmax = 0.10-87 ng/g) compared to average continental crustal background levels remains one of the most important discoveries in the Earth sciences. Since then, similar anomalies have been detected in more than 120 Cretaceous-Paleogene (K-Pg) boundary sites worldwide. Highly elevated Ir and other siderophile element abundances in roughly chondritic ratios are considered strong indicators for the presence of a meteoritic contribution in impact-related lithologies (melt rocks, impact ejecta material, etc.), delivered when an extraterrestrial object strikes Earth. The presented work adds 113 unpublished PGE analyses of 38 K-Pg sections worldwide to the existing literature. The analytical protocol relied on for this purpose consisted of a combination of a nickel-sulfide fire assay pre-concentration technique and subsequent trace metal determination via inductively coupled plasma-mass spectrometry (ICP-MS). Through repeated determination of key siderophile elements (i.e., Cr, Co, Ni, and PGE), the importance of sampling, nugget effects, and analytical methodologies applied becomes more apparent. Even more critically, these analytical effects are superimposed by the local syn- and post-depositional conditions that have affected the pristine meteoritic signature of the K-Pg impactor, including potential fractionation during vaporization and condensation, dissimilar PGE carrier phases, terrestrial PGE input, sedimentation rate, reworking, diagenesis, bioturbation, and chemical diffusion. While chondrite-normalized PGE patterns of individual sites appear relatively flat (i.e., chondritic), strong variations in siderophile element content and inter-element ratios exist between K-Pg locations, inter-laboratory measurements, and replicate analyses, hampering a precise projectile identification using (highly) siderophile elements. Only when considering improved databases of siderophile element concentrations in meteorites, in combination with linear regression analysis to calculate inter-element ratios from a large suite of ejecta deposit sites, the nature of the K-Pg projectile can be resolved. Application of this methodology to an extensive data set of continental and marine sites, very proximal to distal to the Chicxulub impact structure, supports a carbonaceous chondritic impactor (type CM or CO).

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

    PubMed

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

    2007-10-01

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

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

    PubMed

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

    2016-01-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ(15)N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ(15)N and δ(13)C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ(15)N values span the entire gradient, soil δ(15)N values show a positive correlation with aridity as expected. In contrast, foliar δ(15)N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ(15)N values. PMID:26956399

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

    NASA Astrophysics Data System (ADS)

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

    2016-03-01

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

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

    PubMed Central

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

    2016-01-01

    Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ15N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ15N and δ13C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ15N values span the entire gradient, soil δ15N values show a positive correlation with aridity as expected. In contrast, foliar δ15N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ15N values. PMID:26956399

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

    NASA Astrophysics Data System (ADS)

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

    2016-01-01

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

  12. The origin of nitrogen on Jupiter and Saturn from the 15N/14N ratio

    NASA Astrophysics Data System (ADS)

    Fletcher, Leigh N.; Greathouse, T. K.; Orton, G. S.; Irwin, P. G. J.; Mousis, O.; Sinclair, J. A.; Giles, R. S.

    2014-08-01

    The Texas Echelon cross Echelle Spectrograph (TEXES), mounted on NASA’s Infrared Telescope Facility (IRTF), was used to map mid-infrared ammonia absorption features on both Jupiter and Saturn in February 2013. Ammonia is the principle reservoir of nitrogen on the giant planets, and the ratio of isotopologues (15N/14N) can reveal insights into the molecular carrier (e.g., as N2 or NH3) of nitrogen to the forming protoplanets, and hence the source reservoirs from which these worlds accreted. We targeted two spectral intervals (900 and 960 cm-1) that were relatively clear of terrestrial atmospheric contamination and contained close features of 14NH3 and 15NH3, allowing us to derive the ratio from a single spectrum without ambiguity due to radiometric calibration (the primary source of uncertainty in this study). We present the first ground-based determination of Jupiter’s 15N/14N ratio (in the range from 1.4×10-3 to 2.5×10-3), which is consistent with both previous space-based studies and with the primordial value of the protosolar nebula. On Saturn, we present the first upper limit on the 15N/14N ratio of no larger than 2.0×10-3 for the 900-cm-1 channel and a less stringent requirement that the ratio be no larger than 2.8×10-3 for the 960-cm-1 channel (1σ confidence). Specifically, the data rule out strong 15N-enrichments such as those observed in Titan’s atmosphere and in cometary nitrogen compounds. To the extent possible with ground-based radiometric uncertainties, the saturnian and jovian 15N/14N ratios appear indistinguishable, implying that 15N-enriched ammonia ices could not have been a substantial contributor to the bulk nitrogen inventory of either planet. This result favours accretion of primordial N2 on both planets, either in the gas phase from the solar nebula, or as ices formed at very low temperatures. Finally, spatially-resolved TEXES observations are used to derive zonal contrasts in tropospheric temperatures, phosphine and 14NH3 on both planets, allowing us to relate thermal conditions and chemical compositions to phenomena observed at visible wavelengths in 2013 (e.g., Jupiter’s faint equatorial red colouration event and wave activity in the equatorial belts, plus the remnant warm band on Saturn following the 2010-11 springtime storm).

  13. Determining Degradation and Synthesis Rates of Arabidopsis Proteins Using the Kinetics of Progressive 15N Labeling of Two-dimensional Gel-separated Protein Spots*

    PubMed Central

    Li, Lei; Nelson, Clark J.; Solheim, Cory; Whelan, James; Millar, A. Harvey

    2012-01-01

    The growth and development of plant tissues is associated with an ordered succession of cellular processes that are reflected in the appearance and disappearance of proteins. The control of the kinetics of protein turnover is central to how plants can rapidly and specifically alter protein abundance and thus molecular function in response to environmental or developmental cues. However, the processes of turnover are largely hidden during periods of apparent steady-state protein abundance, and even when proteins accumulate it is unclear whether enhanced synthesis or decreased degradation is responsible. We have used a 15N labeling strategy with inorganic nitrogen sources coupled to a two-dimensional fluorescence difference gel electrophoresis and mass spectrometry analysis of two-dimensional IEF/SDS-PAGE gel spots to define the rate of protein synthesis (KS) and degradation (KD) of Arabidopsis cell culture proteins. Through analysis of MALDI-TOF/TOF mass spectra from 120 protein spots, we were able to quantify KS and KD for 84 proteins across six functional groups and observe over 65-fold variation in protein degradation rates. KS and KD correlate with functional roles of the proteins in the cell and the time in the cell culture cycle. This approach is based on progressive 15N labeling that is innocuous for the plant cells and, because it can be used to target analysis of proteins through the use of specific gel spots, it has broad applicability. PMID:22215636

  14. Study of the 15N(p,γ)16O at LUNA

    NASA Astrophysics Data System (ADS)

    Caciolli, Antonio; LUNA Collaboration

    2012-02-01

    The 15N(p,γ)16O reaction links the first CNO cycle with the second one allowing the production of the oxygen and fluorine isotopes during the hydrogen burning in novae and AGB and RGB stars. In recent years, this reaction has been studied deeply by LUNA (Laboratory for Underground Nuclear Astrophysics). As a matter of fact three different experiments have been run to measure the 15N(p,γ)16O S factor down to the energy of the novae and ABG hydrogen burning reaching for the first time the 72.8 keV in the center of mass. In this contribution the results of the first two experiments are summarized and the results of the recently performed experiment, which is dedicated to the study at very low energies, are presented.

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

    PubMed

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

    2005-03-01

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

  16. Field evidence of differential food utilization of phytal harpacticoids collected from Fucus serratus indicated by δ 13C and δ 15N stable isotopes

    NASA Astrophysics Data System (ADS)

    Steinarsdóttir, M. B.; Ingólfsson, A.; Ólafsson, E.

    2010-06-01

    The present study is the first to compare trophic relationships of several co-occurring phytal harpacticoid species, in their natural habitat, using both δ 13C and δ 15N signatures. Three phytal harpacticoid species/taxa ( Zaus spinatus, Tisbe spp., and Parathalestris cf. intermedia) all collected from the alga Fucus serratus, at different times of the year, were analyzed. The results indicated that the harpacticoids were utilizing food sources differently. Specific food sources of the three species/taxa could not be accurately pinpointed, but there were strong indications that F. serratus and fragments from it did contribute significantly to the diet of P. cf. intermedia and Tisbe spp. Both of these harpacticoid species overlapped in δ 13C and δ 15N values with some of the macrofaunal species, collected from the same site in Hvassahraun, Iceland, while no overlap was seen for Z. spinatus. The signatures for Z. spinatus indicated that its food sources changed seasonally.

  17. SEARCH FOR AN 80-ms SPONTANEOUS FISSION ACTIVITY IN BOMBARDMENTS OF 249Bk WITH 15N

    SciTech Connect

    Nitschke, J.M.; Fowler, M.; Ghiorso, A.; Leber, R.E.; Nurmia, M.J.; Somerville, L.P.; Williams, K.E.; Hulet, E.K.; Landrum, J.H.; Lougheed, R.W.; Wild, J.F.; Bemis, Jr., C.E.; Silva, R.J.; Eskola, P.

    1980-01-01

    A rotating drum system was used to search for an 80-ms spontaneous fission (sf) activity in the reaction of {sup 15}N with {sup 249}Bk. No such activity was found beyond a cross section limit of 0.3 {+-} 0.3 nb. A sf activity with a half-life of about 20 ms and a formation cross section of 12 nb at 82 MeV was observed. The identity of this activity has not been determined.

  18. δ15N in the turtle grass from the Mexican Caribbean

    NASA Astrophysics Data System (ADS)

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

    2013-05-01

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

  19. Food webs of two intermittently open estuaries receiving 15N-enriched sewage effluent

    NASA Astrophysics Data System (ADS)

    Hadwen, Wade L.; Arthington, Angela H.

    2007-01-01

    Carbon and nitrogen stable isotope signatures were used to assess the response of food webs to sewage effluent discharged into two small intermittently open estuaries in northern New South Wales, Australia. One of these systems, Tallows Creek, has a history of direct sewage inputs, whilst the other, Belongil Creek, receives wastewater via an extensive wetland treatment system. The food webs of both systems were driven by algal sources of carbon, reflecting high autotrophic productivity in response to the nutrients entering the system from sewage effluent. All aquatic biota collected from Tallows Creek had significantly enriched δ15N signatures relative to their conspecifics from Belongil Creek, indicating that sewage nitrogen had been assimilated and transferred throughout the Tallows Creek food web. These δ15N values were higher than those reported from studies in permanently open estuaries receiving sewage effluent. We suggest that these enriched signatures and the transfer of nitrogen throughout the entire food web reflect differences in hydrology and associated nitrogen cycling processes between permanently open and intermittently open estuaries. Although all organisms in Tallows Creek were generally 15N-enriched, isotopically light (less 15N-enriched) individuals of estuary perchlet ( Ambassis marianus) and sea mullet ( Mugil cephalus) were also collected. These individuals were most likely recent immigrants into Tallows Creek, as this system had only recently been opened to the ocean. This isotopic discrimination between resident (enriched) and immigrant (significantly less enriched) individuals can provide information on fish movement patterns and the role of heavily polluted intermittently open estuaries in supporting commercially and recreationally valuable estuarine species.

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

    USGS Publications Warehouse

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

    2006-01-01

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

  1. Community 15N isoscapes to resolve plant-plant-interactions at the spatial scale

    NASA Astrophysics Data System (ADS)

    Hellmann, Christine; Rascher, Katherine G.; Máguas, Cristina; Werner, Christiane

    2014-05-01

    Isoscapes have greatly improved our ability to understand biogeochemical processes on continental to global scales. However, the isoscapes framework may also have significant potential to resolve the spatial component of within-community interactions. For example, exotic plant invaders often exert strong impacts on ecosystem functioning, particularly regarding water-, carbon- and nutrient-cycles, but the spatial extent of such alterations is largely unknown. Here we show that massive N input by the N2-fixing exotic invasive Acacia longifolia to a Portuguese dune system can be traced using spatially resolved information on native plants' leaf δ15N. We found isotopic signatures of N to differ strongly between the native system (δ15N c. -10 o) and the atmospherically derived N in A. longifolia phyllodes (δ15N c. 0 o). Thus, sources of N for native plants could be readily distinguished. Leaf δ15N of a native, non-fixing species was increasingly enriched the closer the plant grew to the invader, indicating uptake of fixed N provided by A. longifolia. The enrichment was evident far beyond the stands of the invader, demonstrating that A. longifolia affected N budgets of native species up to a distance of 8 m exceeding the margin of the canopy. Furthermore, using the isoscapes approach, we were able to quantify the total area of N enrichment and could thus show that the area affected by invasion was at least 3.5 times larger than the area actually occupied by the invader. However, a native N2-fixing species had no such effects. Thus, downscaling isoscapes to the community level opens new frontiers in quantifying the spatial dimension of functional changes associated with plant invasions. Moreover, considering the feasibility and applicability of this approach, it may provide a promising tool to identify, quantify and monitor different types of functional plant-plant interactions within communities at a spatially explicit scale.

  2. 15N and 13C NMR Determination of Allantoin Metabolism in Developing Soybean Cotyledons 1

    PubMed Central

    Coker, George T.; Schaefer, Jacob

    1985-01-01

    The metabolism of allantoin by immature cotyledons of soybean (Glycine max L. cv Elf) grown in culture was investigated using solid state 13C and 15N nuclear magnetic resonance. All of the nitrogens of allantoin were incorporated into protein in a manner similar to that of each other and to the amide nitrogen of glutamine. The C-2 of allantoin was not incorporated into cellular material; presumably it was lost as CO2. About 50% of the C-5 of allantoin was incorporated into cellular material as a methylene carbon; the other 50% was presumably also lost as CO2. The 13C-15N bonds of [5-13C;1-15N] and [2-13C;1,3-15N]allantoin were broken prior to the incorporation of the nitrogens into protein. These data are consistent with allantoin's degradation to two molecules of urea and one two-carbon fragment. Cotyledons grown on allantoin as a source of nitrogen accumulated 21% of the nitrogen of cotyledons grown on glutamine. Only 50% of the nitrogen of the degraded allantoin was incorporated into the cotyledon as organic nitrogen; the other 50% was recovered as NH4+ in the media in which the cotyledons had been grown. The latter results suggests that the lower accumulation of nitrogen by cotyledons grown on allantoin was in part due to failure to assimilate NH4+ produced from allantoin. The seed coats had a higher activity of glutamine synthetase and a higher rate of allantoin degradation than cotyledons indicating that seed coats play an important role in the assimilation and degradation of allantoin. PMID:16663995

  3. Affordable uniform isotope labeling with (2)H, (13)C and (15)N in insect cells.

    PubMed

    Sitarska, Agnieszka; Skora, Lukasz; Klopp, Julia; Roest, Susan; Fernández, César; Shrestha, Binesh; Gossert, Alvar D

    2015-06-01

    For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80% can be achieved for (15)N and (13)C with yields comparable to expression in full media. For (2)H,(15)N and (2)H,(13)C,(15)N labeling, incorporation is only slightly lower with 75 and 73%, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins. PMID:25929326

  4. Enzymatic Synthesis and Structural Characterization of 13C, 15N - Poly(ADP-ribose)

    PubMed Central

    Schultheisz, Heather L.; Szymczyna, Blair R.; Williamson, James R.

    2009-01-01

    Poly(ADP-ribose) is a significant nucleic acid polymer involved with diverse functions in eukaryotic cells, yet no structural information is available. A method for the synthesis of 13C, 15N- poly(ADP-ribose) (PAR) has been developed to allow characterization of the polymer using multidimensional nuclear magnetic resonance (NMR) spectroscopy. Successful integration of pentose phosphate, nicotinamide adenine dinucleotide biosynthesis, and cofactor recycling pathways with poly(ADP-ribose) polymerase-1 permitted labeling of PAR from 13C-glucose and 13C, 15N- ATP in a single pot reaction. The scheme is efficient, yielding ~ 400 nmoles of purified PAR from 5 μmoles ATP, and the behavior of the synthetic PAR is similar to data from PAR synthesized by cell extracts. The resonances for 13C, 15N-PAR were unambiguously assigned, but the polymer appears to be devoid of inherent regular structure. PAR may form an ordered macromolecular structure when interacting with proteins, and due to the extensive involvement of PAR in cell function and disease, further studies of PAR structure will be required. The labeled PAR synthesis reported here will provide an essential tool for the future study of PAR-protein complexes. PMID:19757771

  5. Impact of seaweed beachings on dynamics of δ(15)N isotopic signatures in marine macroalgae.

    PubMed

    Lemesle, Stéphanie; Mussio, Isabelle; Rusig, Anne-Marie; Menet-Nédélec, Florence; Claquin, Pascal

    2015-08-15

    A fine-scale survey of δ(15)N, δ(13)C, tissue-N in seaweeds was conducted using samples from 17 sampling points at two sites (Grandcamp-Maisy (GM), Courseulles/Mer (COU)) along the French coast of the English Channel in 2012 and 2013. Partial triadic analysis was performed on the parameter data sets and revealed the functioning of three areas: one estuary (EstA) and two rocky areas (GM(∗), COU(∗)). In contrast to oceanic and anthropogenic reference points similar temporal dynamics characterized δ(15)N signatures and N contents at GM(∗) and COU(∗). Nutrient dynamics were similar: the N-concentrations in seawater originated from the River Seine and local coastal rivers while P-concentrations mainly from these local rivers. δ(15)N at GM(∗) were linked to turbidity suggesting inputs of autochthonous organic matter from large-scale summer seaweed beachings made up of a mixture of Rhodophyta, Phaeophyta and Chlorophyta species. This study highlights the coupling between seaweed beachings and nitrogen sources of intertidal macroalgae. PMID:26095988

  6. Incorporation of (15)N from spiked cattle dung pats into soil under two moorland plant communities.

    PubMed

    Shepherd, M J; Anderson, J M; Bol, R; Allen, D K

    2000-01-01

    The rate and depth of cattle dung incorporation into moorland soil may be an important factor influencing plant community dynamics through its effects on soil nutrient availability. This study traces the incorporation of (15)N-labelled dung into a moorland soil under two vegetation types in Dartmoor National Park, UK. Cores of treated and control soil 10 cm deep were collected at 2, 4, 8 and 16 week intervals and divided into 2 cm depth increments. Soil samples were freeze-dried, ground and analysed for atom% (15)N and %N content using continuous-flow isotope-ratio mass spectrometry. The contribution of dung N to the soil N pool was estimated by changes in atom% (15)N of the soil. The incorporation of dung dry matter into the soil was also calculated. The labile component of the dung N was incorporated deeper and more rapidly into soil under grass than under heather vegetation. The implications of these processes for the dynamics of upland plant communities are considered in relation to the ability of plants to compete for nutrients. PMID:10920356

  7. Thousand Year Archives of the Bulk and Compound-Specific δ15N of Export Production From the North Pacific Subtropical Gyre Indicate Increasing Nitrogen Fixation Over the Past 150 Years

    NASA Astrophysics Data System (ADS)

    Sherwood, O.; Batista, F. C.; Brown, J. T.; Guilderson, T. P.; McCarthy, M.

    2012-12-01

    Stable nitrogen isotopic analysis of amino acids (δ15N-AA) preserved in proteins has emerged as a powerful new tool to explore trophic levels and nutrient cycling in nature. To date, little has been done to explore δ15N-AA in paleo-studies of the marine nitrogen cycle. We analysed the bulk and AA-specific δ15N in the long-lived, deep-sea, proteinaceous coral Gerardia. By feeding on sinking particulate organic matter, proteinaceous corals integrate the biogeochemical signature of recently exported production within discrete skeletal growth layers. Sub-decadal resolution time-series records spanning the time period 1000 AD to present were generated from specimens of Gerardia collected from the main Hawaiian Islands, Cross Seamount, and French Frigate Shoals in the North Pacific Subtropical Gyre (NPSG). Records of bulk δ15N from the three different locations, geographically separated by up to 1000 km, showed remarkably similar long term trends. Bulk δ15N remained relatively stable from ~1000-1850 years AD, and then decreased by a total of 2 ‰ from ~1850 AD to the present. The δ15N-AA of the "trophic" group of amino acids indicated no significant changes in trophic level or microbial re-synthesis of export production over this time period. The δ15N of "source" amino acids was significantly correlated with corresponding values of bulk δ15N, with the δ15N of phenylalanine decreasing from 4.2 to 2.1‰. The latter value is similar to recent measurements of subsurface nitrate δ15N near Hawaii, suggesting that the δ15N of phenylalanine may be used to quantitatively track changes in the isotopic signature of nitrate at the base of the food web. Using a simple isotopic mass balance between upwelled nitrate and nitrogen fixation we calculate a 30% increase in nitrogen fixation in the NPSG since ~1850. These results provide invaluable long-term context for recent observations, and highlight profound changes in the marine biogeochemical cycling of nitrogen over the past 150 years in this climatically-sensitive area of the world ocean.

  8. Abundance coefficients, a new method for measuring microorganism relative abundance

    USGS Publications Warehouse

    Forester, R.M.

    1977-01-01

    A new method of measuring the relative abundance of microorganisms by using a set of interrelated coefficients, termed 'abundance coefficients' or 'AC', is proposed. These coefficients provide a means of recording abundance for geometric density categories, and each density measurement represents an approximation of the Poisson parameter ??t. The AC is the natural logarithm of a 'characteristic value,' which is a particular number for each geometric density category. The 'characteristic values' are based upon a probabilistic error statement derived from the Poisson formula, and they present evidence for separation of the geometric category boundaries by e = 2.71828. The proposed AC provide a means for recording species abundance in a manner suitable for arithmetic manipulation, for population structure studies, and for the determination of practical limits for defining the presence or absence of a species. Further, these coefficients provide for both intrasample and intersample abundance comparisons. ?? 1977 Plenum Publishing Corporation.

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

    NASA Astrophysics Data System (ADS)

    Fogel, M. L.; Wooller, M. J.; Cheeseman, J.; Smallwood, B. J.; Roberts, Q.; Romero, I.; Meyers, M. J.

    2008-12-01

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

  10. Controls on Hyporheic Nitrate Removal: Assessing Transport and Substrate Limitations with 15N Tracer Studies (Invited)

    NASA Astrophysics Data System (ADS)

    Zarnetske, J. P.; Haggerty, R.; Wondzell, S. M.; Baker, M. A.

    2010-12-01

    We examined transport time and substrate controls on hyporheic (HZ) nitrification and denitrification in an upland agricultural stream with a series of 15N tracer studies - whole-stream and in situ well-to-well steady-state 15NO3- and conservative tracer (Cl-) addition experiments. For the whole-stream experiment, we measured relevant solute, 15N isotope, and hydraulic transport conditions of the reach and along HZ flowpaths of an instrumented gravel bar. HZ exchange was observed across the entire gravel bar with flowpath lengths up to 4.2m and corresponding median residence times greater than 28.5h. The HZ transitioned from a net nitrification environment at its head (small residence times, <6.9h) to a net denitrification environment at its tail (large residence times, 6.9-28.5h). HZ denitrification was confirmed as 15N2 was produced across the entire gravel bar. Production of 15N2 across all observed flowpaths and residence times indicated that denitrification microsites are present even where net nitrification occurred. At large residence times, the rate of denitrification decreased despite persistent anoxic conditions, indicating substrate (NO3- and carbon) limitations. Consequently, we conducted in situ 15NO3-, conservative tracers (Cl- and Br), and acetate injection experiments to determine how the availability of labile dissolved organic carbon (DOC) as acetate influences microbial denitrification in the HZ, especially along anoxic flowpaths with large residence times. The acetate addition to the HZ stimulated significant increases in 15N2 production by factors of 2.7 to 26.1 in all receiving wells, and significant decreases of NO3- and DOC aromaticity in the wells most hydrologically connected to the injection. Further, 100% of acetate was retained in the HZ, a portion of which is due to biological consumption. These studies demonstrate that: 1. the HZ is an active nitrogen sink in this study system, 2. the distinction between net nitrification and denitrification in the HZ is a function of residence time and exhibits threshold behavior, and 3. microbial denitrification in anaerobic portions of the HZ can be limited by labile DOC supply.

  11. Denitrification in nitrate-rich streams: Application of N2:Ar and 15N-tracer methods in intact cores

    USGS Publications Warehouse

    Smith, L.K.; Voytek, M.A.; Böhlke, J.K.; Harvey, J.W.

    2006-01-01

    Rates of benthic denitrification were measured using two techniques, membrane inlet mass spectrometry (MIMS) and isotope ratio mass spectrometry (IRMS), applied to sediment cores from two NO3--rich streams draining agricultural land in the upper Mississippi River Basin. Denitrification was estimated simultaneously from measurements of N 2:Ar (MIMS) and 15N[N2] (IRMS) after the addition of low-level 15NO3- tracer ( 15N:N = 0.03-0.08) in stream water overlying intact sediment cores. Denitrification rates ranged from about 0 to 4400 lmol N??m -2??h-1 in Sugar Creek and from 0 to 1300 ??mol N??m-2??h-1 in Iroquois River, the latter of which possesses greater streamflow discharge and a more homogeneous streambed and water column. Within the uncertainties of the two techniques, there is good agreement between the MIMS and IRMS results, which indicates that the production of N2 by the coupled process of nitrification/denitrification was relatively unimportant and surface-water NO3- was the dominant source of NO3- for benthic denitrification in these streams. Variation in stream NO3- concentration (from about 20 ??mol/L during low discharge to 1000 ??mol/L during high discharge) was a significant control of benthic denitrification rates, judging from the more abundant MIMS data. The interpretation that NO3- concentration directly affects denitrification rate was corroborated by increased rates of denitrification in cores amended with NO 3-. Denitrification in Sugar Creek removed ???11% per day of the instream NO3- in late spring and removed roughly 15-20% in late summer. The fraction of NO3- removed in Iroquois River was less than that of Sugar Creek. Although benthic denitrification rates were relatively high during periods of high stream flow, when NO3 concentrations were also high, the increase in benthic denitrification could not compensate for the much larger increase in stream NO3- fluxes during high flow. Consequently, fractional NO3- losses were relatively low during high flow. ?? 2006 by the Ecological Society of America.

  12. Foliar retention of 15N-nitrate and 15N-ammonium by red maple (Acer rubrum) and white oak (Quercus alba) leaves from simulated rain

    SciTech Connect

    Garten Jr, Charles T; Hanson, Paul J

    1990-07-01

    Studies of nitrogen cycling in forests indicate that trees assimilate atmospheric nitrate and ammonium and that differences between atmospheric deposition to the forest canopy and deposition measured in forest throughfall can be attributed to the removal of these ions from rain by tree leaves. Red maple and white oak leaves were exposed to artificial rain solutions (pH 4.1) containing {sup 15}N-labeled nitrate (3.5 {micro}g N/ml) or ammonium (2.2 {micro}g N/ml). At two time intervals after exposure (2 hr and 2 days) an exposed leaf and a control (non-exposed) leaf were removed from replicate seedlings. Based on results from {sup 15}N analysis, most of the nitrate applied to tree leaves was removed by washing with water; the mean per cent removal ({+-} standard error, N = 4) was 87 {+-} 1 and 73 {+-} 4% of the {sup 15}NO-N Applied to red maple and white oak leaves, respectively. Relative retention of {sup 15}NH{sub 4}-N by the leaves was greater than that observed for {sup 15}NO{sub 3}-N. In red maple and white oak leaves, 58 {+-} 9 and 84 {+-} 7% (mean {+-} standard error, N = 4), respectively, of the applied ammonium was not removed by washing treatments. Our results show that the foliar uptake of {sup 15}NH{sub 4}{sup +} from simulated rain by deciduous tree leaves is greater than that for {sup 15}NO{sub 3}{sup -}. Greater retention of NH{sub 4}{sup +} than NO{sub 3}{sup -} ions by red maple and white oak leaves from simulated rainfall is consistent with field observations showing a preferential retention of ammonium from rainfall by forest canopies. As nitrogen chemistry and the relative importance of nitrogen compounds in the atmosphere change in response to proposed emission reductions (and possibly climate change), an improved understanding of the fate of airborne nitrogen compounds in forest biogeochemical cycles will be necessary.

  13. Variable Seawater-Peridotite Interactions - First Insights From ODP Leg 209, MAR 15 N

    NASA Astrophysics Data System (ADS)

    Bach, W.; Garrido, C. J.; Harvey, J.; Paulick, H.; Rosner, M.; Odp Leg 193 Shipboard Science Party

    2003-12-01

    Serpentinization of peridotites at slow-spreading mid-ocean ridges has important consequences for the rheology of the oceanic lithosphere, geochemical budgets of the oceans, and microbial processes within, at, and above the seafloor. ODP Leg 209 recovered peridotites that show a remarkable variability of hydrothermal alteration reactions and intensities, including talc-tremolite alteration of pyroxenes associated with incipient serpentinization of olivine, complete alteration of peridotites to serpentine and magnetite followed by the destruction of magnetite and replacement of serpentine by talc, variable degrees of serpentine-brucite alteration, and replacement of brucite by iowaite. These rocks provide a unique opportunity to calibrate our observations against recent results from experimental/theoretical geochemical studies and further our understanding of serpentinization and its role in tectonic accretion and microbial colonization of oceanic lithosphere at slow and ultraslow spreading ridges. We propose that at temperatures above 250 C, pyroxenes react to form serpentine, talc, and tremolite, releasing Ca, Si, H2, and acidity to the reacting fluids that may cause rodingitization in adjacent gabbro bodies. Overall however, rodingites are rare, which may reflect the depleted nature of the mantle protoliths. In the absence of pyroxenes (in dunites) - or at temperatures below 250 C, where pyroxenes react very slowly - the fluids do not become enriched in Ca and Si and serpentine, magnetite, and brucite will form. Many serpentinites lack brucite, tremolite, and talc, because changes in fluid pH and silica activity of the interacting fluids, following the exhaustion of either olivine or pyroxene, caused reaction of these phases to serpentine. Extensive talc alteration of serpentinites and gabbros is usually observed at the intrusive contacts, but large-scale silica metasomatism (or Mg-loss) must be invoked to explain the overall abundance of talc at Site 1268. Significant pyrite/marcasite/hematite veining at Site 1268 indicates fairly oxidizing conditions consistent with the presence of sulfate in the interacting fluids. The first discovery of iowaite in mid-ocean ridge serpentinites (at Site 1272) and the abundant carbonate/Fe-oxyhydroxide alteration, locally extending down to 90 meters below seafloor, indicate that water-rock reactions continue at low temperatures and under strongly oxidizing conditions. Pore fluids from nontronite-bearing serpentine muds in fault gouges may provide information about the nature of these late-stage circulating fluids and potential microbial activity.

  14. N2O and δ15N-N2O and δ18O-N2O from polar ice cores: interpretable data for interglacials

    NASA Astrophysics Data System (ADS)

    Bock, Michael; Schmitt, Jochen; Seth, Barbara; Beck, Jonas; Fischer, Hubertus

    2014-05-01

    Ice cores provide a wealth of information on climate change. For instance, the history of the atmospheric greenhouse gas N2O can be reconstructed using air entrapped in polar ice cores. N2O has several sources in both terrestrial and marine ecosystems, predominantly wetland soils and oxygen minimum zones in the ocean. N2O records generally follow the climatic changes during the glacial-interglacial cycles with higher N2O mixing ratios during warmer climate stages. However, the underlying processes driving these changes are difficult to identify from N2O mixing ratios alone. Additional information on the individual sources and sinks are provided by stable isotope measurements. The emission fluxes of the dominant N2O sources are ascribed to several pathways (nitrification, denitrification), with characteristic fractionation factors for the nitrogen and oxygen isotope signatures of the generated N2O (δ15N-N2O and δ18O-N2O). In the end, the individual proportions of pathways are responsible for distinct δ15N-N2O and δ18O-N2O for the average terrestrial and marine sources. Here, we present new ice core measurements of δ15N-N2O and δ18O-N2O covering the Holocene, MIS 5 and MIS 11. For the past 15 kyrs the δ15N-N2O record shows a continuous decrease starting at 15 kyrs to about 6 kyrs; during the past 6 kyrs δ15N-N2O remains rather constant. The resemblance with a recently published global reconstruction of bulk δ15N is remarkable (McLauchlan et al. 2013, Nature). Taken at face value this could mean that mainly the terrestrial source signature changed rather than a shift in the relative proportions of the terrestrial and marine source. The integrity of N2O ice core records relies on the assumption that the measurements truly represent the past atmosphere. However, comparative analyses of different ice cores from the same age intervals show offsets in the N2O mixing ratios among the records. One likely assumption is that higher mixing ratios are due to in-situ production since N2O should not be lost in the ice. In situ production appears to be evident at least for glacial sections of some ice cores.

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

    NASA Astrophysics Data System (ADS)

    Fogel, M. L.; Wooller, M. J.; Cheeseman, J.; Smallwood, B. J.; Roberts, Q.; Romero, I.; Jacobsen Meyers, M.

    2008-02-01

    Extremes in (δ15N values in mangrove tissues and lichens (range = +4 to -22‰) were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, atmospheric ammonia, mangrove leaves, roots, stems, and wood, and lichens, were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Porewater ammonium concentrations had little to no relationship to N isotopic fractionation in mangrove tissues. The δ15N of fine and coarse roots was 9‰ more positive than leaf tissue from the same tree. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year to a &delta:15N closer to the &delta:15N of porewater ammonium (δ15N=+4‰). Isotopically negative ammonia in the atmosphere (δ15N=-18‰) and in rainwater (δ15N=-9‰) were found on Twin Cays and may be sources of available N for isotopically depleted mangrove trees and lichens. In highly stressed, severely P limited trees, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-03-01

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

  18. Use of protein trans-splicing to produce active and segmentally 2H, 15N labeled mannuronan C5-epimerase AlgE4

    PubMed Central

    Buchinger, Edith; Aachmann, Finn L; Aranko, A Sesilja; Valla, Svein; Skjåk-BræK, Gudmund; Iwaï, Hideo; Wimmer, Reinhard

    2010-01-01

    Alginate epimerases are large multidomain proteins capable of epimerising C5 on β-d-mannuronic acid (M) turning it into α-l-guluronic acid (G) in a polymeric alginate. Azotobacter vinelandii secretes a family of seven epimerases, each of which is capable of producing alginates with characteristic G distribution patterns. All seven epimerases consist of two types of modules, denoted A and R, in varying numbers. Attempts to study these enzymes with solution-state NMR are hampered by their size—the smallest epimerase, AlgE4, consisting of one A- and one R-module, is 58 kDa, resulting in heavy signal overlap impairing the interpretation of NMR spectra. Thus we obtained segmentally 2H, 15N labeled AlgE4 isotopomeres (A-[2H, 15N]-R and [2H, 15N]-A-R) by protein trans-splicing using the naturally split intein of Nostoc punctiforme. The NMR spectra of native AlgE4 and the ligated versions coincide well proving the conservation of protein structure. The activity of the ligated AlgE4 was verified by two different enzyme activity assays, demonstrating that ligated AlgE4 displays the same catalytic activity as wild-type AlgE4. PMID:20552686

  19. Retrieving nitrogen isotopic signatures from fresh leaf reflectance spectra: disentangling δ15N from biochemical and structural leaf properties

    PubMed Central

    Hellmann, Christine; Große-Stoltenberg, André; Lauströ, Verena; Oldeland, Jens; Werner, Christiane

    2015-01-01

    Linking remote sensing methodology to stable isotope ecology provides a promising approach to study ecological processes from small to large spatial scales. Here, we show that δ15N can be detected in fresh leaf reflectance spectra of field samples along a spatial gradient of increasing nitrogen input from an N2-fixing invasive species. However, in field data it is unclear whether δ15N directly influences leaf reflectance spectra or if the relationship is based on covariation between δ15N and foliar nitrogen content or other leaf properties. Using a 15N-labeling approach, we experimentally varied δ15N independently of any other leaf properties in three plant species across different leaf developmental and physiological states. δ15N could successfully be modeled by means of partial least squares (PLSs) regressions, using leaf reflectance spectra as predictor variables. PLS models explained 53–73% of the variation in δ15N within species. Several wavelength regions important for predicting δ15N were consistent across species and could furthermore be related to known absorption features of N-containing molecular bonds. By eliminating covariation with other leaf properties as an explanation for the relationship between reflectance and δ15N, our results demonstrate that 15N itself has an inherent effect on leaf reflectance spectra. Thus, our study substantiates the use of spectroscopic measurements to retrieve isotopic signatures for ecological studies and encourages future development. Furthermore, our results highlight the great potential of optical measurements for up-scaling isotope ecology to larger spatial scales. PMID:25983740

  20. Why so low? Making sense of 15N-depleted nitrogen isotope values in the Late Cretaceous.

    NASA Astrophysics Data System (ADS)

    Junium, C. K.; Meyers, S. R.; Arthur, M. A.

    2014-12-01

    Sedimentary organic matter from Cretaceous Oceanic Anoxic Events is characterized by universally low δ15N values. This observation has generated significant curiosity in the paleoceanographic community and has resulted in several conceptual models that aim to explain the 15N-depletion. The unifying element in these hypotheses is widespread anoxia and a series of nitrogen and/or metal cycling feedbacks. To address these hypotheses I will examine an interval in the middle Turonian (92.0 to 90.8 Ma) that demonstrates the impacts of redox and climate variability on nitrogen cycle dynamics. New N-isotope measurements from ODP Site 1259, Demerara Rise (DR) reveal δ15N values that range from +0.2‰ to -3.5‰ and oscillate by 1.5‰ to 3‰ over 1.6 Ma. A new astronomical time scale reveals a strong ~100 ka cyclicity in δ15N with little variance at ~400 ka through the mid-Turonian. The highest δ15N values and the largest amplitude ~100 ka cycles are found within the proposed glacial interval and cooler sea surface temperatures. The persistence of the ~100 ka δ15N cycle and δ15N maxima suggests a strong link between oceanic ventilation/circulation, climate, and the oscillations in δ15N are readily explained by variations in oxidation and expansion in the extent of denitrification within anoxic intermediate waters. These data confirm the strong link between water column redox variability and low δ15N values, but the exact mechanism of the 15N-depletion remains elusive.

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

    NASA Astrophysics Data System (ADS)

    Schleuß, Per-Marten; Kuzyakov, Yakov

    2014-05-01

    The Kobresia Pygmea ecotone covers approximately 450.000 km2 and is of large global and regional importance due several socio-ecological aspects. For instance Kobresia pastures store high amounts of carbon, nitrogen and other nutrients, represent large grazing areas for herbivores, provide a fast regrowth after grazing events and protect against mechanical degradation and soil erosion. However, Kobresia pastures are assumed to be a grazing induced and are accompanied with distinct root mats varying in thickness between 5-30 cm. Yet, less is known about the morphology and the functions of this root mats, especially in the background of a progressing degradation due to changes of climate and management. Thus we aimed to identify the importance of single soil layers for plant nutrition. Accordingly, nitrogen uptake from different soil depths and its remain in above-ground biomass (AGB), belowground biomass (BGB) and soil were determined by using a 15N pulse labeling approach during the vegetation period in summer 2012. 15N urea was injected into six different soil depths (0.5 cm, 2.5 cm, 7.5 cm, 12.5 cm, 17.5 cm, 22.5 cm / for each 4 replicates) and plots were sampled 45 days after the labeling. For soil and BGB samples were taken in strict sample intervals of 0-1 cm, 1-5 cm, 5-10 cm, 10-15 cm, 15-20 cm, 20-25 cm. Results indicate that total recovery (including AGB, BGB and soil) was highest, if tracer was injected into the top 5 cm and subsequently decreased with decreasing injection depth. This is especially the case for the 15N recovery of BGB, which is clearly attributed to the root density and strongly decreased with soil depth. In contrast, the root activity derived from the 15N content of roots increased with soil depth, which is primary associated to a proportionate increase of living roots related to dead roots. However, most 15N was captured in plant biomass (67.5-85.3 % of total recovery), indicating high 15N uptake efficiency possibly due to N limitation of Kobresia ecosystems. Considering only the nitrogen uptake of AGB hardly any differences appeared between the six injection depths. Nevertheless, it could be shown, that 50.4 % percent of total variance of AGB nitrogen uptake could be explained by combining root density and root activity. Concluding, from the upper root mat horizons highest amounts of nitrogen were taken up by plants, because root densities are correspondingly high. However, in deeper root mat layers the root activity increases and accordingly plays a key role for plant nitrogen supply in this depth. Underlying causes for increasing root activities may be better soil moisture conditions, lower variation of soil temperature and/or a higher access to plant available nitrogen in deeper soil layers.Please fill in your abstract text.

  2. Resonance strengths in the {sup 14}N(p,gamma){sup 15}O and {sup 15}N(p,alphagamma){sup 12}C reactions

    SciTech Connect

    Marta, Michele; Trompler, Erik; Bemmerer, Daniel; Beyer, Roland; Grosse, Eckart; Hannaske, Roland; Junghans, Arnd R.; Nair, Chithra; Schwengner, Ronald; Wagner, Andreas; Yakorev, Dmitry; Broggini, Carlo; Caciolli, Antonio; Erhard, Martin; Menegazzo, Roberto; Fueloep, Zsolt; Gyuerky, Gyoergy; Szuecs, Tamas; Vezzu, Simone

    2010-05-15

    The {sup 14}N(p,gamma){sup 15}O reaction is the slowest reaction of the carbon-nitrogen-oxygen cycle of hydrogen burning in stars. As a consequence, it determines the rate of the cycle. The {sup 15}N(p,alphagamma){sup 12}C reaction is frequently used in inverse kinematics for hydrogen depth profiling in materials. The {sup 14}N(p,gamma){sup 15}O and {sup 15}N(p,alphagamma){sup 12}C reactions have been studied simultaneously, using titanium nitride targets of natural isotopic composition and a proton beam. The strengths of the resonances at E{sub p} = 1058 keV in {sup 14}N(p,gamma){sup 15}O and at E{sub p} = 897 and 430 keV in {sup 15}N(p,alphagamma){sup 12}C have been determined with improved precision, relative to the well-known resonance at E{sub p} = 278 keV in {sup 14}N(p,gamma){sup 15}O. The new recommended values are omegagamma=0.353+-0.018, 362+-20, and 21.9+-1.0 eV for their respective strengths. In addition, the branching ratios for the decay of the E{sub p} = 1058 keV resonance in {sup 14}N(p,gamma){sup 15}O have been redetermined. The data reported here should facilitate future studies of off-resonant capture in the {sup 14}N(p,gamma){sup 15}O reaction that are needed for an improved R-matrix extrapolation of the cross section. In addition, the data on the 430 keV resonance in {sup 15}N(p,alphagamma){sup 12}C may be useful for hydrogen depth profiling.

  3. Foliar uptake of wet-deposited nitrogen by norway spruce. an experiment using 15N

    NASA Astrophysics Data System (ADS)

    Wilson, E. J.; Tiley, C.

    High rates of atmospheric nitrogen deposition have been shown to cause forest decline in some areas. A number of soil-mediated damage mechanisms are well-characterised but much less is known about the role of direct uptake of N by the canopy. The stable 15N isotope has been used in this experiment to quantify the assimilation of wet-deposited N by foliage and branches of 5 yr old Norway spruce trees. The effect of nutrient status (P and K) on N uptake was also investigated. Simulated rain solutions containing 99 atom% 15NH +4-N or 99 atom% 15NO -3-N at 15 mg N ℓ -1 were applied as a fine mist for 0.5 h. Woody twigs, current needles and current +1 year needles were sampled before and after misting and analysed for 15N enrichment and total N by automated mass spectrometry. All three tissue types showed some capacity for uptake of both 15NO -3-N and 15NH +4-N, although in nitrate treatments, N absorption was only statistically significant in woody twigs. Uptake rates were significantly higher in twigs compared with needles and in ammonium treatments compared with nitrate treatments, but only in PK fertilised trees. The concentrations of P and K in spruce foliage did not significantly affect the amount of N taken up by the canopy. Extrapolation of these data to mature forests suggests that foliar uptake is unlikely to exceed about 5% of annual N requirements, and will only make a small contribution to "N-saturation" of vegetation. The results of this experiment indicate that absorption of N by branches and twigs could potentially make an important contribution to total N requirements, although uptake may have been overestimated as it was not possible to differentiate between biological uptake of 15N and retention in the bark.

  4. Site-specific 15N isotopic signatures of abiotically produced N2O

    NASA Astrophysics Data System (ADS)

    Heil, Jannis; Wolf, Benjamin; Brüggemann, Nicolas; Emmenegger, Lukas; Tuzson, Béla; Vereecken, Harry; Mohn, Joachim

    2014-08-01

    Efficient nitrous oxide (N2O) mitigation strategies require the identification of the main source and sink processes and their contribution to total soil N2O production. Several abiotic reactions of nitrification intermediates leading to N2O production are known, but their contribution to total N2O production in soils is uncertain. As the site preference (SP) of 15N in N2O is a promising tool to give more insight into N2O production processes, we investigated the SP of N2O produced by different abiotic reactions in a laboratory study. All reactions involved the nitrification intermediate hydroxylamine (NH2OH) in combination with nitrite (NO2-), Fe3+, Fe2+ and Cu2+, reactants commonly or potentially found in soils, at different concentrations and pH values. N2O production and its four main isotopic species (14N14N16O, 15N14N16O, 14N15N16O, and 14N14N18O) were quantified simultaneously and online at high temporal resolution using quantum cascade laser absorption spectroscopy. Thereby, our study presents the first continuous analysis of δ18O in N2O. The experiments revealed the possibility of purely abiotic reactions over a wide range of acidity (pH 3-8) by different mechanisms. All studied abiotic pathways produced N2O with a characteristic SP in the range of 34-35‰, unaffected by process conditions and remaining constant over the course of the experiments. These findings reflect the benefit of continuous N2O isotopic analysis by laser spectroscopy, contribute new information to the challenging source partitioning of N2O emissions from soils, and emphasize the potentially significant role of coupled biotic-abiotic reactions in soils.

  5. Nitrate Reduction in a Groundwater Microcosm Determined by 15N Gas Chromatography-Mass Spectrometry

    PubMed Central

    Bengtsson, Gran; Annadotter, Helne

    1989-01-01

    Aerobic and anaerobic groundwater continuous-flow microcosms were designed to study nitrate reduction by the indigenous bacteria in intact saturated soil cores from a sandy aquifer with a concentration of 3.8 mg of NO3?-N liter?1. Traces of 15NO3? were added to filter-sterilized groundwater by using a Darcy flux of 4 cm day?1. Both assimilatory and dissimilatory reduction rates were estimated from analyses of 15N2, 15N2O, 15NH4+, and 15N-labeled protein amino acids by capillary gas chromatography-mass spectrometry. N2 and N2O were separated on a megabore fused-silica column and quantified by electron impact-selected ion monitoring. NO3? and NH4+ were analyzed as pentafluorobenzoyl amides by multiple-ion monitoring and protein amino acids as their N-heptafluorobutyryl isobutyl ester derivatives by negative ion-chemical ionization. The numbers of bacteria and their [methyl-3H]thymidine incorporation rates were simultaneously measured. Nitrate was completely reduced in the microcosms at a rate of about 250 ng g?1 day?1. Of this nitrate, 80 to 90% was converted by aerobic denitrification to N2, whereas only 35% was denitrified in the anaerobic microcosm, where more than 50% of NO3? was reduced to NH4+. Assimilatory reduction was recorded only in the aerobic microcosm, where N appeared in alanine in the cells. The nitrate reduction rates estimated for the aquifer material were low in comparison with rates in eutrophic lakes and coastal sediments but sufficiently high to remove nitrate from an uncontaminated aquifer of the kind examined in less than 1 month. PMID:16348048

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

    SciTech Connect

    Wilson, Jennifer E.; Sussman, Aviva Joy

    2015-05-22

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

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

    USGS Publications Warehouse

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

    2007-01-01

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

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

    USGS Publications Warehouse

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

    2007-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2012-12-01

    Bioretention stormwater management systems are engineered ecosystems that capture urban stormwater in order to reduce the harmful effects of stormwater pollution on receiving waters. Bioretention systems have been shown to be effective at reducing the volume of runoff, and thereby reduce the nutrient loading to receiving waters from urban areas. However, little work has been done to evaluate the treatment processes that are responsible for reductions in effluent nitrogen (N). We hypothesize that the pulses of inorganic nitrogen associated with urban runoff events are captured in the plat tissues within these systems and not adsorbed to the soil media, thus creating a long-term, sustainable treatment approach to reducing the total nutrient loading to receiving waters. Nitrogen treatment performance was tested on two bioretention systems in Salt Lake City, UT: 1) an upland native community that does not require irrigation in semi-arid climates, and 2) a wetland community that requires 250 l of daily irrigation to offset the relatively high evaporative demand in the region. Each cell is sized to treat a 2.5 cm storm from a 140 m2 impervious surface: the area of the bioretention system is 10 m2. To test the N removal performance of each system, runoff events were simulated to represent an average precipitation regime using a synthetic stormwater blend starting in January, 2012. Effluent was collected from an underdrain and analyzed for total nitrogen (TN); mass removal was calculated for each month by subtracting the TN mass added to the garden minus the TN mass that flowed out of the garden. To test the hypothesis that plants assimilate stormwater N, 4 g of 100 atom% 15N NH4NO3 tracer was used as the N source in the synthetic stormwater during the first 2,000 l synthetic storm event in May. This isotopic label was calculated to enrich the total N pool of each garden to 100‰ 15N/14Nair. New growth was harvested from each plant in both cells and analyzed for 15N before the isotopic label was introduced and weekly thereafter. In May 2012, the upland garden captured 6.2 grams of TN from the added stormwater (55% of TN added), and the wetland garden captured 7.1 grams of TN from the added stormwater (67% of TN added). Within two weeks of adding the label, the 15N ratio increased 500‰ to 3,000‰ in all plant tissues tested in both systems. The results of the isotopic labeling experiment support the hypothesis that the plants used in both vegetated bioretention systems directly contribute to stormwater N treatment through N assimilation.

  10. Hyporheic Denitrification in an Upland Agricultural Stream: a 15N Tracer Study

    NASA Astrophysics Data System (ADS)

    Zarnetske, J. P.; Haggerty, R.; Wondzell, S. M.; Baker, M. A.

    2008-12-01

    We used whole-stream steady-state 15N-labeled nitrate (15NO3-) and conservative tracer (Cl-) additions to investigate the hydraulic and physiochemical factors controlling denitrification in the hyporheic zone (HZ) of an upland agricultural stream. We measured solute concentrations (15NO3-, 15N2(g), as well as NO3-, DOC, DO, Cl-), and hydraulic transport parameters (head, flow rates, flowpaths, and residence time distributions) of the reach and along HZ flowpaths of an instrumented gravel bar. HZ exchange was observed across the entire gravel bar with nominal flowpath lengths up to 6.1 m and corresponding mean residence times greater than 21.5 hr. Along sampled HZ flowpaths from gravel bar head to distal well, NO3-N decreased from 0.34 to 0.02 mg/L, DO declined from 8.31 to 0.59 mg/L, and DOC dropped from 3.0 to 1.7 mg/L. The rate of the NO3-, DO, and DOC removal was greatest during the first 2 m of the flowpaths but removal continued across the entire gravel bar. Conversely, the surface water chemical and nutrient conditions were more uniform across the length of the reach (NO3-N: 0.31 - 0.34 mg/L, DO: 8.1 - 8.5 mg/L, DOC: 2.8 - 3.4 mg/L). 15NO3- tracing confirmed that the removal of the NO3- along the HZ flowpaths was primarily due to denitrification as 15N2 was produced across the entire gravel bar HZ (in all 12 wells). Relative to the HZ, surface water N2 concentrations were similar, but minimal 15N2-enrichment was observed. These findings demonstrate that the HZ is an active sink of nitrogen in this system. Overall, results support our hypothesis that HZ denitrification is an important nitrogen sink in mid-network streams with significant NO3-, because denitrification in these systems will be less limited by HZ biogeochemical and physical transport conditions compared to streams above and below them in the network.

  11. Characteristics of the δ ^{15} N_{NO_3 } distribution and its drivers in the Changjiang River estuary and adjacent waters

    NASA Astrophysics Data System (ADS)

    Wang, Wentao; Yu, Zhiming; Song, Xiuxian; Wu, Zaixing; Yuan, Yongquan; Zhou, Peng; Cao, Xihua

    2016-05-01

    In this study, we conducted investigations in the Changjiang (Yangtze) River estuary and adjacent waters (CREAW) in June and November of 2014. We collected water samples from different depths to analyze the nitrogen isotopic compositions of nitrate, nutrient concentrations (including inorganic N, P, and Si), and other physical and biological parameters, along with the vertical distribution and seasonal variations of these parameters. The compositions of nitrogen isotope in nitrate were measured with the denitrifier method. Results show that the Changjiang River diluted water (CDW) was the main factor aff ecting the shallow waters (above 10 m) of the CREAW, and CDW tended to influence the northern areas in June and the southern areas in November. δ ^{15} N_{NO_3 } values in CDW ranged from 3.21‰-3.55‰. In contrast, the deep waters (below 30 m) were aff ected by the subsurface water of the Kuroshio Current, which intruded into the waters near 31°N in June. The δ ^{15} N_{NO_3 } values of these waters were 6.03‰-7.6‰, slightly higher than the values of the Kuroshio Current. Nitrate assimilation by phytoplankton in the shallow waters of the study area varied seasonally. Because of the favorable temperature and nutrient conditions in June, abundant phytoplankton growth resulted in harmful algae blooms (HABs). Therefore, nitrate assimilation was strong in June and weak in November. The δ ^{15} N_{NO_3 } fractionations caused by assimilation of phytoplankton were 4.57‰ and 4.41‰ in the shallow waters in June and November, respectively. These results are consistent with previous laboratory cultures and in situ investigations. Nitrification processes were observed in some deep waters of the study area, and they were more apparent in November than in June. The fractionation values of nitrification ranged from 24‰-25‰, which agrees with results for Nitrosospira tenuis reported by previous studies.

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

    NASA Astrophysics Data System (ADS)

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

    2010-12-01

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

  13. The coral δ15N record of terrestrial nitrate loading varies with river catchment land use

    NASA Astrophysics Data System (ADS)

    Yamazaki, A.; Watanabe, T.; Tsunogai, U.; Hasegawa, H.; Yamano, H.

    2015-03-01

    We analysed the nitrogen isotopes in two coral cores (δ15Ncoral) from the mouth of the Todoroki River, Ishigaki Island, Japan, to examine whether the δ15Ncoral reflects the run-off of nitrate related to the land use in the river catchment. The two coral cores were used to examine the seasonal variation in δ15Ncoral for 14 years (CORE1; 1993-2007) and the annual variation of δ15Ncoral for 52 years (CORE2; 1958-2010). In CORE1, the 5-month running mean of δ15Ncoral was positively correlated with that of monthly precipitation, excluding all strong precipitation events (>150 mm d-1). In CORE2, the δ15Ncoral mean in the earlier period (1958-1980) was 1.0 ‰ greater than that in the later period (1981-2010). The annual averages of δ15Ncoral are positively correlated with the total precipitation in the rainy season (May-June) for both time periods. The difference in the δ15Ncoral between the earlier and later periods is probably caused by the land use changed from paddy fields with 15N-rich manure to sugar cane fields in the early 1980s. Although some uncertainties still remain regarding the precision of δ15N coral proxy records, this study emphasises the clear potential for their use in reconstructing terrestrial nitrate discharge records from corals.

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

    PubMed

    Schubert; Smalla; Schmieder; Oschkinat

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective (1)H-(15)N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH(2) or XH(3) (X can be (15)N or (13)C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains. Copyright 1999 Academic Press. PMID:10527741

  15. Experimental assessment of the bioconcentration of (15)N-tamoxifen in Pseudokirchneriella subcapitata.

    PubMed

    Orias, Frédéric; Simon, Laurent; Perrodin, Yves

    2015-03-01

    Nowadays, pharmaceutical compounds (PC) are ubiquitous in aquatic ecosystems. In addition to direct ecotoxicity, the bioconcentration of PC in organisms is a phenomenon which could have an impact on the whole ecosystem. In order to study this phenomenon, we exposed unicellular algae (Pseudokirchneriella subcapitata) to (15)N-tamoxifen, an anticancer drug labelled with a stable nitrogen isotope used as a tracer. By measuring (15)N enrichment over time, we were able to measure the increase of tamoxifen content in algae. This enrichment was measured by an elemental analyser coupled with an isotopic ratio mass spectrometer (EA-IRMS). Algal cells were exposed for 7d to 3 concentrations of tamoxifen: 1, 10 and 100μgL(-1). Our result shows a high bioconcentration in algae from the first minutes of contact. The highest bioconcentration factor measured is around 26500. We also observe that bioconcentration is not linked to the exposure concentration. This study is the first to use stable isotopes in order to monitor PCs in aquatic organisms such as algae. The use of stable isotopes in ecotoxicology offers interesting perspectives in the field of contaminant transfer in organisms and along the trophic web. PMID:25522849

  16. Precursor discrimination of designer drug benzylpiperazine using δ13C and δ15N stable isotopes.

    PubMed

    Beckett, Nicola M; Grice, Darren I; Carter, James F; Cresswell, Sarah L

    2015-01-01

    Advances in analytical technology and emerging techniques have resulted in the increased exploitation of chemical and isotopic profiling for source linkage/discrimination of illicit drugs for forensic purposes. Although not routinely used for illicit drug investigations, such information has been obtained and its application demonstrated through the use of isotope ratio mass spectrometry (IRMS). There is a solid platform of research available relating to the isotopic analysis of methylenedioxymethamphetamine (MDMA) and methamphetamine (MA), however with the recently flourishing designer drug market it was of interest to examine the isotopic profiles of the popular 'party drug' benzylpiperazine hydrochloride (BZP·HCl). A preliminary analysis of δ13C and δ15N isotopic ratios in BZP·HCl products and corresponding synthetic intermediates (piperazine·HCl) synthesized in-house from three different precursor suppliers was conducted using IRMS. Analysis of the δ13C and δ15N isotopic data indicated that discrimination and correct grouping of all the intermediates and some of the product samples examined in this study were achievable. PMID:25577008

  17. MUSIC in Triple-Resonance Experiments: Amino Acid Type-Selective 1H- 15N Correlations

    NASA Astrophysics Data System (ADS)

    Schubert, Mario; Smalla, Maika; Schmieder, Peter; Oschkinat, Hartmut

    1999-11-01

    Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective 1H-15N correlations are presented. They are based on novel coherence transfer schemes, the MUSIC pulse sequence elements, that replace the initial INEPT transfer and are selective for XH2 or XH3 (X can be 15N or 13C). The desired amino acid type is thereby selected based on the topology of the side chain. Experiments for Gly (G-HSQC); Ala (A-HSQC); Thr, Val, Ile, and Ala (TAVI-HSQC); Thr and Ala (TA-HSQC), as well as Asn and Gln (N-HSQC and QN-HSQC), are described. The new experiments are recorded as two-dimensional experiments and therefore need only small amounts of spectrometer time. The performance of the experiments is demonstrated with the application to two protein domains.

  18. Covalent binding of aniline to humic substances. 2. 15N NMR studies of nucleophilic addition reactions

    USGS Publications Warehouse

    Thorn, K.A.; Pettigrew, P.J.; Goldenberg, W.S.; Weber, E.J.

    1996-01-01

    Aromatic amines are known to undergo covalent binding with humic substances in the environment. Although previous studies have examined reaction conditions and proposed mechanisms, there has been no direct spectroscopic evidence for the covalent binding of the amines to the functional groups in humic substances. In order to further elucidate the reaction mechanisms, the Suwannee River and IHSS soil fulvic and humic acids were reacted with 15N-labeled aniline at pH 6 and analyzed using 15N NMR spectrometry. Aniline underwent nucleophilic addition reactions with the quinone and other carbonyl groups in the samples and became incorporated in the form of anilinohydroquinone, anilinoquinone, anilide, imine, and heterocyclic nitrogen, the latter comprising 50% or more of the bound amine. The anilide and anilinohydroquinone nitrogens were determined to be susceptible to chemical exchange by ammonia. In the case of Suwannee River fulvic acid, reaction under anoxic conditions and pretreatment with sodium borohydride or hydroxylamine prior to reaction under oxic conditions resulted in a decrease in the proportion of anilinohydroquinone nitrogen incorporated. The relative decrease in the incorporation of anilinohydroquinone nitrogen with respect to anilinoquinone nitrogen under anoxic conditions suggested that inter- or intramolecular redox reactions accompanied the nucleophilic addition reactions.

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

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  20. Discrimination against 15N among recombinant inbred lines of Phaseolus vulgaris L. contrasting in phosphorus use efficiency for nitrogen fixation.

    PubMed

    Lazali, Mohamed; Bargaz, Adnane; Carlsson, Georg; Ounane, Sidi Mohamed; Drevon, Jean Jacques

    2014-02-15

    Although isotopic discrimination processes during nitrogen (N) transformations influence the outcome of (15)N based quantification of N2 fixation in legumes, little attention has been given to the effects of genotypic variability and environmental constraints such as phosphorus (P) deficiency, on discrimination against (15)N during N2 fixation. In this study, six Phaseolus vulgaris recombinant inbred lines (RILs), i.e. RILs 115, 104, 34 (P deficiency tolerant) and 147, 83, 70 (P deficiency sensitive), were inoculated with Rhizobium tropici CIAT899, and hydroaeroponically grown with P-sufficient (250 μmol P plant(-1) week(-1)) versus P-deficient (75 μmol P plant(-1) week(-1)) supply. Two harvests were done at 15 (before nodule functioning) and 42 (flowering stage) days after transplanting. Nodulation, plant biomass, P and N contents, and the ratios of (15)N over total N content ((15)N/Nt) for shoots, roots and nodules were determined. The results showed lower (15)N/Nt in shoots than in roots, both being much lower than in nodules. P deficiency caused a larger decrease in (15)N/Nt in shoots (-0.18%) than in nodules (-0.11%) for all of the genotypes, and the decrease in shoots was greatest for RILs 34 (-0.33%) and 104 (-0.25%). Nodule (15)N/Nt was significantly related to both the quantity of N2 fixed (R(2)=0.96***) and the P content of nodules (R(2)=0.66*). We conclude that the discrimination against (15)N in the legume N2-fixing symbiosis of common bean with R. tropici CIAT899 is affected by P nutrition and plant genotype, and that the (15)N/Nt in nodules may be used to screen for genotypic variation in P use efficiency for N2 fixation. PMID:24035519

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

    NASA Astrophysics Data System (ADS)

    Gurwell, Mark A.

    2004-11-01

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

  2. 15N depleted nitrogen isotope values in Cretaceous black shales: paleoceanographic event or diagenesis.

    NASA Astrophysics Data System (ADS)

    Junium, C. K.; Arthur, M. A.

    2004-12-01

    Nitrogen isotopic values of bulk sediment samples in black shales are almost exclusively near 0 ‰ and C/N ratios are high (20-35). Sequential extraction of exchangeable and non-exchangeable N fractions demonstrates that the inorganic N fraction is negligible and bulk sediment data reflect the organic N fraction. The trend in \\delta$15N and C/N has been observed in numerous localities and depositional environments in the mid-Cretaceous (Demarara Rise, DSDP Sites 367, 603B and 530, the Cretaceous Western Interior Seaway, Wunstorf, Germany, and Bahloul, Tunisia) as well as the Toarcian of England and in Quaternary Mediterranean Sapropels. Three explanations are considered: 1) That primary production during black shale deposition was dominated by a unique community composed of nitrogen-fixing cyanobacteria; or 2) utilization of a 15N depleted ammonium source by another set of biota; or 3) that the values are the result of diagenetic loss of N prevailing in Corg-rich strata with a low capacity for N adsorption. The depleted nitrogen isotopic values suggest that nitrogen fixation or utilization of a depleted nitrogen source (e.g. ammonium) may have been important which is plausible in consideration of nitrogen deficiencies that might characterize widespread deep-water anoxia. Secular variations in nitrogen isotope values across the Cenomanian-Turonian Boundary of ODP Site 1261, Demarara Rise, show a shift from -1 to -3‰ . One interpretation of the origin of the excursion could be that a fraction of the organic matter was produced utilizing a 15N depleted ammonium source, assuming that the nitrogen isotopic composition of fixed oceanic nitrogen does not change. However, it should be noted that large (greater than 1‰ ) variations in nitrogen isotope values are observed above and below the Cenomanian-Turonian Boundary Event and have no known paleoceanographic forcing. The variations in nitrogen isotope values are matched by antithetic variations in C/N indicating diagenetic N-loss that could be interpreted as the primary control on the nitrogen isotope variations. The high C/N ratios probably result from selective removal of N-enriched compounds in the water column and during burial diagenesis. Clay-poor, Corg-rich sediments have a low sorptive capacity, allowing diffusion of dissolved N out of the Corg-rich sediments resulting in higher C/N ratios. Pore water ammonium concentrations are mM, indicating low sorption and high rates of diffusion into overlying strata. Coupled C/N and N-isotope variations occur by the removal of a 15N enriched fraction. Protein degradation has been suggested to result in negative isotopic shifts but would not result in large increases in C/N and the isotopic shifts are limited by the protein richness of the primary organic matter and the internal isotopic heterogeneity of the organic matter pools. Although we favor an explanation that involves a primary signature, diagenesis cannot be excluded in low δ 15N values.

  3. The Contamination of Commercial 15N2 Gas Stocks with 15N–Labeled Nitrate and Ammonium and Consequences for Nitrogen Fixation Measurements

    PubMed Central

    Dabundo, Richard; Lehmann, Moritz F.; Treibergs, Lija; Tobias, Craig R.; Altabet, Mark A.; Moisander, Pia H.; Granger, Julie

    2014-01-01

    We report on the contamination of commercial 15-nitrogen (15N) N2 gas stocks with 15N-enriched ammonium, nitrate and/or nitrite, and nitrous oxide. 15N2 gas is used to estimate N2 fixation rates from incubations of environmental samples by monitoring the incorporation of isotopically labeled 15N2 into organic matter. However, the microbial assimilation of bioavailable 15N-labeled N2 gas contaminants, nitrate, nitrite, and ammonium, is liable to lead to the inflation or false detection of N2 fixation rates. 15N2 gas procured from three major suppliers was analyzed for the presence of these 15N-contaminants. Substantial concentrations of 15N-contaminants were detected in four Sigma-Aldrich 15N2 lecture bottles from two discrete batch syntheses. Per mole of 15N2 gas, 34 to 1900 µmoles of 15N-ammonium, 1.8 to 420 µmoles of 15N-nitrate/nitrite, and ≥21 µmoles of 15N-nitrous oxide were detected. One 15N2 lecture bottle from Campro Scientific contained ≥11 µmoles of 15N-nitrous oxide per mole of 15N2 gas, and no detected 15N-nitrate/nitrite at the given experimental 15N2 tracer dilutions. Two Cambridge Isotopes lecture bottles from discrete batch syntheses contained ≥0.81 µmoles 15N-nitrous oxide per mole 15N2, and trace concentrations of 15N-ammonium and 15N-nitrate/nitrite. 15N2 gas equilibrated cultures of the green algae Dunaliella tertiolecta confirmed that the 15N-contaminants are assimilable. A finite-differencing model parameterized using oceanic field conditions typical of N2 fixation assays suggests that the degree of detected 15N-ammonium contamination could yield inferred N2 fixation rates ranging from undetectable, <0.01 nmoles N L−1 d−1, to 530 nmoles N L−1 d−1, contingent on experimental conditions. These rates are comparable to, or greater than, N2 fixation rates commonly detected in field assays. These results indicate that past reports of N2 fixation should be interpreted with caution, and demonstrate that the purity of commercial 15N2 gas must be ensured prior to use in future N2 fixation rate determinations. PMID:25329300

  4. Suppression of Diagonal Peaks in TROSY-Type 1H NMR NOESY Spectra of 15N-Labeled Proteins

    NASA Astrophysics Data System (ADS)

    Meissner, Axel; Sørensen, Ole Winneche

    1999-10-01

    A novel method for suppression of diagonal peaks in the amide region of NOESY NMR spectra of 15N-labeled proteins is presented. The method is particularly useful for larger proteins at high magnetic fields where interference between dipolar and chemical shift anisotropy relaxation mechanisms results in large TROSY effects, i.e., large differences in 1HN linewidths depending on the spin state of attached 15N nuclei. In this limit the new TROSY NOESY method does not compromise sensitivity. It is demonstrated using a perdeuterated 15N-labeled protein sample, Neural Cell Adhesion Molecule 213-308 (NCAM) from rat, in H2O at 800 MHz.

  5. Mammalian DNA δ15N exhibits 40‰ intramolecular variation and is unresponsive to dietary protein level

    PubMed Central

    Strable, Maggie S.; Tschanz, Carolyn L.; Varamini, Behzad; Chikaraishi, Yoshito; Ohkouchi, Naohiko; Brenna, J. Thomas

    2014-01-01

    We report the first high precision characterization of molecular and intramolecular δ15N of nucleosides derived from mammalian DNA. The influence of dietary protein level on brain amino acids and deoxyribonucleosides was determined to investigate whether high protein turnover would alter amino acid 15N or 13C. Pregnant guinea pig dams were fed control diets, or high or low levels of dietary protein throughout gestation, and all pups were fed control diets. Cerebellar DNA of offspring was extracted at 2 and 120 days of life, nucleosides isolated and δ15N and δ13C characterized. Mean diet δ15N = 0.45±0.33‰, compared to cerebellar whole tissue and DNA δ15N = +4.1±0.7‰ and −4.5±0.4‰, respectively. Cerebellar deoxythymidine (dT), deoxycytidine (dC), deoxyadenosine (dA), and deoxyguanosine (dG) δ15N were +1.4±0.4, −2.1±0.9, −7.2±0.3, and −10.4±0.5‰, respectively. There were no changes in amino acid or deoxyribonucleoside δ15N due to dietary protein level. Using known metabolic relationships, we developed equations to calculate the intramolecular δ15N originating from aspartate (asp) in purines (pur) or pyrimidines (pyr), glutamine (glu), and glycine (gly) to be δ15NASP-PUR, δ15NASP-PYR, δ15NGLN, and δ15NGLY +11.9±2.3‰, +7.0±2.0‰, −9.1±2.4‰, and −31.8±8.9‰, respectively. A subset of twelve amino acids from food and brain had mean δ15N of 4.3±3.2‰ and 13.8±3.1‰, respectively, and δ15N for gly and asp were 12.6±2.2‰ and 15.2±0.8‰, respectively. A separate isotope tracer study detected no significant turnover of cerebellar DNA in the first six months of life. The large negative δ15N difference between gly and cerebellar purine N at the gly (7) position implies either that there is a major isotope effect during DNA synthesis, or that in utero gly has a different isotope ratio during rapid growth and metabolism than in adult life. Our data show that cerebellar nucleoside intramolecular δ15N vary over more than 40‰ and are not influenced by dietary protein level or age. PMID:22095504

  6. Abundance, Natural Infection with Trypanosomes, and Food Source of an Endemic Species of Triatomine, Panstrongylus howardi (Neiva 1911), on the Ecuadorian Central Coast

    PubMed Central

    Villacís, Anita G.; Ocaña-Mayorga, Sofía; Lascano, Mauricio S.; Yumiseva, César A.; Baus, Esteban G.; Grijalva, Mario J.

    2015-01-01

    The elimination of domestic triatomines is the foundation of Chagas disease control. Regional initiatives are eliminating introduced triatomine species. In this scenario, endemic triatomines can occupy the ecological niches left open and become a threat to long-term Chagas disease control efforts. This study determined the abundance, colonization, and Trypanosoma cruzi infection rate of the endemic Panstrongylus howardi in 10 rural communities located in Ecuador's Manabí Province. In total, 518 individuals of P. howardi were collected. Infestation indices of 1.4% and 6.6% were found in the domestic and peridomestic environments, respectively. We determined a T. cruzi infection rate of 53.2% (N = 47) in this species. P. howardi has a high capacity to adapt to different habitats, especially in the peridomicile. This implies a considerable risk of transmission because of the frequency of intradomicile invasion. Therefore, this species needs to be taken into account in Chagas control and surveillance efforts in the region. PMID:25385867

  7. Expression and purification of 15N- and 13C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.

    PubMed

    Long, Fei; Cho, Wonhwa; Ishii, Yoshitaka

    2011-09-01

    Amyloid fibrils of Alzheimer's ?-amyloid peptide (A?) are a primary component of amyloid plaques, a hallmark of Alzheimer's disease (AD). Enormous attention has been given to the structural features and functions of A? in amyloid fibrils and other type of aggregates in associated with development of AD. This report describes an efficient protocol to express and purify high-quality 40-residue A?(1-40), the most abundant A? in brains, for structural studies by NMR spectroscopy. Over-expression of A?(1-40) with glutathione S-transferase (GST) tag connected by a Factor Xa recognition site (IEGR(?)) in Escherichia coli resulted in the formation of insoluble inclusion bodies even with the soluble GST tag. This problem was resolved by efficient recovery of the GST-A? fusion protein from the inclusion bodies using 0.5% (w/v) sodium lauroyl sarcosinate as solubilizing agent and subsequent purification by affinity chromatography using a glutathione agarose column. The removal of the GST tag by Factor Xa enzymatic cleavage and purification by HPLC yielded as much as ?7 mg and ?1.5mg of unlabeled A?(1-40) and uniformly (15)N- and/or (13)C-protein A?(1-40) from 1L of the cell culture, respectively. Mass spectroscopy of unlabeled and labeled A? and (1)H/(15)N HSQC solution NMR spectrum of the obtained (15)N-labeled A? in the monomeric form confirmed the expression of native A?(1-40). It was also confirmed by electron micrography and solid-state NMR analysis that the purified A?(1-40) self-assembles into ?-sheet rich amyloid fibrils. To the best of our knowledge, our protocol offers the highest yields among published protocols for production of recombinant A?(1-40) samples that are amendable for an NMR-based structural analysis. The protocol may be applied to efficient preparation of other amyloid-forming proteins and peptides that are (13)C- and (15)N-labeled for NMR experiments. PMID:21640828

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

    NASA Astrophysics Data System (ADS)

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

    1996-11-01

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

  9. Nuclear magnetic resonance studies on huwentoxin-XI from the Chinese bird spider Ornithoctonus huwena: 15N labeling and sequence-specific 1H, 15N nuclear magnetic resonance assignments.

    PubMed

    Peng, Kuan; Lin, Ying; Liang, Song-Ping

    2006-07-01

    Huwentoxin-XI purified from the Chinese bird spider Ornithoctonus huwena is a toxin with both antiprotease activity and potassium channel blocking activity. To determine its solution structure, huwentoxin-XI was expressed in a yeast eukaryotic expression system and studied by NMR. The 15N labeling strategy was used to facilitate the process of resonance assignments. The nearly complete sequence-specific assignments of proton and nitrogen resonances were obtained by analyzing a series of two-dimensional (2D) and three-dimensional (3D) spectra, including DQF-COSY, TOCSY, NOESY, 15N-1H HSQC, 15N-1H HNHA, 15N-1H HNHB, 15N-1H TOCSY-HSQC and 15N-1H NOESY-HSQC spectra. Secondary structure analysis of huwentoxin-XI showed that it mainly contains an N-terminal 310-helix from Thr3 to Arg5 and a C-terminal alpha-helix from Gln45 to Cys52, plus a triple-stranded antiparallel beta-sheet of Glu18-Asn23, Thr26-Ile31 and Asn40-Lys41. These studies provide a solid basis for the final structure determination of huwentoxin-XI. PMID:16820861

  10. Robust Abundance Estimation in Animal Abundance Surveys with Imperfect Detection

    EPA Science Inventory

    Surveys of animal abundance are central to the conservation and management of living natural resources. However, detection uncertainty complicates the sampling process of many species. One sampling method employed to deal with this problem is depletion (or removal) surveys in whi...

  11. Sample Artefacts in δ15N and δ18O of Nitrate in the atmosphere

    NASA Astrophysics Data System (ADS)

    Seguin, A.; Norman, A. L.

    2009-12-01

    Nitrate is one of the major inorganic anions in aerosols. Recently nitrogen and oxygen isotopes in nitrates have been used to trace sources and chemical processes in the atmosphere. High volume samplers are used in order to gain enough material for isotope analysis, but artefacts can occur when measuring with this technique. Besides retaining NO3 from aerosol, gaseous HNO3 can be absorbed on the filter increasing the reported values of nitrate reported. Alternatively; when elevated temperatures are present in the atmosphere, nitrate is volatized in the form of NH3NO3 which would lead to an underestimation of nitrogen on the filter [Schaap et al., 2004]. The artefacts from high volume sampling may cause inaccuracies in the reported isotope values due to isotope fractionation or different initial isotope values between gaseous HNO3 and particulate nitrate. High volume samplers were deployed on board a ship in the summer of 2003 jointly with the Canadian Surface Ocean - Lower Atmosphere Study (SOLAS). A quartz filter was positioned on a high volume sampler. A cellulose acetate filter treated with potassium carbonate and glycerol was placed underneath the quartz filter. The cellulose acetate filter is designed to catch sticky gaseous (such as SO2) but was also found to catch a nitrogen gas species (most probable HNO3 and/or NH3NO3). Analysis of isotopic composition of nitrate for both nitrogen and oxygen was carried out on selected samples by the denitrifier method. Values ranged between -4 and +1‰ for δ15N and between +49 and +66‰ for δ18O for quartz filter. The nitrogen gas species had ranges between -11 and -0‰ for δ15N and between +44 and +62‰ for δ18O. Average differences between the aerosol (caught on the quartz filter) and the nitrogen gas species (on the cellulose acetate filter) were 3±4 ‰ for δ15N and 10±7 ‰ for δ18O. These values are examined to find insights in possible fractionation in the nitrate collection method currently used in other studies. Schaap, M., G. Spindler, M. Schulz, K. Acker, W. Maenhaut, A. Berner, W. Wieprecht, N. Streit, K. Müller, E. Brüggemann, X. Chi, J.-P. Putaud, R. Hitzenberger, H. Puxbaum, U. Baltensperger and H. ten Brink (2004), Artefacts in the sampling of nitrate studied in the “INTERCOMP” campaigns of EUROTRAC-AEROSOL, Atmospheric Environment, 38, 6487-6496.

  12. Nitrous oxide nitrification and denitrification 15N enrichment factors from Amazon forest soils.

    PubMed

    Pérez, Tibisay; Garcia-Montiel, Diana; Trumbore, Susan; Tyler, Stanley; de Camargo, Plínio; Moreira, Marcelo; Piccolo, Marisa; Cerri, Carlos

    2006-12-01

    The isotopic signatures of 15N and 18O in N2O emitted from tropical soils vary both spatially and temporally, leading to large uncertainty in the overall tropical source signature and thereby limiting the utility of isotopes in constraining the global N2O budget. Determining the reasons for spatial and temporal variations in isotope signatures requires that we know the isotope enrichment factors for nitrification and denitrification, the two processes that produce N2O in soils. We have devised a method for measuring these enrichment factors using soil incubation experiments and report results from this method for three rain forest soils collected in the Brazilian Amazon: soil with differing sand and clay content from the Tapajos National Forest (TNF) near Santarém, Pará, and Nova Vida Farm, Rondônia. The 15N enrichment factors for nitrification and denitrification differ with soil texture and site: -111 per thousand +/- 12 per thousand and -31 per thousand +/- 11 per thousand for a clay-rich Oxisol (TNF), -102 per thousand +/- 5 per thousand and -45 per thousand +/- 5 per thousand for a sandier Ultisol (TNF), and -10.4 per thousand +/- 3.5 per thousand (enrichment factor for denitrification) for another Ultisol (Nova Vida) soil, respectively. We also show that the isotopomer site preference (delta15Nalpha - delta15Nbeta, where alpha indicates the central nitrogen atom and beta the terminal nitrogen atom in N2O) may allow differentiation between processes of production and consumption of N2O and can potentially be used to determine the contributions of nitrification and denitrification. The site preferences for nitrification and denitrification from the TNF-Ultisol incubated soils are: 4.2 per thousand +/- 8.4 per thousand and 31.6 per thousand +/- 8.1 per thousand, respectively. Thus, nitrifying and denitrifying bacteria populations under the conditions of our study exhibit significantly different 15N site preference fingerprints. Our data set strongly suggests that N2O isotopomers can be used in concert with traditional N2O stable isotope measurements as constraints to differentiate microbial N2O processes in soil and will contribute to interpretations of the isotopic site preference N2O values found in the free troposphere. PMID:17205894

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

    NASA Astrophysics Data System (ADS)

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

    2015-09-01

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

  14. A suite of Mathematica notebooks for the analysis of protein main chain 15N NMR relaxation data.

    PubMed

    Spyracopoulos, Leo

    2006-12-01

    A suite of Mathematica notebooks has been designed to ease the analysis of protein main chain 15N NMR relaxation data collected at a single magnetic field strength. Individual notebooks were developed to perform the following tasks: nonlinear fitting of 15N-T1 and -T2 relaxation decays to a two parameter exponential decay, calculation of the principal components of the inertia tensor from protein structural coordinates, nonlinear optimization of the principal components and orientation of the axially symmetric rotational diffusion tensor, model-free analysis of 15N-T1, -T2, and {1H}-15N NOE data, and reduced spectral density analysis of the relaxation data. The principle features of the notebooks include use of a minimal number of input files, integrated notebook data management, ease of use, cross-platform compatibility, automatic visualization of results and generation of high-quality graphics, and output of analyses in text format. PMID:17061025

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

    SciTech Connect

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

    2015-09-25

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

  16. Comparison of five soil organic matter decomposition models using data from a 14C and 15N labeling field experiment

    NASA Astrophysics Data System (ADS)

    Pansu, Marc; Bottner, Pierre; Sarmiento, Lina; Metselaar, Klaas

    2004-12-01

    Five alternatives of the pre