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Sample records for 18o isotopic labeling

  1. Primary Productivity Rates at Station ALOHA Determined by 18O Labeling and the Triple Isotope Composition of Dissolved Oxygen

    NASA Astrophysics Data System (ADS)

    Juranek, L. W.; Quay, P. D.; Karl, D. M.

    2002-12-01

    Although knowledge of accurate Primary Productivity (PPr) rates is essential to the understanding of ocean carbon cycling, the standard method of determining ocean productivity, 14C labeling, often yields uncertain results. Typically, 14C-derived PPr rates fall ambiguously between gross and net productivity because the method is sensitive to recycling of a relatively small POC pool. Bottle incubations using labeled oxygen produced from 18O-enriched water have shown promise in giving a more consistent measure of gross productivity, since the pool of dissolved oxygen is less sensitive to recycling than POC. Typically this method gives gross PPr rates that are 2-3 times 14C-derived rates. Recently Luz and Barkan (2001) have pioneered a new technique to determine PPr rates using the triple isotope composition of dissolved oxygen as an in situ tracer. This relies on the observation that a signature of mass-independent fractionation originating in the stratosphere and imparted to the surface ocean by air-sea exchange is diminished by biological oxygen production. In February 2002 we measured gross productivity using both the 18O-labeling and triple isotope in situ methods at Hawaii Ocean Time-Series station ALOHA in the N. Pacific subtropical gyre. We found the in situ oxygen isotope method yielded double the 14C-derived PPr rates while 18O bottle incubations yielded similar rates as 14C. In addition, comparison of in situ isotope measurements with the biological oxygen saturation state indicate that community respiration is approximately equal to gross photosynthesis in the upper 60 m while from 80-200 m respiration exceeds photosynthesis by at most 10 %. We will present these results along with new results from upcoming measurements at station ALOHA.

  2. Altered Retinoic Acid Metabolism in Diabetic Mouse Kidney Identified by 18O Isotopic Labeling and 2D Mass Spectrometry

    PubMed Central

    Starkey, Jonathan M.; Zhao, Yingxin; Sadygov, Rovshan G.; Haidacher, Sigmund J.; LeJeune, Wanda S.; Dey, Nilay; Luxon, Bruce A.; Kane, Maureen A.; Napoli, Joseph L.; Denner, Larry; Tilton, Ronald G.

    2010-01-01

    Background Numerous metabolic pathways have been implicated in diabetes-induced renal injury, yet few studies have utilized unbiased systems biology approaches for mapping the interconnectivity of diabetes-dysregulated proteins that are involved. We utilized a global, quantitative, differential proteomic approach to identify a novel retinoic acid hub in renal cortical protein networks dysregulated by type 2 diabetes. Methodology/Principal Findings Total proteins were extracted from renal cortex of control and db/db mice at 20 weeks of age (after 12 weeks of hyperglycemia in the diabetic mice). Following trypsinization, 18O- and 16O-labeled control and diabetic peptides, respectively, were pooled and separated by two dimensional liquid chromatography (strong cation exchange creating 60 fractions further separated by nano-HPLC), followed by peptide identification and quantification using mass spectrometry. Proteomic analysis identified 53 proteins with fold change ≥1.5 and p≤0.05 after Benjamini-Hochberg adjustment (out of 1,806 proteins identified), including alcohol dehydrogenase (ADH) and retinaldehyde dehydrogenase (RALDH1/ALDH1A1). Ingenuity Pathway Analysis identified altered retinoic acid as a key signaling hub that was altered in the diabetic renal cortical proteome. Western blotting and real-time PCR confirmed diabetes-induced upregulation of RALDH1, which was localized by immunofluorescence predominantly to the proximal tubule in the diabetic renal cortex, while PCR confirmed the downregulation of ADH identified with mass spectrometry. Despite increased renal cortical tissue levels of retinol and RALDH1 in db/db versus control mice, all-trans-retinoic acid was significantly decreased in association with a significant decrease in PPARβ/δ mRNA. Conclusions/Significance Our results indicate that retinoic acid metabolism is significantly dysregulated in diabetic kidneys, and suggest that a shift in all-trans-retinoic acid metabolism is a novel feature in

  3. 18O labeling of chlorophyll d in Acaryochloris marina reveals that chlorophyll a and molecular oxygen are precursors.

    PubMed

    Schliep, Martin; Crossett, Ben; Willows, Robert D; Chen, Min

    2010-09-10

    The cyanobacterium Acaryochloris marina was cultured in the presence of either H(2)(18)O or (18)O(2), and the newly synthesized chlorophylls (Chl a and Chl d) were isolated using high performance liquid chromatography and analyzed by mass spectroscopy. In the presence of H(2)(18)O, newly synthesized Chl a and d, both incorporated up to four isotopic (18)O atoms. Time course H(2)(18)O labeling experiments showed incorporation of isotopic (18)O atoms originating from H(2)(18)O into Chl a, with over 90% of Chl a (18)O-labeled at 48 h. The incorporation of isotopic (18)O atoms into Chl d upon incubation in H(2)(18)O was slower compared with Chl a with approximately 50% (18)O-labeled Chl d at 115 h. The rapid turnover of newly synthesized Chl a suggested that Chl a is the direct biosynthetic precursor of Chl d. In the presence of (18)O(2) gas, one isotopic (18)O atom was incorporated into Chl a with approximately the same kinetic incorporation rate observed in the H(2)(18)O labeling experiment, reaching over 90% labeling intensity at 48 h. The incorporation of two isotopic (18)O atoms derived from molecular oxygen ((18)O(2)) was observed in the extracted Chl d, and the percentage of double isotopic (18)O-labeled Chl d increased in parallel with the decrease of non-isotopic-labeled Chl d. This clearly indicated that the oxygen atom in the C3(1)-formyl group of Chl d is derived from dioxygen via an oxygenase-type reaction mechanism.

  4. Multi-isotope labelling of organic matter by diffusion of 2H/18O-H2O vapour and 13C-CO2 into the leaves and its distribution within the plant

    NASA Astrophysics Data System (ADS)

    Studer, M. S.; Siegwolf, R. T. W.; Leuenberger, M.; Abiven, S.

    2015-03-01

    Isotope labelling is a powerful tool to study elemental cycling within terrestrial ecosystems. Here we describe a new multi-isotope technique to label organic matter (OM). We exposed poplars (Populus deltoides × nigra) for 14 days to an atmosphere enriched in 13CO2 and depleted in 2H218O. After 1 week, the water-soluble leaf OM (δ13C = 1346 ± 162‰) and the leaf water were strongly labelled18O = -63 ± 8, δ2H = -156 ± 15‰). The leaf water isotopic composition was between the atmospheric and stem water, indicating a considerable back-diffusion of vapour into the leaves (58-69%) in the opposite direction to the net transpiration flow. The atomic ratios of the labels recovered (18O/13C, 2H/13C) were 2-4 times higher in leaves than in the stems and roots. This could be an indication of the synthesis of more condensed compounds in roots and stems (e.g. lignin vs. cellulose) or might be the result of O and H exchange and fractionation processes during phloem transport and biosynthesis. We demonstrate that the three major OM elements (C, O, H) can be labelled and traced simultaneously within the plant. This approach could be of interdisciplinary interest in the fields of plant physiology, palaeoclimatic reconstruction or soil science.

  5. Direct Synthesis of ESBO Derivatives-18O Labelled with Dioxirane

    PubMed Central

    Tommasi, Immacolata; Fusco, Caterina

    2013-01-01

    This work addresses a new approach developed in our laboratory, consisting in the application of isolated dimethyldioxirane (DDO, 1a) labelled with 18O for synthesis of epoxidized glyceryl linoleate (Gly-LLL, 2). We expect that this work could contribute in improving analytical methods for the determination of epoxidized soybean oil (ESBO) in complex food matrices by adopting an 18O-labelled-epoxidized triacylglycerol as an internal standard. PMID:24163617

  6. 18O Labeling of Chlorophyll d in Acaryochloris marina Reveals That Chlorophyll a and Molecular Oxygen Are Precursors*

    PubMed Central

    Schliep, Martin; Crossett, Ben; Willows, Robert D.; Chen, Min

    2010-01-01

    The cyanobacterium Acaryochloris marina was cultured in the presence of either H218O or 18O2, and the newly synthesized chlorophylls (Chl a and Chl d) were isolated using high performance liquid chromatography and analyzed by mass spectroscopy. In the presence of H218O, newly synthesized Chl a and d, both incorporated up to four isotopic 18O atoms. Time course H218O labeling experiments showed incorporation of isotopic 18O atoms originating from H218O into Chl a, with over 90% of Chl a 18O-labeled at 48 h. The incorporation of isotopic 18O atoms into Chl d upon incubation in H218O was slower compared with Chl a with ∼50% 18O-labeled Chl d at 115 h. The rapid turnover of newly synthesized Chl a suggested that Chl a is the direct biosynthetic precursor of Chl d. In the presence of 18O2 gas, one isotopic 18O atom was incorporated into Chl a with approximately the same kinetic incorporation rate observed in the H218O labeling experiment, reaching over 90% labeling intensity at 48 h. The incorporation of two isotopic 18O atoms derived from molecular oxygen (18O2) was observed in the extracted Chl d, and the percentage of double isotopic 18O-labeled Chl d increased in parallel with the decrease of non-isotopic-labeled Chl d. This clearly indicated that the oxygen atom in the C31-formyl group of Chl d is derived from dioxygen via an oxygenase-type reaction mechanism. PMID:20610399

  7. Evidence of the chemical reaction of (18)O-labelled nitrite with CO2 in aqueous buffer of neutral pH and the formation of (18)OCO by isotope ratio mass spectrometry.

    PubMed

    Tsikas, Dimitrios; Böhmer, Anke; Gros, Gerolf; Endeward, Volker

    2016-05-01

    Inorganic nitrite (NO2(-), ON-O(-) ←→ (-)O-NO) is the autoxidation product of nitric oxide (NO). Nitrite can also be formed from inorganic nitrate (ONO2(-)), the major oxidation product of NO in erythrocytes, by the catalytic action of bacterial nitrate reductase in gut and oral microflora. Nitrite can be reduced to NO by certain cellular proteins and enzymes, as well as in the gastric juice under acidic conditions. Hemoglobin, xanthine oxidoreductase and carbonic anhydrase (CA) have been reported to convert nitrite to NO. Renal CA isoforms are involved in the reabsorption of nitrite and may, therefore, play an important role in NO homeostasis. Yet, the mechanisms underlying the action of CA on nitrite are incompletely understood. The nitrate/nitrite system is regarded as a reservoir of NO. We have recently shown that nitrite reacts chemically with carbon dioxide (CO2), the regular substrate of CA. The present communication reports a stable isotope ratio mass spectrometry (IRMS) study on the reaction of NO2(-) and CO2 performed in 50 mM HEPES buffer of pH 7.4 at 37 °C. By using (18)O-labelled nitrite ((18)ON-O(-)/(-18)O-NO) and CO2 we observed formation of (18)O-labelled CO2. This finding is an unequivocal evidence of the chemical reaction of (18)ON-O(-)/(-18)O-NO with CO2. The reaction is rapid and involves nucleophilic attack of the negatively charged nitrite via one of its oxygen atoms on the partially positively charged CO2 molecule to form the putative intermediate (18)ON-O-CO2(-)/(-)O2C-(18)O-NO. The by far largest fraction of this intermediate decomposes back to (18)ON-O(-)/(-18)O-NO and CO2. A very small fraction of the intermediate, however, rearranges and finally decomposes to form (18)OCO and nitrite. This reaction is slower in the presence of an isolated erythrocytic CA isoform II. In summary, NO2(-), CO2 and CA are ubiquitous. The chemical reaction of NO2(-) with CO2 and its modulation by CA isoforms may play important roles in the transport of

  8. Measurements of 18O18O and 17O18O in the atmosphere and the role of isotope-exchange reactions

    NASA Astrophysics Data System (ADS)

    Yeung, Laurence Y.; Young, Edward D.; Schauble, Edwin A.

    2012-09-01

    Of the six stable isotopic variants of O2, only three are measured routinely. Observations of natural variations in 16O18O/16O16O and 16O17O/16O16O ratios have led to insights in atmospheric, oceanographic, and paleoclimate research. Complementary measurements of the exceedingly rare 18O18O and 17O18O isotopic variants might therefore broaden our understanding of oxygen cycling. Here we describe a method to measure natural variations in these multiply substituted isotopologues of O2. Its accuracy is demonstrated by measuring isotopic effects for Knudsen diffusion and O2 electrolysis in the laboratory that are consistent with theoretical predictions. We then report the first measurements of 18O18O and 17O18O proportions relative to the stochastic distribution of isotopes (i.e., Δ36 and Δ35 values, respectively) in tropospheric air. Measured enrichments in 18O18O and 17O18O yield Δ36 = 2.05 ± 0.24‰ and Δ35 = 1.4 ± 0.5‰ (2σ). Based on the results of our electrolysis experiment, we suggest that autocatalytic O(3P) + O2 isotope exchange reactions play an important role in regulating the distribution of 18O18O and 17O18O in air. We constructed a box model of the atmosphere and biosphere that includes the effects of these isotope exchange reactions, and we find that the biosphere exerts only a minor influence on atmospheric Δ36 and Δ35 values. O(3P) + O2 isotope exchange in the stratosphere and troposphere is therefore expected to govern atmospheric Δ36 and Δ35 values on decadal timescales. These results suggest that the `clumped' isotopic composition of atmospheric O2in ice core records is sensitive to past variations in atmospheric dynamics and free-radical chemistry.

  9. 18O-Labeled Proteome Reference as Global Internal Standards for Targeted Quantification by Selected Reaction Monitoring-Mass Spectrometry

    SciTech Connect

    Kim, Jong Seo; Fillmore, Thomas L.; Liu, Tao; Robinson, Errol W.; Hossain, Mahmud; Champion, Boyd L.; Moore, Ronald J.; Camp, David G.; Smith, Richard D.; Qian, Weijun

    2011-10-11

    Selected reaction monitoring-mass spectrometry (SRM-MS) is an emerging technology for high throughput targeted protein quantification and verification in biological and biomarker discovery studies; however, the cost associated with the use of stable isotope labeled synthetic peptides as internal standards is prohibitive for quantitatively screening large numbers of candidate proteins as often required in the pre-verification phase of biomarker discovery. Herein we present the proof-of-concept experiments of using an 18O-labeled 'universal' reference as comprehensive internal standards for quantitative SRM-MS analysis. With an 18O-labeled whole proteome sample as reference, every peptide of interest will have its own corresponding heavy isotope labeled internal standard, thus providing an ideal approach for quantitative screening of a large number of candidates using SRM-MS. Our results showed that the 18O incorporation efficiency using a recently improved protocol was >99.5% for most peptides investigated, a level comparable to 13C/15N labeled synthetic peptides in terms of heavy isotope incorporation. The accuracy, reproducibility, and linear dynamic range of quantification were further assessed based on known ratios of standard proteins spiked into mouse plasma with an 18O-labeled mouse plasma reference. A dynamic range of four orders of magnitude in relative concentration was obtained with high reproducibility (i.e., coefficient of variance <10%) based on the 16O/18O peak area ratios. Absolute and relative quantification of C-reactive protein and prostate-specific antigen were demonstrated by coupling an 18O-labeled reference with standard additions of protein standards. Collectively, our results demonstrated that the use of 18O-labeled reference provides a convenient and effective strategy for quantitative SRM screening of large number of candidate proteins.

  10. Large-scale multiplexed quantitative discovery proteomics enabled by the use of an (18)O-labeled "universal" reference sample.

    PubMed

    Qian, Wei-Jun; Liu, Tao; Petyuk, Vladislav A; Gritsenko, Marina A; Petritis, Brianne O; Polpitiya, Ashoka D; Kaushal, Amit; Xiao, Wenzhong; Finnerty, Celeste C; Jeschke, Marc G; Jaitly, Navdeep; Monroe, Matthew E; Moore, Ronald J; Moldawer, Lyle L; Davis, Ronald W; Tompkins, Ronald G; Herndon, David N; Camp, David G; Smith, Richard D

    2009-01-01

    The quantitative comparison of protein abundances across a large number of biological or patient samples represents an important proteomics challenge that needs to be addressed for proteomics discovery applications. Herein, we describe a strategy that incorporates a stable isotope (18)O-labeled "universal" reference sample as a comprehensive set of internal standards for analyzing large sample sets quantitatively. As a pooled sample, the (18)O-labeled "universal" reference sample is spiked into each individually processed unlabeled biological sample and the peptide/protein abundances are quantified based on (16)O/(18)O isotopic peptide pair abundance ratios that compare each unlabeled sample to the identical reference sample. This approach also allows for the direct application of label-free quantitation across the sample set simultaneously along with the labeling-approach (i.e., dual-quantitation) since each biological sample is unlabeled except for the labeled reference sample that is used as internal standards. The effectiveness of this approach for large-scale quantitative proteomics is demonstrated by its application to a set of 18 plasma samples from severe burn patients. When immunoaffinity depletion and cysteinyl-peptide enrichment-based fractionation with high resolution LC-MS measurements were combined, a total of 312 plasma proteins were confidently identified and quantified with a minimum of two unique peptides per protein. The isotope labeling data was directly compared with the label-free (16)O-MS intensity data extracted from the same data sets. The results showed that the (18)O reference-based labeling approach had significantly better quantitative precision compared to the label-free approach. The relative abundance differences determined by the two approaches also displayed strong correlation, illustrating the complementary nature of the two quantitative methods. The simplicity of including the (18)O-reference for accurate quantitation makes this

  11. Using power spectrum analysis to evaluate (18)O-water labeling data acquired from low resolution mass spectrometers.

    PubMed

    Sadygov, Rovshan G; Zhao, Yingxin; Haidacher, Sigmund J; Starkey, Jonathan M; Tilton, Ronald G; Denner, Larry

    2010-08-06

    We describe a method for ratio estimations in (18)O-water labeling experiments acquired from low resolution isotopically resolved data. The method is implemented in a software package specifically designed for use in experiments making use of zoom-scan mode data acquisition. Zoom-scan mode data allow commonly used ion trap mass spectrometers to attain isotopic resolution, which makes them amenable to use in labeling schemes such as (18)O-water labeling, but algorithms and software developed for high resolution instruments may not be appropriate for the lower resolution data acquired in zoom-scan mode. The use of power spectrum analysis is proposed as a general approach that may be uniquely suited to these data types. The software implementation uses a power spectrum to remove high-frequency noise and band-filter contributions from coeluting species of differing charge states. From the elemental composition of a peptide sequence, we generate theoretical isotope envelopes of heavy-light peptide pairs in five different ratios; these theoretical envelopes are correlated with the filtered experimental zoom scans. To automate peptide quantification in high-throughput experiments, we have implemented our approach in a computer program, MassXplorer. We demonstrate the application of MassXplorer to two model mixtures of known proteins and to a complex mixture of mouse kidney cortical extract. Comparison with another algorithm for ratio estimations demonstrates the increased precision and automation of MassXplorer.

  12. Isotope 18 O 16 O Ratio Measurements of Water Vapor by use of Photoacoustic Spectroscopy

    NASA Astrophysics Data System (ADS)

    Matsumi, Yutaka; Kishigami, Masahiro; Tanaka, Noriyuki; Kawasaki, Masahiro; Inoue, Gen

    1998-09-01

    We applied a photoacoustic spectroscopy technique to isotope ratio measurements of 16 O and 18 O in water-vapor samples, using a pulsed tunable dye laser pumped by a Nd:YAG laser. The fourth overtone bands (4 OH ) of water molecules near 720 nm were investigated. We identified the absorption lines of H 2 16 O and H 2 18 O in the photoacoustic spectra that we measured by using an 18 O-enriched water sample and the HITRAN database. We measured the difference in the 18 O 16 O isotope ratios for normal distilled water and Antarctic ice, using the photoacoustic method. The value obtained for the difference between the two samples is 18 O 32 16 , where the indicated deviation was a 1 value among 240-s measurements, whereas the value measured with a conventional isotope mass spectrometer was 18 O 28 2 . This method is demonstrated to have the potential of a transportable system for in situ and quick measurements of the H 2 18 O H 2 16 O ratio in the environment.

  13. Dentine oxygen isotopes (δ (18)O) as a proxy for odontocete distributions and movements.

    PubMed

    Matthews, Cory J D; Longstaffe, Fred J; Ferguson, Steven H

    2016-07-01

    Spatial variation in marine oxygen isotope ratios (δ (18)O) resulting from differential evaporation rates and precipitation inputs is potentially useful for characterizing marine mammal distributions and tracking movements across δ (18)O gradients. Dentine hydroxyapatite contains carbonate and phosphate that precipitate in oxygen isotopic equilibrium with body water, which in odontocetes closely tracks the isotopic composition of ambient water. To test whether dentine oxygen isotope composition reliably records that of ambient water and can therefore serve as a proxy for odontocete distribution and movement patterns, we measured δ (18)O values of dentine structural carbonate (δ (18) OSC) and phosphate (δ (18) OP) of seven odontocete species (n = 55 individuals) from regional marine water bodies spanning a surface water δ (18)O range of several per mil. Mean dentine δ (18) OSC (range +21.2 to +25.5‰ VSMOW) and δ (18) OP (+16.7 to +20.3‰) values were strongly correlated with marine surface water δ (18)O values, with lower dentine δ (18) OSC and δ (18) OP values in high-latitude regions (Arctic and Eastern North Pacific) and higher values in the Gulf of California, Gulf of Mexico, and Mediterranean Sea. Correlations between dentine δ (18) OSC and δ (18) OP values with marine surface water δ (18)O values indicate that sequential δ (18)O measurements along dentine, which grows incrementally and archives intra- and interannual isotopic composition over the lifetime of the animal, would be useful for characterizing residency within and movements among water bodies with strong δ (18)O gradients, particularly between polar and lower latitudes, or between oceans and marginal basins.

  14. Isotope parameters (δD, δ18O) and sources of freshwater input to Kara Sea

    NASA Astrophysics Data System (ADS)

    Dubinina, E. O.; Kossova, S. A.; Miroshnikov, A. Yu.; Fyaizullina, R. V.

    2017-01-01

    The isotope characteristics (δD, δ18O) of Kara Sea water were studied for quantitative estimation of freshwater runoff at stations located along transect from Yamal Peninsula to Blagopoluchiya Bay (Novaya Zemlya). Freshwater samples were studied for glaciers (Rose, Serp i Molot) and for Yenisei and Ob estuaries. As a whole, δD and δ18O are higher in glaciers than in river waters. isotope composition of estuarial water from Ob River is δD =-131.4 and δ18O =-17.6‰. Estuarial waters of Yenisei River are characterized by compositions close to those of Ob River (-134.4 and-17.7‰), as well as by isotopically "heavier" compositions (-120.7 and-15.8‰). Waters from studied section of Kara Sea can be product of mixing of freshwater (δD =-119.4, δ18O =-15.5) and seawater (S = 34.9, δD = +1.56, δ18O = +0.25) with a composition close to that of Barents Sea water. isotope parameters of water vary significantly with salinity in surface layer, and Kara Sea waters are desalinated along entire studied transect due to river runoff. concentration of freshwater is 5-10% in main part of water column, and <5% at a depth of >100 m. maximum contribution of freshwater (>65%) was recorded in surface layer of central part of sea.

  15. Microwave-assisted 18O-labeling of proteins catalyzed by formic acid.

    PubMed

    Liu, Ning; Wu, Hanzhi; Liu, Hongxia; Chen, Guonan; Cai, Zongwei

    2010-11-01

    Oxygen exchange may occur at carboxyl groups catalyzed by acid. The reaction, however, takes at least several days at room temperature. The long-time exchanging reaction often prevents its application from protein analysis. In this study, an (18)O-labeling method utilizing microwave-assisted acid hydrolysis was developed. After being dissolved in (16)O/(18)O (1:1) water containing 2.5% formic acid, protein samples were exposed to microwave irradiation. LC-MS/MS analysis of the resulted peptide mixtures indicated that oxygen in the carboxyl groups from glutamic acid, aspartic acid, and the C-terminal residues could be efficiently exchanged with (18)O within less than 15 min. The rate of back exchange was so slow that no detectable back exchange could be found during the HPLC run.

  16. sup 13 C and sup 18 O isotopic disequilibrium in biological carbonates: I. Patterns

    SciTech Connect

    McConnaughey, T. )

    1989-01-01

    Biological carbonates frequently precipitate out of {sup 18}O and {sup 13}C equilibrium with ambient waters. Two patterns of isotopic disequilibrium are particularly common. Kinetic disequilibria, so designated because they apparently result from kinetic isotope effects during CO{sub 2} hydration and hydroxylation, involve simultaneous depletions of {sup 18}O and {sup 13}C as large as 4{per thousand} and 10 to 15{per thousand}, respectively. Rapid skeletogenesis favors strong kinetic effects, and approximately linear correlations between skeletal {delta}{sup 18}O and {delta}{sup 13}C are common in carbonates showing mainly the kinetic pattern. Metabolic effects involve additional positive or negative modulation of skeletal {delta}{sup 13}C, reflecting changes in the {delta}{sup 13}C of dissolved inorganic carbon, caused mainly by photosynthesis and respiration. Kinetic isotope disequilibria tend to be fairly consistent in rapidly growing parts of photosynthetic corals, and time dependent isotopic variations therefore reflect changes in environmental conditions. {delta}{sup 18}O variations from Galapagos corals yields meaningful clues regarding seawater temperature, while {delta}{sup 13}C variations reflect changes in photosynthesis, modulated by cloudiness.

  17. Quantitative analysis of cell surface membrane proteins using membrane-impermeable chemical probe coupled with 18O labeling.

    PubMed

    Zhang, Haizhen; Brown, Roslyn N; Qian, Wei-Jun; Monroe, Matthew E; Purvine, Samuel O; Moore, Ronald J; Gritsenko, Marina A; Shi, Liang; Romine, Margaret F; Fredrickson, James K; Pasa-Tolić, Ljiljana; Smith, Richard D; Lipton, Mary S

    2010-05-07

    We report a mass spectrometry-based strategy for quantitative analysis of cell surface membrane proteome changes. The strategy includes enrichment of surface membrane proteins using a membrane-impermeable chemical probe followed by stable isotope (18)O labeling and LC-MS analysis. We applied this strategy for enriching membrane proteins expressed by Shewanella oneidensis MR-1, a Gram-negative bacterium with known metal-reduction capability via extracellular electron transfer between outer membrane proteins and extracellular electron receptors. LC/MS/MS analysis resulted in the identification of about 400 proteins with 79% of them being predicted to be membrane localized. Quantitative aspects of the membrane enrichment were shown by peptide level (16)O and (18)O labeling of proteins from wild-type and mutant cells (generated from deletion of a type II secretion protein, GspD) prior to LC-MS analysis. Using a chemical probe labeled pure protein as an internal standard for normalization, the quantitative data revealed reduced abundances in Delta gspD mutant cells of many outer membrane proteins including the outer membrane c-type cytochromes OmcA and MtrC, in agreement with a previous report that these proteins are substrates of the type II secretion system.

  18. Carbonate clumped isotope constraints on Silurian ocean temperature and seawater δ18O

    NASA Astrophysics Data System (ADS)

    Cummins, Renata C.; Finnegan, Seth; Fike, David A.; Eiler, John M.; Fischer, Woodward W.

    2014-09-01

    Much of what we know about the history of Earth's climate derives from the chemistry of carbonate minerals in the sedimentary record. The oxygen isotopic compositions (δ18O) of calcitic marine fossils and cements have been widely used as a proxy for past seawater temperatures, but application of this proxy to deep geologic time is complicated by diagenetic alteration and uncertainties in the δ18O of seawater in the past. Carbonate clumped isotope thermometry provides an independent estimate of the temperature of the water from which a calcite phase precipitated, and allows direct calculation of the δ18O of the water. The clumped isotope composition of calcites is also highly sensitive to recrystallization and can help diagnose different modes of diagenetic alteration, enabling evaluation of preservation states and identification of the most pristine materials from within a sample set-critical information for assessing the quality of paleoproxy data generated from carbonates. We measured the clumped isotope composition of a large suite of calcitic fossils (primarily brachiopods and corals), sedimentary grains, and cements from Silurian (ca. 433 Ma) stratigraphic sections on the island of Gotland, Sweden. Substantial variability in clumped isotope temperatures suggests differential preservation with alteration largely tied to rock-buffered diagenesis, complicating the generation of a stratigraphically resolved climate history through these sections. Despite the generally high preservation quality of samples from these sections, micro-scale observations of calcite fabric and trace metal composition using electron backscatter diffraction and electron microprobe analysis suggest that only a subset of relatively pristine samples retain primary clumped isotope signatures. These samples indicate that Silurian tropical oceans were likely warm (33 ± 7 °C) and similar in oxygen isotopic composition to that estimated for a "modern" ice-free world (δ18OVSMOW of -1.1 ± 1

  19. Elemental and isotopic ( 29Si and 18O) tracing of glass alteration mechanisms

    NASA Astrophysics Data System (ADS)

    Valle, Nathalie; Verney-Carron, Aurélie; Sterpenich, Jérôme; Libourel, Guy; Deloule, Etienne; Jollivet, Patrick

    2010-06-01

    To better understand glass alteration mechanisms, especially alteration layers formation, leaching experiments of a borosilicate glass (SON68) doped with a different rare earth element (La, Ce, or Nd) with solutions rich in 29Si and 18O were carried out. The coupled analyses of glass, alteration products, and solution led to a complete elemental and isotopic ( 29Si and 18O) budget. They revealed different behaviours of elements that depend not only on their structural role in the glass, but also on their affinity for alteration products (gel, phyllosilicates, phosphates). However, analyses of both glass and solution are not sufficient to describe the real exchanges occurring between glass and solution. The use of 29Si and 18O tracers gives new insights on the formation of alteration layers. During alteration the phyllosilicates records the isotopic variations of the leaching solution. Their isotopic signatures highlight a mechanism of dissolution/precipitation, which implies equilibrium between the secondary phases and the solution. On the other hand the gel isotopic signature, that is intermediate between the glass and the solution, substantiates the hypothesis that the gel is formed by hydrolysis/condensation reactions. This mechanism can thus explain the influence of the gel formation conditions (alteration conditions, solution saturation) on the structure (reorganisation) and texture (porosity) of this layer and on its protective effect. These hydrolysis/condensation reactions are also certainly involved in the aluminosilicate glass alteration and in the formation of palagonite.

  20. Growth of 18O isotopically enriched ZnO nanorods by two novel VPT methods

    NASA Astrophysics Data System (ADS)

    Gray, Ciarán; Trefflich, Lukas; Röder, Robert; Ronning, Carsten; Henry, Martin O.; McGlynn, Enda

    2017-02-01

    We have developed two novel vapour phase transport methods to grow ZnO nanorod arrays isotopically enriched with 18O. Firstly, a three-step process used to grow natural and Zn-enriched ZnO nanorods has been further modified, by replacing the atmospheric O2 with enriched 18O2, in order to grow 18O-enriched ZnO nanorods using this vapour-solid method on chemical bath deposited buffer layers. In addition, 18O-enriched ZnO nanorods were successfully grown using 18O isotopically enriched ZnO source powders in a vapour-liquid-solid growth method. Scanning electron microscopy studies confirmed the success of both growth methods in terms of nanorod morphology, although in the case of the vapour-liquid-solid samples, the nanorods' c-axes were not vertically aligned due to the use of a non-epitaxial substrate. Raman and PL studies indicated clearly that O-enrichment was successful in both cases, although the results indicate that the enrichment is at a lower level in our samples compared to previous reports with the same nominal enrichment levels. The results of our studies also allow us to comment on both levels of enrichment achieved and on novel effects of the high temperature growth environment on the nanorod growth, as well as suggesting possible mechanisms for such effects. Very narrow photoluminescence line widths, far narrower than those reported previously in the literature for isotopically enriched bulk ZnO, are seen in both the vapour-solid and vapour-liquid-solid nanorod samples demonstrating their excellent optical quality and their potential for use in detailed optical studies of defects and impurities using low temperature photoluminescence.

  1. Quantitative Proteomic Approach for MicroRNA Target Prediction Based on 18O/16O Labeling

    PubMed Central

    Ma, Xuepo; Zhu, Ying; Huang, Yufei; Tegeler, Tony; Gao, Shou-Jiang; Zhang, Jianqiu

    2015-01-01

    MOTIVATION Among many large-scale proteomic quantification methods, 18O/16O labeling requires neither specific amino acid in peptides nor label incorporation through several cell cycles, as in metabolic labeling; it does not cause significant elution time shifts between heavy- and light-labeled peptides, and its dynamic range of quantification is larger than that of tandem mass spectrometry-based quantification methods. These properties offer 18O/16O labeling the maximum flexibility in application. However, 18O/16O labeling introduces large quantification variations due to varying labeling efficiency. There lacks a processing pipeline that warrants the reliable identification of differentially expressed proteins (DEPs). This motivates us to develop a quantitative proteomic approach based on 18O/16O labeling and apply it on Kaposi sarcoma-associated herpesvirus (KSHV) microRNA (miR) target prediction. KSHV is a human pathogenic γ-herpesvirus strongly associated with the development of B-cell proliferative disorders, including primary effusion lymphoma. Recent studies suggest that miRs have evolved a highly complex network of interactions with the cellular and viral transcriptomes, and relatively few KSHV miR targets have been characterized at the functional level. While the new miR target prediction method, photoactivatable ribonucleoside-enhanced cross-linking and immunoprecipitation (PAR-CLIP), allows the identification of thousands of miR targets, the link between miRs and their targets still cannot be determined. We propose to apply the developed proteomic approach to establish such links. METHOD We integrate several 18O/16O data processing algorithms that we published recently and identify the messenger RNAs of downregulated proteins as potential targets in KSHV miR-transfected human embryonic kidney 293T cells. Various statistical tests are employed for picking DEPs, and we select the best test by examining the enrichment of PAR-CLIP-reported targets with

  2. 13C-18O isotope signatures and ‘clumped isotope’ thermometry in foraminifera and coccoliths

    NASA Astrophysics Data System (ADS)

    Tripati, Aradhna K.; Eagle, Robert A.; Thiagarajan, Nivedita; Gagnon, Alexander C.; Bauch, Henning; Halloran, Paul R.; Eiler, John M.

    2010-10-01

    Accurate constraints on past ocean temperatures and compositions are critical for documenting climate change and resolving its causes. Most proxies for temperature are not thermodynamically based, appear to be subject to biological processes, require regional calibrations, and/or are influenced by fluid composition. As a result, their interpretation becomes uncertain when they are applied in settings not necessarily resembling those in which they were empirically calibrated. Independent proxies for past temperature could provide an important means of testing and/or expanding on existing reconstructions. Here we report measurements of abundances of stable isotopologues of calcitic and aragonitic benthic and planktic foraminifera and coccoliths, relate those abundances to independently estimated growth temperatures, and discuss the possible scope of equilibrium and kinetic isotope effects. The proportions of 13C- 18O bonds in these samples exhibits a temperature dependence that is generally similar to that previously been reported for inorganic calcite and other biologically precipitated carbonate-containing minerals (apatite from fish, reptile, and mammal teeth; calcitic brachiopods and molluscs; aragonitic coral and mollusks). Most species that exhibit non-equilibrium 18O/ 16O (δ 18O) and 13C/ 12C (δ 13C) ratios are characterized by 13C- 18O bond abundances that are similar to inorganic calcite and are generally indistinguishable from apparent equilibrium, with possible exceptions among benthic foraminiferal samples from the Arctic Ocean where temperatures are near-freezing. Observed isotope ratios in biogenic carbonates can be explained if carbonate minerals generally preserve a state of ordering that reflects the extent of isotopic equilibration of the dissolved inorganic carbon species.

  3. Tracing atmospheric moisture from precipitation δ18O to climate proxy using an isotope enabled land surface model

    NASA Astrophysics Data System (ADS)

    Kanner, L.; Buenning, N. H.; Stott, L. D.; Timmermann, A.

    2013-12-01

    A paleoclimate interpretation of a terrestrial hydrologic proxy such as the δ18O of tree cellulose or speleothem calcite may be biased or misinterpreted if the isotopic composition of the soil water from which the proxy originated undergoes isotopic exchange or fractionation. In this study, we use a global isotope-enabled land surface model (IsoLSM) to investigate how the δ18O of precipitation may be altered in a soil column due to evaporation and vertical moisture flux. In order to assess how precipitation and evaporation contribute the soil water isotopic variability, we compare seasonal and interannual changes in simulated xylem water δ18O within a control simulation and in a suite of experiments where the effect of precipitation δ18O, water vapor δ18O, and ground water evaporation are independently removed. The simulations, carried out for 1979 to 2004, reveal that in semi-arid regions, such as the southwest United States, the seasonal cycle in xylem water δ18O is strongly affected by evaporative loss during the dry season and this can constitute as much as 50% of the interannual δ18O variance. Additional simulations, including soil water tagging experiments, indicate that upward fluxes of soil water occur during drier periods. For soil water δ18O profiles that are isotopically more depleted in 18O at depth, this imparts a low isotopic signature to xylem water δ18O during such dry intervals. Hence, without taking into account moisture flux processes, an isotopic proxy could be misinterpreted as wet conditions (due to decreased evaporative enrichment) for low δ18O years when instead drier conditions are equally as likely. Using IsoLSM simulated xylem water and leaf water δ18O, offline calculations of cellulose δ18O compare well with observations in diverse climatic regimes. Thus, the driving mechanisms on soil water δ18O identified in this study, and in particular the important role of evaporation on seasonal and interannual timescales, may

  4. Mechanisms linking metabolism of Helicobacter pylori to (18)O and (13)C-isotopes of human breath CO2.

    PubMed

    Som, Suman; De, Anulekha; Banik, Gourab Dutta; Maity, Abhijit; Ghosh, Chiranjit; Pal, Mithun; Daschakraborty, Sunil B; Chaudhuri, Sujit; Jana, Subhra; Pradhan, Manik

    2015-06-03

    The gastric pathogen Helicobacter pylori utilize glucose during metabolism, but the underlying mechanisms linking to oxygen-18 ((18)O) and carbon-13 ((13)C)-isotopic fractionations of breath CO2 during glucose metabolism are poorly understood. Using the excretion dynamics of (18)O/(16)O and (13)C/(12)C-isotope ratios of breath CO2, we found that individuals with Helicobacter pylori infections exhibited significantly higher isotopic enrichments of (18)O in breath CO2 during the 2h-glucose metabolism regardless of the isotopic nature of the substrate, while no significant enrichments of (18)O in breath CO2 were manifested in individuals without the infections. In contrast, the (13)C-isotopic enrichments of breath CO2 were significantly higher in individuals with Helicobacter pylori compared to individuals without infections in response to (13)C-enriched glucose uptake, whereas a distinguishable change of breath (13)C/(12)C-isotope ratios was also evident when Helicobacter pylori utilize natural glucose. Moreover, monitoring the (18)O and (13)C-isotopic exchange in breath CO2 successfully diagnosed the eradications of Helicobacter pylori infections following a standard therapy. Our findings suggest that breath (12)C(18)O(16)O and (13)C(16)O(16)O can be used as potential molecular biomarkers to distinctively track the pathogenesis of Helicobacter pylori and also for eradication purposes and thus may open new perspectives into the pathogen's physiology along with isotope-specific non-invasive diagnosis of the infection.

  5. SVM model for quality assessment of medium resolution mass spectra from 18O-water labeling experiments.

    PubMed

    Nefedov, Alexey V; Gilski, Miroslaw J; Sadygov, Rovshan G

    2011-04-01

    We describe a method for assessing the quality of mass spectra and improving reliability of relative ratio estimations from (18)O-water labeling experiments acquired from low resolution mass spectrometers. The mass profiles of heavy and light peptide pairs are often affected by artifacts, including coeluting contaminant species, noise signal, instrumental fluctuations in measuring ion position and abundance levels. Such artifacts distort the profiles, leading to erroneous ratio estimations, thus reducing the reliability of ratio estimations in high throughput quantification experiments. We used support vector machines (SVMs) to filter out mass spectra that deviated significantly from expected theoretical isotope distributions. We built an SVM classifier with a decision function that assigns a score to every mass profile based on such spectral features as mass accuracy, signal-to-noise ratio, and differences between experimental and theoretical isotopic distributions. The classifier was trained using a data set obtained from samples of mouse renal cortex. We then tested it on protein samples (bovine serum albumin) mixed in five different ratios of labeled and unlabeled species. We demonstrated that filtering the data using our SVM classifier results in as much as a 9-fold reduction in the coefficient of variance of peptide ratios, thus significantly improving the reliability of ratio estimations.

  6. sup 13 C and sup 18 O isotopic disequilibrium in biological carbonates: II. In vitro simulation of kinetic isotope effects

    SciTech Connect

    McConnaughey, T. )

    1989-01-01

    Biological carbonates are built largely from CO{sub 2}, which diffuses across the skeletogenic membrane and reacts to form HCO{sub 3}{sup {minus}}. Kinetic discrimination against the heavy isotopes {sup 18}O and {sup 13}C during CO{sub 2} hydration and hydroxylation apparently causes most of the isotopic disequilibrium observed in biological carbonates. These kinetic isotope effects are expressed when the extracytosolic calcifying solution is thin and alkaline, and HCO{sub 3}{sup {minus}} precipitates fairly rapidly as CaCO{sub 3}. In vitro simulation of the calcifying environment produced heavy isotope depletions qualitatively similar to, but somewhat more extreme than, those seen in biological carbonates. Isotopic equilibration during biological calcification occurs through CO{sub 2} exchange across the calcifying membrane and by admixture ambient waters (containing HCO{sub 3}{sup {minus}}) into the calcifying fluids. Both mechanisms tend to produce linear correlations between skeletal {delta}{sup 13}C and {delta}{sup 18}O.

  7. Terahertz spectroscopy of N18O and isotopic invariant fit of several nitric oxide isotopologs

    NASA Astrophysics Data System (ADS)

    Müller, Holger S. P.; Kobayashi, Kaori; Takahashi, Kazumasa; Tomaru, Kazuko; Matsushima, Fusakazu

    2015-04-01

    A tunable far-infrared laser sideband spectrometer was used to investigate a nitric oxide sample enriched in 18O between 0.99 and 4.75 THz. Regular, electric dipole transitions were recorded between 0.99 and 2.52 THz, while magnetic dipole transitions between the 2Π1/2 and 2Π3/2 spin-ladders were recorded between 3.71 and 4.75 THz. These data were combined with lower frequency data of N18 O (unlabeled atoms refer to 14 N and 16 O, respectively), with rotational data of NO, 15 NO, N17 O, and 15 N18 O, and with heterodyne infrared data of NO to be subjected to one isotopic invariant fit. Rotational, fine and hyperfine structure parameters were determined along with vibrational, rotational, and Born-Oppenheimer breakdown corrections. The resulting spectroscopic parameters permit prediction of rotational spectra suitable for the identification of various nitric oxide isotopologs especially in the interstellar medium by means of rotational spectroscopy.

  8. Mechanisms linking metabolism of Helicobacter pylori to 18O and 13C-isotopes of human breath CO2

    PubMed Central

    Som, Suman; De, Anulekha; Banik, Gourab Dutta; Maity, Abhijit; Ghosh, Chiranjit; Pal, Mithun; Daschakraborty, Sunil B.; Chaudhuri, Sujit; Jana, Subhra; Pradhan, Manik

    2015-01-01

    The gastric pathogen Helicobacter pylori utilize glucose during metabolism, but the underlying mechanisms linking to oxygen-18 (18O) and carbon-13 (13C)-isotopic fractionations of breath CO2 during glucose metabolism are poorly understood. Using the excretion dynamics of 18O/16O and 13C/12C-isotope ratios of breath CO2, we found that individuals with Helicobacter pylori infections exhibited significantly higher isotopic enrichments of 18O in breath CO2 during the 2h-glucose metabolism regardless of the isotopic nature of the substrate, while no significant enrichments of 18O in breath CO2 were manifested in individuals without the infections. In contrast, the 13C-isotopic enrichments of breath CO2 were significantly higher in individuals with Helicobacter pylori compared to individuals without infections in response to 13C-enriched glucose uptake, whereas a distinguishable change of breath 13C/12C-isotope ratios was also evident when Helicobacter pylori utilize natural glucose. Moreover, monitoring the 18O and 13C-isotopic exchange in breath CO2 successfully diagnosed the eradications of Helicobacter pylori infections following a standard therapy. Our findings suggest that breath 12C18O16O and 13C16O16O can be used as potential molecular biomarkers to distinctively track the pathogenesis of Helicobacter pylori and also for eradication purposes and thus may open new perspectives into the pathogen’s physiology along with isotope-specific non-invasive diagnosis of the infection. PMID:26039789

  9. Glycolate metabolism in low and high CO sub 2 -grown chlorella pyrenoidosa and Pavlova lutheri as determined by sup 18 O-labeling

    SciTech Connect

    de Veau, E.J.; Burris, J.E. )

    1989-11-01

    Photorespiration in Chlorella pyrenoidosa Chick. was assayed by measuring {sup 18}O-labeled intermediates of the glycolate pathway. Glycolate, glycine, serine, and excreted glycolate were isolated and analyzed on a gas chromatograph/mass spectrometer to determine isotopic enrichment. Rates of glycolate synthesis were determined from {sup 18}O-labeling kinetics of the intermediates, pool sizes, derived rate equations, and nonlinear regression techniques. Glycolate synthesis was higher in high CO{sub 2}-grown cells than in air-grown cells when both were assayed under the same O{sub 2} and CO{sub 2} concentrations. Synthesis of glycolate, for both types of cells, was stimulated by high O{sub 2} levels and inhibited by high CO{sub 2} levels. Glycolate synthesis in 1.5% CO{sub 2}-grown Chlorella, when exposed to a 0.035% CO{sub 2} atmosphere, increased from about 41 to 86 nanomoles per milligram chlorophyll per minute when the O{sub 2} concentration was increased from 21 to 40%. Glycolate synthesis in air-grown cells increased from 2 to 6 nanomoles per milligram chlorophyll per minute under the same gas levels. Synthesis was undetectable when either the O{sub 2} concentration was lowered to 2% or the CO{sub 2}-concentration was raised to 1.5%. Glycolate excretion was also sensitive to O{sub 2} and CO{sub 2} concentrations in 1.5% CO{sub 2}-grown cells and the glycolate that was excreted was {sup 18}O-labeled. Air-grown cells did not excrete glycolate under any experimental condition. Indirect evidence indicated that glycolate may be excreted as a lactone in Chlorella. Photorespiratory {sup 18}O-labeling kinetics were determined for Pavlova lutheri, which unlike Chlorella and higher plants did not directly synthesize glycine and serine from glycolate. This alga did excrete a significant proportion of newly synthesized glycolate into the media.

  10. Integrated platform with a combination of online digestion and (18)O labeling for proteome quantification via an immobilized trypsin microreactor.

    PubMed

    Zhang, Shen; Yuan, Huiming; Zhao, Baofeng; Zhou, Yuan; Jiang, Hao; Zhang, Lihua; Liang, Zhen; Zhang, Yukui

    2015-08-07

    A novel automated integrated platform for quantitative proteome analysis was established with a combination of online digestion of proteins and in situ(18)O labeling by an immobilized enzyme reactor (IMER); digests were captured and desalted by a C18 trap column, and peptides were analyzed by nanoRPLC-ESI-MS/MS. Bovine serum albumin (BSA) was used to evaluate the performance of the developed platform. Compared with traditional offline methods, not only the digestion and labeling time was shortened from 36 h to just 1 h, but also the labeling efficiency was improved from 95% to 99%. Furthermore, the back-exchange from (18)O to (16)O could also be efficiently avoided by the use of IMER. The platform was further evaluated by the quantitative analysis of 100 ng (18)O and (16)O online labeled yeast sample with a mixing ratio of 1 : 1, and the results showed significantly improved sensitivity and reproducibility, as well as improved quantitative accuracy than offline method. With these advantages, the integrated platform was finally applied to the quantitative profiling of 100 ng proteins extracted from two mouse hepatocarcinoma ascites syngeneic cell lines with high and low lymph node metastases rates, and ten differentially expressed proteins were successfully found, most of which were related to tumorigenesis and tumor metastasis. All these results demonstrate that the developed integrated platform can provide a new way for high efficiency (18)O labeling and the quantitative analysis of trace amounts of sample with high accuracy and high reproducibility.

  11. Combined 34S, 33S and 18O isotope fractionations record different intracellular steps of microbial sulfate reduction

    NASA Astrophysics Data System (ADS)

    Antler, Gilad; Turchyn, Alexandra V.; Ono, Shuhei; Sivan, Orit; Bosak, Tanja

    2017-04-01

    Several enzymatic steps in microbial sulfate reduction (MSR) fractionate the isotope ratios of 33S/32S, 34S/32S and 18O/16O in extracellular sulfate, but the effects of different intracellular processes on the isotopic composition of residual sulfate are still not well quantified. We measured combined multiple sulfur (33S/32S, 34S/32S) and oxygen (18O/16O) isotope ratios of sulfate in pure cultures of a marine sulfate reducing bacterium Desulfovibrio sp. DMSS-1 grown on different organic substrates. These measurements are consistent with the previously reported correlations of oxygen and sulfur isotope fractionations with the cell-specific rate of MSR: faster reduction rates produced smaller isotopic fractionations for all isotopes. Combined isotope fractionation of oxygen and multiple sulfur isotopes are also consistent with the relationship between the rate limiting step during microbial sulfate reduction and the availability of the DsrC subunit. These experiments help reconstruct and interpret processes that operate in natural pore waters characterized by high 18O/16O and moderate 34S/32S ratios and suggest that some multiple isotope signals in the environment cannot be explained by microbial sulfate reduction alone. Instead, these signals support the presence of active, but slow sulfate reduction as well as the reoxidation of sulfide.

  12. Stable oxygen isotopes (delta18(O)) in Austrocedrus chilensis tree rings reflect climate variability in northwestern Patagonia, Argentina.

    PubMed

    Roig, F A; Siegwolf, R; Boninsegna, J A

    2006-11-01

    The stable oxygen isotope (delta (18)O) composition of Austrocedrus chilensis (D. Don) Endl. (Cupressaceae) tree rings potentially provide retrospective views of changes in environment and climate in the semi-arid lands of Patagonia. We report the development of the first annually resolved delta (18)O tree-ring chronology obtained from natural forests of the foothills of the northwestern Patagonian Andes. The isotope record spans between 1890 and 1994 AD. We explore the probable links between this record and the climate of the region. Air temperatures during summer conditions are significantly, but not strongly, inversely correlated with annual delta (18)O values from Austrocedrus tree rings. The strongest correlations are between the southern oscillation index (SOI) and the tree rings. The existence of millennial-age Austrocedrus trees in northern Patagonia provides interesting possibilities for examining these climate-related isotopic signals over most of the last 1,000 years.

  13. Catalytic gasification: Isotopic labeling and transient reaction

    SciTech Connect

    Saber, J.M.; Falconer, J.L.; Brown, L.F.

    1985-01-01

    Temperature-programmed reaction was used with labeled isotopes (/sup 13/C and /sup 18/O) to study interactions between carbon black and potassium carbonate in pure He and 10% CO/sub 2//90% He atmospheres. Catalytic gasification precursor complexes were observed. Carbon and oxygen-bearing carbon surface groups interacted with the carbonate above 500 K to form surface complexes. Between 500 K and 950 K, and in the presence of gaseous carbon dioxide, the complexes promoted carbon and oxygen exchange between the gas-phase CO/sub 2/ and the surface. Oxygen exchanged between the surface complexes; but carbon did not exchange between the carbonate and the carbon black. As the temperature rose, the complexes decomposed to produce carbon dioxide, and catalytic gasification then began. Elemental potassium formed, and the active catalyst appears to alternate between potassium metal and a potassium-oxygen-carbon complex.

  14. The evolution of Phanerozoic seawater - Isotope paleothermometry finds consensus on Early Paleozoic warmth and constant seawater δ18O

    NASA Astrophysics Data System (ADS)

    Grossman, E. L.; Henkes, G. A.; Passey, B. H.; Shenton, B.; Yancey, T. E.; Perez-Huerta, A.

    2015-12-01

    Evolution of metazoan life is closely linked to the Phanerozoic evolution of ocean temperatures and chemistry. Oxygen isotopic evidence for early Phanerozoic paleotemperatures has been equivocal, with decreasing δ18O values with age being interpreted as warmer early oceans, decreasing seawater δ18O with age, or increasing diagenetic alteration in older samples. Here we compare an updated compilation of oxygen isotope data for carbonate and phosphate fossils and microfossils (Grossman, 2012, Geol. Time Scale, Elsevier, 195-220) with a compilation of new and existing clumped isotope data. Importantly, these data are curated based on sample preservation with special consideration given to screening techniques, and tectonic and burial history. Burial history is critical in the preservation of carbonate clumped isotope temperatures in particular, which can undergo reordering in the solid state. We use a model derived for reordering kinetics (Henkes et al., 2014, Geochim. Cosmochim. Acta 139:362-382) to screen clumped isotope data for the effects of solid-state burial alteration. With minor but significant exceptions (Late Cretaceous, Early Triassic), average δ18O values (4 m.y. window, 2 m.y. steps) for post-Devonian brachiopods, belemnites, and foraminifera, representing tropical-subtropical surface ocean conditions, yield average isotopic temperatures below 30°C (assuming a seawater δ18O value [ -1‰ VSMOW] of an "ice-free" world). In contrast, Ordovician to Devonian data show sustained temperatures of 35-40°C. Likewise, isotopic paleotemperatures from conodont apatite, known to be resistant to isotopic exchange, follow the same pattern. Clumped isotope data derived from Paleozoic brachiopod shells that experienced minimal burial (< 100 °C) and <1% reordering according to the taxon-specific clumped isotope reordering model yield typical temperatures of 25-30°C for the Carboniferous, and 35-40°C for the Ordovician-Silurian. Inserting clumped temperatures and

  15. Stable Isotope (18O, 2H) and Arsenic Distribution in the Shallow Aquifers in Araihazar, Bangladesh

    NASA Astrophysics Data System (ADS)

    Zheng, Y.; Datta, S.; Stute, M.; Dhar, R.; Hoque, M. A.; Rahman, M. W.; Ahmed, K. M.; Schlosser, P.; van Geen, A.

    2005-12-01

    Recent estimates indicate that in Bangladesh alone, an estimated 50 million people have been exposed to Arsenic levels that exceed the WHO guideline of 10 μgL-1 for drinking water by up to two orders of magnitude. There is still debate on what processes control the spatial heterogeneity of dissolved As concentrations. One recent suggestion has been that surface waters enriched in labile organic matter and transferred to greater depths by irrigation pumping may be an important factor. We have monitored for a year the oxygen and hydrogen isotopic composition of precipitation in Dhaka, Bangladesh, and of surface waters and groundwaters in a 25 km2 study area in Araihazar, 20 km east of Dhaka. The data show a large spatial and temporal heterogeneity, with δ18O covering a range of up to 12 ‰. The isotopic composition of precipitation falls on the global meteoric water line (GMWL), while most surface waters collected from rivers, ponds and irrigated rice fields plot below and to the right of the meteoric water line, suggesting that evaporation is an important mechanism in this system. Surface waters show a strong evaporative enrichment during the dry season of up to 10 ‰ in δ18O and then show increased mixing with precipitation during the wet season. The groundwater isotopic composition obtained at 6 multi level well sites covers the range between the GMWL and moderately evaporated surface waters. These data indicate that some groundwaters are recharged directly by precipitation while others show evidence of recharge from evaporated surface waters during the wet and at the beginning of the dry season. For several well nests, the sources of groundwater vary in a systematic way as a function of depth. Highly evaporated irrigation water from rice fields in the dry season does not seem to contribute much to groundwater recharge. The degree of evaporation expressed as deuterium excess does not correlate with As concentrations in the groundwater samples. This finding

  16. Validation of the doubly-labeled water (H/sup 3/H/sup 18/O) method for measuring water flux and energy metabolism in tenebrionid beetles

    SciTech Connect

    Cooper, P.D.

    1981-01-01

    Doubly-labeled water (H/sup 3/H/sup 18/O) has been used to determine water flux and energy metabolism in a variety of vertebrates. This study examines the applicability of this technique to arthropods. The theory of the technique depends upon the assumption that doubly-labeled water introduced into the animal's body water equilibrates with water and carbon dioxide by the action of carbonic anhydrase. Tritium (/sup 3/H) is lost from the animal only with water while oxygen-18 is lost with both water and carbon dioxide. The difference bwtween the rates of loss of the two isotopes is proportional to CO/sub 2/ loss rate. Validation of the use of tritiated water for measuring water flux was accomplished by comparing gravimetric measurements of water gain with flux rates determined by loss of tritiated water. At room humidity, an overestimate for influx calculated from labeled water calculations was found, averaging 12 mg H/sub 2/O (g.d)/sup -1/. Comparison of CO/sub 2/ loss rate determined isotopically with rates of CO/sub 2/ loss determined by standard metabolic rates also yielded overestimates for the isotopic technique, overestimates ranging between 20 and 30%. The relevance of this for studies using labeled water for studying water fluxes and free metabolism of free-ranging arthropods is discussed.

  17. Isolating relative humidity: dual isotopes d18O and dD as deuterium deviations from the global meteoric water line

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cellulose d18O and dD can provide insights on climates and hydrological cycling in the distant past and how these factors differ spatially. However, most studies of plant cellulose have used only one isotope, most commonly d18O, resulting in difficulties partitioning variation in d18O of precipitati...

  18. Oxygen isotopes in tree rings record variation in precipitation δ18O and amount effects in the south of Mexico

    PubMed Central

    Brienen, Roel J W; Hietz, Peter; Wanek, Wolfgang; Gloor, Manuel

    2013-01-01

    [1] Natural archives of oxygen isotopes in precipitation may be used to study changes in the hydrological cycle in the tropics, but their interpretation is not straightforward. We studied to which degree tree rings of Mimosa acantholoba from southern Mexico record variation in isotopic composition of precipitation and which climatic processes influence oxygen isotopes in tree rings (δ18Otr). Interannual variation in δ18Otr was highly synchronized between trees and closely related to isotopic composition of rain measured at San Salvador, 710 km to the southwest. Correlations with δ13C, growth, or local climate variables (temperature, cloud cover, vapor pressure deficit (VPD)) were relatively low, indicating weak plant physiological influences. Interannual variation in δ18Otr correlated negatively with local rainfall amount and intensity. Correlations with the amount of precipitation extended along a 1000 km long stretch of the Pacific Central American coast, probably as a result of organized storm systems uniformly affecting rainfall in the region and its isotope signal; episodic heavy precipitation events, of which some are related to cyclones, deposit strongly 18O-depleted rain in the region and seem to have affected the δ18Otr signal. Large-scale controls on the isotope signature include variation in sea surface temperatures of tropical north Atlantic and Pacific Ocean. In conclusion, we show that δ18Otr of M. acantholoba can be used as a proxy for source water δ18O and that interannual variation in δ18Oprec is caused by a regional amount effect. This contrasts with δ18O signatures at continental sites where cumulative rainout processes dominate and thus provide a proxy for precipitation integrated over a much larger scale. Our results confirm that processes influencing climate-isotope relations differ between sites located, e.g., in the western Amazon versus coastal Mexico, and that tree ring isotope records can help in disentangling the processes

  19. Oxygen isotopes in tree rings record variation in precipitation δ(18)O and amount effects in the south of Mexico.

    PubMed

    Brienen, Roel J W; Hietz, Peter; Wanek, Wolfgang; Gloor, Manuel

    2013-12-01

    [1] Natural archives of oxygen isotopes in precipitation may be used to study changes in the hydrological cycle in the tropics, but their interpretation is not straightforward. We studied to which degree tree rings of Mimosa acantholoba from southern Mexico record variation in isotopic composition of precipitation and which climatic processes influence oxygen isotopes in tree rings (δ(18)Otr). Interannual variation in δ(18)Otr was highly synchronized between trees and closely related to isotopic composition of rain measured at San Salvador, 710 km to the southwest. Correlations with δ(13)C, growth, or local climate variables (temperature, cloud cover, vapor pressure deficit (VPD)) were relatively low, indicating weak plant physiological influences. Interannual variation in δ(18)Otr correlated negatively with local rainfall amount and intensity. Correlations with the amount of precipitation extended along a 1000 km long stretch of the Pacific Central American coast, probably as a result of organized storm systems uniformly affecting rainfall in the region and its isotope signal; episodic heavy precipitation events, of which some are related to cyclones, deposit strongly (18)O-depleted rain in the region and seem to have affected the δ(18)Otr signal. Large-scale controls on the isotope signature include variation in sea surface temperatures of tropical north Atlantic and Pacific Ocean. In conclusion, we show that δ(18)Otr of M. acantholoba can be used as a proxy for source water δ(18)O and that interannual variation in δ(18)Oprec is caused by a regional amount effect. This contrasts with δ(18)O signatures at continental sites where cumulative rainout processes dominate and thus provide a proxy for precipitation integrated over a much larger scale. Our results confirm that processes influencing climate-isotope relations differ between sites located, e.g., in the western Amazon versus coastal Mexico, and that tree ring isotope records can help in

  20. Oxygen isotopes in tree rings record variation in precipitation δ18O and amount effects in the south of Mexico

    NASA Astrophysics Data System (ADS)

    Brienen, Roel J. W.; Hietz, Peter; Wanek, Wolfgang; Gloor, Manuel

    2013-12-01

    Natural archives of oxygen isotopes in precipitation may be used to study changes in the hydrological cycle in the tropics, but their interpretation is not straightforward. We studied to which degree tree rings of Mimosa acantholoba from southern Mexico record variation in isotopic composition of precipitation and which climatic processes influence oxygen isotopes in tree rings (δ18Otr). Interannual variation in δ18Otr was highly synchronized between trees and closely related to isotopic composition of rain measured at San Salvador, 710 km to the southwest. Correlations with δ13C, growth, or local climate variables (temperature, cloud cover, vapor pressure deficit (VPD)) were relatively low, indicating weak plant physiological influences. Interannual variation in δ18Otr correlated negatively with local rainfall amount and intensity. Correlations with the amount of precipitation extended along a 1000 km long stretch of the Pacific Central American coast, probably as a result of organized storm systems uniformly affecting rainfall in the region and its isotope signal; episodic heavy precipitation events, of which some are related to cyclones, deposit strongly 18O-depleted rain in the region and seem to have affected the δ18Otr signal. Large-scale controls on the isotope signature include variation in sea surface temperatures of tropical north Atlantic and Pacific Ocean. In conclusion, we show that δ18Otr of M. acantholoba can be used as a proxy for source water δ18O and that interannual variation in δ18Oprec is caused by a regional amount effect. This contrasts with δ18O signatures at continental sites where cumulative rainout processes dominate and thus provide a proxy for precipitation integrated over a much larger scale. Our results confirm that processes influencing climate-isotope relations differ between sites located, e.g., in the western Amazon versus coastal Mexico, and that tree ring isotope records can help in disentangling the processes

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

    DOEpatents

    Goldblatt, M.; McInteer, B.B.

    1974-01-29

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

  2. Correction of MS data for naturally occurring isotopes in isotope labelling experiments.

    PubMed

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

    2014-01-01

    Mass spectrometry (MS) in combination with isotope labelling experiments is widely used for investigations of metabolism and other biological processes. Quantitative applications-e.g., (13)C metabolic flux analysis-require correction of raw MS data (isotopic clusters) for the contribution of all naturally abundant isotopes. This chapter describes how to perform such correction using the software IsoCor. This flexible, user-friendly software can be used to exploit any isotopic tracer, from well-known ((13)C, (15)N, (18)O, etc.) to unusual ((57)Fe, (77)Se, etc.) isotopes. It also provides options-e.g., correction for the isotopic purity of the tracer-to improve the accuracy of quantitative isotopic studies, and allows automated correction of large datasets that can be collected with modern MS methods.

  3. Isotope Labeling in Insect Cells

    PubMed Central

    Saxena, Krishna; Dutta, Arpana; Klein-Seetharaman, Judith

    2011-01-01

    Recent years have seen remarkable progress in applying nuclear magnetic resonance (NMR) spectroscopy to proteins that have traditionally been difficult to study due to issues with folding, posttranslational modification, and expression levels or combinations thereof. In particular, insect cells have proved useful in allowing large quantities of isotope-labeled, functional proteins to be obtained and purified to homogeneity, allowing study of their structures and dynamics by using NMR. Here, we provide protocols that have proven successful in such endeavors. PMID:22167667

  4. Isotopic (18O) characteristics of weekly precipitation collected across the USA: an initial analysis with application to water source studies

    NASA Astrophysics Data System (ADS)

    Welker, J. M.

    2000-06-01

    A portion of the precipitation samples collected and stored by the National Atmospheric Deposition Program (NADP) are shown to be useful for analysis of isotopes in precipitation. The potential problems with evaporation are small based on deuterium excess analyses and comparisons with the Global Meteroic Water Line. Presented here are the 18O values of precipitation collected from nine NADP sites during 1989, 1990 and 1991. The trends in the isotopic (18O) characteristics of recent precipitation are in agreement with findings from previous International Atomic Energy Agency (IAEA) sites in the USA. The findings are also in agreement with several major isotope-environment relationships, further supporting the use of these samples for a modern global data base on the isotopes in precipitation being developed by IAEA, called GNIP (Global Network for Isotopes in Precipitation) and for use by research groups in the hydrological modelling, palaeoclimate and ecological communities.As expected, the average 18O values of precipitation that is derived from the Gulf of Mexico (-3) and from the Pacific North-west are isotopically distinct (-7). In addition, using the NADP network, isotopic depletion in the 18O values of precipitation in the range of 8 was observed from coastal to inland locations either in the Pacific North-west or along the east side of the Rocky Mountains, from Texas to Eastern Montana. In central USA, especially at high elevation, there is a strong seasonal variation in the 18O values of precipitation, differing by almost 25 between January and August, whereas at coastal locations the seasonal variation in the 18O values of precipitation was minimal. Comparisons between the average 18

  5. Stable isotope variations (δ18O and δD) in modern waters across the Andean Plateau

    NASA Astrophysics Data System (ADS)

    Bershaw, John; Saylor, Joel E.; Garzione, Carmala N.; Leier, Andrew; Sundell, Kurt E.

    2016-12-01

    Environmental parameters that influence the isotopic composition of meteoric water (δ18O and δD) are well characterized up the windward side of mountains, where orographic precipitation results in a predictable relationship between the isotopic composition of precipitation and elevation. The topographic and climatic evolution of the Andean Plateau and surrounding regions has been studied extensively by exploiting this relationship through the use of paleowater proxies. However, interpretation on the plateau itself is challenged by a poor understanding of processes that fractionate isotopes during vapor transport and rainout, and by the relative contribution of unique moisture sources. Here, we present an extensive dataset of modern surface water samples for the northern Andean Plateau and surrounding regions to elucidate patterns and causes of isotope fractionation in this continental environment. These data show a progressive increase in δ18O of stream water west of the Eastern Cordillera (∼1‰/70 km), almost identical to the rate observed across the Tibetan Plateau, attributed to a larger fraction of recycled water in precipitation and/or increased evaporative enrichment downwind. This may lead to underestimates of paleoelevation, particularly for sites deep into the rainshadow of the Eastern Cordilleran crest. That said, elevation is a primary control on the isotopic composition of surface waters across the entire Andean Plateau and its flanks when considering the most negative δ18O values, highlighting the need for sufficiently large datasets to distinguish minimally evaporated samples. There is a general increase in δ18O on the plateau from north to south, concomitant with an increase in aridity and decrease in convective moistening (amount effect). Lastly, stable isotope and seasonal precipitation patterns suggest easterlies provide the vast majority of moisture that falls as precipitation across the Andean Plateau and Western Cordillera, from Peru to

  6. A Two-year Record of Daily Rainfall Isotopes from Fiji: Implications for Reconstructing Precipitation from Speleothem δ18O

    NASA Astrophysics Data System (ADS)

    Brett, M.; Mattey, D.; Stephens, M.

    2015-12-01

    Oxygen isotopes in speleothem provide opportunities to construct precisely dated records of palaeoclimate variability, underpinned by an understanding of both the regional climate and local controls on isotopes in rainfall and groundwater. For tropical islands, a potential means to reconstruct past rainfall variability is to exploit the generally high correlation between rainfall amount and δ18O: the 'amount effect'. The GNIP program provides δ18O data at monthly resolution for several tropical Pacific islands but there are few data for precipitation isotopes at daily resolution, for investigating the amount effect over different timescales in a tropical maritime setting. Timescales are important since meteoric water feeding a speleothem has undergone storage and mixing in the aquifer system and understanding how the isotope amount effect is preserved in aquifer recharge has fundamental implications on the interpretation of speleothem δ18O in terms of palaeo-precipitation. The islands of Fiji host speleothem caves. Seasonal precipitation is related to the movement of the South Pacific Convergence Zone, and interannual variations in rainfall are coupled to ENSO behaviour. Individual rainfall events are stratiform or convective, with proximal moisture sources. We have daily resolution isotope data for rainfall collected at the University of the South Pacific in Suva, covering every rain event in 2012 and 2013. δ18O varies between -18‰ and +3‰ with the annual weighted averages at -7.6‰ and -6.8‰ respectively, while total recorded rainfall amount is similar in both years. We shall present analysis of our data compared with GNIP, meteorological data and back trajectory analyses to demonstrate the nature of the relationship between rainfall amount and isotopic signatures over this short timescale. Comparison with GNIP data for 2012-13 will shed light on the origin of the amount effect at monthly and seasonal timescales in convective, maritime, tropical

  7. Observation of Isotope Ratios (δ2H, δ18O, 87Sr/86Sr) of Tap Water in Urban Environments

    NASA Astrophysics Data System (ADS)

    Mancuso, C. J.; Tipple, B. J.; Ehleringer, J. R.

    2014-12-01

    Urban environments are centers for rapidly growing populations. In order to meet the culinary water needs of these areas, municipal water departments use water from multiple locations and/or sources, often piped differentially to different locations within a municipality. This practice creates isotopically distinct locations within an urban area and therefore provides insight to urban water management practices. In our study we selected urban locations in the Salt Lake Valley, UT (SLV) and San Francisco Bay Area, CA (SFB) where we hypothesized geographically distinct water isotopic ratio differences existed. Within the SLV, municipal waters come from the same mountainous region, but are derived from different geologically distinct watersheds. In contrast, SFB waters are derived from regionally distinct water sources. We hypothesized that the isotope ratios of tap waters would differ based upon known municipal sources. To test this, tap water samples were collected throughout the urban regions in SLV and SFB and analyzed for δ2H, δ18O and 87Sr/86Sr isotope ratios. Seasonal collections were also made to assess if isotope ratios differed throughout the year. Within SLV and SFB, different regions were characterized by distinct paired δ18O and 87Sr/86Sr values. These different realms also agreed with known differences in municipal water supplies within the general geographic region. Waters from different cities within Marin County showed isotopic differences, consistent with water derived from different local reservoirs. Seasonal variation was observed in paired δ18O and 87Sr/86Sr values of tap water for some locations within SLV and SFB, indicating management decisions to shift from one water source to another depending on demand and available resources. Our study revealed that the δ18O and 87Sr/86Sr values of tap waters in an urban region can exhibit significant differences despite close spatial proximity if districts differ in their use of local versus

  8. Stable isotope time-series in mammalian teeth: In situ δ18O from the innermost enamel layer

    NASA Astrophysics Data System (ADS)

    Blumenthal, Scott A.; Cerling, Thure E.; Chritz, Kendra L.; Bromage, Timothy G.; Kozdon, Reinhard; Valley, John W.

    2014-01-01

    Stable carbon and oxygen isotope ratios in mammalian tooth enamel are commonly used to understand the diets and environments of modern and fossil animals. Isotope variation during the period of enamel formation can be recovered by intra-tooth microsampling along the direction of growth. However, conventional sampling of the enamel surface provides highly time-averaged records in part due to amelogenesis. We use backscattered electron imaging in the scanning electron microscope (BSE-SEM) to evaluate enamel mineralization in developing teeth from one rodent and two ungulates. Gray levels from BSE-SEM images suggest that the innermost enamel layer, <20 μm from the enamel-dentine junction, is highly mineralized early in enamel maturation and therefore may record a less attenuated isotopic signal than other layers. We sampled the right maxillary incisor from a woodrat subjected to an experimentally induced water-switch during the period of tooth development, and demonstrate that secondary ion mass spectrometry (SIMS) can be used to obtain δ18O values with 4-5-μm spots from mammalian tooth enamel. We also demonstrate that SIMS can be used to discretely sample the innermost enamel layer, which is too narrow for conventional microdrilling or laser ablation. An abrupt δ18O switch of 16.0‰ was captured in breath CO2, a proxy for body water, while a laser ablation enamel surface intra-tooth profile of the left incisor captured a δ18O range of 12.1‰. The innermost enamel profile captured a δ18O range of 15.7‰, which approaches the full magnitude of δ18O variation in the input signal. This approach will likely be most beneficial in taxa such as large mammalian herbivores, whose teeth are characterized by less rapid mineralization and therefore greater attenuation of the enamel isotope signal.

  9. Sauna, sweat and science - quantifying the proportion of condensation water versus sweat using a stable water isotope ((2)H/(1)H and (18)O/(16)O) tracer experiment.

    PubMed

    Zech, Michael; Bösel, Stefanie; Tuthorn, Mario; Benesch, Marianne; Dubbert, Maren; Cuntz, Matthias; Glaser, Bruno

    2015-01-01

    Most visitors of a sauna appreciate the heat pulse that is perceived when water is poured on the stones of a sauna stove. However, probably only few bathers are aware that this pleasant heat pulse is caused by latent heat being released onto our skin due to condensation of water vapour. In order to quantify the proportion of condensation water versus sweat to dripping water of test persons we conducted sauna experiments using isotopically labelled (δ(18)O and δ(2)H) thrown water as tracer. This allows differentiating between 'pure sweat' and 'condensation water'. Two ways of isotope mass balance calculations were applied and yielded similar results for both water isotopes. Accordingly, condensation contributed considerably to dripping water with mean proportions of 52 ± 12 and 54 ± 7% in a sauna experiment in winter semester 2011/12 and 30 ± 13 and 33 ± 6% in a sauna experiment in winter semester 2012/13, respectively, depending on the way of calculating the isotope mass balance. It can be concluded from the results of our dual isotope labelling sauna experiment that it is not all about sweat in the sauna.

  10. A novel methodological approach for δ(18)O analysis of sugars using gas chromatography-pyrolysis-isotope ratio mass spectrometry.

    PubMed

    Zech, Michael; Saurer, Matthias; Tuthorn, Mario; Rinne, Katja; Werner, Roland A; Siegwolf, Rolf; Glaser, Bruno; Juchelka, Dieter

    2013-01-01

    Although the instrumental coupling of gas chromatography-pyrolysis-isotope ratio mass spectrometry (GC-Py-IRMS) for compound-specific δ(18)O analysis has been commercially available for more than a decade, this method has been hardly applied so far. Here we present the first GC-Py-IRMS δ(18)O results for trimethylsilyl-derivatives of plant sap-relevant sugars and a polyalcohol (glucose, fructose, sucrose, raffinose and pinitol). Particularly, we focus on sucrose, which is assimilated in leaves and which is the most important transport sugar in plants and hence of utmost relevance in plant physiology and paleoclimate studies. Replication measurements of sucrose standards and concentration series indicate that the GC-Py-IRMS δ(18)O measurements are not stable over time and that they are amount (area) dependent. We, therefore, suggest running sample batch replication measurements in alternation with standard concentration series of reference material. This allows for carrying out (i) a drift correction, (ii) a calibration against reference material and (iii) an amount (area) correction. Tests with (18)O-enriched water do not provide any evidence for oxygen isotope exchange reactions affecting sucrose and raffinose. We present the first application of GC-Py-IRMS δ(18)O analysis for sucrose from needle extract (soluble carbohydrate) samples. The obtained δ(18)Osucrose/ Vienna Standard Mean Ocean Water (VSMOW) values are more positive and vary in a wider range (32.1-40.1 ‰) than the δ(18)Obulk/ VSMOW values (24.6-27.2 ‰). Furthermore, they are shown to depend on the climate parameters maximum day temperature, relative air humidity and cloud cover. These findings suggest that δ(18)Osucrose of the investigated needles very sensitively reflects the climatically controlled evaporative (18)O enrichment of leaf water and thus highlights the great potential of GC-Py-IRMS δ(18)Osucrose analysis for plant physiology and paleoclimate studies.

  11. Periodicity analysis of δ18O in precipitation over Central Europe: Time-frequency considerations of the isotopic 'temperature' effect

    NASA Astrophysics Data System (ADS)

    Salamalikis, V.; Argiriou, A. A.; Dotsika, E.

    2016-03-01

    In this paper the periodic patterns of the isotopic composition of precipitation (δ18O) for 22 stations located around Central Europe are investigated through sinusoidal models and wavelet analysis over a 23 years period (1980/01-2002/12). The seasonal distribution of δ18O follows the temporal variability of air temperature providing seasonal amplitudes ranging from 0.94‰ to 4.47‰; the monthly isotopic maximum is observed in July. The isotopic amplitude reflects the geographical dependencies of the isotopic composition of precipitation providing higher values when moving inland. In order to describe the dominant oscillation modes included in δ18O time series, the Morlet Continuous Wavelet Transform is evaluated. The main periodicity is represented at 12-months (annual periodicity) where the wavelet power is mainly concentrated. Stations (i.e. Cuxhaven, Trier, etc.) with limited seasonal isotopic effect provide sparse wavelet power areas at the annual periodicity mode explaining the fact that precipitation has a complex isotopic fingerprint that cannot be examined solely by the seasonality effect. Since temperature is the main contributor of the isotopic variability in mid-latitudes, the isotope-temperature effect is also investigated. The isotope-temperature slope ranges from 0.11‰/°C to 0.47‰/°C with steeper values observed at the southernmost stations of the study area. Bivariate wavelet analysis is applied in order to determine the correlation and the slope of the δ18O - temperature relationship over the time-frequency plane. High coherencies are detected at the annual periodicity mode. The time-frequency slope is calculated at the annual periodicity mode ranging from 0.45‰/°C to 0.83‰/°C with higher values at stations that show a more distinguishable seasonal isotopic behavior. Generally the slope fluctuates around a mean value but in certain cases (sites with low seasonal effect) abrupt slope changes are derived and the slope becomes

  12. On the use of stable oxygen isotope18O) measurements for tracking avian movements in North America

    PubMed Central

    Hobson, Keith A; Koehler, Geoff

    2015-01-01

    Tracking migratory animals has benefitted using measurements of naturally occurring stable isotopes of hydrogen (δ2H) in keratinous tissues such as hair and feathers to link animal origins to continental patterns or isoscapes of δ2H in precipitation. However, for most taxa, much less information exists on the use of stable oxygen isotope ratios (δ18O) despite the fact that δ2H and δ18O are strongly linked in environmental waters through the meteoric relationship and the possibility of using both isotopes to infer greater information on origins and climatic conditions where tissues are grown. A fundamental requirement of using stable isotopes to assign individuals and populations to origins is the development of a rescaling function linking environmental food web signals to the tissue of interest and for birds, this has not been carried out. Here, we derived the relationship between H and O isotopes in known source feathers of 104 individuals representing 11 species of insectivorous passerines sampled across the strong precipitation isoscape of North America. We determined again a strong expected relationship between feather δ2H (δ2Hf) and long-term amount-weighted precipitation δ2H (δ2Hp; r2 = 0.77), but the corresponding relationship between δ18Of and δ18Op was poor (r2 = 0.32) for the same samples. This suggests that δ2H measurements are currently more useful for assignment of insectivorous songbirds to precipitation isoscapes but does not preclude other uses of the δ18Of data. Currently, mechanisms responsible for the decoupling of H and O isotopes in food webs is poorly known, and we advocate a much broader sampling of both isotopes in the same keratinous tissues across precipitation isotope gradients and across taxa to resolve this issue and to increase the power of using water isotopes to track migratory animals. PMID:25691999

  13. Isotopic tracing (D, 18O and 29Si) to understand the alteration on historic glass

    NASA Astrophysics Data System (ADS)

    Verney-Carron, Aurélie; Saheb, Mandana; Valle, Nathalie; Mangin, Denis; Remusat, Laurent; Loisel, Claudine

    2015-04-01

    In order to better preserve historic glasses, e.g. stained glass windows, the understanding of their alteration mechanisms and of what controls the kinetics corresponding to each process is required. The ancient stained glasses are characterized by thick alteration layers, continuous or as pits, that are cracked or lost. Therefore, if a passivating role of the alteration layer has been proved on some other kinds of glass (such as basaltic or nuclear glass) in aqueous medium, the issue can be addressed for low durable stained glass weathered in varying atmospheric conditions. The mechanism of alteration layer formation was first investigated by performing dynamic and static experiments on model medieval glasses altered with a solution doped in 29Si at different concentrations (or saturation degrees). Solid analyses were carried out by SIMS and solution by HR-ICP-MS. Medieval stained glass has mainly a potash-lime-silica composition with a low content in alumina. The alkaline and alkaline-earth elements have thus a modifier role in the glassy network. This structural difference compared to boro- or alumino-silicate glasses could induce differences in the alteration mechanisms. However, the analysis of the Si isotopic signature of the gel layer highlighted that diffusion, but also hydrolysis/condensation reactions, are also involved in the gel layer formation process, leading to a structural and textural reorganization. The second objective was to determine the kinetic role of the alteration layer, and especially to trace the circulation of water once the altered layer is formed. For that, ancient glasses were exposed to simulated rainfall events / drying periods cycles during 3 months by using a solution doped in D and 18O. NanoSIMS analyses have shown that the transport in the alteration layer is mainly driven by diffusion in the porosity despite the presence of cracks that could have been preferential ways of circulation. This demonstrates also a potential

  14. Isotopic effects in the ( π±, 2N) reactions on 16O and 18O

    NASA Astrophysics Data System (ADS)

    Altman, A.; Ashery, D.; Piasetzky, E.; Lichtenstadt, J.; Yavin, A. I.; Bertl, W.; Felawka, L.; Walter, H. K.; Powers, R. J.; Winter, R. G.; v. d. Pluym, J.

    1984-09-01

    The ( π+, 2p), ( π+, pn) and ( π-, pn) reactions on 16O and 18O were studied at 165 MeV. The cross section for the ( π+, 2p) reaction on 18O is larger than that for 16O be only 5% ± 3%, while the total π+ absorption cross section is larger by 17% ± 5%. This supports the assumption that two-nucleon absorption occurs mainly on nucleons in the same shell. It is further concluded that Δ++n → pp is not only absorption mechanism that couples strongly to the nucleon knock out reactions.

  15. Directly Measured Clumped Isotope Temperatures From Known And Proposed Paleozoic Glacial Intervals Suggest That Oceans Were Depleted in 18O

    NASA Astrophysics Data System (ADS)

    Petrizzo, D. A.; Runnegar, B.; Ivany, L.; Young, E. D.

    2011-12-01

    Oceans enriched in 13C are thought to result from atmospheric CO2 drawdown and concomitant global cooling resulting from increased burial of organic matter. We investigated ocean temperatures during two times when the oceans were exceptionally heavy in 13C, the Lau Event of the late Silurian and the Late Paleozoic Ice Age (LPIA), using both δ18O and "directly measured" clumped isotope (Δ47) temperatures. We report a tropical ocean temperature of 16 ± 3°C at the peak of the Lau Event, confirm seasonality at a high-latitude LPIA site, and raise the possibility that some degradation of 13C-18O bonds may be widespread in apparently unaltered carbonates that have seen temperatures higher than 100-150°C. Silurian conodonts from Gotland, Sweden, are almost unaltered (CAI ~ 0) indicating burial temperatures of <50°C. We measured atrypid brachiopod calcite from the peak of the late Silurian Lau event, the largest positive carbon isotope excursion (+8%) since the Cambrian, and obtained Δ47 = 0.687 ± 0.014, giving a low latitude water temperature of 16 ± 3°C. This is significantly cooler than tropical temperatures reported from pentamerid brachiopod calcite of the early Silurian greenhouse period (35°C, Came et al., 2007) and those derived from rugose corals during the Hirnantian (Ordovician) positive carbon isotope excursion (+5%) and accompanying glaciation (27-32°C, Finnegan et al., 2010). We also measured Δ47 in two shells of the Australian Permian bivalve Eurydesma, a circumpolar genus associated with cold water indicators. Ivany and Runnegar (2010) found high-amplitude annual cycles in δ18O in one of these specimens but the calculated temperatures seemed too warm for the periglacial conditions indicated by approximately coeval dropstones and glendonites unless Permian ocean water δ18O was lighter than ~ -3%. Our Δ47 results also give unrealistically warm winter (~12°C) and summer (~23°C) temperatures for this high-latitude site, raising the possibility

  16. Correction algorithm for online continuous flow δ13C and δ18O carbonate and cellulose stable isotope analyses

    NASA Astrophysics Data System (ADS)

    Evans, M. N.; Selmer, K. J.; Breeden, B. T.; Lopatka, A. S.; Plummer, R. E.

    2016-09-01

    We describe an algorithm to correct for scale compression, runtime drift, and amplitude effects in carbonate and cellulose oxygen and carbon isotopic analyses made on two online continuous flow isotope ratio mass spectrometry (CF-IRMS) systems using gas chromatographic (GC) separation. We validate the algorithm by correcting measurements of samples of known isotopic composition which are not used to estimate the corrections. For carbonate δ13C (δ18O) data, median precision of validation estimates for two reference materials and two calibrated working standards is 0.05‰ (0.07‰); median bias is 0.04‰ (0.02‰) over a range of 49.2‰ (24.3‰). For α-cellulose δ13C (δ18O) data, median precision of validation estimates for one reference material and five working standards is 0.11‰ (0.27‰); median bias is 0.13‰ (-0.10‰) over a range of 16.1‰ (19.1‰). These results are within the 5th-95th percentile range of subsequent routine runtime validation exercises in which one working standard is used to calibrate the other. Analysis of the relative importance of correction steps suggests that drift and scale-compression corrections are most reliable and valuable. If validation precisions are not already small, routine cross-validated precision estimates are improved by up to 50% (80%). The results suggest that correction for systematic error may enable these particular CF-IRMS systems to produce δ13C and δ18O carbonate and cellulose isotopic analyses with higher validated precision, accuracy, and throughput than is typically reported for these systems. The correction scheme may be used in support of replication-intensive research projects in paleoclimatology and other data-intensive applications within the geosciences.

  17. Seawater intrusion into groundwater aquifer through a coastal lake - complex interaction characterised by water isotopes (2)H and (18)O.

    PubMed

    Gemitzi, Alexandra; Stefanopoulos, Kyriakos; Schmidt, Marie; Richnow, Hans H

    2014-01-01

    The present study investigates the complex interactions among surface waters, groundwaters and a coastal lake in northeastern Greece, using their stable isotopic composition (δ(18)O, δ(2)H) in combination with hydrogeological and hydrochemical data. Seasonal and spatial trends of water isotopes were studied and revealed that all water bodies in the study area interact. It was also shown that the aquifer's increased salinity is not due to fossil water from past geological periods, but is attributed to brackish lake water intrusion into the aquifer induced by the extensive groundwater pumping for irrigation purposes. Quantification of the contribution of the lake to the aquifer was achieved using the simple dilution formula. The isotopic signatures of the seawater and the groundwaters are considerably different, so there is a very little possibility of direct seawater intrusion into the aquifer.

  18. Evaluation of a High Intensity Focused Ultrasound-Immobilized Trypsin Digestion and 18 O-Labeling Method for Quantitative Proteomics

    SciTech Connect

    Lopez-Ferrer, Daniel; Hixson, Kim K.; Smallwood, Heather S.; Squier, Thomas C.; Petritis, Konstantinos; Smith, Richard D.

    2009-08-01

    A new method that uses immobilized trypsin concomitant with ultrasonic irradiation results in ultra-rapid digestion and thorough 18O labeling for quantitative protein comparisons. The reproducible and highly efficient method provided effective digestions in <1 min and minimized the amount of enzyme required compared to traditional methods. This method was demonstrated for digestion of both simple and complex protein mixtures, including bovine serum albumin, a global proteome extract from bacteria Shewanella oneidensis, and mouse plasma, as well as for the labeling of complex protein mixtures, which validated the application of this method for differential proteomic measurements. This approach is simple, reproducible, cost effective, and rapid, and thus well-suited for automation.

  19. Quantitative Proteomic Analysis of Mouse Embryonic Fibroblasts and Induced Pluripotent Stem Cells Using 16O /18O labeling

    SciTech Connect

    Huang, Xin; Tian, Changhai; Liu, Miao; Wang, Yongxiang; Tolmachev, Aleksey V.; Sharma, Seema; Yu, Fang; Fu, Kai; Zheng, Jialin; Ding, Shi-Jian

    2012-04-06

    Induced pluripotent stem cells (iPSC) hold great promise for regenerative medicine as well as for investigations into the pathogenesis and treatment of various diseases. Understanding of key intracellular signaling pathways and protein targets that control development of iPSC from somatic cells is essential for designing new approaches to improve reprogramming efficiency. Here we report the development and application of an integrated quantitative proteomics platform for investigating differences in protein expressions between mouse embryonic fibroblasts (MEF) and MEF-derived iPSC. This platform consists of 16O/18O labeling, multidimensional peptide separation coupled with tandem mass spectrometry, and data analysis with UNiquant software. Using this platform a total of 2,481 proteins were identified and quantified from the 16O/18O-labeled MEF-iPSC proteome mixtures with a false discovery rate of 0.01. Among them, 218 proteins were significantly upregulated, while 247 proteins were significantly downregulated in iPSC compared to MEF. Many nuclear proteins, including Hdac1, Dnmt1, Pcna, Ccnd1, Smarcc1, and subunits in DNA replication and RNA polymerase II complex were found to be enhanced in iPSC. Protein network analysis revealed that Pcna functions as a hub orchestrating complicated mechanisms including DNA replication, epigenetic inheritance (Dnmt1) and chromatin remodeling (Smarcc1) to reprogram MEF and maintain stemness of iPSC.

  20. Isotopic composition (δ18O and δD) of the shallow groundwater in the Poyang Lake basin

    NASA Astrophysics Data System (ADS)

    Soldatova, E. A.; Sun, Z.; Guseva, N. V.

    2016-03-01

    The article is focused on the identification of evaporation and other natural processes which affect the isotopic composition of shallow groundwater in the Poyang Lake basin, such as water-rock interaction and mixing of the shallow groundwater and surface water. For this purpose the dual isotope approach (δD-H2O and δ18O-H2O) was used. The samples were collected from domestic wells around the Poyang Lake. The value of δD obtained for the shallow groundwater ranges from -21.5 to -42.6∘/∘∘. The δ18O value varies from -3.5 to -7.1∘/∘∘. It was found that the shallow groundwater of the Poyang Lake catchment is of meteoric origin. The influence of evaporation on the isotopic composition of shallow groundwater is negligible and observed mainly during the dry season. The deviation from the local meteoric water line, especially during the rainy season, may be explained by the processes in the water-rock system, but this issue is required further research.

  1. An assessment of the isotopic (2H/18O) integrity of water samples collected and stored by unattended precipitation totalizers

    NASA Astrophysics Data System (ADS)

    Terzer, Stefan; Wassenaar, Leonard I.; Douence, Cedric; Araguas-Araguas, Luis

    2016-04-01

    The IAEA-WMO Global Network of Isotopes in Precipitation (GNIP) provides worldwide δ18O and δ2H data for numerous hydrological and climatological studies. The traditional GNIP sample collection method relies on weather station operators to accumulate precipitation obtained from manual rain gauges. Over the past decades, widespread weather station automatization resulted in the increased use of unattended precipitation totalizers that accumulate and store the rainwater in the field for up to one month. Several low-tech measures were adopted to prevent in situ secondary evaporative isotopic enrichment (SEE) of totalized water samples (i.e. disequilibrium isotopic fractionation after precipitation is stored in the collection device). These include: (a) adding a 0.5-1 cm floating layer of paraffin oil to the totalizer bottle, (b) using an intake tube leading from the collection funnel and submerged to the bottom of the totalizer bottle, or (c) placing a table tennis ball in the funnel aiming to reduce evaporation of the collected water from the receiving bottle to the atmosphere. We assessed the isotopic integrity of stored rainwater samples for three totalizers under controlled settings: each aforementioned totalizer was filled with a 100 or 500 mL of isotopically known water and installed in the field with the intake funnels sheltered to prevent rainwater collection. Potential evapotranspiration (PET) was obtained from on-site meteorological recordings. Stored evaporative loss from each totalizer was evaluated on a monthly basis; gravimetrically and by analysing δ18O and δ2H of the stored water, for a period of 6 months and a cumulative PET of ˜500 mm. The gravimetric and isotope results revealed that for smaller water volumes (100 ml, corresponding to ca. 5 mm of monthly precipitation), negligible isotope enrichment (δ18O) was observed in the paraffin-oil based totalizer, whereas unacceptable evaporative isotope effects were observed for the ball

  2. Ab initio path integral simulation study on 16O/ 18O isotope effect in water and hydronium ion

    NASA Astrophysics Data System (ADS)

    Tachikawa, Masanori; Shiga, Motoyuki

    2005-05-01

    An ab initio path integral molecular dynamics simulation has been performed to study the 16O and 18O isotopomers for a water molecule and a hydronium ion at temperature 300 K. The average O-H bond length of H 218O molecule is slightly shorter than that of H 216O molecule, while that of H 318O + is slightly longer than that of H 316O +. For hydronium ions, the Walden inversion of H 318O +, as well as D 316O +, is found to be more restrained than that of H 316O +. The isotope effect in the electronic structure and thermochemical properties for these isotopomers are also shown.

  3. Investigating human geographic origins using dual-isotope (87Sr/86Sr, δ18O) assignment approaches

    PubMed Central

    Sonnemann, Till F.; Shafie, Termeh; Hofman, Corinne L.; Brandes, Ulrik; Davies, Gareth R.

    2017-01-01

    Substantial progress in the application of multiple isotope analyses has greatly improved the ability to identify nonlocal individuals amongst archaeological populations over the past decades. More recently the development of large scale models of spatial isotopic variation (isoscapes) has contributed to improved geographic assignments of human and animal origins. Persistent challenges remain, however, in the accurate identification of individual geographic origins from skeletal isotope data in studies of human (and animal) migration and provenance. In an attempt to develop and test more standardized and quantitative approaches to geographic assignment of individual origins using isotopic data two methods, combining 87Sr/86Sr and δ18O isoscapes, are examined for the Circum-Caribbean region: 1) an Interval approach using a defined range of fixed isotopic variation per location; and 2) a Likelihood assignment approach using univariate and bivariate probability density functions. These two methods are tested with enamel isotope data from a modern sample of known origin from Caracas, Venezuela and further explored with two archaeological samples of unknown origin recovered from Cuba and Trinidad. The results emphasize both the potential and limitation of the different approaches. Validation tests on the known origin sample exclude most areas of the Circum-Caribbean region and correctly highlight Caracas as a possible place of origin with both approaches. The positive validation results clearly demonstrate the overall efficacy of a dual-isotope approach to geoprovenance. The accuracy and precision of geographic assignments may be further improved by better understanding of the relationships between environmental and biological isotope variation; continued development and refinement of relevant isoscapes; and the eventual incorporation of a broader array of isotope proxy data. PMID:28222163

  4. Modeling Interannual Variability of δ^1^8O of Atmospheric CO2 and its Dependence on Humidity and Isotope Hydrology

    NASA Astrophysics Data System (ADS)

    Buenning, N. H.; Noone, D. C.; Still, C. J.; Riley, W. J.; Randerson, J. T.; Welp, L. R.; White, J. W.; Vaughn, B.; Miller, J. B.; Tans, P. P.

    2006-12-01

    Measurements of the δ^1^8O value of CO2 at the NOAA/ESRL baseline observatories showed a gradual downward trend from the early 1990s until 1997. The cause of this trend is not well understood, although it is likely due to a change in the isotopic composition of the terrestrial water pools with which CO2 interacts during photosynthesis and respiration, particularly in the tropics, where the largest isotope forcing occurs. There are a number of factors that affect the isotopic composition of soil and leaf water, however, studies have indicated that relative humidity has a strong impact on the water pools. Humidity records at several stations in Southeast Asia show an upward trend during the 1990s, which is consistent with the expected trend in the δ^1^8O value of atmospheric CO2. While an increase in humidity would increase stomatal conductance and in turn increase biospheric productivity, it also will allow leaves to take in more of the isotopically light water vapor, causing the leaf water to become less enriched with ^1^8O isotope. Using the isotopic version of the NCAR Land Surface Model (ISOLSM) and Community Atmosphere Model (CAM), the interannual variability of simulated δ^1^8O of CO2 were examined from 1979 to 2002. ISOLSM was forced with interannually varying meteorological data from the NCEP reanalysis. Computed fluxes from ISOLSM for each month of the 24-year simulation were used in CAM to simulate the seasonal cycle and trends in δ^1^8O values of CO2. Experiments were constructed to determine the impact on interannual variability in the δ^1^8O value of CO2 of humidity, δ^1^8O of precipitation, and δ^1^8O of water vapor. To demonstrate the affect of humidity, two experiments were constructed whereby relative humidity (1) is gradually increased by 0.5% per year from 1990 to 1997 (as is seen in some of the humidity records in Southeast Asia during the early 1990s yet this trend does not appear in the NCEP Reanalysis) and (2) assigned long-term monthly

  5. Water - Isotope - Map (δ 18O, δ 2H, 3H) of Austria: Applications, Extremes and Trends

    NASA Astrophysics Data System (ADS)

    Wyhlidal, Stefan; Kralik, Martin; Benischke, Ralf; Leis, Albrecht; Philippitsch, Rudolf

    2016-04-01

    The isotopic ratios of oxygen and hydrogen in water (2H/1H and 18O/16O) are important tools to characterise waters and their cycles. This starts in the atmosphere as rain or snow and continues in surface water and ends in shallow groundwater as well as in deep groundwater. Tritium formed by natural cosmic radiation in the upper atmosphere and in the last century by tests of thermonuclear bombs in the atmosphere, is characterised by its radioactive decay with a half-life of 12.32 years and is an ideal age-marker during the last 60 years. To determine the origin and mean age of waters in many projects concerning water supply, engineering and scientific projects in the last 45 years on more than 1,350 sites, more than 40,000 isotope measurements were performed in Austria. The median value of all sites of oxygen-18 is δ 18O -10.7 ‰ and for hydrogen-2 δ 2H -75 ‰. As the fractionation is mainly temperature dependent the lowest negative values are observed in winter precipitation (oxygen-18 as low as δ 18O -23 ‰) and in springs in the mountain regions (δ 18O -15.1 ‰). In contrast the highest values were observed in summer precipitation (up to δ 18O - 0.5 ‰) and in shallow lakes in the Seewinkel (up to δ 18O + 5 ‰). The isotopic ratios of the Austrian waters are also influenced by the origin of the evaporated water masses. Therefore the precipitation in the region south of the main Alpine crest (East-Tyrol, Carinthia and South-East Styria) is approximately 1 ‰ higher in δ 18O-values than sites at the same altitude in the northern part. This is most probably caused by the stronger influence of precipitation from the mediterranean area. The median value of all 1,120 sampling sites of decay corrected (2015) tritium measurements is 6.2 tritium units (TU). This is somewhat smaller than the median value of all precipitation stations with 7.2 TU. This can be explained by the fact that in most cases in groundwater the median value has been reduced by decay

  6. The use of δ(2)H and δ(18)O isotopic analyses combined with chemometrics as a traceability tool for the geographical origin of bell peppers.

    PubMed

    de Rijke, E; Schoorl, J C; Cerli, C; Vonhof, H B; Verdegaal, S J A; Vivó-Truyols, G; Lopatka, M; Dekter, R; Bakker, D; Sjerps, M J; Ebskamp, M; de Koster, C G

    2016-08-01

    Two approaches were investigated to discriminate between bell peppers of different geographic origins. Firstly, δ(18)O fruit water and corresponding source water were analyzed and correlated to the regional GNIP (Global Network of Isotopes in Precipitation) values. The water and GNIP data showed good correlation with the pepper data, with constant isotope fractionation of about -4. Secondly, compound-specific stable hydrogen isotope data was used for classification. Using n-alkane fingerprinting data, both linear discriminant analysis (LDA) and a likelihood-based classification, using the kernel-density smoothed data, were developed to discriminate between peppers from different origins. Both methods were evaluated using the δ(2)H values and n-alkanes relative composition as variables. Misclassification rates were calculated using a Monte-Carlo 5-fold cross-validation procedure. Comparable overall classification performance was achieved, however, the two methods showed sensitivity to different samples. The combined values of δ(2)H IRMS, and complimentary information regarding the relative abundance of four main alkanes in bell pepper fruit water, has proven effective for geographic origin discrimination. Evaluation of the rarity of observing particular ranges for these characteristics could be used to make quantitative assertions regarding geographic origin of bell peppers and, therefore, have a role in verifying compliance with labeling of geographical origin.

  7. SAIL--stereo-array isotope labeling.

    PubMed

    Kainosho, Masatsune; Güntert, Peter

    2009-11-01

    Optimal stereospecific and regiospecific labeling of proteins with stable isotopes enhances the nuclear magnetic resonance (NMR) method for the determination of the three-dimensional protein structures in solution. Stereo-array isotope labeling (SAIL) offers sharpened lines, spectral simplification without loss of information and the ability to rapidly collect and automatically evaluate the structural restraints required to solve a high-quality solution structure for proteins up to twice as large as before. This review gives an overview of stable isotope labeling methods for NMR spectroscopy with proteins and provides an in-depth treatment of the SAIL technology.

  8. Clumped Isotope Verification of δ18O-Based Freshwater Mussel Shell Growth Chronology for a High-Resolution Climate and River Discharge Record

    NASA Astrophysics Data System (ADS)

    VanPlantinga, A.; Grossman, E. L.; Passey, B. H.; Randklev, C.

    2015-12-01

    Isotope profiles in freshwater mussel shells can be used to reconstruct climate, water source, and river discharge, but problems arise from variable water temperature and δ18O. To resolve this complexity and expand the application of isotope sclerochronology to the study of past river systems, we measured δ18O and Δ47 in two common freshwater mussel species from the Brazos River in Texas. To compare the environmental record with the shell record and develop a sclerochronology, weekly water temperature and δ18O data were collected from the Brazos River near College Station from January 2012 to August 2013. The river data reveal complex, irregular patterns for predicted aragonite δ18O. Comparing δ18O profiles from micromilled transects (70-200 µm increments) of coeval shell growth within and between shells yielded consistent patterns. Shell δ18O can be accurately matched to predicted δ18O, providing a chronology of shell growth. However, without a water temperature and δ18O record, interpreting a sclerochronology would be impossible. Shell Δ47 can potentially provide a seasonal chronology to verify the δ18O sclerochronology, which would be invaluable for the use of δ18O sclerochronology in historical and ancient shells. For Δ47 analyses, samples were taken at 0.5 mm resolution in presumed seasonal dark and light growth bands. Clumped temperatures range between 21 and 35 ± 4˚C (Henkes et al., 2013) and track the river temperature record, supporting the interpreted shell δ18O chronology. Shell Δ47-calculated water δ18O values range from -1.2 to 1.5 ± 0.9‰ and match river δ18O. High-resolution shell δ18O profiles combined with Δ47 temperatures can reconstruct a weekly history of water δ18O, and with the observed river discharge vs. water δ18O relation, produce a qualitative record of river discharge. These analytical techniques applied to a historical Brazos River mussel shell collected prior to dam construction reveal weekly records of

  9. Growth Rates, Stable Oxygen Isotopes (18O), and Strontium (Sr/Ca) Composition in Two Species of Pacific Sclerosponges (Acanthocheatetes wellsi and Astrosclera willeyana) with 18O Calibration and Application to Paleoceanography

    SciTech Connect

    Grottoli, A.; Adkins, J; Panero, W; Reaman, D; Moots, K

    2010-01-01

    The isotopic and elemental composition of sclerosponge skeletons is used to reconstruct paleoceanographic records. Yet few studies have systematically examined the natural variability in sclerosponge skeletal {delta}{sup 18}O, growth, and Sr/Ca, and how that may influence the interpretation of sclerosponge proxy records. Here, we analyzed short records in seven specimens of Acanthocheatetes wellsi (high-Mg calcite, 21 mol% Mg) from Palau, four A. wellsi (high-Mg calcite, 21 mol% Mg) from Saipan, and three Astrosclera willeyana (aragonite) sclerosponges from Saipan, as well as one long record in an A. wellsi specimen from Palau spanning 1945-2001.5. In Saipan, species-specific and mineralogical effects appear to have a negligible effect on sclerosponge {delta}{sup 18}O, facilitating the direct comparison of {delta}{sup 18}O records between species at a given location. At both sites, A. wellsi {delta}{sup 18}O and growth rates were sensitive to environmental conditions, but Sr/Ca was not sensitive to the same conditions. High-resolution {delta}{sup 18}O analyses confirmed this finding as both A. wellsi and A. willeyana deposited their skeleton in accordance with the trends in isotopic equilibrium with seawater, though with a 0.27{per_thousand} offset in the case of A. willeyana. In the high-Mg-calcite species A. wellsi, Mg may be interfering with Sr incorporation into the skeleton. On multidecadal timescales, A. wellsi sclerosponge {delta}{sup 18}O in Palau tracked the Southern Oscillation Index variability post-1977, but not pre-1977, coincident with the switch in the Pacific Decadal Oscillation (PDO) at {approx}1976. This suggests that water mass circulation in the region is influenced by El Nino-Southern Oscillation variability during positive PDO phases, but not during negative ones.

  10. Determining Carbonate Concretion Formation Temperatures and Pore Water δ18O Values Using the Clumped Isotope Approach

    NASA Astrophysics Data System (ADS)

    Loyd, S. J.; Corsetti, F. A.; Tripati, A. K.

    2010-12-01

    The porosity/permeability of siliciclastic strata is affected by post-depositional cementation, but determining at what depth and under what conditions cementation occurs is difficult with standard techniques. The oxygen isotopic composition of solid phase carbonate cements (δ18Ocarb) can be related to temperature (and by extension depth) of formation, and thus has been widely used in diagenetic studies. However, δ18Ocarb paleothermometry requires the prediction or assumption of pore water δ18O (δ18Opw), a parameter that is poorly constrained in past diagenetic environments (for convenience δ18Opw is usually assumed to be 0‰ VSMOW). Here, we use clumped isotope thermometry (CIT)—a fluid δ18O-independent temperature proxy—to avoid the often ambiguous yet necessary δ18Opw assumption applied to δ18Ocarb paleothermometery and reevaluate the temperature of carbonate concretion formation in the Miocene Monterey Formation (dolomite) and the Cretaceous Holz Shale (calcite) of southern California. CIT analysis of Monterey Formation concretions produced slightly increased temperatures of formation versus traditional δ18Ocarb paleothermometry, whereas the Holz Shale concretions produced significantly decreased temperatures. Inputting the CIT-derived temperature into the associated δ18Ocarb-temperature equation allows the calculation of the ancient δ18Opw. Calculated δ18Opw values range from ~ -8 to +2‰ VSMOW, significantly different from coeval seawater. δ18Opw less than 0‰ can be generated by a number of processes including the influx of non-marine fluids and/or hydrate formation, whereas δ18Opw greater than 0‰ can be produced by silicate diagenesis, influx of evaporative brines, or hydrate dissolution. These data demonstrate that pore water modifying diagenetic processes were operating in past environments and emphasize that the formation temperatures of diagenetic carbonates should be calculated using a fluid δ18O-independent approach, such as

  11. Oxygen isotopes in nitrate: New reference materials for 18O:17O:16O measurements and observations on nitrate-water equilibration

    USGS Publications Warehouse

    Böhlke, J.K.; Mroczkowski, S.J.; Coplen, T.B.

    2003-01-01

    Despite a rapidly growing literature on analytical methods and field applications of O isotope-ratio measurements of NO3- in environmental studies, there is evidence that the reported data may not be comparable because reference materials with widely varying ?? 18O values have not been readily available. To address this problem, we prepared large quantities of two nitrate salts with contrasting O isotopic compositions for distribution as reference materials for O isotope-ratio measurements: USGS34 (KNO3) with low ??18O and USGS35 (NaNO3) with high ??18O and 'mass-independent' ??17O. The procedure used to produce USGS34 involved equilibration of HNO3 with 18O-depleted meteoric water. Nitric acid equilibration is proposed as a simple method for producing laboratory NO3- reference materials with a range of ??18O values and normal (mass-dependent) 18O: 17O:16O variation. Preliminary data indicate that the equilibrium O isotope-fractionation factor (??) between [NO 3-] and H2O decreases with increasing temperature from 1.0215 at 22??C to 1.0131 at 100??C. USGS35 was purified from the nitrate ore deposits of the Atacama Desert in Chile and has a high 17O:18O ratio owing to its atmospheric origin. These new reference materials, combined with previously distributed NO3- isotopic reference materials IAEA-N3 (=IAEA-NO-3) and USGS32, can be used to calibrate local laboratory reference materials for determining offset values, scale factors, and mass-independent effects on N and O isotope-ratio measurements in a wide variety of environmental NO 3- samples. Preliminary analyses yield the following results (normalized with respect to VSMOW and SLAP, with reproducibilities of ??0.2-0.3???, 1??): IAEA-N3 has ??18O = +25.6??? and ??17O = +13.2??? USGS32 has ?? 18O = +25.7??? USGS34 has ??18O = -27. 9??? and ??17O = -14.8??? and USGS35 has ?? 18O = +57.5??? and ??17O = +51.5???.

  12. ZoomQuant: an application for the quantitation of stable isotope labeled peptides.

    PubMed

    Halligan, Brian D; Slyper, Ronit Y; Twigger, Simon N; Hicks, Wayne; Olivier, Michael; Greene, Andrew S

    2005-03-01

    The main goal of comparative proteomics is the quantitation of the differences in abundance of many proteins between two different biological samples in a single experiment. By differentially labeling the peptides from the two samples and combining them in a single analysis, relative ratios of protein abundance can be accurately determined. Protease catalyzed (18)O exchange is a simple method to differentially label peptides, but the lack of robust software tools to analyze the data from mass spectra of (18)O labeled peptides generated by common ion trap mass spectrometers has been a limitation. ZoomQuant is a stand-alone computational tool that analyzes the mass spectra of (18)O labeled peptides from ion trap instruments and determines relative abundance ratios between two samples. Starting with a filtered list of candidate peptides that have been successfully identified by Sequest, ZoomQuant analyzes the isotopic forms of the peptides using high-resolution zoom scan spectrum data. The theoretical isotope distribution is determined from the peptide sequence and is used to deconvolute the peak areas associated with the unlabeled, partially labeled, and fully labeled species. The ratio between the labeled and unlabeled peptides is then calculated using several different methods. ZoomQuant's graphical user interface allows the user to view and adjust the parameters for peak calling and quantitation and select which peptides should contribute to the overall abundance ratio calculation. Finally, ZoomQuant generates a summary report of the relative abundance of the peptides identified in the two samples.

  13. Larix decidua δ(18)O tree-ring cellulose mainly reflects the isotopic signature of winter snow in a high-altitude glacial valley of the European Alps.

    PubMed

    Leonelli, Giovanni; Battipaglia, Giovanna; Cherubini, Paolo; Saurer, Matthias; Siegwolf, Rolf T W; Maugeri, Maurizio; Stenni, Barbara; Fusco, Stella; Maggi, Valter; Pelfini, Manuela

    2017-02-01

    We analyzed the chronologies of cellulose stable isotopes (δ(13)C and δ(18)O) and tree-ring widths from European larch (Larix decidua) in a high-altitude site (2190ma.s.l.) at the bottom of a glacial valley in the Italian Alps, and investigated their dependence on monthly meteorological variables and δ(18)O precipitation values. The δ(18)O of tree-ring cellulose appears to be strongly driven by the δ(18)O of winter snowfall (November to March), which suggests that larch trees mostly use the snow-melt water of the previous winter during the growing season. This water, which also comes from the slope streams and from the underground flow of nearby steep slopes, infiltrates the soil in the valley bottom. The tree-ring cellulose δ(18)O values were also found to be influenced by the August precipitation δ(18)O and mean temperature. The associated regression model shows that the δ(18)O chronology from the tree rings explains up to 34% of the variance in the winter precipitation δ(18)O record, demonstrating the potential for reconstructing the δ(18)O isotopic composition of past winter precipitation in the study region. Unlike most other tree-ring studies that focus on growing season signals, in our study the summer signal was small and the winter signal dominant due to the special conditions of the glacial valley. Site topography, geomorphology and soil characteristics in particular influence the stable isotope signal in tree-ring cellulose.

  14. Targeting erythrocyte carbonic anhydrase and 18O-isotope of breath CO2 for sorting out type 1 and type 2 diabetes

    PubMed Central

    Ghosh, Chiranjit; Mandal, Santanu; Banik, Gourab D.; Maity, Abhijit; Mukhopadhyay, Prabuddha; Ghosh, Shibendu; Pradhan, Manik

    2016-01-01

    The inability to envisage the acute onset and progression of type 1 diabetes (T1D) has been a major clinical stumbling block and an important area of biomedical research over the last few decades. Therefore there is a pressing need to develop a new and an effective strategy for early detection of T1D and to precisely distinguish T1D from type 2 diabetes (T2D). Here we describe the precise role of the enzymatic activity of carbonic anhydrase (CA) in erythrocytes in the pathogenesis of T1D and T2D. We show that CA activities are markedly altered during metabolism of T1D and T2D and this facilitates to the oxygen-18 (18O) isotopic fractionations of breath CO2. In our observations, T1D exhibited considerable depletions of 18O-isotopes of CO2, whereas T2D manifested isotopic enrichments of 18O in breath CO2, thus unveiling a missing link of breath18O-isotopic fractionations in T1D and T2D. Our findings suggest that the alterations in erythrocytes CA activities may be the initial step of altered metabolism of T1D and T2D, and breath 18O-isotope regulated by the CA activity is a potential diagnostic biomarker that can selectively and precisely distinguish T1D from T2D and thus may open a potential unifying strategy for treating these diseases. PMID:27767104

  15. Factors Influencing the Stable Oxygen and Hydrogen Isotopic Composition (δ 18O and δ D) of a Subarctic Freshwater Lake Ecosystem

    NASA Astrophysics Data System (ADS)

    Wang, Y.; Wooller, M. J.

    2005-12-01

    Previous studies have shown that the stable oxygen and hydrogen isotopic compositions (δ 18O and δD) in various animal tissues can be used to examine past climates and animal migration pattern. Little attention has been paid to the relative roles of diet and water influencing the overall δ 18O and δD of animal tissues in freshwater ecosystems. It is unclear whether different trophic levels in a freshwater lake ecosystem have an identical relationship to the water that surrounds them. The δ18O and δD values of animal tissues may be controlled by numerous different factors, including metabolic and biosynthetic isotopic fractionation and variations of δ 18O and δD in the food available. We began to examine these issues by analyzing the δ 18O and δD throughout a freshwater aquatic ecosystem at Smith Lake in Alaska. We collected samples representing primary producers and consumers (primary and secondary). Samples included green algae, various aquatic plants, such as Nuphar variegatum (water lily), Polygonum amphibium (water smartweed), Carex utriculata (sedge), Utricularia vulgaris (common bladderwort), Typha latifolia (common cattail), and a range of aquatic invertebrates, including Chironomus. sp (midge), Zygoptera (damselfly), Anisoptera (dragonfly), Dytiscidae (diving beetle) and Euhirudinea (leeches). The δ 18O and δD of Smith Lake water were ~-13.5e and -129.0e, respectively, and we present the δ 18O and δD of the rest of the ecosystem relative to these data. For instance, the δ 18O of chironomus sp. was ~12.1, which is greater than the of the lake water. Preliminary results suggest the extent of the fractionation between δ 18O of chironomids vs. lake water δ 18O is consistent with previous studies. Our data provide an insight into the range of variations that could be expected within a single freshwater ecosystem.

  16. Analysis of Hydrogen Isotopic Exchange: Lava Creek Tuff Ash and Isotopically Labeled Water

    NASA Astrophysics Data System (ADS)

    Ross, A. M.; Seligman, A. N.; Bindeman, I. N.; Nolan, G. S.

    2015-12-01

    Nolan and Bindeman (2013) placed secondarily hydrated ash from the 7.7 ka eruption of Mt. Mazama (δD=-149‰, 2.3wt% H2Ot) in isotopically labeled water (+650 ‰ δD, +56 ‰ δ18O) and observed that the H2Ot and δ18O values remained constant, but the δD values of ash increased with the surrounding water at 20, 40 and 70 °C. We expand on this work by conducting a similar experiment with ash from the 640 ka Lava Creek Tuff (LCT, δD of -128 ‰; 2.1 wt.% H2Ot) eruption of Yellowstone to see if significantly older glass (with a hypothesized gel layer on the surface shielding the interior from alteration) produces the same results. We have experiments running at 70, 24, and 5 °C, and periodically remove ~1.5 mg of glass to measure the δD (‰) and H2Ot (wt.%) of water extracted from the glass on a TC/EA MAT 253 continuous flow system. After 600 hours, the δD of the samples left at 5 and 24 °C remains at -128 ‰, but increased 8‰ for the 70 °C run series. However, there is no measurable change in wt.% of H2Ot, indicating that hydrogen exchange is not dictated by the addition of water. We are measuring and will report further progress of isotope exchange. We also plan to analyze the water in the LCT glass for δ18O (‰) to see if, as is the case for the Mt. Mazama glass, the δ18O (‰) remains constant. We also analyzed Mt. Mazama glass from the Nolan and Bindeman (2013) experiments that have now been sitting in isotopically labeled water at room temperature for ~5 years. The water concentration is still unchanged (2.3 wt.% H2Ot), and the δD of the water in the glass is now -111 ‰, causing an increase of 38 ‰. Our preliminary results show that exchange of hydrogen isotopes of hydrated glass is not limited by the age of the glass, and that the testing of hydrogen isotopes of secondarily hydrated glass, regardless of age, may not be a reliable paleoclimate indicator.

  17. Ontogeny and habitat change in Mesozoic cephalopods revealed by stable isotopes ( δ18O, δ13C)

    NASA Astrophysics Data System (ADS)

    Lukeneder, Alexander; Harzhauser, Mathias; Müllegger, Stefan; Piller, Werner E.

    2010-07-01

    Stable isotope ( δ18O and δ13C) ratios were measured in successive aragonitic shell sequences of ammonoids (class Cephalopoda) to determine whether their depth distributions changed within ontogeny and whether stable isotope values differ in various morphological groups (e.g. Leiostraca vs. Trachyostraca). We concentrate mainly on δ18O for temperature results and added δ13C data to obtain information on the ontogenetic history, for which full spiral measurements were undertaken for the first time. To obtain valid stable isotope data from ammonoid shells, we measured ontogenetic sequences (full shell) within different genera. Data sets from the Jurassic ( Cadoceras) and Cretaceous ( Hypacanthoplites, Nowakites) were chosen due to the pure primary aragonitic shell preservation. The study was designed to extract better information on the habitat and life cycle of fossil cephalopods (e.g. ammonoids) in comparison with recent cephalopods (e.g. Nautilus, Spirula, Sepia) possessing equivalent or comparable hard parts. The data from three genera suggest different modes of life in at least two morphological groups. We detected and established two main groups with different ontogenetic strategies based on the δ18O data. The wcw-type (warm-cool-warm type) of Cadoceras resembles strategies in Nautilus and Sepia, which migrate from shallow into deeper environments and back in ontogeny ( wc-type, warm-cool-type), and the cw-type (cool-warm type) of Hypacanthoplites resembling the first two migration phases of Spirula ( cwc-type), which migrates from deeper into shallower and back again into deeper habitats. The main (three) phases revealed by both δ18O and δ13C data sets most probably reflect diet changes in juvenile to mid-aged individuals, followed by a habitat change for spawning adults. In Cadoceras the temperatures range from 21.2 °C for juveniles down to 12.1 °C for mid-aged individuals and back up 16.9 °C in adults. The cw- type strategy of Hypacanthoplites

  18. The stable isotopic composition of a phosphorite deposit: δ13C, δ34S, and δ18O

    USGS Publications Warehouse

    Piper, D.Z.; Kolodny, Y.

    1987-01-01

    The stable isotopes of carbon and sulfur in a major marine sedimentary phosphate deposit from the northwestern United States (the Phosphoria Formation of Permian age) characterize the chemical properties of the depositional environment. The δ34S and δ13C analyses suggest deposition under conditions of variable redox from a solution the acidity of which was controlled by reaction with carbonate rocks and exchange with seawater. The δ18O concentration of apetite indicates phosphatization in a shallow sea, during three glacial and intervening interglacial stages. These data tend to corroborate the interpretation of field studies by others, that the apatite formed on a continental shelf in an area of intense oceanic upwelling during several episodes of sea level change. 

  19. Optimal design of isotope labeling experiments.

    PubMed

    Yang, Hong; Mandy, Dominic E; Libourel, Igor G L

    2014-01-01

    Stable isotope labeling experiments (ILE) constitute a powerful methodology for estimating metabolic fluxes. An optimal label design for such an experiment is necessary to maximize the precision with which fluxes can be determined. But often, precision gained in the determination of one flux comes at the expense of the precision of other fluxes, and an appropriate label design therefore foremost depends on the question the investigator wants to address. One could liken ILE to shadows that metabolism casts on products. Optimal label design is the placement of the lamp; creating clear shadows for some parts of metabolism and obscuring others.An optimal isotope label design is influenced by: (1) the network structure; (2) the true flux values; (3) the available label measurements; and, (4) commercially available substrates. The first two aspects are dictated by nature and constrain any optimal design. The second two aspects are suitable design parameters. To create an optimal label design, an explicit optimization criterion needs to be formulated. This usually is a property of the flux covariance matrix, which can be augmented by weighting label substrate cost. An optimal design is found by using such a criterion as an objective function for an optimizer. This chapter uses a simple elementary metabolite units (EMU) representation of the TCA cycle to illustrate the process of experimental design of isotope labeled substrates.

  20. Contributions of evaporation, isotopic non-steady state transpiration and atmospheric mixing on the delta18O of water vapour in Pacific Northwest coniferous forests.

    PubMed

    Lai, Chun-Ta; Ehleringer, James R; Bond, Barbara J; Paw U, Kyaw Tha

    2006-01-01

    Changes in the 2H and 18O of atmospheric water vapour provide information for integrating aspects of gas exchange within forest canopies. In this study, we show that diurnal fluctuations in the oxygen isotope ratio (delta 18O) as high as 4% per hundred were observed for water vapour (delta (18)Ovp) above and within an old-growth coniferous forest in the Pacific Northwest region of the United States. Values of delta 18Ovp decreased in the morning, reached a minimum at midday, and recovered to early-morning values in the late afternoon, creating a nearly symmetrical diurnal pattern for two consecutive summer days. A mass balance budget was derived and assessed for the 18O of canopy water vapour over a 2-d period by considering the 18O-isoflux of canopy transpiration, soil evaporation and the air entering the canopy column. The budget was used to address two questions: (1) do delta 18O values of canopy water vapour reflect the biospheric influence, or are such signals swamped by atmospheric mixing? and (2) what mechanisms drive temporal variations of delta 18Ovp? Model calculations show that the entry of air into the canopy column resulted in an isotopically depleted 18O-isoflux in the morning of day 1, causing values of delta 18Ovp, to decrease. An isotopically enriched 18O-isoflux resulting from transpiration then offset this decreased delta 18Ovp later during the day. Contributions of 18O-isoflux from soil evaporation were relatively small on day 1 but were more significant on day 2, despite the small H2(16)O fluxes. From measurements of leaf water volume and sapflux, we determined the turnover time of leaf water in the needles of Douglas-fir trees as approximately 11 h at midday. Such an extended turnover time suggests that transpiration may not have occurred at the commonly assumed isotopic steady state. We tested a non-steady state model for predicting delta 18O of leaf water. Our model calculations show that assuming isotopic steady state increased isoflux of

  1. The Kuo-Brown effective interaction: From 18O to the Sn isotopes

    NASA Astrophysics Data System (ADS)

    Engeland, Torgeir; Hjorth-Jensen, Morten; Kartamyshev, Maxim; Osnes, Eivind

    2014-08-01

    After briefly reviewing the pioneering work on effective interactions by Gerry Brown and his group, and the developments which followed, we apply present-day effective interactions to large-scale shell-model calculations on the entire range of Sn isotopes from 102Sn to 132Sn. We have made explorative calculations starting from three different nucleon-nucleon potentials (Argonne V18, CD-Bonn, and N3LO) and evaluated the higher-order contributions to the effective interaction from both G-matrix and Vlowk interactions. Further, we have checked the convergence of intermediate-state excitations up to 10ħω harmonic oscillator energy. Final extensive calculations were made of binding energies, excitation energies and B(E2) transition rates using an effective interaction based on a G-matrix evaluated from the chiral N3LO potential and including intermediate excitations up to 10ħω harmonic oscillator energy. The energy spectra are well reproduced throughout the region while overbinding of the ground states emerges as valence nucleons are added. The B(E2) rates agree well for the heavy isotopes, while they seem too low for the lighter ones.

  2. North American precipitation isotope18O) zones revealed in time series modeling across Canada and northern United States

    NASA Astrophysics Data System (ADS)

    Delavau, C.; Chun, K. P.; Stadnyk, T.; Birks, S. J.; Welker, J. M.

    2015-02-01

    Delineating spatial patterns of precipitation isotopes ("isoscapes") is becoming increasingly important to understand the processes governing the modern water isotope cycle and their application to migration forensics, climate proxy interpretation, and ecohydrology of terrestrial systems. However, the extent to which these patterns can be empirically predicted across Canada and the northern United States has not been fully articulated, in part due to a lack of time series precipitation isotope data for major regions of North America. In this study, we use multiple linear regressions of CNIP, GNIP, and USNIP observations alongside climatological variables, teleconnection indices, and geographic indicators to create empirical models that predict the δ18O of monthly precipitation (δ18Oppt) across Canada and the northern United States. Five regionalization approaches are used to separate the study domain into isotope zones to explore the effect of spatial grouping on model performance. Stepwise regression-derived parameterizations quantified by permutation testing indicate the significance of precipitable water content and latitude as predictor variables. Within the Canadian Arctic and eastern portion of the study domain, models from all regionalizations capture the interannual and intraannual variability of δ18Oppt. The Pacific coast and northwestern portions of the study domain show less agreement between models and poorer model performance, resulting in higher uncertainty in simulations throughout these regions. Long-term annual average δ18Oppt isoscapes are generated, highlighting the uncertainty in the regionalization approach as it compounds over time. Additionally, monthly time series simulations are presented at various locations, and model structure uncertainty and 90% bootstrapped prediction bounds are detailed for these predictions.

  3. EFFECT OF RAPID SHALLOW BREATHING ON THE DISTRIBUTION OF 18-O-LABELED OZONE REACTION PRODUCT IN THE RESPIRATORY TRACT OF THE RAT

    EPA Science Inventory

    We examined the effect of breathing pattern on ozone reaction product content within the respiratory tract. Thirty-four anesthetized, maleWistar rats were exposed to oxygen-18 (18O)-labeled ozone at 1.0 ppm for 2 h using a dual-chamber, negative-pressure ventilation system. Fre...

  4. Effect of 2H and 18O water isotopes in kinesin-1 gliding assay

    PubMed Central

    Herskowitz, Lawrence J.; Koch, Steven J.

    2014-01-01

    We show for the first time the effects of heavy-hydrogen water (2H2O) and heavy-oxygen water (H218O) on the gliding speed of microtubules on kinesin-1 coated surfaces. Increased fractions of isotopic waters used in the motility solution decreased the gliding speed of microtubules by a maximum of 21% for heavy-hydrogen and 5% for heavy-oxygen water. We also show that gliding microtubule speed returns to its original speed after being treated with heavy-hydrogen water. We discuss possible interpretations of these results and the importance for future studies of water effects on kinesin and microtubules. We also discuss the implication for using heavy waters in biomolecular devices incorporating molecular motors. PMID:24711961

  5. Measurement of Whole-Body CO2 Production in Birds Using Real-Time Laser-Derived Measurements of Hydrogen (δ(2)H) and Oxygen (δ(18)O) Isotope Concentrations in Water Vapor from Breath.

    PubMed

    Mitchell, G W; Guglielmo, C G; Hobson, K A

    2015-01-01

    The doubly labeled water (DLW) method is commonly used to measure energy expenditure in free-living wildlife and humans. However, DLW studies involving animals typically require three blood samples, which can affect behavior and well-being. Moreover, measurement of H (δ(2)H) and O (δ(18)O) isotope concentrations in H2O derived from blood using conventional isotope ratio mass spectrometry is technically demanding, time-consuming, and often expensive. A novel technique that would avoid these constraints is the real-time measurement of δ(2)H and δ(18)O in the H2O vapor of exhaled breath using cavity ring-down (CRD) spectrometry, provided that δ(2)H and δ(18)O from body H2O and breath were well correlated. Here, we conducted a validation study with CRD spectrometry involving five zebra finches (Taeniopygia guttata), five brown-headed cowbirds (Molothrus ater), and five European starlings (Sturnus vulgaris), where we compared δ(2)H, δ(18)O, and rCO2 (rate of CO2 production) estimates from breath with those from blood. Isotope concentrations from blood were validated by comparing dilution-space estimates with measurements of total body water (TBW) obtained from quantitative magnetic resonance. Isotope dilution-space estimates from δ(2)H and δ(18)O values in the blood were similar to and strongly correlated with TBW measurements (R(2) = 0.99). The (2)H and (18)O (ppm) in breath and blood were also highly correlated (R(2) = 0.99 and 0.98, respectively); however, isotope concentrations in breath were always less enriched than those in blood and slightly higher than expected, given assumed fractionation values between blood and breath. Overall, rCO2 measurements from breath were strongly correlated with those from the blood (R(2) = 0.90). We suggest that this technique will find wide application in studies of animal and human energetics in the field and laboratory. We also provide suggestions for ways this technique could be further improved.

  6. Stable Isotope Values of the Mesoamerican Monsoon: δ18O and δ2H Values Reveal Climate Controls on Summer Rainfall Amount

    NASA Astrophysics Data System (ADS)

    Bernal, J. P.; Lachniet, M. S.; Rosales Lagarde, L.; Morales Puente, P.; Cienfuegos, E.

    2014-12-01

    Paleoclimate reconstructions using δ18O as a proxy for the isotopic composition of rainfall are based upon the mostly untested assumption that either rainfall amount or equilibration temperature are the main drivers modulating the isotopic composition of pluvial precipitation. Whilst a broad correlation between geographical location and driving mechanisms has been long recognized (i.e. amount effect is pervasive in tropical areas), further tests are required to determine the effect that different sources of moisture might impose on the isotopic composition of precipitation, particularly in areas where contributions from different ocean-basins might be significant, such as south Mexico. Here, we present the δ18O and δ2H composition of summer rainfall collected throughout south, central and western Mexico, particularly from Veracruz, Puebla, Guerrero, Morelos, Mexico City, Jalisco, Michoacán and Querétaro states. The geographical and temporal extent of our sampling (2004, 2005, 2007, 2008, 2011) results in a large dataset comprising more than 600 samples and represents the base data to understand the atmospheric mechanisms modulating the isotopic composition of rainfall in Mexico. Our data span a range of 30‰ in δ18O, from high values nearest the Gulf of Mexico coast and during weak rainfall events, to lowest values in high-altitude central Mexico and during heavy rainfall events associated with tropical cyclones. Values on the Pacific Coast are intermediate, and likely reflect a contribution of both Gulf of Mexico and Pacific sources. Our data define a meteoric water line of δ2H = 7.92 × δ18O + 9.48, which indicate that most precipitation values formed close to isotopic equilibrium with water vapor. The two primary physiographic variables controlling δ18O values are distance from the Gulf of Mexico and altitude, which together explain about 70% of the variation in spatial δ18O values.

  7. Stable Isotope Labeling Strategy for Curcumin Metabolite Study in Human Liver Microsomes by Liquid Chromatography-Tandem Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Gao, Dan; Chen, Xiaowu; Yang, Xiaomei; Wu, Qin; Jin, Feng; Wen, Hongliang; Jiang, Yuyang; Liu, Hongxia

    2015-04-01

    The identification of drug metabolites is very important in drug development. Nowadays, the most widely used methods are isotopes and mass spectrometry. However, the commercial isotopic labeled reagents are usually very expensive, and the rapid and convenient identification of metabolites is still difficult. In this paper, an 18O isotope labeling strategy was developed and the isotopes were used as a tool to identify drug metabolites using mass spectrometry. Curcumin was selected as a model drug to evaluate the established method, and the 18O labeled curcumin was successfully synthesized. The non-labeled and 18O labeled curcumin were simultaneously metabolized in human liver microsomes (HLMs) and analyzed by liquid chromatography/mass spectrometry (LC-MS). The two groups of chromatograms obtained from metabolic reaction mixture with and without cofactors were compared and analyzed using Metabolynx software (Waters Corp., Milford, MA, USA). The mass spectra of the newly appearing chromatographic peaks in the experimental sample were further analyzed to find the metabolite candidates. Their chemical structures were confirmed by tandem mass spectrometry. Three metabolites, including two reduction products and a glucuronide conjugate, were successfully detected under their specific HLMs metabolic conditions, which were in accordance with the literature reported results. The results demonstrated that the developed isotope labeling method, together with post-acquisition data processing using Metabolynx software, could be used for fast identification of new drug metabolites.

  8. Stable isotope labeling strategy for curcumin metabolite study in human liver microsomes by liquid chromatography-tandem mass spectrometry.

    PubMed

    Gao, Dan; Chen, Xiaowu; Yang, Xiaomei; Wu, Qin; Jin, Feng; Wen, Hongliang; Jiang, Yuyang; Liu, Hongxia

    2015-04-01

    The identification of drug metabolites is very important in drug development. Nowadays, the most widely used methods are isotopes and mass spectrometry. However, the commercial isotopic labeled reagents are usually very expensive, and the rapid and convenient identification of metabolites is still difficult. In this paper, an (18)O isotope labeling strategy was developed and the isotopes were used as a tool to identify drug metabolites using mass spectrometry. Curcumin was selected as a model drug to evaluate the established method, and the (18)O labeled curcumin was successfully synthesized. The non-labeled and (18)O labeled curcumin were simultaneously metabolized in human liver microsomes (HLMs) and analyzed by liquid chromatography/mass spectrometry (LC-MS). The two groups of chromatograms obtained from metabolic reaction mixture with and without cofactors were compared and analyzed using Metabolynx software (Waters Corp., Milford, MA, USA). The mass spectra of the newly appearing chromatographic peaks in the experimental sample were further analyzed to find the metabolite candidates. Their chemical structures were confirmed by tandem mass spectrometry. Three metabolites, including two reduction products and a glucuronide conjugate, were successfully detected under their specific HLMs metabolic conditions, which were in accordance with the literature reported results. The results demonstrated that the developed isotope labeling method, together with post-acquisition data processing using Metabolynx software, could be used for fast identification of new drug metabolites.

  9. Down Core Oxygen Isotopic Measurements Of Diatom δ18O From The Guaymas Basin, Gulf Of California

    NASA Astrophysics Data System (ADS)

    Menicucci, A. J.; Spero, H. J.; Thunell, R.

    2015-12-01

    The Guaymas Basin (GB), Gulf of California (27º53'N, 111º40'W ), is an evaporative basin, with sea surface temperatures (SST) varying between ~30oC (summer) and ~15oC (winter). Productivity is controlled mostly by seasonal upwelling starting in fall (early November) and extending into spring. We are currently analyzing δ18Odiatom from a boxcore (BC-43) using microfluorination (Menicucci, et al. 2013). This boxcore was previously analyzed for UK '37 alkenones and 210Pb activity (Goni, et al. 2001). Residual BC-43 material was sampled at ~2cm intervals. Samples were cleaned to isolate diatoms from other sediments, then equilibrated in water with δ18Owater = +85‰ for 70 hours at 21oC prior to vacuum dehydroxylation and microfluorination. The latter equilibration was done to account for fractionation between covalently bound O and OH- groups during vacuum dehydroxylation, preserving the original δ18Odiatom value. We present δ18Odiatom data from BC-43 samples covering 27cm, equivalent to >225 years of sediment accumulation. δ18O data are converted to temperature (T) based on an existing calibration (Leclerc and Labeyrie 1987). Our data suggest δ18Odiatom values record a T range of 22-18oC, corresponding to the mixed layer depth and the chlorophyll maximum during the fall bloom. These T values are offset from SST data by a mean of 5oC for the same sample intervals. However, δ18Odiatom values from the most recent samples suggest a ~2oC increase in diatom T relative to SST during the last 35 years. This subsurface warming may be due to decreased fall upwelling, increased mixed layer and chlorophyll maximum depths, and/or the timing of the peak diatom bloom. Such correlations are being investigated and the latest results will be presented. Goni, M. A., et al. (2001). Oceanographic considerations for the application of the alkenone-based paleotemperature U-37(K ') index in the Gulf of California. Geochimica Et Cosmochimica Acta 65: 545-557. Leclerc, A. J. and L

  10. USGS48 Puerto Rico precipitation - A new isotopic reference material for δ2H and δ18O measurements of water

    USGS Publications Warehouse

    Qi, Haiping; Coplen, Tyler B.; Tarbox, Lauren V.; Lorenz, Jennifer M.; Scholl, Martha A.

    2014-01-01

    A new secondary isotopic reference material has been prepared from Puerto Rico precipitation, which was filtered, homogenised, loaded into glass ampoules, sealed with a torch, autoclaved to eliminate biological activity, and calibrated by dual-inlet isotope-ratio mass spectrometry. This isotopic reference material, designated as USGS48, is intended to be one of two isotopic reference waters for daily normalisation of stable hydrogen (δ2H) and stable oxygen (δ18O) isotopic analysis of water with a mass spectrometer or a laser absorption spectrometer. The δ2H and δ18O values of this reference water are−2.0±0.4 and−2.224±0.012 ‰, respectively, relative to Vienna Standard Mean Ocean Water on scales normalised such that the δ2H and δ18O values of Standard Light Antarctic Precipitation reference water are−428 and−55.5 ‰, respectively. Each uncertainty is an estimated expanded uncertainty (U=2uc) about the reference value that provides an interval that has about a 95 % probability of encompassing the true value. This isotopic reference water is available by the case of 144 glass ampoules containing 5 mL of water in each ampoule.

  11. Matching isotopic distributions from metabolically labeled samples

    PubMed Central

    McIlwain, Sean; Page, David; Huttlin, Edward L.; Sussman, Michael R.

    2008-01-01

    Motivation: In recent years stable isotopic labeling has become a standard approach for quantitative proteomic analyses. Among the many available isotopic labeling strategies, metabolic labeling is attractive for the excellent internal control it provides. However, analysis of data from metabolic labeling experiments can be complicated because the spacing between labeled and unlabeled forms of each peptide depends on its sequence, and is thus variable from analyte to analyte. As a result, one generally needs to know the sequence of a peptide to identify its matching isotopic distributions in an automated fashion. In some experimental situations it would be necessary or desirable to match pairs of labeled and unlabeled peaks from peptides of unknown sequence. This article addresses this largely overlooked problem in the analysis of quantitative mass spectrometry data by presenting an algorithm that not only identifies isotopic distributions within a mass spectrum, but also annotates matches between natural abundance light isotopic distributions and their metabolically labeled counterparts. This algorithm is designed in two stages: first we annotate the isotopic peaks using a modified version of the IDM algorithm described last year; then we use a probabilistic classifier that is supplemented by dynamic programming to find the metabolically labeled matched isotopic pairs. Such a method is needed for high-throughput quantitative proteomic metabolomic experiments measured via mass spectrometry. Results: The primary result of this article is that the dynamic programming approach performs well given perfect isotopic distribution annotations. Our algorithm achieves a true positive rate of 99% and a false positive rate of 1% using perfect isotopic distribution annotations. When the isotopic distributions are annotated given ‘expert’ selected peaks, the same algorithm gets a true positive rate of 77% and a false positive rate of 1%. Finally, when annotating using

  12. Plasma proteome response to severe burn injury revealed by 18O-labeled "universal" reference-based quantitative proteomics.

    PubMed

    Qian, Wei-Jun; Petritis, Brianne O; Kaushal, Amit; Finnerty, Celeste C; Jeschke, Marc G; Monroe, Matthew E; Moore, Ronald J; Schepmoes, Athena A; Xiao, Wenzhong; Moldawer, Lyle L; Davis, Ronald W; Tompkins, Ronald G; Herndon, David N; Camp, David G; Smith, Richard D

    2010-09-03

    A burn injury represents one of the most severe forms of human trauma and is responsible for significant mortality worldwide. Here, we present the first quantitative proteomics investigation of the blood plasma proteome response to severe burn injury by comparing the plasma protein concentrations of 10 healthy control subjects with those of 15 severe burn patients at two time-points following the injury. The overall analytical strategy for this work integrated immunoaffinity depletion of the 12 most abundant plasma proteins with cysteinyl-peptide enrichment-based fractionation prior to LC-MS analyses of individual patient samples. Incorporation of an 18O-labeled "universal" reference among the sample sets enabled precise relative quantification across samples. In total, 313 plasma proteins confidently identified with two or more unique peptides were quantified. Following statistical analysis, 110 proteins exhibited significant abundance changes in response to the burn injury. The observed changes in protein concentrations suggest significant inflammatory and hypermetabolic response to the injury, which is supported by the fact that many of the identified proteins are associated with acute phase response signaling, the complement system, and coagulation system pathways. The regulation of approximately 35 proteins observed in this study is in agreement with previous results reported for inflammatory or burn response, but approximately 50 potentially novel proteins previously not known to be associated with burn response or inflammation are also found. Elucidating proteins involved in the response to severe burn injury may reveal novel targets for therapeutic interventions as well as potential predictive biomarkers for patient outcomes such as multiple organ failure.

  13. Accuracy of delta 18O isotope ratio measurements on the same sample by continuous-flow isotope-ratio mass spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The doubly labeled water method is considered the reference method to measure energy expenditure. Conventional mass spectrometry requires a separate aliquot of the same sample to be prepared and analyzed separately. With continuous-flow isotope-ratio mass spectrometry, the same sample could be analy...

  14. A first Late Glacial and Early Holocene coupled 18O and 2H biomarker isotope record from Gemuendener Maar, Germany

    NASA Astrophysics Data System (ADS)

    Zech, Michael; Bromm, Tobias; Hepp, Johannes; Benesch, Marianne; Sirocko, Frank; Glaser, Bruno; Zech, Roland

    2015-04-01

    During the last years, we developed a method for compound-specific d18O analyses of hemicellulose-derived sugars from plants, soils and sediment archives (Zech and Glaser, 2009; Zech et al., 2014). The coupling of respective d18O sugar results with d2H alkane results from paleosol and sediment climate archives proved to be a valuable innovative approach towards quantitative paleoclimate reconstruction (Hepp et al., 2014; Zech et al., 2013). Here we present a first coupled d18O sugar and d2H alkane biomarker record obtained for Late Glacial and Early Holocene sediments from the Gemuendener Maar in the Eifel, Germany. The d18O sugar biomarker record resembles the d18O ice core records of Greenland. The coupling with the d2H alkane biomarker results allows drawing further more quantitative paleocimate information in terms of (i) paleohumidity and (ii) d18O of paleoprecipitation.

  15. Controls on the Nitrogen and Oxygen Isotopic Composition (δ 15N, δ 18O, δ 17O) of Atmospheric Nitrate in Princeton, NJ

    NASA Astrophysics Data System (ADS)

    Hastings, M. G.; Malcolm, E.; Kaiser, J.; Sigman, D. M.

    2004-12-01

    The oxygen isotopic composition of atmospheric nitrate reflects the oxidative mechanisms that convert NOx to HNO3, while the nitrogen isotopic composition of atmospheric nitrate may reflect different NOx source signatures and/or fractionations related to NOx chemistry [Michalski et al., 2003; Hastings et al., 2003; Freyer et al., 1993]. New analysis techniques are capable of determining the 15N/14N, 18O/16O and 17O/16O isotope ratios in samples at the nanomolar level [Sigman et al., 2001; Casciotti et al., 2002; see Kaiser et al., session H38]. This allows for the analysis of short-term variations in the isotopes of HNO3 with the potential to diagnose causal relationships by comparing the isotopic data with other features of atmospheric deposition. The 15N/14N, 18O/16O and 17O/16O of nitrate were analyzed from precipitation samples collected on an event-basis in Princeton, NJ between December 2002 and 2003. The nitrate concentration in Princeton rain ranges from 2.5 to 99.7 μ M (mean=21.1 μ M, n=61), similar to that found in other urban areas of New Jersey by the National Atmospheric Deposition Program. The isotopes of nitrate fall in the wide range reported for various environments with the δ 15N ranging from -4.0 to 9.5‰ (vs. air), and the δ 18O and δ 17O ranging from 57.2 to 90.5‰ and 50.7 to 77.8‰ (vs. VSMOW), respectively. The correlation between nitrate and sulfate concentration (R2=0.66) and the lack of a relationship between these major ions and the isotopes of nitrate supports the conclusion that below cloud scavenging is not the dominant control on the isotopic variations observed. Seasonal variations are observed in both the nitrogen and oxygen isotopes of nitrate. Overall the δ 15N is not correlated with either δ 18O or δ 17O, although both the δ 15N and δ 18O average lowest in the summer and highest in the winter. δ 18O is highly correlated with δ 17O of nitrate with anomalous enrichment in 17O relative to 18O (Δ 17O ranges from 19

  16. Biscayne aquifer drinking water (USGS45): a new isotopic reference material for δ2H and δ18O measurements of water

    USGS Publications Warehouse

    Lorenz, Jennifer M.; Tarbox, Lauren V.; Buck, Bryan; Qi, Haiping; Coplen, Tyler B.

    2014-01-01

    RATIONALE As a result of the scarcity of isotopic reference waters for daily use, a new secondary isotopic reference material for international distribution has been prepared from drinking water collected from the Biscayne aquifer in Ft. Lauderdale, Florida. METHODS This isotopic reference water was filtered, homogenized, loaded into glass ampoules, sealed with a torch, autoclaved to eliminate biological activity, and measured by dual-inlet isotope-ratio mass spectrometry. This reference material is available by the case of 144 glass ampoules containing either 4 mL or 5 mL of water in each ampoule. RESULTS The δ2H and δ18O values of this reference material are –10.3 ± 0.4 ‰ and –2.238 ± 0.011 ‰, respectively, relative to VSMOW, on scales normalized such that the δ2H and δ18O values of SLAP reference water are, respectively, –428 and –55.5 ‰. Each uncertainty is an estimated expanded uncertainty (U = 2uc) about the reference value that provides an interval that has about a 95 % probability of encompassing the true value. CONCLUSIONS This isotopic reference material, designated as USGS45, is intended as one of two isotopic reference waters for daily normalization of stable hydrogen and oxygen isotopic analysis of water with an isotope-ratio mass spectrometer or a laser absorption spectrometer. 

  17. Analysis of stable isotope ratios (δ18O and δ2H) in precipitation of the Verde River watershed, Arizona 2013 through 2014

    USGS Publications Warehouse

    Beisner, Kimberly R.; Paretti, Nicholas V.; Tucci, Rachel S.

    2016-04-25

    Stable isotope delta values (δ18O and δ2H) of precipitation can vary with elevation, and quantification of the precipitation elevation gradient can be used to predict recharge elevation within a watershed. Precipitation samples were analyzed for stable isotope delta values between 2003 and 2014 from the Verde River watershed of north-central Arizona. Results indicate a significant decrease in summer isotopic values overtime at 3,100-, 4,100-, 6,100-, 7,100-, and 8,100-feet elevation. The updated local meteoric water line for the area is δ2H = 7.11 δ18O + 3.40. Equations to predict stable isotopic values based on elevation were updated from previous publications in Blasch and others (2006), Blasch and Bryson (2007), and Bryson and others (2007). New equations were separated for samples from the Camp Verde to Flagstaff transect and the Prescott to Chino Valley transect. For the Camp Verde to Flagstaff transect, the new equations for winter precipitation are δ18O = -0.0004z − 8.87 and δ2H = -0.0029z − 59.8 (where z represents elevation in feet) and the summer precipitation equations were not statistically significant. For the Prescott to Chino Valley transect, the new equations for summer precipitation are δ18O = -0.0005z − 3.22 and δ2H = -0.0022z − 27.9; the winter precipitation equations were not statistically significant and, notably, stable isotope values were similar across all elevations. Interpretation of elevation of recharge contributing to surface and groundwaters in the Verde River watershed using the updated equations for the Camp Verde to Flagstaff transect will give lower elevation values compared with interpretations presented in the previous studies. For waters in the Prescott and Chino Valley area, more information is needed to understand local controls on stable isotope values related to elevation.

  18. Treatment methods for the determination of delta2H and delta18O of hair keratin by continuous-flow isotope-ratio mass spectrometry.

    PubMed

    Bowen, Gabriel J; Chesson, Lesley; Nielson, Kristine; Cerling, Thure E; Ehleringer, James R

    2005-01-01

    The structural proteins that comprise approximately 90% of animal hair have the potential to record environmentally and physiologically determined variation in delta2H and delta18O values of body water. Broad, systematic, geospatial variation in stable hydrogen and oxygen isotopes of environmental water and the capacity for rapid, precise measurement via methods such as high-temperature conversion elemental analyzer/isotope ratio mass spectrometry (TC/EA-IRMS) make these isotope systems particularly well suited for applications requiring the geolocation of hair samples. In order for such applications to be successful, however, methods must exist for the accurate determination of hair delta2H and delta18O values reflecting the primary products of biosynthesis. Here, we present the results of experiments designed to examine two potential inaccuracies affecting delta2H and delta18O measurements of hair: the contribution of non-biologic hydrogen and oxygen to samples in the form of sorbed molecular water, and the exchange of hydroxyl-bound hydrogen between hair keratin and ambient water vapor. We show that rapid sorption of molecular water from the atmosphere can have a substantial effect on measured delta2H and delta18O values of hair (comprising approximately 7.7% of the measured isotopic signal for H and up to approximately 10.6% for O), but that this contribution can be effectively removed through vacuum-drying of samples for 6 days. Hydrogen exchange between hair keratin and ambient vapor is also rapid (reaching equilibrium within 3-4 days), with 9-16% of the total hydrogen available for exchange at room temperature. Based on the results of these experiments, we outline a recommended sample treatment procedure for routine measurement of delta2H and delta18O in mammal hair.

  19. NMR studies of isotopically labeled RNA

    SciTech Connect

    Pardi, A.

    1994-12-01

    In summary, the ability to generate NMR quantities of {sup 15}N and {sup 13}C-labeled RNAs has led to the development of heteronuclear multi-dimensional NMR techniques for simplifying the resonance assignment and structure determination of RNAs. These methods for synthesizing isotopically labeled RNAs are only several years old, and thus there are still relatively few applications of heteronuclear multi-dimensional NMR techniques to RNA. However, given the critical role that RNAs play in cellular function, one can expect to see an increasing number of NMR structural studies of biologically active RNAs.

  20. USGS46 Greenland ice core water – A new isotopic reference material for δ2H and δ18O measurements of water

    USGS Publications Warehouse

    Coplen, Tyler B.; Qi, Haiping; Tarbox, Lauren V.; Lorenz, Jennifer M.; Buck, Bryan

    2015-01-01

    Ice core from Greenland was melted, filtered, homogenised, loaded into glass ampoules, sealed, autoclaved to eliminate biological activity, and calibrated by dual-inlet isotope-ratio mass spectrometry. This isotopic reference material (RM), USGS46, is intended as one of two secondary isotopic reference waters for daily normalisation of stable hydrogen (δ2H) and stable oxygen (δ18O) isotopic analysis of water with a mass spectrometer or a laser absorption spectrometer. The measured δ2H and δ18O values of this reference water were −235.8 ± 0.7‰ and −29.80 ± 0.03‰, respectively, relative to VSMOW on scales normalised such that the δ2H and δ18O values of SLAP reference water are, respectively, −428 and −55.5‰. Each uncertainty is an estimated expanded uncertainty (U = 2uc) about the reference value that provides an interval that has about a 95-percent probability of encompassing the true value. This reference water is available in cases containing 144 glass ampoules that are filled with either 4 ml or 5 ml of water per ampoule.

  1. A novel isotopic fractionation during dissolved oxygen consumption in mesopelagic waters inferred from observation and model simulation of dissolved oxygen δ18O in open oceanic regions

    NASA Astrophysics Data System (ADS)

    Nakayama, N.; Oka, A.; Gamo, T.

    2012-12-01

    Oxygen isotopic ratio (δ18O) of dissolved oxygen is a useful for bioactive tracer of the subsurface aphotic (mesopelagic) ocean since it varies nonlinearly related to oxygen consumption via stoichiometry of organic matter decomposition. Therefore, along with global circulation model (GCM), observed δ18O and their vertical/geographical distribution can be effectively used to quantitatively determine how marine biological and ocean physical processes contribute to varying dissolved oxygen (DO) concentration in the ocean, in particular mesopelagic zone where pronounced biological activity alters DO concentration significantly. In the central north Pacific Ocean and Indian Ocean, including Arabian Sea, one of the few regions in the open ocean which has oxygen minimum zone (OMZ, a layer with severely depleted DO), vertical profiles of DO and δ18O were observed. These observed data are compared with a GCM simulation in which a constant isotopic fractionation factor of DO by marine biological respiration and a fixed Redfield molar ratio between P and O are assumed. Even in the Arabian Sea OMZ, relationship between DO and δ18O was found to be similar to those observed in other open oceans, indicating that no specific oxygen consumption process occurred in the OMZ. Using the GCM model, we attempted to reproduce the observed overall relationship between DO and δ18O, but it failed when we adopted the previously reported isotopic fractionation factor: Discrepancy became larger when oxygen saturation level decreased, in particular in thermocline water (at 20% oxygen saturation level, modeled δ18O was heavier than observed values by +7‰). Sensitivity simulations with the GCM model revealed that (1) simply changing the intensity of oxygen consumption by respiration/organic matter decomposition nor physical processes (diffusion and/or advection) could explain the observed relationship between DO and δ18O, (2) applying a smaller isotopic fractionation for deep waters

  2. Quasiclassical trajectory studies of 18O(3P) + NO2 isotope exchange and reaction to O2 + NO on D0 and D1 potentials

    NASA Astrophysics Data System (ADS)

    Fu, Bina; Zhang, Dong H.; Bowman, Joel M.

    2013-07-01

    We report quasiclassical trajectory calculations for the bimolecular reaction 18O(3P) + NO2 on the recent potential energy surfaces of the ground (D0) and first excited (D1) states of NO3 [B. Fu, J. M. Bowman, H. Xiao, S. Maeda, and K. Morokuma, J. Chem. Theory. Comput. 9, 893 (2013)], 10.1021/ct3009792. The branching ratio of isotope exchange versus O2 + NO formation, as well as the product angular distributions and energy and rovibrational state distributions are presented. The calculations are done at the collision energy of relevance to recent crossed beam experiments [K. A. Mar, A. L. Van Wyngarden, C.-W. Liang, Y. T. Lee, J. J. Lin, and K. A. Boering, J. Chem. Phys. 137, 044302 (2012)], 10.1063/1.4736567. Very good agreement is achieved between the current calculations and these experiments for the branching ratio and final translational energy and angular distributions of isotope exchange products 16O(3P) + NO2 and O2 + NO formation products. The reactant 18O atom results in 18O16O but not N18O for the O2 + NO formation product channel, consistent with the experiment. In addition, the detailed vibrational and rotational state information of diatomic molecules calculated currently for the 34O2 + NO formation channel on D0 and D1 states are in qualitative agreement with the previous experimental and theoretical results of the photodissociation of NO3 and are consistent with older thermal bimolecular kinetics measurements.

  3. Stable isotope18O and δ2H) data for precipitation, stream water, and groundwater in Puerto Rico

    USGS Publications Warehouse

    Scholl, Martha A.; Torres-Sanchez, Angel; Rosario-Torres, Manuel

    2014-01-01

    , hurricanes, and cold fronts, although frequent low-intensity orographic showers occur throughout the year in the mountains. The stable isotope signatures of rainfall (δ2H and δ18O) are broadly correlated with the weather type that produced the rainfall (Scholl and others, 2009; Scholl and Murphy, 2014).

  4. Oxygen isotope evolution of the Lake Owyhee volcanic field, Oregon, and implications for low-δ18O magmas of the Snake River Plain - Yellowstone hotspot

    NASA Astrophysics Data System (ADS)

    Blum, T.; Kitajima, K.; Nakashima, D.; Valley, J. W.

    2013-12-01

    The Snake River Plain - Yellowstone (SRP-Y) hotspot trend is one of the largest known low-δ18O magmatic provinces, yet the timing and distribution of hydrothermal alteration relative to hotspot magmatism remains incompletely understood. Existing models for SRP-Y low-δ18O magma genesis differ regarding the timing of protolith alteration (e.g. Eocene vs. present), depth at which alteration occurs (e.g. 15 km vs. <5 km), and physical controls on the extent of alteration (e.g. caldera collapse, crustal scale fluid flow, etc.). We expand the existing oxygen isotope data set for zircon in the Lake Owyhee volcanic field (LOVF) of east central Oregon to further identify magmatic oxygen isotope trends within the field. These data offer insight into the timing of alteration and the extent of the greater SRP-Y low-δ18O province, as well as the conditions that generate large low-δ18O provinces. 16-14 Ma silicic volcanism in the LOVF is linked to the pre-14 Ma SRP-Y hotspot, with volcanism partially overlapping extension in the north-south trending Oregon-Idaho Graben (OIG). Ion microprobe analyses of zircons from 16 LOVF silicic lavas and tuffs reveal homogeneous zircons on both the single grain and hand sample scales: individual samples have 2 S.D. for δ18O ranging from 0.27 to 0.96‰ (SMOW), and sample averages ranging from 1.8 to 6.0‰, excluding texturally chaotic and/or porous zircons which have δ18O values as low as 0.0‰. All low-δ18O LOVF magmas, including the caldera-forming Tuff of Leslie Gulch and Tuff of Spring Creek, are confined to the OIG, although not all zircons from within the OIG have low δ18O values. The presence and sequence of low-δ18O magmas in the LOVF and adjacent central Snake River Plain (CSRP) cannot be explained by existing caldera subsidence or pre-hotspot source models. These data, however, combined with volumetrically limited low-δ18O material in the adjacent Idaho Batholith and Basin and Range, are consistent with low-δ18O magmas

  5. Winter Precipitation Isotope Gradients (δ18O) of the Contiguous USA and Their Relationship to the Pacific/North American (PNA) Pattern

    NASA Astrophysics Data System (ADS)

    Liu, Z.; Bowen, G. J.; Welker, J. M.

    2011-12-01

    This study investigates the synoptic-dynamic relationship between Pacific/North American (PNA) pattern and winter precipitation isotopes of the contiguous USA using 2-year (1990 and 1992) USNIP (the United States Network for Isotopes in Precipitation) dataset. We find that patterns in the spatial gradient of precipitation isotope values reflect the position of the polar jet stream and juxtaposition of air masses associated with variation in the PNA pattern. During the positive PNA winter, a southward shift of zones of steep δ18O gradients in the eastern USA coincides with southward displacement of the polar jet stream, which leads to a greater frequency of polar air masses and typically depleted δ18O values in the region. A coincident eastward shift in high-gradient zones in the western USA is related to more frequent penetration of tropical air masses, which in juxtaposition with polar air in the mid-continent leads to higher gradient values in the western region. Our findings highlight the importance of PNA pattern in determining spatial patterns of precipitation isotopes, with implications for interpretations of paleo-water isotope values and isotopic applications to study modern hydrological processes.

  6. Oxygen isotope ratios (18O/16O) of hemicellulose-derived sugar biomarkers in plants, soils and sediments as paleoclimate proxy I: Insight from a climate chamber experiment

    NASA Astrophysics Data System (ADS)

    Zech, Michael; Mayr, Christoph; Tuthorn, Mario; Leiber-Sauheitl, Katharina; Glaser, Bruno

    2014-02-01

    The oxygen isotopic composition of cellulose is a valuable proxy in paleoclimate research. However, its application to sedimentary archives is challenging due to extraction and purification of cellulose. Here we present compound-specific δ18O results of hemicellulose-derived sugar biomarkers determined using gas chromatography-pyrolysis-isotope ratio mass spectrometry, which is a method that overcomes the above-mentioned analytical challenges. The biomarkers were extracted from stem material of different plants (Eucalyptus globulus, Vicia faba and Brassica oleracea) grown in climate chamber experiments under different climatic conditions. The δ18O values of arabinose and xylose range from 31.4‰ to 45.9‰ and from 28.7‰ to 40.8‰, respectively, and correlate highly significantly with each other (R = 0.91, p < 0.001). Furthermore, δ18Ohemicellulose (mean of arabinose and xylose) correlate highly significantly with δ18Oleaf water (R = 0.66, p < 0.001) and significantly with modeled δ18Ocellulose (R = 0.42, p < 0.038), as well as with relative air humidity (R = -0.79, p < 0.001) and temperature (R = -0.66, p < 0.001). These findings confirm that the hemicellulose-derived sugar biomarkers, like cellulose, reflect the oxygen isotopic composition of plant source water altered by climatically controlled evapotranspirative 18O enrichment of leaf water. While relative air humidity controls most rigorously the evapotranspirative 18O enrichment, the direct temperature effect is less important. However, temperature can indirectly exert influence via plant physiological reactions, namely by influencing the transpiration rate which affects δ18Oleaf water due to the Péclet effect. In a companion paper (Tuthorn et al., this issue) we demonstrate the applicability of the hemicellulose-derived sugar biomarker δ18O method to soils and provide evidence from a climate transect study confirming that relative air humidity exerts the dominant control on evapotranspirative 18O

  7. General statistical framework for quantitative proteomics by stable isotope labeling.

    PubMed

    Navarro, Pedro; Trevisan-Herraz, Marco; Bonzon-Kulichenko, Elena; Núñez, Estefanía; Martínez-Acedo, Pablo; Pérez-Hernández, Daniel; Jorge, Inmaculada; Mesa, Raquel; Calvo, Enrique; Carrascal, Montserrat; Hernáez, María Luisa; García, Fernando; Bárcena, José Antonio; Ashman, Keith; Abian, Joaquín; Gil, Concha; Redondo, Juan Miguel; Vázquez, Jesús

    2014-03-07

    The combination of stable isotope labeling (SIL) with mass spectrometry (MS) allows comparison of the abundance of thousands of proteins in complex mixtures. However, interpretation of the large data sets generated by these techniques remains a challenge because appropriate statistical standards are lacking. Here, we present a generally applicable model that accurately explains the behavior of data obtained using current SIL approaches, including (18)O, iTRAQ, and SILAC labeling, and different MS instruments. The model decomposes the total technical variance into the spectral, peptide, and protein variance components, and its general validity was demonstrated by confronting 48 experimental distributions against 18 different null hypotheses. In addition to its general applicability, the performance of the algorithm was at least similar than that of other existing methods. The model also provides a general framework to integrate quantitative and error information fully, allowing a comparative analysis of the results obtained from different SIL experiments. The model was applied to the global analysis of protein alterations induced by low H₂O₂ concentrations in yeast, demonstrating the increased statistical power that may be achieved by rigorous data integration. Our results highlight the importance of establishing an adequate and validated statistical framework for the analysis of high-throughput data.

  8. Long-Term Precipitation Isotope Ratios (δ18O, δ2H, d-excess) in the Northeast US Reflect Atlantic Ocean Warming and Shifts in Moisture Sources

    NASA Astrophysics Data System (ADS)

    Puntsag, T.; Welker, J. M.; Mitchell, M. J.; Klein, E. S.; Campbell, J. L.; Likens, G.

    2014-12-01

    The global water cycle is exhibiting dramatic changes as global temperatures increase resulting in increases in: drought extremes, flooding, alterations in storm track patterns with protracted winter storms, and greater precipitation variability. The mechanisms driving these changes can be difficult to assess, but the spatial and temporal patterns of precipitation water isotopes18O, δ2H, d-excess) provide a means to help understand these water cycle changes. However, extended temporal records of isotope ratios in precipitation are infrequent, especially in the US. In our study we analyzed precipitation isotope ratio data from the Hubbard Brook Experimental Forest in New Hampshire that has the longest US precipitation isotope record, to determine: 1) the monthly composited averages and trends from 1967 to 2012 (45 years); ; 2) the relationships between abiotic properties such as local temperatures, precipitation type, storm tracks and isotope ratio changes; and 3) the influence of regional shifts in moisture sources and/or changes in N Atlantic Ocean water conditions on isotope values. The seasonal variability of Hubbard Brook precipitation isotope ratios is consistent with other studies, as average δ18O values are ~ -15‰ in January and ~ -5 ‰ in July. However, over the 45 year record there is a depletion trend in the δ 18O values (becoming isotopically lighter with a greater proportion of 16O), which coupled with less change in δ 2H leads to increases in d-excess values from ~ -10‰ around 1970 to greater than 10‰ in 2009. These changes occurred during a period of warming as opposed to cooling local temperatures indicating other processes besides temperature are controlling long-term water isotope traits in this region. We have evidence that these changes in precipitation isotope traits are controlled in large part by an increases in moisture being sourced from a warming N Atlantic Ocean that is providing evaporated, isotopically

  9. Characterization of biodegradation intermediates of nonionic surfactants by MALDI-MS. 2. Oxidative biodegradation profiles of uniform octylphenol polyethoxylate in 18O-labeled water.

    PubMed

    Sato, Hiroaki; Shibata, Atsushi; Wang, Yang; Yoshikawa, Hiromichi; Tamura, Hiroto

    2003-01-01

    This paper reports the characterization of the biodegradation intermediates of octylphenol octaethoxylate (OP(8)EO) by means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The biodegradation test study was carried out in a pure culture (Pseudomonas putida S-5) under aerobic conditions using OP(8)EO as the sole carbon source and (18)O-labeled water as an incubation medium. In the MALDI-MS spectra of biodegraded samples, a series of OP(n)EO molecules with n = 2-8 EO units and their corresponding carboxylic acid products (OP(n)EC) were observed. The use of purified OP(8)EO enabled one to distinguish the shortened OPEO molecules as biodegradation intermediates. Furthermore, the formation of OP(8)EC (the oxidized product of OP(8)EO) supported the notion that terminal oxidation is a step in the biodegradation process. When biodegradation study was carried out in (18)O-labeled water, incorporation of (18)O atoms into the carboxyl group was observed for OPEC, while no incorporation was observed for the shortened OPEO products. These results could provide some rationale to the biodegradation mechanism of alkylphenol polyethoxylates.

  10. The stable isotopic composition of Daphnia ephippia reflects changes in δ13C and δ18O values of food and water

    NASA Astrophysics Data System (ADS)

    Schilder, J.; Tellenbach, C.; Möst, M.; Spaak, P.; van Hardenbroek, M.; Wooller, M. J.; Heiri, O.

    2015-06-01

    The stable isotopic composition of fossil resting eggs (ephippia) of Daphnia spp. is being used to reconstruct past environmental conditions in lake ecosystems. However, the underlying assumption that the stable isotopic composition of the ephippia reflects the stable isotopic composition of the parent Daphnia, of their diet and of the environmental water have yet to be confirmed in a controlled experimental setting. We performed experiments with Daphnia pulicaria cultures, which included a control treatment conducted at 12 °C in filtered lake water and with a diet of fresh algae and three treatments in which we manipulated the stable carbon isotopic composition (δ13C value) of the algae, stable oxygen isotopic composition (δ18O value) of the water and the water temperature, respectively. The stable nitrogen isotopic composition (δ15N value) of the algae was similar for all treatments. At 12 °C, differences in algal δ13C values and in δ18O values of water were reflected in those of Daphnia. The differences between ephippia and Daphnia stable isotope ratios were similar in the different treatments (δ13C: +0.2 ± 0.4 ‰ (standard deviation); δ15N: -1.6 ± 0.4 ‰; δ18O: -0.9 ± 0.4 ‰), indicating that changes in dietary δ13C values and in δ18O values of water are passed on to these fossilizing structures. A higher water temperature (20 °C) resulted in lower δ13C values in Daphnia and ephippia than in the other treatments with the same food source and in a minor change in the difference between δ13C values of ephippia and Daphnia (to -1.3 ± 0.3 ‰). This may have been due to microbial processes or increased algal respiration rates in the experimental containers, which may not affect Daphnia in natural environments. There was no significant difference in the offset between δ18O and δ15N values of ephippia and Daphnia between the 12 and 20 °C treatments, but the δ18O values of Daphnia and ephippia were on average 1.2 ‰ lower at 20 °C than at

  11. sup 18 O/ sup 16 O and sup 13 C/ sup 12 C in lower Paleozoic articulate brachiopods: Implications for the isotopic composition of seawater

    SciTech Connect

    Wadleigh, M.A. ); Veizer, J. Ruhr Univ., Bochum )

    1992-01-01

    Three hundred and eleven articulate brachiopods, with extensive geographic coverage, spanning the Cambrian to Silurian stratigraphic interval, were analyzed for oxygen and carbon isotopic composition. Cambrian samples have {delta}{sup 18}O {le} {minus}7{per thousand}, Ordovician samples {le} {minus}2.4{per thousand}, and Silurian samples {le} {minus}1.9{per thousand}, confirming the previously established trend towards lighter isotopic compositions with increasing age throughout the Lower Paleozoic. Forty-nine best preserved' Ordovician and Silurian samples were identified based on scanning electron microscopy and trace element analysis. They were found to bracket the isotopic compositions of over 85% of specimens from these stratigraphic intervals supporting widespread preservation of the isotopic signal. Some latest Ordovician and Lower Ludlovian samples associated with shale sequences are apparently enriched' in {sup 18}O. These are interpreted as an environmental phenomenon, perhaps related to water temperature and glaciation. A number of Silurian samples of varying genera and stratigraphic levels are highly enriched in {sup 13}C, up to +6{per thousand}. Some are shale related, but some are associated with carbonate-bearing basins. These are also thought to represent near-original' compositions, but a single environmental cause is unknown. The present data show that luminescence is not a decisive criterion for evaluating the degree of brachiopod preservation. Whole-shell values were isotopically similar to their nonluminescent portions for both oxygen and carbon.

  12. Reconstructing lake evaporation history and the isotopic composition of precipitation by a coupled δ18O-δ2H biomarker approach

    NASA Astrophysics Data System (ADS)

    Hepp, Johannes; Tuthorn, Mario; Zech, Roland; Mügler, Ines; Schlütz, Frank; Zech, Wolfgang; Zech, Michael

    2015-10-01

    Over the past decades, δ18O and δ2H analyses of lacustrine sediments became an invaluable tool in paleohydrology and paleolimnology for reconstructing the isotopic composition of past lake water and precipitation. However, based on δ18O or δ2H records alone, it can be challenging to distinguish between changes of the precipitation signal and changes caused by evaporation. Here we propose a coupled δ18O-δ2H biomarker approach that provides the possibility to disentangle between these two factors. The isotopic composition of long chain n-alkanes (n-C25, n-C27, n-C29, n-C31) were analyzed in order to establish a 16 ka Late Glacial and Holocene δ2H record for the sediment archive of Lake Panch Pokhari in High Himalaya, Nepal. The δ2Hn-alkane record generally corroborates a previously established δ18Osugar record reporting on high values characterizing the deglaciation and the Older and the Younger Dryas, and low values characterizing the Bølling and the Allerød periods. Since the investigated n-alkane and sugar biomarkers are considered to be primarily of aquatic origin, they were used to reconstruct the isotopic composition of lake water. The reconstructed deuterium excess of lake water ranges from +57‰ to -85‰ and is shown to serve as proxy for the evaporation history of Lake Panch Pokhari. Lake desiccation during the deglaciation, the Older Dryas and the Younger Dryas is affirmed by a multi-proxy approach using the Hydrogen Index (HI) and the carbon to nitrogen ratio (C/N) as additional proxies for lake sediment organic matter mineralization. Furthermore, the coupled δ18O and δ2H approach allows disentangling the lake water isotopic enrichment from variations of the isotopic composition of precipitation. The reconstructed 16 ka δ18Oprecipitation record of Lake Panch Pokhari is well in agreement with the δ18O records of Chinese speleothems and presumably reflects the Indian Summer Monsoon variability.

  13. Can tree-ring isotopes18O and δ13C) improve our understanding of hydroclimate variability in the Columbia River Basin?

    NASA Astrophysics Data System (ADS)

    Csank, A. Z.; Wise, E.; McAfee, S. A.

    2015-12-01

    The trajectory of incoming storms from the Pacific Ocean has a strong impact on hydroclimate in the Pacific Northwest. Shifts between zonal and meridional flow are a key influence on drought and pluvial regimes in both the PNW and the western United States as a whole. Circulation-dependent variability in the isotopic composition of precipitation can be recorded and potentially reconstructed using δ18O records derived from tree-rings. Here we present isotopic records of δ18O and δ13C from ponderosa pine (Pinus ponderosa) for the period 1950-2013 from six sites located in the lee of the Cascades in eastern Washington. Because of the orientation of the Cascades, zonal flow will result in an intensified rain shadow whereas meridional flow allows moisture to penetrate at a lower elevation leading to a lower rainout effect. This means zonal flow results in drier conditions in eastern Washington and the converse for meridional flow. We hypothesized that more depleted precipitation δ18O values will occur with periods of more zonal flow across the PNW and will be recorded by trees at our sites. Results show a strong relationship between our δ18O chronologies and winter precipitation (R = -0.50; p<0.001). δ13C chronologies from the same trees showed a relationship to prior fall/winter (pOct-pDec) precipitation (R = -0.46; p<0.005) suggesting a possible link to antecedent moisture conditions. With a focus on years with clear zonal and meridional flow regimes, we regressed the tree-ring δ18O anomaly against the instrumental record of total precipitation and compared the residual series to records of storm track for the period 1978-2008, and we found a detectable signal where the most depleted δ18O was generally associated with zonal flow and the most enriched δ18O with meridional flow. However, there are still some years where the relationship is unclear. Further work is aimed at understanding these anomalous years and extending our record beyond the instrumental

  14. What can Δ 15N and Δ 18O isotopes tell us about sources, transport, and fate of nitrate in the Mississippi River Basin?

    NASA Astrophysics Data System (ADS)

    Battaglin, W. A.

    2003-12-01

    Water and nutrients, primarily nitrate (NO3) in Mississippi River discharge, affect the size and severity of the Gulf of Mexico hypoxic (depleted dissolved oxygen) zone. Approximately 120 water samples were collected from 16 sites on small streams and 6 sites on large rivers within the Mississippi River Basin in 1997-98 to see if NO3 sources and transformations can be identified using the stable isotopic ratios Δ 15N and Δ 18O. Results from Lagrangian sampling at the large river sites indicate that nitrate mass decreases slightly, while Δ 15N and Δ 18O isotope ratios are unchanged in the 1500 river kilometers between the Upper Mississippi-Ohio River confluence and the Gulf of Mexico. Results also show that Δ 15N and Δ 18O values from small streams draining lands dominated by row crops or livestock tended to be distinct from those dominated by urban or undeveloped land. Mean Δ 15N values at the 16 sites on small streams were most strongly correlated (Pearson's r) with manure production rate (0.64), percent residential land use (-0.45), and urea use rate (0.43). The best multiple linear regression (MLR) model for mean Δ 15N values (r2=0.69) used manure production rate and ammonium nitrate use rate as explanatory variables. Mean Δ 18O values were most strongly correlated with percent wetlands (0.72), mean NO3 concentration (-0.71), and percent residential land use (0.58). The best MLR model for mean Δ 18O values (r2=0.85) used percent residential land use, percent wetlands, and ammonium nitrate use rate as explanatory variables. Mean NO3 concentrations were most strongly correlated with percent row-crops land use (0.84), nitrogen-fertilizer use rate (0.74), and hog-manure production rate (0.66). The best MLR model for mean NO3 concentration (r2=0.85) used percent row-crops land use and percent grain-crops land use as explanatory variables. MLR equations developed from the 16 smaller streams were used to predict mean Δ 15N and Δ 18O values and NO3

  15. Extraction of CO2 from air samples for isotopic analysis and limits to ultra high precision delta18O determination in CO2 gas.

    PubMed

    Werner, R A; Rothe, M; Brand, W A

    2001-01-01

    The determination of delta18O values in CO2 at a precision level of +/-0.02 per thousand (delta-notation) has always been a challenging, if not impossible, analytical task. Here, we demonstrate that beyond the usually assumed major cause of uncertainty - water contamination - there are other, hitherto underestimated sources of contamination and processes which can alter the oxygen isotope composition of CO2. Active surfaces in the preparation line with which CO2 comes into contact, as well as traces of air in the sample, can alter the apparent delta18O value both temporarily and permanently. We investigated the effects of different surface materials including electropolished stainless steel, Duran glass, gold and quartz, the latter both untreated and silanized. CO2 frozen with liquid nitrogen showed a transient alteration of the 18O/16O ratio on all surfaces tested. The time to recover from the alteration as well as the size of the alteration varied with surface type. Quartz that had been ultrasonically cleaned for several hours with high purity water (0.05 microS) exhibited the smallest effect on the measured oxygen isotopic composition of CO2 before and after freezing. However, quartz proved to be mechanically unstable with time when subjected to repeated large temperature changes during operation. After several days of operation the gas released from the freezing step contained progressively increasing trace amounts of O2 probably originating from inclusions within the quartz, which precludes the use of quartz for cryogenically trapping CO2. Stainless steel or gold proved to be suitable materials after proper pre-treatment. To ensure a high trapping efficiency of CO2 from a flow of gas, a cold trap design was chosen comprising a thin wall 1/4" outer tube and a 1/8" inner tube, made respectively from electropolished stainless steel and gold. Due to a considerable 18O specific isotope effect during the release of CO2 from the cold surface, the thawing time had to

  16. Use of 2H and 18O stable isotopes to investigate water sources for different ages of Populus euphratica along the lower Heihe River

    USGS Publications Warehouse

    Shubao Liu,; Yaning Chen,; Yapeng Chen,; Friedman, Jonathan M.; Gonghuan Fan,; Hati, Jarre Heng A.

    2015-01-01

    Investigation of the water sources used by trees of different ages is essential to formulate a conservation strategy for the riparian tree, P. euphratica. This study addressed the contributions of different potential water sources to P. euphratica based on levels of stable oxygen and hydrogen isotopes18O, δ2H) in the xylem of different aged P. euphratica, as well as in soil water and groundwater along the lower Heihe River. We found significant differences in δ18O values in the xylem of different aged P. euphratica. Specifically, the δ18O values of young, mature and over-mature forests were −5.368(±0.252) ‰, −6.033(± 0.185) ‰ and −6.924 (± 0.166) ‰, respectively, reflecting the reliance of older trees on deeper sources of water with a δ18O value closer to that of groundwater. Different aged P. euphratica used different water sources, with young forests rarely using groundwater (mean <15 %) and instead primarily relying on soil water from a depth of 0–50 cm (mean >45 %), and mature and over-mature forests using water from deeper than 100 cm derived primarily from groundwater.

  17. Stable Oxygen (δ 18O) and Carbon (δ 13C) Isotopes in the Skeleton of Bleached and Recovering Corals From Hawaii

    NASA Astrophysics Data System (ADS)

    Rodrigues, L. J.; Grottoli, A. G.

    2004-12-01

    Coral skeletal stable oxygen isotopes18O) reflect changes in seawater temperature and salinity, while stable carbon isotopes (δ 13C) reflect a combination of both metabolic (photosynthesis and feeding) and kinetic fractionation. Together, the two isotopic signatures may be used as a proxy for past bleaching events. During bleaching, increased seawater temperatures often contribute to a decline in zooxanthellae and/or chlorophyll concentrations, resulting in a decrease in photosynthesis. We experimentally investigated the effect of bleaching and subsequent recovery on the δ 13C and δ 18O values of coral skeleton. Fragments from two coral species (Montipora capitata and Porites compressa) from Kaneohe Bay, Hawaii were bleached in outdoor tanks by raising the seawater temperature to 30° C. Additional fragments from the same parent colonies were maintained at ambient seawater temperatures (27° C) in separate tanks as controls. After one month in the tanks, a subset of the fragments was frozen and all remaining fragments were placed back on the reef to recover. All coral fragments were analyzed for their skeletal δ 13C and δ 18O compositions at five time intervals: before, immediately after, 1.5, 4, and 8 months after bleaching. In addition, rates of photosynthesis, calcification, and heterotrophy were also measured. Immediately after bleaching, δ 18O decreased in bleached M. capitata relative to controls, reflecting their exposure to increased seawater temperatures. During recovery, δ 18O values in the treatment M. capitata were not different from the controls. In P. compressa, δ 18O did not significantly differ in bleached and control corals at any time during the experiment. Immediately after bleaching, δ 13C decreased in the bleached fragments of both species relative to controls reflecting decreased photosynthetic rates. However, during recovery δ 13C in both species was greater in bleached than control fragments despite photosynthesis remaining

  18. Chemical weathering and the role of sulfuric and nitric acids in carbonate weathering: Isotopes (13C, 15N, 34S, and 18O) and chemical constraints

    NASA Astrophysics Data System (ADS)

    Li, Cai; Ji, Hongbing

    2016-05-01

    Multiple isotopes (13C-DIC, 34S and 18O-SO42-, 15N and 18O-NO3-) and water chemistry were used to evaluate weathering rates and associated CO2 consumption by carbonic acid and strong acids (H2SO4 and HNO3) in a typical karst watershed (Wujiang River, Southwest China). The dual sulfate isotopes indicate that sulfate is mainly derived from sulfide oxidation in coal stratum and sulfide-containing minerals, and dual nitrate isotopes indicate that nitrate is mainly derived from soil N and nitrification. The correlation between isotopic compositions and water chemistry suggests that sulfuric and nitric acids, in addition to carbonic acid, are involved in carbonate weathering. The silicate and carbonate weathering rates are 7.2 t km-2 yr-1 and 76 t km-2 yr-1, respectively. In comparison with carbonate weathering rates (43 t km-2 yr-1) by carbonic acid alone, the subsequent increase in rates indicates significant enhancement of weathering when combined with sulfuric and nitric acids. Therefore, the role of sulfuric and nitric acids in the rock weathering should be considered in the global carbon cycle.

  19. Autonomous Instrumentation for Fast, Continuous and Accurate Isotopic Measurements of Water Vapor (δ18O, δ 2H, H2O) in the Field

    NASA Astrophysics Data System (ADS)

    Liem, J. S.; Dong, F.; Owano, T. G.; Baer, D. S.

    2010-12-01

    Stable isotopes of water vapor are powerful tracers to investigate the hydrological cycle and ecological processes. Therefore, continuous, in-situ and accurate measurements of δ18O and δ2H are critical to advance the understanding of water-cycle dynamics worldwide. Furthermore, the combination of meteorological techniques and high-frequency isotopic water measurements can provide detailed time-resolved information on the eco-physiological performance of plants and enable improved understanding of water fluxes at ecosystem scales. In this work, we present recent development and field deployment of a novel Water Vapor Isotope Measurement System (WVIMS) capable of simultaneous in situ measurements of δ18O and δ2H and water mixing ratio (H2O) with high precision, accuracy and speed (up to 10 Hz measurement rate). The WVIMS consists of an Analyzer (Water Vapor Isotope Analyzer), based on cavity enhanced laser absorption spectroscopy, and a Standard Source (Water Vapor Isotope Standard Source), based on quantitative evaporation of a liquid water standard (with known isotopic content), and operates in a dual-inlet configuration. The WVIMS automatically controls the entire sample and data collection, data analysis and calibration process to allow for continuous, autonomous unattended long-term operation. The WVIMS has been demonstrated for accurate (i.e. fully calibrated) measurements ranging from 500 ppmv (typical of arctic environments) to over 30,000 ppmv (typical of tropical environments) in air. Dual-inlet operation, which involves regular calibration with isotopic water vapor reference standards, essentially eliminates measurement drift, ensures data reliability, and allows operation over an extremely wide ambient temperature range (5-45C). This presentation will include recent measurements recorded using the WVIMS in plant growth chambers and in arctic environments. The availability of this new instrumentation provides new opportunities for detailed continuous

  20. Coordinated Isotopic and Mineral Characterization of Highly Fractionated 18O-Rich Silicates in the Queen Alexandra Range 99177 CR3 Chondrite

    NASA Technical Reports Server (NTRS)

    Nguyen, A. N.; Keller, L. P.; Messenger, S.; Rahman, Z.

    2016-01-01

    Carbonaceous chondrites contain a mixture of solar system condensates, pre-solar grains, and primitive organic matter. Each of these materials record conditions and processes in different regions of the solar nebula, on the meteorite parent body, and beyond the solar system. Oxygen isotopic studies of meteorite components can trace interactions of distinct oxygen isotopic reservoirs in the early solar system and secondary alteration processes. The O isotopic compositions of the earliest solar system condensates fall along a carbonaceous chondrite anhydrous mineral (CCAM) line of slope approximately 1 in a plot of delta 17O against delta 18O. This trend is attributed to mixing of material from 16O-poor and 16O-rich reservoirs. Secondary processing can induce mass-dependent fractionation of the O isotopes, shifting these compositions along a line of slope approximately 0.52. Substantial mass-dependent fractionation of O isotopes has been observed in secondary minerals in CAIs, calcite, and FUN inclusions. These fractionations were caused by significant thermal or aqueous alteration. We recently reported the identification of four silicate grains with extremely fractionated O isotopic ratios (delta 18O equals 37 - 55 per mille) in the minimally altered CR3 chondrite QUE 99177. TEM analysis of one grain indicates it is a nebular condensate that did not experience substantial alteration. The history of these grains is thus distinct from those of the aforementioned fractionated materials. To constrain the origin of the silicate grains, we conducted further Mg and Fe isotopic studies and TEM analyses of two grains.

  1. Sm-Nd, Rb-Sr, and /sup 18/O//sup 16/O isotopic systematics in an oceanic crustal section: Evidence from the Samial ophiolite

    SciTech Connect

    McCulloch, M.T.; Gregory, R.T.; Wasserburg, G.J.; Taylor, H.P. Jr.

    1981-04-10

    The Sm-Nd, Rb-Sr, and /sup 18/O//sup 16/O isotopic systems have been used to distinguish between the effects of seafloor hydrothermal alteration and primary magmatic isotopic variations. The Sm-Nd isotopic system is essentially unaffected by seawater alteration, while the Rb-Sr and /sup 18/O//sup 16/O systems are sensitive to hydrothermal interactions with seawater. Sm-Nd mineral isochrons from the cumulate gabbros of the Samail ophiolite have an initial /sup 143/Nd//sup 144/Nd ratio of e/sub Nd/ = 7.8 +- 0.3, which clearly substantiates the oceanic affinity of this complex. The initial /sup 143/Nd//sup 144/Nd ratios for the harzburgite, plagiogranite, sheeted diabase dikes, and basalt units have a limited range in e/sub Nd/ of from 7.5 to 8.6, indicating that all the lithologies have distinctive oceanic affinities, although there is also some evidence for small isotopic heterogeneities in the magma reservoirs. The Sm-Nd mineral isochrons give crystallization ages of 128 +- 20 m.y. and 150 +- 40 m.y. from Ibra and 100 +- 20 m.y. from Wadi Fizh, which is approximately 300 km NW of Ibra. These crystallization ages are interpreted as the time of formation of the oceanic crust. The /sup 87/Sr//sup 86/Sr initial ratios on the same rocks have an extremely large range of from 0.7030 to 0.7065 and the d/sup 18/O values vary from 2.6 to 12.7. These large variations clearly demonstrate hydrothermal interaction of oceanic crust with seawater.

  2. Prediction of plant vulnerability to salinity increase in a coastal ecosystem by stable isotopic composition (δ18O) of plant stem water: a model study

    USGS Publications Warehouse

    Zhai, Lu; Jiang, Jiang; DeAngelis, Don; Sternberg, Leonel d.S.L

    2016-01-01

    Sea level rise and the subsequent intrusion of saline seawater can result in an increase in soil salinity, and potentially cause coastal salinity-intolerant vegetation (for example, hardwood hammocks or pines) to be replaced by salinity-tolerant vegetation (for example, mangroves or salt marshes). Although the vegetation shifts can be easily monitored by satellite imagery, it is hard to predict a particular area or even a particular tree that is vulnerable to such a shift. To find an appropriate indicator for the potential vegetation shift, we incorporated stable isotope 18O abundance as a tracer in various hydrologic components (for example, vadose zone, water table) in a previously published model describing ecosystem shifts between hammock and mangrove communities in southern Florida. Our simulations showed that (1) there was a linear relationship between salinity and the δ18O value in the water table, whereas this relationship was curvilinear in the vadose zone; (2) hammock trees with higher probability of being replaced by mangroves had higher δ18O values of plant stem water, and this difference could be detected 2 years before the trees reached a tipping point, beyond which future replacement became certain; and (3) individuals that were eventually replaced by mangroves from the hammock tree population with a 50% replacement probability had higher stem water δ18O values 3 years before their replacement became certain compared to those from the same population which were not replaced. Overall, these simulation results suggest that it is promising to track the yearly δ18O values of plant stem water in hammock forests to predict impending salinity stress and mortality.

  3. Oxygen isotope evolution of the Lake Owyhee volcanic field, Oregon, and implications for the low-δ18O magmatism of the Snake River Plain-Yellowstone hotspot and other low-δ18O large igneous provinces

    NASA Astrophysics Data System (ADS)

    Blum, Tyler B.; Kitajima, Kouki; Nakashima, Daisuke; Strickland, Ariel; Spicuzza, Michael J.; Valley, John W.

    2016-11-01

    The Snake River Plain-Yellowstone (SRP-Y) hotspot track represents the largest known low-δ18O igneous province; however, debate persists regarding the timing and distribution of meteoric hydrothermal alteration and subsequent melting/assimilation relative to hotspot magmatism. To further constrain alteration relations for SRP-Y low-δ18O magmatism, we present in situ δ18O and U-Pb analyses of zircon, and laser fluorination δ18O analyses of phenocrysts, from the Lake Owyhee volcanic field (LOVF) of east-central Oregon. U-Pb data place LOVF magmatism between 16.3 and 15.4 Ma, and contain no evidence for xenocrystic zircon. LOVF δ18O(Zrc) values demonstrate (1) both low-δ18O and high-δ18O caldera-forming and pre-/post-caldera magmas, (2) relative increases in δ18O between low-δ18O caldera-forming and post-caldera units, and (3) low-δ18O magmatism associated with extension of the Oregon-Idaho Graben. The new data, along with new compilations of (1) in situ zircon δ18O data for the SRP-Y, and (2) regional δ18O(WR) and δ18O(magma) patterns, further constrain the thermal and structural associations for hydrothermal alteration in the SRP-Y. Models for low-δ18O magmatism must be compatible with (1) δ18O(magma) trends within individual SRP-Y eruptive centers, (2) along axis trends in δ18O(magma), and (3) the high concentration of low-δ18O magmas relative to the surrounding regions. When considered with the structural and thermal evolution of the SRP-Y, these constraints support low-δ18O magma genesis originating from syn-hotspot meteoric hydrothermal alteration, driven by hotspot-derived thermal fluxes superimposed on extensional tectonics. This model is not restricted to continental hotspot settings and may apply to several other low-δ18O igneous provinces with similar thermal and structural associations.

  4. Experimental assessment of the purity of α-cellulose produced by variations of the Brendel method: Implications for stable isotope (δ13C, δ18O) dendroclimatology

    NASA Astrophysics Data System (ADS)

    Brookman, Tom; Whittaker, Thomas

    2012-09-01

    Stable isotope dendroclimatology using α-cellulose has unique potential to deliver multimillennial-scale, sub-annually resolved, terrestrial climate records. However, lengthy processing and analytical methods often preclude such reconstructions. Variants of the Brendel extraction method have reduced these limitations, providing fast, easy methods of isolating α-cellulose in some species. Here, we investigate application of Standard Brendel (SBrendel) variants to resinous soft-woods by treating samples of kauri (Agathis australis), ponderosa pine (Pinus ponderosa) and huon pine (Lagarastrobus franklinii), varying reaction vessel, temperature, boiling time and reagent volume. Numerous samples were visibly `under-processed' and Fourier Transform infrared spectroscopic (FTIR) investigation showed absorption peaks at 1520 cm-1 and ˜1600 cm-1 in those fibers suggesting residual lignin and retained resin respectively. Replicate analyses of all samples processed at high temperature yielded consistent δ13C and δ18O despite color and spectral variations. Spectra and isotopic data revealed that α-cellulose δ13C can be altered during processing, most likely due to chemical contamination from insufficient acetone removal, but is not systematically affected by methodological variation. Reagent amount, temperature and extraction time all influence δ18O, however, and our results demonstrate that different species may require different processing methods. FTIR prior to isotopic analysis is a fast and cost effective way to determine α-cellulose extract purity. Furthermore, a systematic isotopic test such as we present here can also determine sensitivity of isotopic values to methodological variables. Without these tests, isotopic variability introduced by the method could obscure or `create' climatic signals within a data set.

  5. UV-laser microdissection system - A novel approach for the preparation of high-resolution stable isotope records (δ13C/δ18O) from tree rings

    NASA Astrophysics Data System (ADS)

    Schollaen, Karina; Helle, Gerhard

    2013-04-01

    Intra-annual stable isotope (δ13C and δ18O) studies of tree rings at various incremental resolutions have been attempting to extract valuable seasonal climatic and environmental information or assessing plant ecophysiological processes. For preparing high-resolution isotope samples normally wood segments or cores are mechanically divided in radial direction or cut in tangential direction. After mechanical dissection, wood samples are ground to a fine powder and either cellulose is extracted or bulk wood samples are analyzed. Here, we present a novel approach for the preparation of high-resolution stable isotope records from tree rings using an UV-laser microdissection system. Firstly, tree-ring cellulose is directly extracted from wholewood cross-sections largely leaving the wood anatomical structure intact and saving time as compared to the classical procedure. Secondly, micro-samples from cellulose cross-sections are dissected with an UV-Laser dissection microscope. Tissues of interest from cellulose cross-sections are identified and marked precisely with a screen-pen and dissected via an UV-laser beam. Dissected cellulose segments were automatically collected in capsules and are prepared for stable isotope (δ13C and δ18O) analysis. The new techniques facilitate inter- and intra-annual isotope analysis on tree-ring and open various possibilities for comparisons with wood anatomy in plant eco-physiological studies. We describe the design and the handling of this novel methodology and discuss advantages and constraints given by the example of intra-annual oxygen isotope analysis on tropical trees.

  6. Localization of groundwater infiltration in the combined sewers of Brussels by stable isotopes measurements (δ18O, δD) by Cavity Ring Down Spectroscopy.

    NASA Astrophysics Data System (ADS)

    De Bondt, Kevin; Claeys, Philippe

    2014-05-01

    In the last 20 years research has been conducted to quantify the infiltration of groundwater into the sewers. This groundwater, called parasitic water, increases the volume of waste-water to be treated and consequently the cost of this treatment. Moreover, in the case of combined sewer systems, the parasitic water also limits the sewer capacity and indirectly increases the risks of combined sewer overflows and floods. The infiltration of groundwater occurs trough cracks, sewer collapses and from direct connections with old springs. Different methods quantify the intrusion of parasitic water. Among these, the use of the stable isotopes of water (δ18O & δD) shows good result in catchments or cities close to Mountainous regions (example from Lyon, Zurich), where isotopic signals vary significantly because of continental and altitude effects. However many cities, such as Brussels, are located in more oceanic settings and theoretically offer less potential for the application of the stable isotopes method. In the case of Brussels, river-water from the Meuse is used to produce domestic-water. The catchment of this river extends into the Ardennes, which are affected by slightly different climatic conditions. δ18O & δD analyzes of groundwater from the main aquifer (Ledo-Paniselian-Brusselian) and domestic-water from the Callois reservoir fed by the Meuse River show sufficient isotopic differences in the south of Brussels, but only during the summer. The discrimination potential is better with δD than with δ18O. The improvement of δD measurements (precision, costs,...) brought by Cavity Ring Down Spectroscopy largely contributes to the potential of using stable isotopes method to trace water in Brussels. The first campaigns in the sewers also show a little enrichment (in heavy isotopes) of the waste-water in comparison with the reservoir waters and tap waters. This increases the potential of the method but constrains the sampling to pure waste-water in sewer segments

  7. Determination of energy expenditure during heavy exercise, normal daily activity, and sleep using the doubly-labelled-water (/sup 2/H/sub 2/ 18O) method

    SciTech Connect

    Stein, T.P.; Hoyt, R.W.; Settle, R.G.; O'Toole, M.; Hiller, W.D.

    1987-03-01

    Energy expenditure of four subjects was measured by the doubly-labelled-water (/sup 2/H/sub 2/ 18O) method to determine if energy expenditure could be determined over short periods. Three subjects were studied while they performed 8 h of heavy exercise in a laboratory environment. Urine and blood samples were taken before and after exercise. Estimated energy expended during 8 h of high-intensity exercise for three subjects was 757 +/- 118 kcal/h by the doubly-labelled-water method using urine and a two-point calculation, which compared favorably with 735 +/- 82 kcal/h obtained by respiratory gas exchange. For the fourth subject, daytime, nighttime, and daily energy expenditure was calculated by both the two-pair method and decay-curve analysis of urine and saliva samples collected in the morning and at night. Daytime and nighttime energy expenditures differed significantly (p less than 0.05).

  8. Quantitative proteomics by 2-DE, 16O/18O labelling and linear ion trap mass spectrometry analysis of lymph nodes from piglets inoculated by porcine circovirus type 2.

    PubMed

    Ramírez-Boo, María; Núnez, Estefania; Jorge, Inmaculada; Navarro, Pedro; Fernandes, Lana T; Segalés, Joaquim; Garrido, Juan J; Vázquez, Jesús; Moreno, Angela

    2011-09-01

    Porcine circovirus type 2 (PCV2) has been identified as the essential causal agent of postweaning multisystemic wasting syndrome. However, little is known regarding the mechanism(s) underlying the pathogenesis of PCV2-induced disease and the interaction of the virus with the host immune system. Here, we present a proteomics study on inguinal lymph nodes of piglets inoculated with PCV2, in order to better understand the pathogenesis of postweaning multisystemic wasting syndrome and the pathways might be affected after infection. We used two proteomics strategies, 2-DE and 1-DE followed by (16)O/(18)O peptide labelling and peptide identification and quantification by MS. More than 100 proteins were found to be differentially regulated and the results obtained by the two strategies were fairly concordant but also complementary, the (18)O labelling approach being a more robust alternative. Analysis of these proteins by systems biology tools revealed the implication of acute phase response and NrF2-mediated oxidative stress, suggesting a putative role for these pathways in the pig immune response. Besides, CD81 was found to be up-regulated, suggesting a possible role in the internalization of the virus. The use of proteomics technologies together with biology analysis systems opens up the way to gain more exhaustive and systematic knowledge of virus-pathogen interactions.

  9. Applications and Advantages of Stable Isotope Phosphate Labeling of RNA in Mass Spectrometry.

    PubMed

    Borland, Kayla; Limbach, Patrick A

    2017-04-01

    Mass spectrometry (MS) has become an enabling technology for the characterization of post-transcriptionally modified nucleosides within ribonucleic acids (RNAs). These modified RNAs tend to be more challenging to completely characterize using conventional genomic-based sequencing technologies. As with many biological molecules, information relating to the presence or absence of a particular compound (i.e., qualitative measurement) is only one step in sample characterization. Additional useful information is found by performing quantitative measurements on the levels of the compound of interest in the sample. Phosphate labeling of modified RNAs has been developed by our laboratory to enhance conventional mass spectrometry techniques. By taking advantage of the mechanism of action of many ribonucleases (RNases), digesting RNA samples in the presence of (18)O-labeled water generates an (18)O-labeled 3'-phosphate in each digestion product. We describe the historical development of this approach, contrast this stable isotope labeling strategy with others used in RNA mass spectrometry, and provide examples of new analytical mass spectrometry methods that are enabled by phosphate labeling in this fashion.

  10. Unveiling stomata 24/7: can we use carbonyl sulfide (COS) and oxygen isotopes (18O) to constrain estimates of nocturnal transpiration across different evolutionary plant forms?

    NASA Astrophysics Data System (ADS)

    Gimeno, Teresa E.; Ogee, Jerome; Bosc, Alexander; Genty, Bernard; Wohl, Steven; Wingate, Lisa

    2015-04-01

    Numerous studies have reported a continued flux of water through plants at night, suggesting that stomata are not fully closed. Growing evidence indicates that this nocturnal flux of transpiration might constitute an important fraction of total ecosystem water use in certain environments. However, because evaporative demand is usually low at night, nocturnal transpiration fluxes are generally an order of magnitude lower than rates measured during the day and perilously close to the measurement error of traditional gas-exchange porometers. Thus estimating rates of stomatal conductance in the dark (gnight) precisely poses a significant methodological challenge. As a result, we lack accurate field estimates of gnight and how it responds to different atmospheric drivers, indicating the need for a different measurement approach. In this presentation we propose a novel method to obtain detectable and robust estimates of gnight. We will demonstrate using mechanistic theory how independent tracers including the oxygen isotope composition of CO2 (δ18O) and carbonyl sulfide (COS) can be combined to obtain robust estimates of gnight. This is because COS and CO18O exchange within leaves are controlled by the light insensitive enzyme carbonic anhydrase. Thus, if plant stomata are open in the dark we will continue to observe COS and CO18O exchange. Using our theoretical model we will demonstrate that the exchange of these tracers can now be measured using advances in laser spectrometry techniques at a precision high enough to determine robust estimates of gnight. We will also present our novel experimental approach designed to measure simultaneously the exchange of CO18O and COS alongside the conventional technique that relies on measuring the total water flux from leaves in the dark. Using our theoretical approach we will additionally explore the feasibility of our proposed experimental design to detect variations in gnight during drought stress and across a variety of plant

  11. The suitability of the dual isotope approach (δ13C and δ18O) in tree ring studies

    NASA Astrophysics Data System (ADS)

    Siegwolf, Rolf; Saurer, Matthias

    2016-04-01

    The use of stable isotopes, complementary to tree ring width data in tree ring research has proven to be a powerful tool in studying the impact of environmental parameters on tree physiology and growth. These three proxies are thus instrumental for climate reconstruction and improve the understanding of underlying causes of growth changes. In various cases, however, their use suggests non-plausible interpretations. Often the use of one isotope alone does not allow the detection of such "erroneous isotope responses". A careful analysis of these deviating results shows that either the validity of the carbon isotope discrimination concept is no longer true (Farquhar et al. 1982) or the assumptions for the leaf water enrichment model (Cernusak et al., 2003) are violated and thus both fractionation models are not applicable. In this presentation we discuss such cases when the known fractionation concepts fail and do not allow a correct interpretation of the isotope data. With the help of the dual isotope approach (Scheidegger et al.; 2000) it is demonstrated, how to detect and uncover the causes for such anomalous isotope data. The fractionation concepts and their combinations before the background of CO2 and H2O gas exchange are briefly explained and the specific use of the dual isotope approach for tree ring data analyses and interpretations are demonstrated. References: Cernusak, L. A., Arthur, D. J., Pate, J. S. and Farquhar, G. D.: Water relations link carbon and oxygen isotope discrimination to phloem sap sugar concentration in Eucalyptus globules, Plant Physiol., 131, 1544-1554, 2003. Farquhar, G. D., O'Leary, M. H. and Berry, J. A.: On the relationship between carbon isotope discrimination and the intercellular carbon dioxide concentration in leaves, Aust. J. Plant Physiol., 9, 121-137, 1982. Scheidegger, Y., Saurer, M., Bahn, M. and Siegwolf, R.: Linking stable oxygen and carbon isotopes with stomatal conductance and photosynthetic capacity: A conceptual model

  12. Oxygen isotope ratios (18O/16O) of hemicellulose-derived sugar biomarkers in plants, soils and sediments as paleoclimate proxy II: Insight from a climate transect study

    NASA Astrophysics Data System (ADS)

    Tuthorn, Mario; Zech, Michael; Ruppenthal, Marc; Oelmann, Yvonne; Kahmen, Ansgar; Valle, Héctor Francisco del; Wilcke, Wolfgang; Glaser, Bruno

    2014-02-01

    The oxygen isotopic composition of precipitation (δ18Oprec) is well known to be a valuable (paleo-)climate proxy. Paleosols and sediments and hemicelluloses therein have the potential to serve as archives recording the isotopic composition of paleoprecipitation. In a companion paper (Zech et al., 2014) we investigated δ18Ohemicellulose values of plants grown under different climatic conditions in a climate chamber experiment. Here we present results of compound-specific δ18O analyses of arabinose, fucose and xylose extracted from modern topsoils (n = 56) along a large humid-arid climate transect in Argentina in order to answer the question whether hemicellulose biomarkers in soils reflect δ18Oprec. The results from the field replications indicate that the homogeneity of topsoils with regard to δ18Ohemicellulose is very high for most of the 20 sampling sites. Standard deviations for the field replications are 1.5‰, 2.2‰ and 1.7‰, for arabinose, fucose and xylose, respectively. Furthermore, all three hemicellulose biomarkers reveal systematic and similar trends along the climate gradient. However, the δ18Ohemicellulose values (mean of the three sugars) do not correlate positively with δ18Oprec (r = -0.54, p < 0.014, n = 20). By using a Péclet-modified Craig-Gordon (PMCG) model it can be shown that the δ18Ohemicellulose values correlate highly significantly with modeled δ18Oleaf water values (r = 0.81, p < 0.001, n = 20). This finding suggests that hemicellulose biomarkers in (paleo-)soils do not simply reflect δ18Oprec but rather δ18Oprec altered by evaporative 18O enrichment of leaf water due to evapotranspiration. According to the modeling results, evaporative 18O enrichment of leaf water is relatively low (˜10‰) in the humid northern part of the Argentinian transect and much higher (up to 19‰) in the arid middle and southern part of the transect. Model sensitivity tests corroborate that changes in relative air humidity exert a dominant

  13. Groundwater discharge and hydrologic partition of the lakes in desert environment: Insights from stable 18O/2H and radium isotopes

    NASA Astrophysics Data System (ADS)

    Luo, Xin; Jiao, Jiu Jimmy; Wang, Xu-sheng; Liu, Kun; Lian, Ergang; Yang, Shouye

    2017-03-01

    Studies of isotope characteristics of lake water in a desert can provide important information on groundwater discharge and hydrologic partition of the lakes in the desert. This paper presents the investigation of 18O and 2H stable isotopes and radiogenic radium of different water endmembers in three representative lakes of Badain-E, Badain-W and Sumujilin-S in the Badain Jaran Desert (BJD), the fourth largest desert in the world. A stable 18O and 2H isotopic buildup model is constructed to classify the hydrologic conditions of the desert lakes by estimating the ratio between groundwater discharge rate (Fin) and lake surface evaporation (E). Then the radium mass balance models are developed to quantify Fin. Based on the obtained Fin/E and Fin, Badain-E, Badain-W and Sumujilin-S are classified as flowing through, terminal and desiccating lakes, respectively, and their hydrologic partition is obtained. The groundwater discharge rate of Badain-E, Badain-W and Sumujilin-S, is estimated to be 8-10 mm d-1, 4-5 mm d-1, and 7-8 mm d-1, respectively. The total groundwater discharge to the lake areas in the BJD is about 1.68 × 105 m3 d-1. The flow-through condition explains the existence of the fresh lakes, while the terminal and desiccating conditions lead to the lake salinization over time. This study represents the first attempt to couple both stable and radium isotopic approaches to investigate the groundwater discharge and hydrologic partition of desert lakes in the BJD and is instructional to lake studies in other deserts in the world.

  14. Investigating the influence of sulphur dioxide (SO 2) on the stable isotope ratios (δ 13C and δ 18O) of tree rings

    NASA Astrophysics Data System (ADS)

    Rinne, K. T.; Loader, N. J.; Switsur, V. R.; Treydte, K. S.; Waterhouse, J. S.

    2010-04-01

    This study reports the influence of a 20th century pollution signal recorded in the δ 13C and δ 18O of absolutely dated tree rings from Quercus robur and Pinus sylvestris from southern England. We identify a correspondence between the inter-relationship and climate sensitivity of stable isotope series that appears to be linked to recent trends in local SO 2 emissions. This effect is most clearly exhibited in the broadleaved trees studied but is also observed in the δ 13C values of the (less polluted) pine site at Windsor. The SO 2 induced stomatal closure leads to a maximum increase of 2.5‰ in the isotope values (δ 13C). The combined physiological response to high pollution levels is less in δ 18O than δ 13C. The SO 2 signal also seems to be present as a period of reduced growth in the two ring-width chronologies. Direct, quantitative correction for the SO 2 effect represents a significant challenge owing to the nature of the records and likely local plant response to environmental pollution. Whilst it appears that this signal is both limited to the late industrial period and demonstrates a recovery in line with improvements in air quality, the role of atmospheric pollution during the calibration period should not be underestimated and adequate consideration needs to be taken when calibrating biological environmental proxies in order to avoid development of biased reconstructions.

  15. Physiological and isotopic (delta(13)C and delta(18)O) responses of three tropical tree species to water and nutrient availability.

    PubMed

    Cernusak, Lucas A; Winter, Klaus; Turner, Benjamin L

    2009-10-01

    Water-use efficiency and stable isotope composition were studied in three tropical tree species. Seedlings of Tectona grandis, Swietenia macrophylla and Platymiscium pinnatum were grown at either high or low water supply, and with or without added fertilizer. These three species previously exhibited low, intermediate and high whole-plant water-use efficiency (TE) when grown at high water supply in unfertilized soil. Responses of TE to water and nutrient availability varied among species. The TE was calculated as experiment-long dry matter production divided by cumulative water use. Species-specific offsets were observed in relationships between TE and whole-plant (13)C discrimination (Delta(13)C(p)). These offsets could be attributed to a breakdown in the relationship between Delta(13)C(p) and the ratio of intercellular to ambient CO(2) partial pressures (c(i)/c(a)) in P. pinnatum, and to variation among species in the leaf-to-air vapour pressure difference (v). Thus, a plot of v.TE against c(i)/c(a) showed a general relationship among species. Relationships between delta(18)O of stem dry matter and stomatal conductance ranged from strongly negative for S. macrophylla to no relationship for T. grandis. Results suggest inter-specific variation among tropical tree species in relationships between stable isotope ratios (delta(13)C and delta(18)O) and the gas exchange processes thought to affect them.

  16. Environmental isotopes (18O, 2H, and 87Sr/86Sr) as a tool in groundwater investigations in the Keta Basin, Ghana

    NASA Astrophysics Data System (ADS)

    Jørgensen, Niels; Banoeng-Yakubo, Bruce

    2001-03-01

    Analyses of environmental isotopes (18O, 2H, and 87Sr/86Sr) are applied to groundwater studies with emphasis on saline groundwater in aquifers in the Keta Basin, Ghana. The 87Sr/86Sr ratios of groundwater and surface water of the Keta Basin primarily reflect the geology and the mineralogical composition of the formations in the catchments and recharge areas. The isotopic compositions of 18O and 2H of deep groundwater have small variations and plot close to the global meteoric water line. Shallow groundwater and surface water have considerably larger variations in isotopic compositions, which reflect evaporation and preservation of seasonal fluctuations. A significant excess of chloride in shallow groundwater in comparison to the calculated evaporation loss is the result of a combination of evaporation and marine sources. Groundwaters from deep wells and dug wells in near-coastal aquifers are characterized by relatively high chloride contents, and the significance of marine influence is evidenced by well-defined mixing lines for strontium isotopes, and hydrogen and oxygen stable isotopes, with isotopic compositions of seawater as one end member. The results derived from environmental isotopes in this study demonstrate that a multi-isotope approach is a useful tool to identify the origin and sources of saline groundwater. Résumé. L'analyse des isotopes du milieu (18O, 2H, et 87Sr/86Sr) a été mise en œuvre pour des études hydrogéologiques portant sur des eaux souterraines salées des aquifères du bassin de Keta (Ghana). Les rapports isotopiques 87Sr/86Sr de l'eau souterraine et de l'eau de surface du bassin de Keta reflètent principalement la géologie et la composition minéralogique des formations des bassins d'alimentation et des zones de recharge. Les compositions isotopiques en 18O et en 2H des eaux souterraines profondes présentent de faibles variations et se placent près de la droite des eaux météoriques mondiales. Les eaux des nappes peu profondes

  17. A reassessment of isotopic equilibrium (Δ47 and δ18O) in the Laghetto Basso pool carbonates

    NASA Astrophysics Data System (ADS)

    Daëron, M.; Drysdale, R.; Blamart, D.; Genty, D.; Zanchetta, G.

    2013-12-01

    To the best of our current knowledge, the overwhelming majority of modern speleothems have Δ47 values which differ significantly from those predicted for thermodynamic equilibrium. The extent to which these differences may vary temporally and spatially is still an open issue, precluding a straightforward application of clumped isotopes paleothermometry to speleothem records. Here we report on the recent results of a reassessment of isotopic equilibrium in the Laghetto Basso pool carbonates (Antro del Corchia cave, NW Italy), which offer excellent a priori conditions for equilibrium carbonate precipitation and provide a continuous or quasi-continuous isotopic record of the past million years.

  18. Assessing sources of nitrate contamination in the Shiraz urban aquifer (Iran) using the δ(15)N and δ(18)O dual-isotope approach.

    PubMed

    Amiri, Haleh; Zare, Mohammad; Widory, David

    2015-01-01

    Nitrate ([Formula: see text]) is one of the major threats to the quality of the drinking water taken from the Shiraz aquifer. This aquifer undergoes high anthropogenic pressures from multiple local urban (including uncontrolled sewage systems), agricultural and industrial activities, resulting in [Formula: see text] concentrations as high as 149 mg L(-1), well above the 50 mg L(-1) guideline defined by the World Health Organisation. We coupled here classical chemical and dual isotope (δ(15)N and δ(18)O of [Formula: see text]) approaches trying to characterize sources and potential processes controlling the budget of this pollutant. Chemical data indicate that nitrate in this aquifer is explained by distinct end-members: while mineral fertilizers isotopically show to have no impact, our isotope approach identifies natural soil nitrification and organic [Formula: see text] (manure and/or septic waste) as the two main contributors. Isotope data suggest that natural denitrification may occur within the aquifer, but this conclusion is not supported by the study of other chemical parameters.

  19. /sup 18/O isotope effect in /sup 13/C nuclear magnetic resonance spectroscopy. Part 9. Hydrolysis of benzyl phosphate by phosphatase enzymes and in acidic aqueous solutions

    SciTech Connect

    Parente, J.E.; Risley, J.M.; Van Etten, R.L.

    1984-12-26

    The /sup 18/O isotope-induced shifts in /sup 13/C and /sup 31/P nuclear magnetic resonance (NMR) spectroscopy were used to establish the position of bond cleavage in the phosphatase-catalyzed and acid-catalyzed hydrolysis reactions of benzyl phosphate. The application of the /sup 18/O-isotope effect in NMR spectroscopy affords a continuous, nondestructive assay method for following the kinetics and position of bond cleavage in the hydrolytic process. The technique provides advantages over most discontinuous methods in which the reaction components must be isolated and converted to volatile derivatives prior to analysis. In the present study, (..cap alpha..-/sup 13/C,ester-/sup 18/O)benzyl phosphate and (ester-/sup 18/O)benzyl phosphate were synthesized for use in enzymatic and nonenzymatic studies. Hydrolysis reactions catalyzed by the alkaline phosphatase from E. coli and by the acid phosphatases isolated from human prostate and human liver were all accompanied by cleavage of the substrate phosphorus-oxygen bond consistent with previously postulated mechanisms involving covalent phosphoenzyme intermediates. An extensive study of the acid-catalyzed hydrolysis of benzyl phosphate at 75/sup 0/C revealed that the site of bond cleavage is dependent on pH. At pH less than or equal to 1.3, the hydrolysis proceeds with C-O bond cleavage; at 1.3 < pH < 2.0, there is a mixture of C-O and P-O bond scission, the latter progressively predominating as the pH is raised; at pH greater than or equal to 2.0, the hydrolysis proceeds with exclusive P-O bond scission. (S)-(+)-(..cap alpha..-/sup 2/H)Benzyl phosphate was also synthesized. Hydrolysis of this chiral benzyl derivative demonstrated that the acid-catalyzed C-O bond scission of benzyl phosphate proceeds by an A-1 (S/sub N/1) mechanism with 70% racemization and 30% inversion at carbon. 37 references, 4 figures, 2 tables.

  20. Isotope-Labeled Amyloids via Synthesis, Expression, and Chemical Ligation for Use in FTIR, 2D IR, and NMR Studies.

    PubMed

    Zhang, Tianqi O; Grechko, Maksim; Moran, Sean D; Zanni, Martin T

    2016-01-01

    This chapter provides protocols for isotope-labeling the human islet amyloid polypeptide (hIAPP or amylin) involved in type II diabetes and γD-crystallin involved in cataract formation. Because isotope labeling improves the structural resolution, these protocols are useful for experiments using Fourier transform infrared (FTIR), two-dimensional infrared (2D IR), and NMR spectroscopies. Our research group specializes in using 2D IR spectroscopy and isotope labeling. 2D IR spectroscopy provides structural information by measuring solvation from 2D diagonal lineshapes and vibrational couplings from cross peaks. Infrared spectroscopy can be used to study kinetics, membrane proteins, and aggregated proteins. Isotope labeling provides greater certainty in the spectral assignment, which enables new structural insights that are difficult to obtain with other methods. For amylin, we provide a protocol for (13)C/(18)O labeling backbone carbonyls at one or more desired amino acids in order to obtain residue-specific structural resolution. We also provide a protocol for expressing and purifying amylin from E. coli, which enables uniform (13)C or (13)C/(15)N labeling. Uniform labeling is useful for measuring the monomer infrared spectrum in an amyloid oligomer or fiber as well as amyloid protein bound to another polypeptide or protein, such as a chaperone or an inhibitor. In addition, our expression protocol results in 2-2.5 mg of amylin peptide per 1 L cell culture, which is a high enough yield to straightforwardly obtain the 2-10 mg needed for high resolution and solid-state NMR experiments. Finally, we provide a protocol to isotope-label either of the two domains of γD-crystallin using expressed protein ligation. Domain labeling makes it possible to resolve the structures of the two halves of the protein in FTIR and 2D IR spectra. With modifications, these strategies and protocols for isotope labeling can be applied to other amyloid polypeptides and proteins.

  1. Following 18O uptake in scCO2–H2O mixtures with Raman spectroscopy

    SciTech Connect

    Windisch, Charles F.; Schaef, Herbert T.; Martin, Paul F.; Owen, Antionette T.; McGrail, B. Peter

    2012-03-01

    The kinetics of 18O/16O isotopic exchange in scCO2 containing liquid water was followed with Raman spectroscopy using a specially designed high-pressure optical cell. Characteristic bands from the C16O18O and C18O2 molecules were identified in the supercritical phase and measured in the spectra as a function of time after introducing liquid H218O into scC16O2. Temporal dependence indicated the isotopic exchange was diffusion-limited in our cell for both molecules, and that the chemical reactions within the liquid phase were comparatively rapid. However, the ratio of concentrations of the 18O-labeled CO2 molecules, C18O2/C16O18O, was much higher than expected in the supercritical phase, suggesting the role of an intermediate step, possibly desorption, in moderating the concentrations of these species in the liquid water phase.

  2. Isotope labeling of proteins in insect cells.

    PubMed

    Skora, Lukasz; Shrestha, Binesh; Gossert, Alvar D

    2015-01-01

    Protein targets of contemporary research are often membrane proteins, multiprotein complexes, secreted proteins, or other proteins of human origin. These are difficult to express in the standard expression host used for most nuclear magnetic resonance (NMR) studies, Escherichia coli. Insect cells represent an attractive alternative, since they have become a well-established expression system and simple solutions have been developed for generation of viruses to efficiently introduce the target protein DNA into cells. Insect cells enable production of a larger fraction of the human proteome in a properly folded way than bacteria, as insect cells have a very similar set of cytosolic chaperones and a closely related secretory pathway. Here, the limited and defined glycosylation pattern that insect cells produce is an advantage for structural biology studies. For these reasons, insect cells have been established as the most widely used eukaryotic expression host for crystallographic studies. In the past decade, significant advancements have enabled amino acid type-specific as well as uniform isotope labeling of proteins in insect cells, turning them into an attractive expression host for NMR studies.

  3. Competition for water between walnut seedlings (Juglans regia) and rye grass (Lolium perenne) assessed by carbon isotope discrimination and delta18O enrichment.

    PubMed

    Picon-Cochard, C; Nsourou-Obame, A; Collet, C; Guehl, J M; Ferhi, A

    2001-02-01

    Container-grown walnut seedlings (Juglans regia L.) were subjected to competition with rye grass (Lolium perenne L.) and to a 2-week soil drying cycle. One and 2 weeks after the beginning of the drought treatment, H2 18O (delta approximately equals +100%) was added to the bottom layer of soil in the plant containers to create a vertical H2 18O gradient. Rye grass competition reduced aboveground and belowground biomass of the walnut seedlings by 60%, whereas drought had no effect. The presence of rye grass reduced the dry weight of walnut roots in the upper soil layer and caused a 50% reduction in lateral root length. Rye grass competition combined with the drought treatment reduced walnut leaf CO2 assimilation rate (A) and leaf conductance (gw) by 20 and 39%, respectively. Transpiration rates in rye grass, both at the leaf level and at the plant or tiller level, were higher than in walnut seedlings. Leaf intrinsic water-use efficiency (A/gw) of walnut seedlings increased in response to drought and no differences were observed between the single-species and mixed-species treatments, as confirmed by leaf carbon isotope discrimination measurements. Measurement of delta18O in soil and in plant xylem sap indicated that the presence of rye grass did not affect the vertical profile of soil water uptake by walnut seedlings. Walnut seedlings and rye grass withdrew water from the top and middle soil layers in well-watered conditions, whereas during the drought treatment, walnut seedlings obtained water from all soil layers, but rye grass took up water from the bottom soil layer only.

  4. USE OF OXYGEN-18 ISOTOPE LABELING FOR MEASUREMENT OF OXIDATIVE STRESS

    EPA Science Inventory

    Oxygen-18 (18-O) labeling provides a sensitive means for quantifying oxygen
    binding that occurs during in vivo oxidations. Oxidants (ozone, nitrogen
    oxides, hydrogen peroxide, etc.) are first synthesized using 18-O, then cells
    or tissues are exposed to the labeled ...

  5. Geology, Petrology and O and H isotope geochemistry of remarkably 18O depleted Paleoproterozoic rocks of the Belomorian Belt, Karelia, Russia, attributed to global glaciation 2.4 Ga

    NASA Astrophysics Data System (ADS)

    Bindeman, I. N.; Serebryakov, N. S.

    2011-06-01

    This paper deals with strongly 18O-depleted (down to - 27.3‰ VSMOW) 1.9Ga Paleoproterozoic mid-grade metamorphic rocks found in the Belomorian Belt of Karelia (E. Baltic Shield). The protolith of these rocks is attributed to have been altered by glacial meltwaters during the world's first 2.4-2.3 Ga Paleoproterozoic "Slushball" glaciation, when Karelia was located near equatorial latitudes. We describe in detail three and report seven new localities with unusually depleted 18O signatures that now span 220 km across the Belomorian Belt. Hydrogen isotope ratios measured in amphibole, biotite and staurolite also display remarkably low values of - 212 to - 235‰. Isotope mapping in the three best exposed localities has allowed us to identify the world's most 18O depleted rock, located at Khitostrov with a δ 18O value - 27‰. In Khitostrov samples, zircons have normal δ 18O detrital cores and low-δ 18O metamorphic rims. Mapping demonstrates that zones of δ 18O depletion occur in a concentric pattern 100-400 m in dimension, and each locality displays significant δ 18O and δD heterogeneity on a meter to centimeter scale, characteristic of meteoric-hydrothermal systems worldwide. The zone of maximum δ 18O depletions usually has the highest concentration of metamorphic corundum, rutile, and zircon and also display doubled concentrations of insoluble trace elements (Zr, Ti, Cr, HREE). These results are explained by elemental enrichment upon mass loss during hydrothermal dissolution in pH-neutral meteoric fluid. Remarkably low-δ 18O and δD values suggest that alteration could have only happened by glacial meltwaters in a subglacial rift zone. Many localities with δ 18O depletions occur inside metamorphozed 2.4 Ga gabbro-noritic intrusions, or near their contact with Belomorian gneisses, implying that the intrusions were driving meteoric hydrothermal systems during the known 2.4 Ga episode of Belomorian rifting. Given that the isotopically-depleted localities now

  6. Paleocene to Early Eocene paleoceanography of the Middle East: The δ13C and δ18O isotopes from foraminiferal calcite

    NASA Astrophysics Data System (ADS)

    Charisi, Stella D.; Schmitz, Birger

    1998-02-01

    Paleocene to early Eocene benthic foraminiferal δ13C and δ18O records from southern Tethyan sections at Ben Gurion, Israel (paleodepth 500-700 m), and Gebel Aweina, Egypt (paleodepth 150-200 m), show generally similar trends but 1-3‰ more negative values than coeval deep-sea isotopic records. In both Tethyan sections a negative δ13C excursion of 2.5-3‰ marks the benthic extinction event in the latest Paleocene. For at least 1 m.y. after this event, δ13C values were 1.5-2‰ more negative on the shelf than at upper bathyal depths, reflecting a deepening of the oxygen minimum zone, possibly related to an increase or spatial shift in upwelling. Benthic δ18O records indicate a 2-4°C temperature gradient between the shelf and upper bathyal depths. Temperature-salinity reconstructions suggest that upwelling was a dominant mechanism for surface water formation in this part of the southern Tethys during the late Paleocene.

  7. Trends in nitrate concentrations and determination of its origin using stable isotopes (18O and 15N) in groundwater of the Western Central Valley, Costa Rica.

    PubMed

    Reynolds-Vargas, Jenny; Fraile-Merino, Julio; Hirata, Ricardo

    2006-08-01

    A study was conducted to evaluate long-term trends in nitrate concentrations and to try to identify the origin of nitrate using stable isotopes (15N(NO3-) and 18O(NO3-)) in the aquifers of the western Central Valley, Costa Rica, where more than 1 million people depend on groundwater to satisfy their daily needs. Data from 20 sites periodically sampled for 4 to 17 years indicate an increasing trend in nitrate concentrations at five sites, which in a period ranging from 10 to 40 years, will exceed recommended maximum concentrations. Results of isotopic analysis indicate a correspondence between land use patterns and the isotopic signature of nitrate in groundwater and suggest that urbanization processes without adequate waste disposal systems, followed by coffee fertilization practices, are threatening water quality in the region. We conclude that groundwater management in this area is not sustainable, and that land use substitution processes from agricultural activity to residential occupation that do not have proper sewage disposal systems may cause a significant increment in the nitrate contaminant load.

  8. Oxygen isotopes of Pacific seawater, 0-40 kyr, based on d18O and Mg/Ca of benthic and planktic foraminifera: relation to deglacial sealevel rise.

    NASA Astrophysics Data System (ADS)

    Mix, A. C.; Klinkhammer, G. P.

    2007-12-01

    Combining high resolution (~200 year sample resolution) oxygen and carbon isotope measurements and Mg/Ca analyses in planktic (G. ruber and N. dutertrei) and benthic foraminifera (Uvigerina sp.; use of infaunal benthics minimizes likely CO3= effects) from the mid-depth eastern Pacific provides for detailed estimates of changing d18O of seawater over the past 40 kyr at ODP Site 1242. The key to this analysis is improved precision of the Mg/Ca analyses based on a new generation of high precision flow-through time-resolved analysis (FT-TRA) (e.g., average internal precision for Mg/Ca is now +/-0.005 benthic, and +/-0.015 planktic). This method is relatively insensitive to mild dissolution of heterogeneous shells, and thus requires no corrections for preservation. The change in seawater d18O (at 1364 m depth) between the Holocene and Last Glacial Maximum is 1.2 +/- 0.04 permil when averaged over the stages, consistent with pore-water diffusion estimates; however, millennial scale events (which the pore-water data cannot detect) bring the total range up to about 1.6 permil. Are such short-term events related to sealevel change, or do they reflect changing watermasses? Measurements of d13C data (C. wuellerstorfi), sensitive to modern subsurface watermass gradients, are not highly correlated to short- term changes in d18Oseawater suggesting a transient response to ice volume changes. Glacial weakening of AAIW (salty, high d18O, high d13C) relative to north Pacific watermasses (fresher, lower d18O, low d13C) suggest that the benthic d18Oseawater may underestimate total local changes related to ice volume (with a caveat regarding proper scaling of benthic Mg/Ca to temperature). Benthic d18Oseawater falls through the deglaciation in steps, starting at 18 cal ka, with maximum rates of change at 14-15 ka, and with secondary rapid steps at 16-17 ka and 10-11 kar. Planktic foraminifera yield smaller glacial-interglacial d18Oseawater values, with Holocene-to-LGM stage- average

  9. Intra-shell d18O in Cultured Benthic Foraminiferan Amphistegina lobifera and the Influence of Seawater Carbonate Chemistry and Temperature on this Isotopic Composition

    NASA Astrophysics Data System (ADS)

    Rollion-Bard, C.; Erez, J.; Zilberman, T.; Segev, E.

    2006-12-01

    Using secondary ion mass spectrometry (SIMS) we looked at the natural variability in the oxygen isotope ratio of the shallow water, symbionts bearing foraminiferan A. lobifera. Live foraminifera were collected in June 2005 in the Gulf of Eilat, Israel. Vertical section exposing the knob area of this species represents the growth history of this species from December 2004 to June 2005. SIMS profile at a resolution of ~ 2 weeks yielded δ^1^8O changes of ~ 1.5 ‰, (from -0.1 ‰ to 1.45 ‰) that are compatible with the known temperature changes for the Gulf of Eilat for this period (20 to 25° C). Natural variability between primary and secondary calcite at the knob area were obtained on horizontal section of the upper knob area. The primary calcite is on average 2 ‰ more negative then the secondary calcite that represents the bulk of the skeleton (more then 95 % by weight). The δ^1^8O in the margin keel area of A. lobifera is also lower compared to the bulk secondary calcite. Specimens that were cultured in the laboratory at a constant temperature and inorganic carbon but at different pH have increased their CaCO3 weight by roughly a factor of 8. Single specimen from each pH (ranging between 7.9 and 8.5) was investigated with the SIMS at the knob area. While there is some variability within each specimen (perhaps related to the primary calcite), the general trend was a decrease in δ^1^8O with increasing pH (or CO32^- concentration), in agreement with previous studies on planktonic foraminifera. Specimens cultured in laboratory at a constant pH, but different temperature were also analysed in the knob area. The temperature range is between 21 and 33° C (experiments at 21, 24, 27 and 33° C). While there is also some variability within each specimen, the trend was a decrease in δ^1^8O with increasing temperature. The decrease measured is 2.7 ± 0.7 ‰ for the entire temperature range, which is completely in agreement with the theoretical value (-0.2 ‰ per

  10. Stereoselective synthesis of stable-isotope-labeled amino acids

    SciTech Connect

    Unkefer, C.J.; Martinez, R.A.; Silks, L.A. III; Lodwig, S.N.

    1994-12-01

    For magnetic resonance and vibrational spectroscopies to reach their full potential, they must be used in combination with sophisticated site-specific stable isotope labeling of biological macromolecules. Labeled amino acids are required for the study of the structure and function of enzymes and proteins. Because there are 20 common amino acids, each with its own distinguishing chemistry, they remain a synthetic challenge. The Oppolzer chiral auxiliary provides a general tool with which to approach the synthesis of labeled amino acids. By using the Oppolzer auxiliary, amino acids can be constructed from several small molecules, which is ideal for stable isotope labeling. In addition to directing the stereochemistry at the {alpha}-carbon, the camphorsultam can be used for stereo-specific isotope labeling at prochiral centers in amino acids. By using the camphorsultam auxiliary we have the potential to synthesize virtually any isotopomer of all of the common amino acids.

  11. Towards a Better Understanding of the Oxygen Isotope Signature of Atmospheric CO2: Determining the 18O-Exchange Between CO2 and H2O in Leaves and Soil On-line with Laser-Based Spectroscopy

    NASA Astrophysics Data System (ADS)

    Gangi, L.; Rothfuss, Y.; Vereecken, H.; Brueggemann, N.

    2013-12-01

    The oxygen isotope signature of carbon dioxide (δ18O-CO2) is a powerful tool to disentangle CO2 fluxes in terrestrial ecosystems, as CO2 attains a contrasting 18O signature by the interaction with isotopically different soil and leaf water pools during soil respiration and photosynthesis, respectively. However, using the δ18O-CO2 signal to quantify plant-soil-atmosphere CO2 fluxes is still challenging due to a lack of knowledge concerning the magnitude and effect of individual fractionation processes during CO2 and H2O diffusion and during CO2-H2O isotopic exchange in soils and leaves, especially related to short-term changes in environmental conditions (non-steady state). This study addresses this research gap by combined on-line monitoring of the oxygen isotopic signature of CO2 and water vapor during gas exchange in soil and plant leaves with laser-based spectroscopy, using soil columns and plant chambers. In both experimental setups, the measured δ18O of water vapor was used to infer the δ18O of liquid water, and, together with the δ18O-CO2, the degree of oxygen isotopic equilibrium between the two species (θ). Gas exchange experiments with different functional plant types (C3 coniferous, C3 monocotyledonous, C3 dicotyledonous, C4) revealed that θ and the influence of the plant on the ambient δ18O-CO2 (CO18O-isoforcing) not only varied on a diurnal timescale but also when plants were exposed to limited water availability, elevated air temperature, and abrupt changes in light intensity (sunflecks). Maximum θ before treatments ranged between 0.7 and 0.8 for the C3 dicotyledonous (poplar) and C3 monocotyledonous (wheat) plants, and between 0.5 and 0.6 for the conifer (spruce) and C4 plant (maize) while maximum CO18O-isoforcing was highest in wheat (0.03 m s-1 ‰), similar in poplar and maize (0.02 m s-1 ‰), and lowest in spruce (0.01 m s-1 ‰). Multiple regression analysis showed that up to 97 % of temporal dynamics in CO18O-isoforcing could be

  12. Conformational changes of recombinant monoclonal antibodies by limited proteolytic digestion, stable isotope labeling, and liquid chromatography-mass spectrometry.

    PubMed

    Ponniah, Gomathinayagam; Nowak, Christine; Kita, Adriana; Cheng, Guilong; Kori, Yekaterina; Liu, Hongcheng

    2016-03-15

    Limited proteolytic digestion is a method with a long history that has been used to study protein domain structures and conformational changes. A method of combining limited proteolytic digestion, stable isotope labeling, and mass spectrometry was established in the current study to investigate protein conformational changes. Recombinant monoclonal antibodies with or without the conserved oligosaccharides, and with or without oxidation of the conserved methionine residues, were used to test the newly proposed method. All of the samples were digested in ammonium bicarbonate buffer prepared in normal water. The oxidized deglycosylated sample was also digested in ammonium bicarbonate buffer prepared in (18)O-labeled water. The sample from the digestion in (18)O-water was spiked into each sample digested in normal water. Each mixed sample was subsequently analyzed by liquid chromatography-mass spectrometry (LC-MS). The molecular weight differences between the peptides digested in normal water versus (18)O-water were used to differentiate peaks from the samples. The relative peak intensities of peptides with or without the C-terminal incorporation of (18)O atoms were used to determine susceptibility of different samples to trypsin and chymotrypsin. The results demonstrated that the method was capable of detecting local conformational changes of the recombinant monoclonal antibodies caused by deglycosylation and oxidation.

  13. Isotope effects in the CO dimer: Millimeter wave spectrum and rovibrational calculations of (12C18O)2

    NASA Astrophysics Data System (ADS)

    Surin, L. A.; Fourzikov, D. N.; Giesen, T. F.; Schlemmer, S.; Winnewisser, G.; Panfilov, V. A.; Dumesh, B. S.; Vissers, G. W. M.; van der Avoird, A.

    2006-09-01

    The millimeter wave spectrum of the isotopically substituted CO dimer, (C12O18)2, was studied with the Orotron jet spectrometer, confirming and extending a previous infrared study [A. R. W. McKellar, J. Mol. Spectrosc. 226, 190 (2004)]. A very dilute gas mixture of CO in Ne was used, which resulted in small consumption of C12O18 sample gas and produced cold and simple spectra. Using the technique of combination differences together with the data from the infrared work, six transitions in the 84-127GHz region have been assigned. They belong to two branches, which connect four low levels of A+ symmetry to three previously unknown levels of A- symmetry. The discovery of the lowest state of A- symmetry, which corresponds to the projection K =0 of the total angular momentum J onto the intermolecular axis, identifies the geared bending mode of the C12O18 dimer at 3.607cm-1. Accompanying rovibrational calculations using a recently developed hybrid potential from ab initio coupled cluster [CCSD(T)] and symmetry-adapted perturbation theory calculations [G. W. M. Vissers et al., J. Chem. Phys. 122, 054306 (2005)] gave very good agreement with experiment. The isotopic dependence of the A+/A- energy splitting, the intermolecular separation R, and the energy difference of two ground state isomers, which change significantly when O18 or C13 are substituted into the normal (C12O16)2 isotopolog [L. A. Surin et al., J. Mol. Spectrosc. 223, 132 (2004)], was explained by these calculations. It turns out that the change in anisotropy of the intermolecular potential with respect to the shifted monomer centers of mass is particularly significant.

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

    PubMed

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

    2013-06-01

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

  15. Selected scientific topics of the 11th International Isotope Symposium on the Synthesis and Applications of Isotopes and Isotopically Labeled Compounds.

    PubMed

    Atzrodt, Jens; Derdau, Volker

    2013-01-01

    This micro-review describes hot topics and new trends in isotope science discussed at the 11th International Isotope Symposium on the Synthesis and Applications of Isotopes and Isotopically Labeled Compounds from a personal perspective.

  16. Energy costs of surgery as measured by the doubly labeled water (/sup 2/H/sub 2//sup 18/O) method

    SciTech Connect

    Novick, W.M.; Nusbaum, M.; Stein, T.P.

    1988-01-01

    Energy expenditure before and after surgery was determined in seven patients by the doubly labeled water (/sup 2/H/sub 2//sup 18/O) method (DLW). The values were compared with values obtained by respiratory gas exchange by means of a metabolic measuring cart (MMC). Patients were maintained on total parenteral nutrition before and after trauma. The principal finding was an increase in the rate of CO/sub 2/ production of 11.9 +/- 5.0% after surgery. This corresponds to a 267 +/- increase in energy expenditure (p less than 0.05). No trauma-associated change in energy expenditure was found with the MMC. The correlation of preoperative values from MMC and DLW was not statistically significant (r = 0.25), nor was the correlation of MMC and the Harris-Benedict equation, but the correlation of DLW with Harris-Benedict equation was statistically significant (r = 0.73, p less than 0.05). We suggest that the discrepancy is because the DLW method measures the cumulative energy expenditure over a period, whereas the MMC gives a spot measurement.

  17. Experimental investigation of rates and mechanisms of isotope exchange (O, H) between volcanic ash and isotopically-labeled water

    NASA Astrophysics Data System (ADS)

    Nolan, Gary S.; Bindeman, Ilya N.

    2013-06-01

    The hydrogen and oxygen isotope ratios in hydrous minerals and volcanic glass are routinely used as paleo-proxies to infer the isotopic values of meteoric waters and thus paleo-climatic conditions. We report a series of long-term exposure experiments of distal 7700 BP Mt. Mazama ash (-149‰ δ2H, +7‰ δ18O, 3.8 wt.% H2O) with isotopically-labeled water (+650‰ δ2H, +56‰ δ18O). Experiments were done at 70, 40 and 20 °C, and ranged in duration from 1 to 14454 h (˜20 months), to evaluate the rates of deuterium and 18O exchange, and investigate the relative role of exchange and diffusion. We also investigate the effect of drying on H2Otot and δ2H in native and reacted ash that can be used in defining the protocols for natural sample preparation. We employ Thermal Conversion Elemental Analyzer (TCEA) mass spectrometry, thermogravimetric analysis and a KBr pellet technique with infrared spectroscopy to measure the evolution of δ2H, total water, and OH water peaks in the course of exposure experiments, and in varying lengths of vacuum drying. Time series experiments aided by infrared measurements demonstrate the following new results: (i) It wasobserved that from 5 to >100‰ δ2H increases with time, with faster deuterium exchange at higher temperatures. Times at 15% of theoretical "full δ2H exchange" are: 15.8 years at 20 °C, 5.2 years at 40 °C, and 0.4 years at 70 °C. (ii) Even at extended exposure durations experiments show no net increase in water weight percent nor in δ18O in ash; water released from ash rapidly by thermal decomposition is not enriched in δ18O. This observation clearly suggests that it is hydrogen exchange, and not water addition or oxygen exchange that characterizes the process. (iii) Our time series drying, Fourier transform infrared (FTIR)-KBr and Thermogravimetric Analyzer (TGA) analyses collectively suggest a simple mechanistic view that there are three kinds of "water" in ash: water (mostly H2O) that is less strongly bonded

  18. A conifer-friendly high-throughput α-cellulose extraction method for δ13C and δ18O stable isotope ratio analysis

    NASA Astrophysics Data System (ADS)

    Lin, W.; Noormets, A.; domec, J.; King, J. S.; Sun, G.; McNulty, S.

    2012-12-01

    Wood stable isotope ratios (δ13C and δ18O) offer insight to water source and plant water use efficiency (WUE), which in turn provide a glimpse to potential plant responses to changing climate, particularly rainfall patterns. The synthetic pathways of cell wall deposition in wood rings differ in their discrimination ratios between the light and heavy isotopes, and α-cellulose is broadly seen as the best indicator of plant water status due to its local and temporal fixation and to its high abundance within the wood. To use the effects of recent severe droughts on the WUE of loblolly pine (Pinus taeda) throughout Southeastern USA as a harbinger of future changes, an effort has been undertaken to sample the entire range of the species and to sample the isotopic composition in a consistent manner. To be able to accommodate the large number of samples required by this analysis, we have developed a new high-throughput method for α-cellulose extraction, which is the rate-limiting step in such an endeavor. Although an entire family of methods has been developed and perform well, their throughput in a typical research lab setting is limited to 16-75 samples per week with intensive labor input. The resin exclusion step in conifersis is particularly time-consuming. We have combined the recent advances of α-cellulose extraction in plant ecology and wood science, including a high-throughput extraction device developed in the Potsdam Dendro Lab and a simple chemical-based resin exclusion method. By transferring the entire extraction process to a multiport-based system allows throughputs of up to several hundred samples in two weeks, while minimizing labor requirements to 2-3 days per batch of samples.

  19. Effect of temperature on the oxygen isotope composition of carbon dioxide (δ18O) prepared from carbonate minerals by reaction with polyphosphoric acid: An example of the rhombohedral CaCO 3-MgCO 3 group minerals

    NASA Astrophysics Data System (ADS)

    Crowley, Stephen F.

    2010-11-01

    Measurement of the ratio of 18O to 16O in CO 2(δ18O) produced from rhombohedral carbonate minerals in the compositional range CaCO 3-MgCO 3 by reaction with polyphosphoric acid (PPA), at temperatures of between 25 and 110 °C, shows that values of δ18O are linearly correlated ( r o > 0.99) with the reciprocal of absolute reaction temperature (K/ T). This observation is consistent with earlier studies documenting the effect of temperature on the kinetic fractionation of oxygen isotopes between parent carbonate and product CO 2 and H 2O during acid decomposition. However, analysis of the resultant data reveals: (1) a progressive increase in dδ18O/dT-1 with increasing Mg content, and (2) a significant variation in dδ18O/dT-1 between individual samples of carbonate of identical lattice symmetry and similar chemical composition. The overall increase in gradient with increasing Mg content is assumed to reflect cation radius dependent factors that control the bonding environment at the interface between the metal cation exposed at the surface of the reacting carbonate solid and a H 2CO 3 transitional species during disproportionation of H 2CO 3 to CO 2 and H 2O ("cluster model" of Guo et al., 2009). Phase-specific variations in dδ18O/dT-1 might result from differences in lattice structure variables (e.g., degree of lattice distortion, extent of positional disorder, and non-ideal mixing of substituent cations where carbonates depart from end-member compositions). Lattice structure variables may be dependent on geochemical conditions pertaining at the time of carbonate precipitation (e.g., biosynthetic versus inorganic precipitates) and suggests that dδ18O/dT-1 has the potential to vary, within limits, in response to both the chemical composition and structure of each carbonate sample. Because the oxygen isotope composition of carbonate minerals (δ18O) measured on the VPDB scale is defined by the oxygen isotope composition of CO 2 prepared from NBS19 (calcite) by

  20. The Po river water from the Alps to the Adriatic Sea (Italy): new insights from geochemical and isotopic (δ(18)O-δD) data.

    PubMed

    Marchina, Chiara; Bianchini, Gianluca; Natali, Claudio; Pennisi, Maddalena; Colombani, Nicolò; Tassinari, Renzo; Knoeller, Kay

    2015-04-01

    Although the Po river is the most important fluvial system of Northern Italy, the systematic geochemical and isotopic investigations of its water are rare and were never reported for the whole basin. The present contribution aims to fill this knowledge gap, reporting a comprehensive data set including oxygen and hydrogen stable isotopes as well as major and trace element concentration of dissolved species for 54 Po river water samples, mainly collected in different hydrological conditions (peak discharge in April, drought in August) at increasing distance from the source, i.e., from the upper part of the catchment to the terminal (deltaic) part of the river at the confluence with the Adriatic Sea. The isotopic compositions demonstrate that the predominant part of the runoff derives from the Alpine sector of the catchment through important tributaries such as the Dora Baltea, Ticino, Adda, and Tanaro rivers, whereas the contribution from the Apennines tributaries is less important. The geochemical and isotopic compositions show that the Po river water attains a homogeneous composition at ca. 100 km from the source. The average composition is characterized by δ(18)O -9.8‰, δD -66.2‰, total dissolved solid (TDS) 268 mg/L, and chloride 17 mg/L and by a general Ca-HCO3 hydrochemical facies, which is maintained for most of the river stream, only varying in the terminal part where the river is diverted in a complex deltaic system affected by more significant evaporation and mixing with saline water evidenced by higher TDS and chloride content (up to 8198 and 4197 mg/L, respectively). Geochemical and isotopic maps have been drawn to visualize spatial gradients, which reflect the evolution of the river water composition at progressive distance from the source; more detailed maps were focused on the deltaic part in order to visualize the processes occurring in the transitional zone toward the Adriatic Sea. The data also highlight anthropogenic contributions, mainly

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2014-01-16

    Tracing rare stable isotopes from plant material through the ecosystem provides the most sensitive information about ecosystem processes; from CO2 fluxes and soil organic matter formation to small-scale stable-isotope biomarker probing. Coupling multiple stable isotopes such as (13)C with (15)N, (18)O or (2)H has the potential to reveal even more information about complex stoichiometric relationships during biogeochemical transformations. Isotope labeled plant material has been used in various studies of litter decomposition and soil organic matter formation(1-4). From these and other studies, however, it has become apparent that structural components of plant material behave differently than metabolic components (i.e. leachable low molecular weight compounds) in terms of microbial utilization and long-term carbon storage(5-7). The ability to study structural and metabolic components separately provides a powerful new tool for advancing the forefront of ecosystem biogeochemical studies. Here we describe a method for producing (13)C and (15)N labeled plant material that is either uniformly labeled throughout the plant or differentially labeled in structural and metabolic plant components. Here, we present the construction and operation of a continuous (13)C and (15)N labeling chamber that can be modified to meet various research needs. Uniformly labeled plant material is produced by continuous labeling from seedling to harvest, while differential labeling is achieved by removing the growing plants from the chamber weeks prior to harvest. Representative results from growing Andropogon gerardii Kaw demonstrate the system's ability to efficiently label plant material at the targeted levels. Through this method we have produced plant material with a 4.4 atom%(13)C and 6.7 atom%(15)N uniform plant label, or material that is differentially labeled by up to 1.29 atom%(13)C and 0.56 atom%(15)N in its metabolic and structural components (hot water extractable and hot water

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

    SciTech Connect

    Hooker, Jacob Matthew; Schonberger, Matthias; Schieferstein, Hanno; Fowler, Joanna S.

    2011-10-04

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

  4. Online Determination of 18O Fractionation Between CO2 and Soil-Water during Soil Dessication by a Novel Mid-Infrared CO2 Isotope Analyzer Coupled to an Dynamic Chamber Incubation System

    NASA Astrophysics Data System (ADS)

    Nowak, A.

    2015-12-01

    The stable oxygen isotope composition of CO2 is an important tracer for quantifying gas interactions between soils and atmosphere. Soils impact atmospheric 18O-CO2 signatures by CO2-H2O equilibration during diffusion of CO2 through the soil column. However, recent research has revealed that also catalytic reactions by carbonic anhydrase, an enzyme used by microorganisms for triggering the conversion of CO2 and water to bicarbonate and protons, is an important factor influencing the oxygen isotopic signature of CO2. In order to study the importance of biotic and abiotic factors for 18O-CO2, we used a novel mid infrared 18O/13C-CO2 analyser coupled to a dynamic chamber system, which allowed us to measure online 18O and 13C of a continuous CO2 stream percolating through soil samples while drying out from fully water saturated to air dry. Our results indicate that changes in CO2- 18O signatures peak at certain soil moistures levels, which is most probably catalysed by the activity of certain microbial groups under optimum growth conditions. More analyses with different soil types and depths, combined with molecular analyses are planned in order to understand the importance of microbial processes and dynamics for influencing soil-CO2 interactions.

  5. High-Precision Tungsten Isotopic Analysis by Multicollection Negative Thermal Ionization Mass Spectrometry Based on Simultaneous Measurement of W and (18)O/(16)O Isotope Ratios for Accurate Fractionation Correction.

    PubMed

    Trinquier, Anne; Touboul, Mathieu; Walker, Richard J

    2016-02-02

    Determination of the (182)W/(184)W ratio to a precision of ± 5 ppm (2σ) is desirable for constraining the timing of core formation and other early planetary differentiation processes. However, WO3(-) analysis by negative thermal ionization mass spectrometry normally results in a residual correlation between the instrumental-mass-fractionation-corrected (182)W/(184)W and (183)W/(184)W ratios that is attributed to mass-dependent variability of O isotopes over the course of an analysis and between different analyses. A second-order correction using the (183)W/(184)W ratio relies on the assumption that this ratio is constant in nature. This may prove invalid, as has already been realized for other isotope systems. The present study utilizes simultaneous monitoring of the (18)O/(16)O and W isotope ratios to correct oxide interferences on a per-integration basis and thus avoid the need for a double normalization of W isotopes. After normalization of W isotope ratios to a pair of W isotopes, following the exponential law, no residual W-O isotope correlation is observed. However, there is a nonideal mass bias residual correlation between (182)W/(i)W and (183)W/(i)W with time. Without double normalization of W isotopes and on the basis of three or four duplicate analyses, the external reproducibility per session of (182)W/(184)W and (183)W/(184)W normalized to (186)W/(183)W is 5-6 ppm (2σ, 1-3 μg loads). The combined uncertainty per session is less than 4 ppm for (183)W/(184)W and less than 6 ppm for (182)W/(184)W (2σm) for loads between 3000 and 50 ng.

  6. Growth decline and divergent tree ring isotopic composition (δ(13) C and δ(18) O) contradict predictions of CO2 stimulation in high altitudinal forests.

    PubMed

    Gómez-Guerrero, Armando; Silva, Lucas C R; Barrera-Reyes, Miguel; Kishchuk, Barbara; Velázquez-Martínez, Alejandro; Martínez-Trinidad, Tomás; Plascencia-Escalante, Francisca Ofelia; Horwath, William R

    2013-06-01

    Human-induced changes in atmospheric composition are expected to affect primary productivity across terrestrial biomes. Recent changes in productivity have been observed in many forest ecosystems, but low-latitude upper tree line forests remain to be investigated. Here, we use dendrochronological methods and isotopic analysis to examine changes in productivity, and their physiological basis, in Abies religiosa (Ar) and Pinus hartwegii (Ph) trees growing in high-elevation forests of central Mexico. Six sites were selected across a longitudinal transect (Transverse Volcanic Axis), from the Pacific Ocean toward the Gulf of Mexico, where mature dominant trees were sampled at altitudes ranging from 3200 to 4000 m asl. A total of 60 Ar and 84 Ph trees were analyzed to describe changes in growth (annual-resolution) and isotopic composition (decadal-resolution) since the early 1900s. Our results show an initial widespread increase in basal area increment (BAI) during the first half of the past century. However, BAI has decreased significantly since the 1950s with accentuated decline after the 1980s in both species and across sites. We found a consistent reduction in atmosphere to wood (13) C discrimination, resulting from increasing water use efficiency (20-60%), coinciding with rising atmospheric CO2 . Changes in (13) C discrimination were not followed, however, by shifts in tree ring δ(18) O, indicating site- and species-specific differences in water source or uptake strategy. Our results indicate that CO2 stimulation has not been enough to counteract warming-induced drought stress, but other stressors, such as progressive nutrient limitation, could also have contributed to growth decline. Future studies should explore the distinct role of resource limitation (water vs. nutrients) in modulating the response of high-elevation ecosystems to atmospheric change.

  7. Stable Isotope Labeling Strategy for Protein-Ligand Binding Analysis in Multi-Component Protein Mixtures

    NASA Astrophysics Data System (ADS)

    DeArmond, Patrick D.; West, Graham M.; Huang, Hai-Tsang; Fitzgerald, Michael C.

    2011-03-01

    Described here is a stable isotope labeling protocol that can be used with a chemical modification- and mass spectrometry-based protein-ligand binding assay for detecting and quantifying both the direct and indirect binding events that result from protein-ligand binding interactions. The protocol utilizes an H{2/16}O2 and H{2/18}O2 labeling strategy to evaluate the chemical denaturant dependence of methionine oxidation in proteins both in the presence and absence of a target ligand. The differential denaturant dependence to the oxidation reactions performed in the presence and absence of ligand provides a measure of the protein stability changes that occur as a result of direct interactions of proteins with the target ligand and/or as a result of indirect interactions involving other protein-ligand interactions that are either induced or disrupted by the ligand. The described protocol utilizes the 18O/16O ratio in the oxidized protein samples to quantify the ligand-induced protein stability changes. The ratio is determined using the isotopic distributions observed for the methionine-containing peptides used for protein identification in the LC-MS-based proteomics readout. The strategy is applied to a multi-component protein mixture in this proof-of-principle experiment, which was designed to evaluate the technique's ability to detect and quantify the direct binding interaction between cyclosporin A and cyclophilin A and to detect the indirect binding interaction between cyclosporin A and calcineurin (i.e., the protein-protein interaction between cyclophilin A and calcineurin that is induced by cyclosporin A binding to cyclophilin A).

  8. Oxygen isotope ratios and rare earth elements in 3.3 to 4.4 Ga zircons: Ion microprobe evidence for high δ 18O continental crust and oceans in the Early Archean

    NASA Astrophysics Data System (ADS)

    Peck, William H.; Valley, John W.; Wilde, Simon A.; Graham, Colin M.

    2001-11-01

    Ion microprobe analyses of oxygen isotope ratios in Early Archean (Hadean) zircons (4.0- to 4.4-Ga) reveal variable magmatic δ 18O values, including some that are high relative to the mantle, suggesting interaction between magmas and already-formed continental crust during the first 500 million yr of Earth's history. The high average δ 18O value of these zircons is confirmed by conventional analysis. A metaconglomerate from the Jack Hills in the Yilgarn Craton (Western Australia) contains detrital zircons with ages > 4.0 Ga (Compston and Pidgeon, 1986) and one crystal that is 4.40-Ga old (Wilde et al., 2001). The newly discovered 4.40-Ga grain is the oldest recognized terrestrial mineral. The Jack Hills metaconglomerate also contains a large 3.3- to 3.6-Ga-old zircon population with an average δ 18O value of 6.3 ± 0.1‰ (1 s.e.,; n = 32 spot analyses). Two 4.15-Ga zircons have an average δ 18O of 5.7 ± 0.2‰ ( n = 13). In addition, a 4.13-Ga zircon has an average δ 18O of 7.2 ± 0.3‰ ( n = 8) and another 4.01-Ga zircon has an average δ 18O of 6.8 ± 0.4‰ ( n = 10). The oldest grain (4.40 Ga) is zoned with respect trace element composition (especially LREE), and intensity of cathodoluminescence, all of which correlate with oxygen isotope ratios (7.4‰ vs. 5.0‰). High LREE and high-δ 18O values from the 4.01- to 4.40-Ga grains are consistent with growth in evolved granitic magmas (δ 18O(WR) = 8.5 to 9.5‰) that had interacted with supracrustal materials. High δ 18O values show that low-temperature surficial processes (i.e., diagenesis, weathering, or low-temperature alteration) occurred before 4.0 Ga, and even before 4.40 Ga, shortly following the hypothesized date of core differentiation and impact of a Mars-sized body to form the Moon at ˜4.45 Ga. This is the first evidence of continental crust as early as 4.40 Ga and suggests differentiation during the period of intense meteorite bombardment of the early Earth. The magnitude of water and rock

  9. The origin of hydrous, high-δ18O voluminous volcanism: diverse oxygen isotope values and high magmatic water contents within the volcanic record of Klyuchevskoy volcano, Kamchatka, Russia

    NASA Astrophysics Data System (ADS)

    Auer, Sara; Bindeman, Ilya; Wallace, Paul; Ponomareva, Vera; Portnyagin, Maxim

    2009-02-01

    Klyuchevskoy volcano, in Kamchatka’s subduction zone, is one of the most active arc volcanoes in the world and contains some of the highest δ18O values for olivines and basalts. We present an oxygen isotope and melt inclusion study of olivine phenocrysts in conjunction with major and trace element analyses of 14C- and tephrochronologically-dated tephra layers and lavas spanning the eruptive history of Klyuchevskoy. Whole-rock and groundmass analyses of tephra layers and lava samples demonstrate that both high-Mg (7-12.5 wt% MgO) and high-Al (17-19 wt% Al2O3, 3-6.5 wt% MgO) basalt and basaltic andesite erupted coevally from the central vent and flank cones. Individual and bulk olivine δ18O range from normal MORB values of 5.1‰ to values as high as 7.6‰. Likewise, tephra and lava matrix glass have high-δ18O values of 5.8-8.1‰. High-Al basalts dominate volumetrically in Klyuchevskoy’s volcanic record and are mostly high in δ18O. High-δ18O olivines and more normal-δ18O olivines occur in both high-Mg and high-Al samples. Most olivines in either high-Al or high-Mg basalts are not in oxygen isotopic equilibrium with their host glasses, and Δ18Oolivine-glass values are out of equilibrium by up to 1.5‰. Olivines are also out of Fe-Mg equilibrium with the host glasses, but to a lesser extent. Water concentrations in olivine-hosted melt inclusions from five tephra samples range from 0.4 to 7.1 wt%. Melt inclusion CO2 concentrations vary from below detection (<50 ppm) to 1,900 ppm. These values indicate depths of crystallization up to ~17 km (5 kbar). The variable H2O and CO2 concentrations likely reflect crystallization of olivine and entrapment of inclusions in ascending and degassing magma. Oxygen isotope and Fe-Mg disequilibria together with melt inclusion data indicate that olivine was mixed and recycled between high-Al and high-Mg basaltic melts and cumulates, and Fe-Mg and δ18O re-equilibration processes were incomplete. Major and trace elements in

  10. Clumped Isotopes, trace elements, and δ18O of stromatolites from the Laney Member of the Green River Formation (Eocene): Implications for paleoenvironments during the Eocene Climatic Optimum

    NASA Astrophysics Data System (ADS)

    Corsetti, F. A.; Miller, H. M.; Asangba, A. E.; Johannessen, K. C.; Wang, D. T.; Petryshyn, V. A.; Tripati, A.; Shapiro, R. S.

    2013-12-01

    The Green River Formation, a large lacustrine deposit located across parts of Utah, Colorado, and Wyoming, was deposited during the Eocene Climatic Optimum (~50 Ma), a period of sustained high temperatures and high atmospheric CO2 levels that may provide a geologic analog for future climate scenarios. Large variations in basin hydrology, water chemistry, and paleotemperatures occurring on time scales of tens of thousands of years or longer have been documented in the sedimentary record. Here, we use stromatolites to investigate much finer-scale resolution of paleoenvironmental changes in the Green River Formation and paleo-Lake Gosiute. We studied the lower LaClede Bed, the base of the Laney Member of the Green River Formation, comprised of cyclic layers of oil shale and carbonate. The lower LaClede Bed represents the filling of the lake following an extended period of closure during deposition of the underlying Wilkins Peak Member. To characterize fluctuations in water chemistry and lake level at greater temporal resolution, we conducted micro-stratigraphic and chemostratigraphic analyses on 24 distinct mm-scale laminae in a single 10 cm carbonate stromatolite bed, including δ13C, δ18O, and trace elemental analyses (Mg, Mn, Fe, Si, K, Na, Al, Sr). Sub-cm-scale correlations between petrographic analyses, elemental composition, and carbonate δ13C and δ18O suggest that this stromatolite records both hydrologically-closed and -open periods in the history of Lake Gosiute. During periods of apparent basin closure, we used two models to investigate lake volume change: 1) a Rayleigh distillation model of water evaporation to estimate lake depth variations and 2) a conservative ion model based on Na incorporation into the stromatolites. In both models, lake depth fluctuated by up to 8 m; this represents up to 40km of shoreline change in Lake Gosiute during the deposition of this stromatolite layer. Interestingly, the modern Great Salt Lake experienced similar

  11. Hydrochemical and isotopic (2H, 18O and 37Cl) constraints on evolution of geothermal water in coastal plain of Southwestern Guangdong Province, China

    NASA Astrophysics Data System (ADS)

    Chen, Liuzhu; Ma, Teng; Du, Yao; Xiao, Cong; Chen, Xinming; Liu, Cunfu; Wang, Yanxin

    2016-05-01

    Geothermal energy is abundant in Guangdong Province of China, however, majority of it is still unexploited. To take full advantage of this energy, it is essential to know the information of geothermal system. Here, physical parameters such as pH and temperature, major ion (Na+, Ca2 +, Mg2 +, Cl-, SO42 - and HCO3-), trace elements (Br-, Sr2 +, Li+ and B3 +) and stable isotopes (2H, 18O and 37Cl) in geothermal water, non-geothermal water (river water, cold groundwater) and seawater were used to identify the origin and evolution of geothermal water in coastal plain of Southwest of Guangdong. Two separate groups of geothermal water have been identified in study area. Group A, located in inland of study area, is characterized by Na+ and HCO3-. Group B, located in coastal area, is characterized by Na+ and Cl-. The relationships of components vs. Cl for different water samples clearly suggest the hydrochemical differences caused by mixing with seawater and water-rock interactions. It's evident that water-rock interactions under high temperature make a significant contribution to hydrochemistry of geothermal water for both Group A and Group B. Besides, seawater also plays an important role during geothermal water evolution for Group B. Mixing ratios of seawater with geothermal water for Group B are calculated by Cl and Br binary diagram, the estimated results show that about < 1% to < 35% of seawater has mixed into geothermal water, and seawater might get into the geothermal system by deep faults. Molar Na/Cl ratios also support these two processes. Geothermal and non-geothermal water samples plot around GMWL in the δ2H vs. δ18O diagram, indicating that these samples have a predominant origin from meteoric water. Most of geothermal water samples display δ37Cl values between those of the non-geothermal water and seawater samples, further reveals three sources of elements supply for geothermal water, including atmospheric deposition, bedrocks and seawater, which show a

  12. Interpreting δD and δ18O isotopic signals of ambient water vapor in PNW coniferous forest using a high frequency CRDS analyzer

    NASA Astrophysics Data System (ADS)

    Allen, S. T.; Bond, B. J.; McDonnell, J. J.; Brooks, J. R.; Thomas, C. K.

    2010-12-01

    Wavelength-Scanned Cavity Ring-Down Spectroscopy provides real-time simultaneous measurement of stable isotopologues of water vapor in natural environments. Continuous, high-frequency sampling provides a new and exciting look at water cycle processes and creates many new possibilities for studying the vapor phase of the hydrologic cycle. However, as with any new tool, the first challenge is to understand the sources of variability in the signal. This includes disentangling potential instrument variability from environmental variability as well as the identification and quantification of environmental end members. We deployed a Picarro L-1102 Liquid / Vapor analyzer at the mouth of a small watershed in the H.J. Andrews Experimental Forest located in the West-Central Oregon Cascades range in November, 2009. The steeply-sloped watershed is covered by a closed-canopied, young-mature Douglas fir forest; it has been used for many previous ecological, hydrological, and meteorological studies. The data reveal very high diel variability in δD in and δ18O as well as δD to δ18O ratios and a strong deviation from the global meteoric water line. A hysteresis effect differs dramatically from one day to the next and confounds apparent trends. To interpret these results, we are conducting controlled tests of instrument performance and we propose a plan to partition individual vapor source contributions. Application of this vapor signature to ecological or hydrological studies requires knowledge of individual end-member contributions to the isotope measurements. We hypothesize that by determining end-member fluxes and in-situ fractionation factors paired with micrometeorological data, we can better understand processes driving these patterns. Combined with meteorological tower data, high frequency data allows the possibility of scaling up from continuous point measurements to ecosystem-scale processes. Previous studies in this watershed have demonstrated the ability to estimate

  13. Proteome Analysis using Selective Incorporation of Isotopically Labeled Amino Acids

    SciTech Connect

    Veenstra, Timothy D.; Martinovic, Suzana; Anderson, Gordon A.; Pasa-Tolic, Liljiana; Smith, Richard D.

    2000-01-01

    A method is described for identifying intact proteins from genomic databases using a combination of accurate molecular mass measurements and partial amino acid content. An initial demonstration was conducted for proteins isolated from Escherichia coli (E. coli) using a multiple auxotrophic strain of K12. Proteins were extracted from the organism grown in natural isotopic abundance minimal medium and also minimal medium containing isotopically labeled leucine (Leu-D10), were mixed and analyzed by capillary isoelectric focusing (CIEF) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICR). The incorporation of the isotopically labeled Leu residue has no effect on the CIEF separation of the protein, therefore both versions of the protein are observed within the same FTICR spectrum. The difference in the molecular mass of the natural isotopic abundance and Leu-D10 isotopically labeled proteins is used to determine the number of Leu residues present in that particular protein. Knowledge of the molecular mass and number of Leu residues present can be used to unambiguously identify the intact protein. Preliminary results show the efficacy of using this method to unambiguously identify proteins isolated from E. coli.

  14. Stable isotopes18O and δ 13C), trace and minor element compositions of Recent scleractinians and Last Glacial bivalves at the Santa Maria di Leuca deep-water coral province, Ionian Sea

    NASA Astrophysics Data System (ADS)

    Correa, Matthias López; Montagna, Paolo; Vendrell-Simón, Begoña; McCulloch, Malcolm; Taviani, Marco

    2010-03-01

    The aragonitic skeletons of bathyal cold-water corals have a high potential as geochemical in situ archives for paleoceanography. Oxygen isotopes and stable carbon isotopes18O and δ 13C) have been analyzed, as well as trace and minor element compositions (e.g. Mg/Ca, Sr/Ca, U/Ca, B/Ca and P/Ca) in Lophelia pertusa, one of the most important frame-builders at the Santa Maria di Leuca (SML) deep-water coral hotspot in the Central Mediterranean. The Apulian Bank is swept by strong currents of the Adriatic Deep Water Outflow. The temperature of 13.9 °C is the highest temperature recorded for L. pertusa and provides an important end-member of environmental conditions for geochemical analyses on living Atlantic and Mediterranean cold-water corals. Temperature and salinity (38.77 PSU) are stable throughout the year, and thus virtually no changes should be observed in the stable oxygen isotope signal—if the coral precipitates its skeleton in equilibrium with seawater. We measured various marine properties, such as the seawater oxygen isotope composition (δ 18O sw), stable carbon isotope composition (δ 13C DIC) of dissolved inorganic carbon (DIC), and dissolved inorganic nutrient concentrations (PO 4, NO 3, NO 2, NH 3 and SiO 2). Bottom water at the coral sites shows a mean oxygen isotope composition of 1.47‰ δ 18O sw-VSMOW, and δ 13C DIC showed a mean of 1.1‰ VPDB. A section of a living L. pertusa with a thick theca calcification was probed with a Merchantek MicroMill at a high spatial sampling resolution with 10 samples per 1 mm. This reduced the signal-smoothing inherent to conventional sampling. The δ 18O ag of coral aragonite ranges between -2.0‰ and +2.8‰ VPDB and the δ 13C ag ranges between -7.77‰ and +1.47‰ VPDB. The Gaussian data distribution for both parameters, including heavy equilibrium values, suggests the completeness of the captured isotopic variability. The strict linear correlation of δ 13C and δ 18O displays a strong 'kinetic

  15. Stable Isotope Labeling for Improved Comparative Analysis of RNA Digests by Mass Spectrometry.

    PubMed

    Paulines, Mellie June; Limbach, Patrick A

    2017-03-01

    Even with the advent of high throughput methods to detect modified ribonucleic acids (RNAs), mass spectrometry remains a reliable method to detect, characterize, and place post-transcriptional modifications within an RNA sequence. Here we have developed a stable isotope labeling comparative analysis of RNA digests (SIL-CARD) approach, which improves upon the original (18)O/(16)O labeling CARD method. Like the original, SIL-CARD allows sequence or modification information from a previously uncharacterized in vivo RNA sample to be obtained by direct comparison with a reference RNA, the sequence of which is known. This reference is in vitro transcribed using a (13)C/(15)N isotopically enriched nucleoside triphosphate (NTP). The two RNAs are digested with an endonuclease, the specificity of which matches the labeled NTP used for transcription. As proof of concept, several transfer RNAs (tRNAs) were characterized by SIL-CARD, where labeled guanosine triphosphate was used for the reference in vitro transcription. RNase T1 digestion products from the in vitro transcript will be 15 Da higher in mass than the same digestion products from the in vivo tRNA that are unmodified, leading to a doublet in the mass spectrum. Singlets, rather than doublets, arise if a sequence variation or a post-transcriptional modification is present that results in a relative mass shift different from 15 Da. Moreover, the use of the in vitro synthesized tRNA transcript allows for quantitative measurement of RNA abundance. Overall, SIL-CARD simplifies data analysis and enhances quantitative RNA modification mapping by mass spectrometry. Graphical Abstract ᅟ.

  16. Stable Isotope Labeling for Improved Comparative Analysis of RNA Digests by Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Paulines, Mellie June; Limbach, Patrick A.

    2017-03-01

    Even with the advent of high throughput methods to detect modified ribonucleic acids (RNAs), mass spectrometry remains a reliable method to detect, characterize, and place post-transcriptional modifications within an RNA sequence. Here we have developed a stable isotope labeling comparative analysis of RNA digests (SIL-CARD) approach, which improves upon the original 18O/16O labeling CARD method. Like the original, SIL-CARD allows sequence or modification information from a previously uncharacterized in vivo RNA sample to be obtained by direct comparison with a reference RNA, the sequence of which is known. This reference is in vitro transcribed using a 13C/15N isotopically enriched nucleoside triphosphate (NTP). The two RNAs are digested with an endonuclease, the specificity of which matches the labeled NTP used for transcription. As proof of concept, several transfer RNAs (tRNAs) were characterized by SIL-CARD, where labeled guanosine triphosphate was used for the reference in vitro transcription. RNase T1 digestion products from the in vitro transcript will be 15 Da higher in mass than the same digestion products from the in vivo tRNA that are unmodified, leading to a doublet in the mass spectrum. Singlets, rather than doublets, arise if a sequence variation or a post-transcriptional modification is present that results in a relative mass shift different from 15 Da. Moreover, the use of the in vitro synthesized tRNA transcript allows for quantitative measurement of RNA abundance. Overall, SIL-CARD simplifies data analysis and enhances quantitative RNA modification mapping by mass spectrometry.

  17. Stable Isotope Labeling for Improved Comparative Analysis of RNA Digests by Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Paulines, Mellie June; Limbach, Patrick A.

    2017-01-01

    Even with the advent of high throughput methods to detect modified ribonucleic acids (RNAs), mass spectrometry remains a reliable method to detect, characterize, and place post-transcriptional modifications within an RNA sequence. Here we have developed a stable isotope labeling comparative analysis of RNA digests (SIL-CARD) approach, which improves upon the original 18O/16O labeling CARD method. Like the original, SIL-CARD allows sequence or modification information from a previously uncharacterized in vivo RNA sample to be obtained by direct comparison with a reference RNA, the sequence of which is known. This reference is in vitro transcribed using a 13C/15N isotopically enriched nucleoside triphosphate (NTP). The two RNAs are digested with an endonuclease, the specificity of which matches the labeled NTP used for transcription. As proof of concept, several transfer RNAs (tRNAs) were characterized by SIL-CARD, where labeled guanosine triphosphate was used for the reference in vitro transcription. RNase T1 digestion products from the in vitro transcript will be 15 Da higher in mass than the same digestion products from the in vivo tRNA that are unmodified, leading to a doublet in the mass spectrum. Singlets, rather than doublets, arise if a sequence variation or a post-transcriptional modification is present that results in a relative mass shift different from 15 Da. Moreover, the use of the in vitro synthesized tRNA transcript allows for quantitative measurement of RNA abundance. Overall, SIL-CARD simplifies data analysis and enhances quantitative RNA modification mapping by mass spectrometry.

  18. Enzymatic synthesis of isotopically labeled isoprenoid diphosphates.

    PubMed

    Christensen, D J; Poulter, C D

    1994-07-01

    Recombinant yeast isopentenyl diphosphate (IPP) isomerase and avian farnesyl diphosphate (FPP) synthase from overproducing strains of Escherichia coli were used to synthesize FPP from IPP and dimethylallyl diphosphate (DMAPP). [2,4,5-13C3]IPP and [2,4,5-13C3]DMAPP were synthesized from ethyl [2-13C]bromoacetate and [1,3-13C2]acetone. Thes compounds were used as substrates for enzymatic synthesis of FPP selectivity labeled at the first or third isoprene residue or at all three.

  19. Using Halogens (Cl, Br, F, I) and Stable Isotopes of Water (δ18O, δ2H) to Trace Hydrological and Biogeochemical Processes in Prairie Wetlands

    NASA Astrophysics Data System (ADS)

    Levy, Z. F.; Lu, Z.; Mills, C. T.; Goldhaber, M. B.; Rosenberry, D. O.; Mushet, D.; Siegel, D. I.; Fiorentino, A. J., II; Gade, M.; Spradlin, J.

    2014-12-01

    Prairie pothole wetlands are ubiquitous features of the Great Plains of North America, and important habitat for amphibians and migratory birds. The salinity of proximal wetlands varies highly due to groundwater-glacial till interactions, which influence wetland biota and associated ecosystem functions. Here we use halogens and stable isotopes of water to fingerprint hydrological and biogeochemical controls on salt cycling in a prairie wetland complex. We surveyed surface, well, and pore waters from a groundwater recharge wetland (T8) and more saline closed (P1) and open (P8) basin discharge wetlands in the Cottonwood Lake Study Area (ND) in August/October 2013 and May 2014. Halogen concentrations varied over a broad range throughout the study area (Cl = 2.2 to 170 mg/L, Br = 13 to 2000 μg/L, F = < 30 (MDL) to 740 μg/L, I = 1 to 538 μg/L). The Cl/Br molar ratios were higher (171 to 574) at the recharge wetland, indicating meteoric sources, and had a tighter and lower range (33 to 320) at the down-gradient sites. The Cl/I molar ratios of waters throughout the site had a wide range (32 to 26,000). Lowest values occurred at the upgradient shore of P1 (32 to 43) due to low Cl concentrations and the center of P1 (196 to 213) where pore water of weathered till underlying 1.2 m of organic-rich sediment and silty clay soil is enriched in I to ~500 µg/L. Stable isotopes of water showed that evaporation-enriched pond water (δ18O = -9.5 to -2.71 ‰) mixes with shallow groundwater in the top 0.6 m of fringing wetland soils and 1.2 m of the substrate in the center of P1. Our results suggest endogenous sources for Br and I within the prairie landscape that may be controlled by biological mechanisms or weathering of shale from glacial till.

  20. Measurement of the D/H, 18O/16O, and 17O/16O Isotope Ratios in Water by Laser Absorption Spectroscopy at 2.73 μm

    PubMed Central

    Wu, Tao; Chen, Weidong; Fertein, Eric; Masselin, Pascal; Gao, Xiaoming; Zhang, Weijun; Wang, Yingjian; Koeth, Johannes; Brückner, Daniela; He, Xingdao

    2014-01-01

    A compact isotope ratio laser spectrometry (IRLS) instrument was developed for simultaneous measurements of the D/H, 18O/16O and 17O/16O isotope ratios in water by laser absorption spectroscopy at 2.73 μm. Special attention is paid to the spectral data processing and implementation of a Kalman adaptive filtering to improve the measurement precision. Reduction of up to 3-fold in standard deviation in isotope ratio determination was obtained by the use of a Fourier filtering to remove undulation structure from spectrum baseline. Application of Kalman filtering enables isotope ratio measurement at 1 s time intervals with a precision (<1‰) better than that obtained by conventional 30 s averaging, while maintaining a fast system response. The implementation of the filter is described in detail and its effects on the accuracy and the precision of the isotope ratio measurements are investigated. PMID:24854363

  1. δ13C and δ18O isotopic composition of CaCO3 measured by continuous flow isotope ratio mass spectrometry: statistical evaluation and verification by application to Devils Hole core DH-11 calcite

    USGS Publications Warehouse

    Revesz, Kinga M.; Landwehr, Jurate M.

    2002-01-01

    A new method was developed to analyze the stable carbon and oxygen isotope ratios of small samples (400 ± 20 µg) of calcium carbonate. This new method streamlines the classical phosphoric acid/calcium carbonate (H3PO4/CaCO3) reaction method by making use of a recently available Thermoquest-Finnigan GasBench II preparation device and a Delta Plus XL continuous flow isotope ratio mass spectrometer. Conditions for which the H3PO4/CaCO3 reaction produced reproducible and accurate results with minimal error had to be determined. When the acid/carbonate reaction temperature was kept at 26 °C and the reaction time was between 24 and 54 h, the precision of the carbon and oxygen isotope ratios for pooled samples from three reference standard materials was ≤0.1 and ≤0.2 per mill or ‰, respectively, although later analysis showed that materials from one specific standard required reaction time between 34 and 54 h for δ18O to achieve this level of precision. Aliquot screening methods were shown to further minimize the total error. The accuracy and precision of the new method were analyzed and confirmed by statistical analysis. The utility of the method was verified by analyzing calcite from Devils Hole, Nevada, for which isotope-ratio values had previously been obtained by the classical method. Devils Hole core DH-11 recently had been re-cut and re-sampled, and isotope-ratio values were obtained using the new method. The results were comparable with those obtained by the classical method with correlation = +0.96 for both isotope ratios. The consistency of the isotopic results is such that an alignment offset could be identified in the re-sampled core material, and two cutting errors that occurred during re-sampling then were confirmed independently. This result indicates that the new method is a viable alternative to the classical reaction method. In particular, the new method requires less sample material permitting finer resolution and allows

  2. Heavy isotope production by multinucleon transfer reactions with /sup 254/Es. [101 MeV /sup 16/O, 98 MeV /sup 18/O, 127 MeV /sup 22/Ne

    SciTech Connect

    Schaedel, M.; Bruechle, W.; Bruegger, M.; Gaeggeler, H.; Moody, K.J.; Schardt, D.; Suemmerer, K.; Hulet, E.K.; Douran, A.D.; Dougan, R.J.

    1985-01-01

    Fast automated on-line and quasi-on-line radiochemical techniques were applied to search for new isotopes, to measure their decay characteristics, and to study the cross sections of the heaviest, most neutron-rich actinide isotopes in reactions of /sup 16,18/O and /sup 22/Ne projectiles with /sup 254/Es as a target. The measured yields for isotopes up to Lr-260 are three or more orders of magnitude higher than in any other reaction used so far. A comparison with data for similar transfers from /sup 248/Cm targets is made. Transfer cross sections are extrapolated for the production of unknown, neutron-rich isotopes of elements 101 through 105, and the unique potential of /sup 254/Es as a target to make these exoctic nuclei accessible is demonstrated. 18 refs., 2 figs., 1 tab.

  3. Complete Measurement of Stable Isotopes in N2O (δ15N, δ15Nα, δ15Nβ, δ18O, δ17O) Using Off-Axis Integrated Cavity Output Spectroscopy (OA-ICOS)

    NASA Astrophysics Data System (ADS)

    Leen, J. B.; Gupta, M.

    2014-12-01

    Nitrate contamination in water is a worldwide environmental problem and source apportionment is critical to managing nitrate pollution. Fractionation caused by physical, chemical and biological processes alters the isotope ratios of nitrates (15N/14N, 18O/16O and 17O/16O) and biochemical nitrification and denitrification impart different intramolecular site preference (15N14NO vs. 14N15NO). Additionally, atmospheric nitrate is anomalously enriched in 17O compared to other nitrate sources. The anomaly (Δ17O) is conserved during fractionation processes, providing a tracer of atmospheric nitrate. All of these effects can be used to apportion nitrate in soil. Current technology for measuring nitrate isotopes is complicated and costly - it involves conversion of nitrate to nitrous oxide (N2O), purification, preconcentration and measurement by isotope ratio mass spectrometer (IRMS). Site specific measurements require a custom IRMS. There is a pressing need to make this measurement simpler and more accessible. Los Gatos Research has developed a next generation mid-infrared Off-Axis Integrated Cavity Output Spectroscopy (OA-ICOS) analyzer to quantify all stable isotope ratios of N2O (δ15N, δ15Nα, δ15Nβ, δ18O, δ17O). We present the latest performance data demonstrating the precision and accuracy of the OA-ICOS based measurement. At an N2O concentration of 322 ppb, the analyzer quantifies [N2O], δ15N, δ15Na, δ15Nb, and δ18O with a precision of ±0.05 ppb, ±0.4 ‰, ±0.45 ‰, and ±0.6 ‰, and ±0.8 ‰ respectively (1σ, 100s; 1σ, 1000s for δ18O). Measurements of gas standards demonstrate accuracy better than ±1 ‰ for isotope ratios over a wide dynamic range (200 - 100,000 ppb). The measurement of δ17O requires a higher concentration (1 - 50 ppm), easily obtainable through conversion of nitrates in water. For 10 ppm of N2O, the instrument achieves a δ17O precision of ±0.05 ‰ (1σ, 1000s). This performance is sufficient to quantify atmospheric

  4. Adaptation of the doubly labeled water method for subjects consuming isotopically enriched water.

    PubMed

    Gretebeck, R J; Schoeller, D A; Socki, R A; Davis-Street, J; Gibson, E K; Schulz, L O; Lane, H W

    1997-02-01

    The use of doubly labeled water (DLW) to measure energy expenditure is subject to error if the background abundance of the oxygen and hydrogen isotope tracers changes during the test period. This study evaluated the accuracy and precision of different methods by which such background isotope changes can be corrected, including a modified method that allows prediction of the baseline that would be achieved if subjects were to consume water from a given source indefinitely. Subjects in this study were eight women (4 test subjects and 4 control subjects) who consumed for 28 days water enriched to resemble drinking water aboard the United States space shuttle. Test subjects and control subjects were given a DLW dose on days 1 and 15, respectively. The change to an enriched water source produced a bias in expenditure calculations that exceeded 2.9 MJ/day (35%), relative to calculations from intake-balance. The proposed correction based on the predicted final abundance of 18O and deuterium after equilibration to the new water source eliminated this bias, as did the traditional use of a control group. This new modified correction method is advantageous under field conditions when subject numbers are limited.

  5. Isotopic labeling for the understanding of the alteration of limestone used in built cultural heritage

    NASA Astrophysics Data System (ADS)

    Saheb, Mandana; Chabas, Anne; Mertz, Jean-Didier; Rozenbaum, Olivier; Verney-Carron, Aurélie

    2015-04-01

    This project belongs to a specific work aiming at developing isotopic tools to better understand the alteration of materials used in the built cultural heritage. It is focused on the study of the alteration of limestone used in the facades of historic buildings subject to atmospheric polluted environment. Actually in the elevated parts of the buildings, water as rainfall (runoff or wet deposition) or in vapor form (condensation or dry deposition) is the main agent of alteration. Thus, the rock/water interactions need to be well understood to propose adapted solution to better preserve the buildings. To identify the water transfer within the porous limestone and locate the reaction preferential sites, two isotopic tracers (D and 18O) are used to monitor the alteration solution (D) and locate the zones containing the secondary phases (18O). The Saint-Maximin limestone used in many monuments in the suburbs of Paris (France) as a building and restoration stone has been specifically studied. Pristine materials, stones from monuments (monuments in the Paris area) and samples altered in laboratory constitute the analytical corpus to compare different stages of alteration. In a first step the stones are characterized at different scales to identify the alteration pattern (SEM-EDS, Raman microspectrometry, XRD, rugosimetry) and study the water transfers (X-ray tomography, mercury porosimetry, imbibition kinetics). The samples are then altered in the laboratory by realistic and controlled wet or dry deposition using isotopically labeled solutions to locate the reaction zones by SIMS. The multiscale characterization of the alteration pattern has allowed proposing alteration mechanisms linked to the properties of the stones and their location inside the building. Moreover, the location of the reactive zones inside the materials determined by the isotopic experiments helps examining the role of the evolution of porosity and formation of alteration products within the material

  6. Isotope labeling for NMR studies of macromolecular structure and interactions

    SciTech Connect

    Wright, P.E.

    1994-12-01

    Implementation of biosynthetic methods for uniform or specific isotope labeling of proteins, coupled with the recent development of powerful heteronuclear multidimensional NMR methods, has led to a dramatic increase in the size and complexity of macromolecular systems that are now amenable to NMR structural analysis. In recent years, a new technology has emerged that combines uniform {sup 13}C, {sup 15}N labeling with heteronuclear multidimensional NMR methods to allow NMR structural studies of systems approaching 25 to 30 kDa in molecular weight. In addition, with the introduction of specific {sup 13}C and {sup 15}N labels into ligands, meaningful NMR studies of complexes of even higher molecular weight have become feasible. These advances usher in a new era in which the earlier, rather stringent molecular weight limitations have been greatly surpassed and NMR can begin to address many central biological problems that involve macromolecular structure, dynamics, and interactions.

  7. Nucleosynthesis in AGB stars traced by oxygen isotopic ratios. I. Determining the stellar initial mass by means of the 17O/18O ratio

    NASA Astrophysics Data System (ADS)

    De Nutte, R.; Decin, L.; Olofsson, H.; Lombaert, R.; de Koter, A.; Karakas, A.; Milam, S.; Ramstedt, S.; Stancliffe, R. J.; Homan, W.; Van de Sande, M.

    2017-03-01

    Aims: We seek to investigate the 17O/18O ratio for a sample of AGB stars containing M-, S-, and C-type stars. These ratios are evaluated in relation to fundamental stellar evolution parameters: the stellar initial mass and pulsation period. Methods: Circumstellar 13C16O, 12C17O, and 12C18O line observations were obtained for a sample of nine stars with various single-dish long-wavelength facilities. Line intensity ratios are shown to relate directly to the surface 17O/18O abundance ratio. Results: Stellar evolution models predict the 17O/18O ratio to be a sensitive function of initial mass and to remain constant throughout the entire TP-AGB phase for stars initially less massive than 5 M⊙. This makes the measured ratio a probe of the initial stellar mass. Conclusions: Observed 17O/18O ratios are found to be well in the range predicted by stellar evolution models that do not consider convective overshooting. From this, accurate initial mass estimates are calculated for seven sources. For the remaining two sources, there are two mass solutions, although there is a larger probability that the low-mass solution is correct. Finally, we present hints at a possible separation between M/S- and C-type stars when comparing the 17O/18O ratio to the stellar pulsation period. The reduced spectra are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (http://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/600/A71

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  9. Using nitrate dual isotopic composition (δ15N and δ18O) as a tool for exploring sources and cycling of nitrate in an estuarine system: Elkhorn Slough, California

    NASA Astrophysics Data System (ADS)

    Wankel, Scott D.; Kendall, Carol; Paytan, Adina

    2009-03-01

    Nitrate (NO3-) concentrations and dual isotopic composition (δ15N and δ18O) were measured during various seasons and tidal conditions in Elkhorn Slough to evaluate mixing of sources of NO3- within this California estuary. We found the isotopic composition of NO3- was influenced most heavily by mixing of two primary sources with unique isotopic signatures, a marine (Monterey Bay) and terrestrial agricultural runoff source (Old Salinas River). However, our attempt to use a simple two end-member mixing model to calculate the relative contribution of these two NO3- sources to the Slough was complicated by periods of nonconservative behavior and/or the presence of additional sources, particularly during the dry season when NO3- concentrations were low. Although multiple linear regression generally yielded good fits to the observed data, deviations from conservative mixing were still evident. After consideration of potential alternative sources, we concluded that deviations from two end-member mixing were most likely derived from interactions with marsh sediments in regions of the Slough where high rates of NO3- uptake and nitrification result in NO3- with low δ15N and high δ18O values. A simple steady state dual isotope model is used to illustrate the impact of cycling processes in an estuarine setting which may play a primary role in controlling NO3- isotopic composition when and where cycling rates and water residence times are high. This work expands our understanding of nitrogen and oxygen isotopes as biogeochemical tools for investigating NO3- sources and cycling in estuaries, emphasizing the role that cycling processes may play in altering isotopic composition.

  10. Radiogenic and stable isotopes of mid-Miocene silicic volcanism in eastern Oregon: Evidence for variable and high Sr / low δ18O domains west of the terrane-cratonic lithosphere transition

    NASA Astrophysics Data System (ADS)

    Jenkins, E. N.; Streck, M. J.; Ramos, F. C.; Bindeman, I. N.

    2013-12-01

    Widespread mid-Miocene rhyolite volcanism of eastern Oregon mostly coeval with flood basalts of the Columbia River Basalt Province allows for mapping crustal domains using radiogenic and stable isotopes. Rhyolites are thought to be derived in large part by partial melting of the crust and thus yield direct information on the composition of the crust. Silicic volcanism is expressed in the form of numerous domes and tuffs exposed over a wide area (~300 km in N-S dimension and ~100 km in E-W dimension) west of the craton boundary, which runs parallel but mostly east of the Oregon-Idaho state border as delineated by geophysical characteristics and isotopic transitions. Here, we mainly focus on initial 87Sr/86Sr ratios and δ18O obtained from mid-Miocene silicic volcanic centers in eastern Oregon. Our data, in combination with data from the literature, indicate variable 87Sr/86Sr mostly along longitudinal sections, yet more similar ratios in latitudinal directions. Except for rare examples on the west side, dispersion of 87Sr/86Sr ratios among both silicic and basaltic rocks occurs eastward of 118.6°W. For example, rhyolites in the Owyhee region between 117.10°W and 117.25°W retain 87Sr/86Sr ratios ranging from 0.70413 to 0.70566. The most radiogenic Sri ratio of 0.70787 in our study is obtained on a plagioclase separate from Buchanan Dome complex located near the western boundary of our study area. Feldspar separates and fresh groundmass of samples from adjacent centers yield similar 87Sr/86Sr ratios. δ18O values for feldspars range from below 2‰ to above 9‰. In addition, there is a crude trend of rhyolites having lower δ18O and more radiogenic 87Sr/86Sr ratios. With one exception, all samples with 87Sr/86Sr above 0.7050 are depleted in 18O18O <5.5‰), while rhyolites with 87Sr/86Sr below 0.7045 are enriched in 18O18O >6‰). The most depleted oxygen ratios (<2‰) come from rhyolites ~80 km west of the cratonic margin reflecting remelting or

  11. Isotopic Labeling of Red Cabbage Anthocyanins with Atmospheric 13-CO2

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Isotopic labeling of plants provides a unique opportunity for understanding metabolic processes. A significant challenge of isotopic labeling during plant growth is that isotopes must be administered without disrupting plant development and at sufficient levels for mass spectral analysis. We describ...

  12. The value of stable Isotope (18O) and electrical conductivity (EC) as tracers for submarine Groundwater exfiltration and density-driven flow infiltration into the aquifer.

    NASA Astrophysics Data System (ADS)

    Müller, Sascha; Engesgaard, Peter; Duque, Carlos; Jessen, Søren; Sonnenborg, Torben; Stau, Joakim; Neilson, Bethany

    2015-04-01

    Saltwater intrusion (SWI) into a freshwater aquifer is a dynamic process due to e.g. natural changes in sea levels (tides) and recharge. Coastal lagoons, on the other hand, are often controlled water bodies where the water level and salinity are managed by the operation of a sluice connecting the lagoon to the ocean. This study describes the seasonal dynamics of the saltwater/freshwater interface and submarine groundwater discharge (SGD) patterns at a coastal lagoon on the West coast of Denmark. Here the salinity of the lagoon is high in the summer period, where recharge is low (favoring SWI) and vice versa in the winter time. SGD was measured over four seasons in 2012 along two transects. 18O and electrical conductivity (EC) were measured at the same time to a depth of 3.5 m with a sample interval of 0.25 m. In September 2014 a transect with 12 piezometers (screening depth between 1.5 and 15 m below surface) and one profile well (with measurements every 1 m down to 15 m) was established across the saltwater/freshwater interface at one of the transects. 18O and EC were measured and each piezometer was equipped with a CTD-diver measuring pressure head, temperature, and EC in the period switching from summer to winter conditions. Although 18O and EC is relatively well correlated (correlation coefficient of 0.8) the use of both tracers are recommend for this type of environment. Salinity (or EC) in the lagoon changes seasonally, whereas 18O in both lagoon water and groundwater is relatively stable within each end- member, suggesting that 18O is the tracer to prefer. However, on the other hand EC is an easy and in-expensive (continuous) measurement allowing a much better resolution in both space and time. The combination of both tracers can improve the explanation of the origin of water with more certainty. Both tracers show a seasonal interplay between freshwater discharge into the lagoon and a density- driven recycling with opposing flow into the aquifer. 18O and EC

  13. Heating Isotopically Labeled Bernal Stacked Graphene: A Raman Spectroscopy Study.

    PubMed

    Ek-Weis, Johan; Costa, Sara; Frank, Otakar; Kalbac, Martin

    2014-02-06

    One of the greatest issues of nanoelectronics today is how to control the heating of the components. Graphene is a promising material in this area, and it is essential to study its thermal properties. Here, the effect of heating a bilayer structure was investigated using in situ Raman spectroscopy. In order to observe the effects on each individual layer, an isotopically labeled bilayer graphene was synthesized where the two layers were composed of different carbon isotopes. Therefore, the frequency of the phonons in the Raman spectra was shifted in relation to each other. This technique was used to investigate the influence of different stacking order. It was found that in bilayer graphene grown by chemical vapor deposition (CVD), the two layers behave very similarly for both Bernal stacking and randomly oriented structures, while for transferred samples, the layers act more independently. This highlights a significant dependence on the sample preparation procedure.

  14. Life history of the individuals buried in the St. Benedict Cemetery (Prague, 15th-18th centuries): insights from (14)C dating and stable isotope (δ(13)C, δ(15)N, δ(18)O) analysis.

    PubMed

    Salesse, Kevin; Dufour, Élise; Castex, Dominique; Velemínský, Petr; Santos, Frédéric; Kuchařová, Hedvika; Jun, Libor; Brůžek, Jaroslav

    2013-06-01

    Funerary practices and bioarchaeological (sex and age) data suggest that a mortality crisis linked to an epidemic episode occurred during the fifth phase of the St. Benedict cemetery in Prague (Czech Republic). To identify this mass mortality episode, we reconstructed individual life histories (dietary and mobility factors), assessed the population's biological homogeneity, and proposed a new chronology through stable isotope analysis (δ(13)C, δ(18)O and δ(15)N) and direct radiocarbon dating. Stable isotope analysis was conducted on the bone and tooth enamel (collagen and carbonate) of 19 individuals from three multiple graves (MG) and 12 individuals from individual graves (IG). The δ(15)N values of collagen and the difference between the δ(13)C values of collagen and bone carbonate could indicate that the IG individuals had a richer protein diet than the MG individuals or different food resources. The human bone and enamel carbonate and δ(18)O values suggest that the majority of individuals from MG and all individuals from IG spent most of their lives outside of the Bohemian region. Variations in δ(18)O values also indicate that all individuals experienced residential mobility during their lives. The stable isotope results, biological (age and sex) data and eight (14)C dates clearly differentiate the MG and IG groups. The present work provides evidence for the reuse of the St. Benedict cemetery to bury soldiers despite the funeral protest ban (1635 AD). The Siege of Prague (1742 AD) by French-Bavarian-Saxon armies is identified as the cause of the St. Benedict mass mortality event.

  15. The oxygen-hafnium isotope paradox in the early post Columbia River Basalt silicic volcanism: Evidence for complex batch assembly of upper crustal, lower crustal and low-δ18O silicic magmas

    NASA Astrophysics Data System (ADS)

    Colon, D.; Bindeman, I. N.; Ellis, B. S.; Schmitt, A. K.; Fisher, C. M.; Vervoort, J. D.

    2013-12-01

    Eruptions of the Columbia River flood basalts were immediately followed by large eruptions of silicic magmas; some may have been coeval, others genetically-linked to the CRB. Among the most voluminous of these eruptions was the Jarbidge Rhyolite, which comprises ~500 km3 of lava erupted from 16.1-15.0 Ma in northern Nevada. Activity at Jarbidge was followed at 15.0 Ma by a series of rhyolitic ignimbrites and lavas in the J-P Desert of Idaho ~50 km NW of the Jarbidge Rhyolite center. To constrain magmatic origins and upper crustal magma storage conditions of these two silicic magmatic systems, we conducted bulk and high spatial resolution analysis of whole rocks and minerals (quartz, feldspar, and zircon). Bulk quartz and plagioclase δ18O values of the J-P Desert units are only moderately lower than mantle values, with δ18O-quartz of 5.0-5.5‰ and plagioclase δ18O of ~3.9-5.8‰, along with slightly unradiogenic Nd and Hf whole rock values (average ɛHf and ɛNd of -13.1 and -10.0, respectively), while quartz from the Jarbidge Rhyolite has normal δ18O (+8.4‰), but very unradiogenic ɛHf-ɛNd (ɛHf = -34.7, ɛNd = -24.0), fingerprinting Archean upper crust. SIMS analysis of J-P Desert zircons reveals considerably diverse δ18O values, ranging from -0.6‰ to +6.5‰ in a single unit. The same zircon spots yielded U-Pb SIMS ages which generally agree with the 40Ar/39Ar eruption ages, with no evidence of inheritance of pre-Miocene zircons. Combined with LA-MC-ICP-MS analysis of Hf isotopes overlapping the earlier SIMS spots, these zircons show a clear near-linear correlation between ɛHf and δ18O values observed in individual zircons. This relationship suggests variable mixing of two distinct silicic magmas prior to eruption of the J-P Desert rhyolites. One of these, characterized by extremely low ɛHf values and normal δ18O values, is likely a mantle magma strongly contaminated with shallow Archean crust, represented by the Jarbidge Rhyolite. The other is

  16. Plan of study to determine if the isotopic ratios [delta]15 N and [delta]18 O can reveal the sources of nitrate discharged by the Mississippi River into the Gulf of Mexico

    USGS Publications Warehouse

    Battaglin, William A.; Kendall, Carol; Goolsby, Donald A.; Boyer, Laurie L.

    1997-01-01

    Nitrate and other nutrients discharged from the Mississippi River basin are suspected of causing a zone of depleted dissolved oxygen (hypoxic zone) in the Gulf of Mexico each summer. The hypoxic zone may have an adverse effect on aquatic life and commercial fisheries. Commercial fertilizers are the dominant source of nitrogen input to the Mississippi basin. Other nitrogen sources include animal waste, fixation of atmospheric nitrogen by legumes, precipitation, domestic and industrial effluent, and the soil. The inputs of nitrogen from most of these sources to the Mississippi basin can be estimated and the outputs in surface water can be measured. However, nitrogen from each source is affected differently by physical, chemical, and biological processes that control nitrogen cycling in terrestrial and aquatic systems. Hence, the relative contributions from the various sources of nitrogen to nitrate load in the Mississippi River are unknown because the different sources may not contribute proportionally to their inputs in the basin. It may be possible to determine the relative contributions of the major sources of nitrate in river water using the stable isotopic ratios d15N and d18O of the nitrate ion. A few researchers have used the d15N and/or d18O isotope ratios to determine sources of nitrate in ground water, headwater catchments, and small rivers, but little is known about the isotopic composition of nitrate in larger rivers. The objective of this study is to measure the isotopic composition of nitrate and suspended organic matter in the Mississippi River and its major tributaries, in discharge to the Gulf of Mexico, and in streamflow from smaller watersheds that have distinct sources of nitrogen (row crops, animal wastes, and urban effluents) or are minimally impacted by man (undeveloped). Samples from seven sites on the Mississippi River and its tributaries and from 17 sites in smaller watersheds within the Mississippi River basin will be analyzed for d15N and

  17. Determination of the 15N/14N, 17O/16O, and 18O/16O ratios of nitrous oxide by using continuous-flow isotope-ratio mass spectrometry.

    PubMed

    Komatsu, Daisuke D; Ishimura, Toyoho; Nakagawa, Fumiko; Tsunogai, Urumu

    2008-05-01

    We developed a rapid, sensitive, and automated analytical system to determine the delta15N, delta18O, and Delta17O values of nitrous oxide (N2O) simultaneously in nanomolar quantities for a single batch of samples by continuous-flow isotope-ratio mass spectrometry (CF-IRMS) without any cumbersome and time-consuming pretreatments. The analytical system consisted of a vacuum line to extract and purify N2O, a gas chromatograph for further purification of N2O, an optional thermal furnace to decompose N2O to O2, and a CF-IRMS system. We also used pneumatic valves and pneumatic actuators in the system so that we could operate it automatically with timing software on a personal computer. The analytical precision was better than 0.12 per thousand for delta15N with >4 nmol N2O injections, 0.25 per thousand for delta18O with >4 nmol N2O injections, and 0.20 per thousand for Delta17O with >20 nmol N2O injections for a single measurement. We were also easily able to improve the precision (standard errors) to better than 0.05 per thousand for delta15N, 0.10 per thousand for delta18O, and 0.10 per thousand for Delta17O through multiple analyses with more than four repetitions with 190 nmol samples using the automated analytical system. Using the system, the delta15N, delta18O, and Delta17O values of N2O can be quantified not only for atmospheric samples, but also for other gas or liquid samples with low N2O content, such as soil gas or natural water. Here, we showed the first ever Delta17O measurements of soil N2O.

  18. Production of isotopically-labeled standards from a uniformly labeled precursor for quantitative volatile metabolomic studies

    PubMed Central

    Gómez-Cortés, Pilar; Brenna, J. Thomas; Sacks, Gavin L.

    2012-01-01

    Optimal accuracy and precision in small molecule profiling by mass spectrometry generally requires isotopically labeled standards chemically representative of all compounds of interest. However, preparation of mixed standards from commercially available pure compounds is often prohibitively expensive and time consuming, and many labeled compounds are not available in pure form. We used a single prototype uniformly labeled [U-13C]-compound to generate [U-13C]-volatile standards for use in subsequent experimental profiling studies. [U-13C]-α-linolenic acid (C18:3n-3, ALA) was thermally oxidized to produce labeled lipid degradation volatiles which were subsequently characterized qualitatively and quantitatively. Twenty-five [U-13C]-labeled volatiles were identified by headspace solid-phase microextraction-gas chromatography-time of flight-mass spectrometry (HS-SPME-GC-TOF-MS) by comparison of spectra with unlabeled volatiles. Using 250 μL starting sample, labeled volatiles were quantified by a reverse isotope dilution procedure. Using the [U-13C]-labeled standards, limits of detection comparable to or better than previous HS-SPME reports were achieved, 0.010–1.04 ng/g. The performance of the [U-13C]-volatile standards was evaluated using a commodity soybean oil (CSO) oxidized at 60°C from 0 to 15 d. Relative responses of n-decane, an unlabeled internal standard otherwise absent from the mixture, and [U-13C]-oxidation products changed by up to 8-fold as the CSO matrix was oxidized, demonstrating that reliance on a single standard in volatile profiling studies yields inaccurate results due to changing matrix effects. The [U-13C]-standard mixture was used to quantify 25 volatiles in oxidized CSO and low-ALA soybean oil with an average relative standard deviation of 8.5%. Extension of this approach to other labeled substrates, e.g., [U-13C]-sugars and amino acids, for profiling studies should be feasible and can dramatically improve quantitative results compared to

  19. Residue-Specific Structural Kinetics of Proteins through the Union of Isotope Labeling, Mid-IR Pulse Shaping, and Coherent 2D IR Spectroscopy

    PubMed Central

    Middleton, Chris T.; Woys, Ann Marie; Mukherjee, Sudipta S.; Zanni, Martin T.

    2010-01-01

    We describe a methodology for studying protein kinetics using a rapid-scan technology for collecting 2D IR spectra. In conjunction with isotope labeling, 2D IR spectroscopy is able to probe the secondary structure and environment of individual residues in polypeptides and proteins. It is particularly useful for membrane and aggregate proteins. Our rapid-scan technology relies on a mid-IR pulse shaper that computer generates the pulse shapes, much like in an NMR spectrometer. With this device, data collection is faster, easier, and more accurate. We describe our 2D IR spectrometer, as well as protocols for 13C=18O isotope labeling, and then illustrate the technique with an application to the aggregation of the human islet amyloid polypeptide form type 2 diabetes. PMID:20472067

  20. Legacy of contaminant N sources to the NO3‑ signature in rivers: a combined isotopic (δ15N-NO3‑, δ18O-NO3‑, δ11B) and microbiological investigation

    NASA Astrophysics Data System (ADS)

    Briand, Cyrielle; Sebilo, Mathieu; Louvat, Pascale; Chesnot, Thierry; Vaury, Véronique; Schneider, Maude; Plagnes, Valérie

    2017-02-01

    Nitrate content of surface waters results from complex mixing of multiple sources, whose signatures can be modified through N reactions occurring within the different compartments of the whole catchment. Despite this complexity, the determination of nitrate origin is the first and crucial step for water resource preservation. Here, for the first time, we combined at the catchment scale stable isotopic tracers (δ15N and δ18O of nitrate and δ11B) and fecal indicators to trace nitrate sources and pathways to the stream. We tested this approach on two rivers in an agricultural region of SW France. Boron isotopic ratios evidenced inflow from anthropogenic waters, microbiological markers revealed organic contaminations from both human and animal wastes. Nitrate δ15N and δ18O traced inputs from the surface leaching during high flow events and from the subsurface drainage in base flow regime. They also showed that denitrification occurred within the soils before reaching the rivers. Furthermore, this study highlighted the determinant role of the soil compartment in nitrate formation and recycling with important spatial heterogeneity and temporal variability.

  1. Legacy of contaminant N sources to the NO3− signature in rivers: a combined isotopic (δ15N-NO3−, δ18O-NO3−, δ11B) and microbiological investigation

    PubMed Central

    Briand, Cyrielle; Sebilo, Mathieu; Louvat, Pascale; Chesnot, Thierry; Vaury, Véronique; Schneider, Maude; Plagnes, Valérie

    2017-01-01

    Nitrate content of surface waters results from complex mixing of multiple sources, whose signatures can be modified through N reactions occurring within the different compartments of the whole catchment. Despite this complexity, the determination of nitrate origin is the first and crucial step for water resource preservation. Here, for the first time, we combined at the catchment scale stable isotopic tracers (δ15N and δ18O of nitrate and δ11B) and fecal indicators to trace nitrate sources and pathways to the stream. We tested this approach on two rivers in an agricultural region of SW France. Boron isotopic ratios evidenced inflow from anthropogenic waters, microbiological markers revealed organic contaminations from both human and animal wastes. Nitrate δ15N and δ18O traced inputs from the surface leaching during high flow events and from the subsurface drainage in base flow regime. They also showed that denitrification occurred within the soils before reaching the rivers. Furthermore, this study highlighted the determinant role of the soil compartment in nitrate formation and recycling with important spatial heterogeneity and temporal variability. PMID:28150819

  2. Determination of wine authenticity and geographical origin by measuring non-exchangeable hydrogen stable isotopes in wine ethanol with EIM-IRMS® methodology in combination with δ18O values obtained from wine water.

    NASA Astrophysics Data System (ADS)

    Smajlovic, Ivan; Glavanovic, Mirko; Sparks, Kimberlee L.; Sparks, Jed P.; Jovic, Slobodan

    2014-05-01

    Wine consumption has grown significantly in the last two decades, with the United States being the leading consumer of wine in the world. It is also the second largest wine producer and importer after the European Union, which consists of 27 European countries. The world has seen a significant increase in production from new world countries, especially the United States, Australia and Chile, and wine imports have grown significantly with this globalization. The quality and authenticity of products have become critical concerns. With the amount of wine being imported the need for verifying wine authenticity and understanding procedures used in wine making has become more important than ever. Understanding the origin of consumed wine in rapidly expanding global economy has become fundamental in order to control quality and protect consumers. In our previous scientific work we have shown that EIM-IRMS®, Ethanol Isotope Measurement - Isotope Ratio Mass Spectrometry (EIM-IRMS®), is capable of providing unique molecular fingerprint that cannot be reproduced or counterfeited. Today we know that δ18O value from the wine water is one of the most important parameters which can give information about wine geographical origin. Earlier we have suggested that grape juice or grape pulp is a closed biochemical system in which all chemical compounds stand in dynamic equilibrium and are in direct connection with each other. Taking that into consideration we have concluded that if system is genuine and if no water, or no sugar has been added to the grape must or grape juice prior to alcoholic fermentation, then ethanol which is made in process of alcoholic fermentation will have specific δD value of non-exchangeable hydrogen stable isotopes which will be in range from -205 to -215 ‰ vs. V-SMOW. In this work we will show that this value, which we named δDn (non-exchangeable hydrogen stable isotopes in ethanol), is very important because it can support or refute conclusions

  3. Water isotope ratios D/H, 18O/16O, 17O/16O in and out of clouds map dehydration pathways.

    PubMed

    Webster, Christopher R; Heymsfield, Andrew J

    2003-12-05

    Water isotope ratios have been measured by laser absorption spectroscopy in and out of cirrus clouds formed in situ and convectively generated in anvils over subtropical regions. Water vapor in the tropical and subtropical upper troposphere shows a wide range of isotopic depletion not observed previously. The range suggests that dehydration of upper tropospheric air occurs both by convective dehydration and by gradual dehydration mechanisms. Twenty-five percent of upper tropospheric water sampled is in ice particles whose isotopic signatures are used to identify those grown in situ from those lofted from below.

  4. Mathematical modeling of isotope labeling experiments for metabolic flux analysis.

    PubMed

    Nargund, Shilpa; Sriram, Ganesh

    2014-01-01

    Isotope labeling experiments (ILEs) offer a powerful methodology to perform metabolic flux analysis. However, the task of interpreting data from these experiments to evaluate flux values requires significant mathematical modeling skills. Toward this, this chapter provides background information and examples to enable the reader to (1) model metabolic networks, (2) simulate ILEs, and (3) understand the optimization and statistical methods commonly used for flux evaluation. A compartmentalized model of plant glycolysis and pentose phosphate pathway illustrates the reconstruction of a typical metabolic network, whereas a simpler example network illustrates the underlying metabolite and isotopomer balancing techniques. We also discuss the salient features of commonly used flux estimation software 13CFLUX2, Metran, NMR2Flux+, FiatFlux, and OpenFLUX. Furthermore, we briefly discuss methods to improve flux estimates. A graphical checklist at the end of the chapter provides a reader a quick reference to the mathematical modeling concepts and resources.

  5. Latest Paleocene benthic extinction event on the southern Tethyan shelf (Egypt): Foraminiferal stable isotopic (δ13C, δ18O) records

    NASA Astrophysics Data System (ADS)

    Schmitz, B.; Speijer, R. P.; Aubry, M.-P.

    1996-04-01

    The dramatic global extinction of 35% 50% of benthic foraminifera species in the deep sea in the latest Paleocene and associated negative excursions in δ13C and δ18O may be related to spreading of warm, saline bottom water from subtropical Tethyan shallow regions over the sea floor worldwide. Our study of neritic sections in Egypt shows that in the southern shallow Tethys, a prominent long-term change in bottom-water chemistry, sedimentation, and benthic foraminifera fauna was initiated at the time when the deep-sea benthic extinction event (BEE) took place. Bottom-water δ13C values on the Tethyan shelf show a sudden 3.0‰ negative shift at this event; however, contrary to the deep sea, in which the δ13C excursion was of short duration, Tethyan δ13C values did not fully return to preboundary values, but remained depressed by ˜1.5‰ for at least 1 m.y. The δ13C values at the Egyptian shelf during the BEE are much lower than would be expected if this was a source region for global deep water. The δ18O values indicate no significant change in bottom-water salinity or temperature at the BEE. The long-lasting environmental changes that began on the Egyptian shelf at the BEE may be related to, for example, gateway reorganization along the Tethyan seaway. Paleogeographic changes possibly also triggered a change in the loci of global deep-water formation; however, these loci must be sought in another part of the Tethys.

  6. Water isotope ratio (δ2H and δ18O) measurements in atmospheric moisture using an optical feedback cavity enhanced absorption laser spectrometer

    NASA Astrophysics Data System (ADS)

    Iannone, Rosario Q.; Romanini, Daniele; Cattani, Olivier; Meijer, Harro A. J.; Kerstel, Erik R. Th.

    2010-05-01

    Water vapor isotopes represent an innovative and excellent tool for understanding complex mechanisms in the atmospheric water cycle over different time scales, and they can be used for a variety of applications in the fields of paleoclimatology, hydrology, oceanography, and ecology. We use an ultrasensitive near-infrared spectrometer, originally designed for use on airborne platforms in the upper troposphere and lower stratosphere, to measure the water deuterium and oxygen-18 isotope ratios in situ, in ground-level tropospheric moisture, with a high temporal resolution (from 300 s down to less than 1 s). We present some examples of continuous monitoring of near-surface atmospheric moisture, demonstrating that our infrared laser spectrometer could be used successfully to record high-concentration atmospheric water vapor mixing ratios in continuous time series, with a data coverage of ˜90%, interrupted only for daily calibration to two isotope ratio mass spectrometry-calibrated local water standards. The atmospheric data show that the water vapor isotopic composition exhibits a high variability that can be related to weather conditions, especially to changes in relative humidity. Besides, the results suggest that observed spatial and temporal variations of the stable isotope content of atmospheric water vapor are strongly related to water vapor transport in the atmosphere.

  7. Using phylogenetic probes for quantification of stable isotope labeling and microbial community analysis

    SciTech Connect

    Brodie, Eoin L; DeSantis, Todd Z; Karaoz, Ulas; Andersen, Gary L

    2014-12-09

    Herein is described methods for a high-sensitivity means to measure the incorporation of stable isotope labeled substrates into RNA following stable isotope probing experiments (SIP). RNA is hybridized to a set of probes such as phylogenetic microarrays and isotope incorporation is quantified such as by secondary ion mass spectrometer imaging (NanoSIMS).

  8. Diatom-inferred δ18O of a Bolivian paleolake during the last deglaciation (18.6-11.7 ka): impact of the paleolake evaporation and water recycling on the isotopic composition of Andean glaciers

    NASA Astrophysics Data System (ADS)

    Quesada, Benjamin; Sylvestre, Florence; Vimeux, Françoise; Black, Jessica; Paillès, Christine; Sonzogni, Corinne; Alexandre, Anne; Blard, Pierre-Henri; Bruneton, Hélène

    2013-04-01

    During the last deglaciation, on the Bolivian Altiplano (~16°S), a wide paleolake covering at least 51,000 km2, named lake Tauca reached its maximum highstand between 16.5 and 15 ka. Overlooking this site, an ice-core from the Sajama ice-cap (covering the last 25,000 years) evidenced an oxygen 18 isotopic excursion of +7‰ matching with the end of the Tauca phase, and more pronounced by about +5‰ compared with the neighboring Andean ice-cores isotopic records. Here we i) provide a new and original experimental use of lacustrine diatoms (n=21) at medium resolution (~300 years) together with ostracods (n=4) for isotopic δ18O paleolake water reconstruction (δ18Olake), ii) detail a simple hydro-isotopical model with constraints given by literature to explain the strong features in the δ18Olake signal and iii) explore whether the Tauca paleolake could contribute as a moisture source to precipitation at Sajama site when it disappears at 14.2 ka. Based on a new chronostratigraphy, the sedimentary sections cover lake Tauca phase (~18.7-14.1 ka) and lake Coipasa phase (~12.6-11.7 ka). On centuries time scale, strong features consistently appear in δ18Olake: an abrupt decrease during lake filling phases immediately followed by an increase during lake level stable phases. The highest variation occurred at ~15.9 ka with a δ18Olake fall of about ~14‰ concomitant with lake Tauca highstand, and followed by a δ18Olake increase of a similar amplitude four centuries later. We also show that this unexpected re-enrichment of δ18Olake can be partly explained with a simple hydro-isotopic model, based on Craig and Gordon's model, with coherent constraints by a re-equilibration of isotopic fluxes in lake steady-state. Based on an evaporative lake model and a simple water stable isotopic balance between two potential moisture sources for Sajama precipitation (eastward advected moisture and lake evaporation), we show that total or partial (from 5 to 60%) evaporation of the lake

  9. Raman spectroscopic and mass spectrometric investigations of the hydrogen isotopes and isotopically labelled methane

    SciTech Connect

    Jewett, J.R., Fluor Daniel Hanford

    1997-02-24

    Suitable analytical methods must be tested and developed for monitoring the individual process steps within the fuel cycle of a fusion reactor and for tritium accountability. The utility of laser-Raman spectroscopy accompanied by mass spectrometry with an Omegatron was investigated using the analysis of all hydrogen isotopes and isotopically labeled methanes as an example. The Omegatron is useful for analyzing all hydrogen isotopes mixed with the stable helium isotopes. The application of this mass spectrometer were demonstrated by analyzing mixtures of deuterated methanes. In addition, it was employed to study the radiochemical Witzbach exchange reaction between tritium and methanes. A laser-Raman spectrometer was designed for analysis of tritium-containing gases and was built from individual components. A tritium-compatible, metal-sealed Raman cuvette having windows with good optical properties and additional means for measuring the stray light was first used successfully in this work. The Raman spectra of the hydrogen isotopes were acquired in the pure rotation mode and in the rotation-vibration mode and were used for on. The deuterated methanes were measured by Raman spectroscopy, the wavenumbers determined were assigned to the corresponding vibrations, and the wavenumbers for the rotational fine-structure were summarized in tables. The fundamental Vibrations of the deuterated methanes produced Witzbach reactions were detected and assigned. The fundamental vibrations of the molecules were obtained with Raman spectroscopy for the first time in this work. The @-Raman spectrometer assembled is well suited for the analysis of tritium- containing gases and is practical in combination with mass spectrometry using an Omegatron, for studying gases used in fusion.

  10. Chironomid δ 18O as a proxy for past lake water δ 18O: a Lateglacial record from Rotsee (Switzerland)

    NASA Astrophysics Data System (ADS)

    Verbruggen, F.; Heiri, O.; Reichart, G.-J.; Lotter, A. F.

    2010-08-01

    We explored whether the stable oxygen isotope composition (δ 18O) of fossil chironomid remains can be used to reconstruct past variations in lake water δ 18O from Lateglacial and early Holocene sediments from Rotsee (Switzerland). A sediment core from the former littoral zone of the lake was examined since it contained both high concentrations of chironomid remains and abundant authigenic carbonates and therefore allowed a direct comparison of chironomid δ 18O with values measured on bulk carbonates. Since carbonate particles adhering to chironomid remains potentially affect 18O measurements we tested two methods to chemically remove residual carbonates. Trials with isotopically heavy and light acid solutions indicated that treatment with hydrochloric acid promoted oxygen exchange between chironomid remains and the water used during pretreatment. In contrast, a buffered 2 M ammonium chloride (NH 4Cl) solution did not seem to affect chironomid δ 18O to a significant extent. Fossil chironomid δ 18O was analyzed for the Rotsee record both using standard palaeoecological methods and after pretreatment with NH 4Cl. Samples prepared using standard techniques showed a poor correlation with δ 18O of bulk carbonate ( r2 = 0.14) suggesting that carbonate contamination of the chironomid samples obscured the chironomid δ 18O signature. Samples pretreated with NH 4Cl correlated well with bulk carbonate δ 18O ( r2 = 0.67) and successfully tracked the well-known Lateglacial changes in δ 18O. Chironomid δ 18O indicated depleted lake water δ 18O during the Oldest Dryas period, the Aegelsee and Gerzensee Oscillations, and the Younger Dryas, whereas enriched δ 18O values were associated with sediments deposited during the Lateglacial interstadial and the early Holocene. Differences in the amplitude of variations in bulk carbonate and chironomid δ 18O are attributed to differential temperature effects on oxygen isotope fractionation during the formation of carbonates and

  11. O-18/O-16 and Si-30/Si-28 studies of some Apollo 15, 16, and 17 samples. [oxygen and silicon isotope ratios

    NASA Technical Reports Server (NTRS)

    Taylor, H. P., Jr.; Epstein, S.

    1973-01-01

    A study of lunar rock samples from eight sites on the near side of the moon showed oxygen isotope abundance variations much smaller than those in meteorites and earth material. The grain-surface coatings of the lunar fines were found to be generally depleted in oxygen relative to silicon. The lunar soils, on the other hand, were somewhat richer in both O18 and Si30 than the lunar crystalline rock.

  12. Variations in [sup 18]O/[sup 16]O ratios of kaolinites within a lateritic profile: Their significance for laterite genesis and isotope paleoclimatology

    SciTech Connect

    Giral, S.; Girard, J.P.; Savin, S.M. . Dept. of Geological Sciences); Nahon, D.B. )

    1992-01-01

    The authors have made an integrated study of the field occurrence, petrology, mineralogy and crystallography, and oxygen isotope geochemistry of an active lateritic profile from about 60 km north of Manaus (Amazonia, Brazil). The parent rock is an arkosic sandstone. The delta O-18 values of kaolinites from the profile are far from uniform. The total range is about 2.4 per mil (18.7 to 21.1 per mil). The calculated delta O-18 value of kaolinite in isotopic equilibrium with local average precipitation and mean annual temperature is 19.6 per mil, within the range of the measured values. Kaolinite of each of several textural occurrences also shows significant isotopic variation both vertically and within a given horizon. Different size fractions of kaolinite of a single textural occurrence within a single horizon also exhibit differences in delta O-18 values. At depths below a few meters, they expect the temperature and the delta O-18 values of the soil water profile to be relatively uniform at any time. If this is so, the variations in delta O-18 values of the kaolinites would suggest that the formation of different populations occurred at different times. They cannot yet distinguish between variations of conditions that were seasonal and variations that occurred on scales of many years. However, it is most important to resolve the causes of these variations before using the delta O-18 values of soil clays for purposes of paleoclimatic reconstruction.

  13. Simultaneous determination of stable isotopic compositions of nitrous oxide (δ15N and δ18O of N2O) and methane (δ13C of CH4) in nanomolar quantities from a single water sample

    NASA Astrophysics Data System (ADS)

    Hirota, A.; Tsunogai, U.; Komatsu, D. D.; Nakagawa, F.

    2010-12-01

    The stable isotopic compositions of nitrous oxide (δ15N of N2O and δ18O of N2O, respectively) and methane (δ13C of CH4) have provided us with some interesting geochemical insights. We have developed a rapid, sensitive, and automated analytical system to simultaneously determine the concentrations and stable isotopic compositions of nanomolar quantities of N2O and CH4 in the environmental water, by combining continuous-flow isotope-ratio mass spectrometry and a He-sparging system to extract and purify the dissolved gases. Our system, which is composed of a sparging bottle, a chemical trap, four cold traps and a capillary gas chromatograph that use ultra-pure helium as the carrier gas, achieves complete extraction of N2O and CH4 in a water sample and separation among N2O, CH4, and the other component gases. The flow path subsequent to gas chromatograph was periodically changed to pass the gases through the combustion furnace to convert CH4 and the other hydrocarbons into CO2, or to bypass the combustion furnace for the direct introduction of eluted N2O into the mass spectrometer, for determining the stable isotopic compositions through monitoring m/z = 44, 45, and 46, on the bases of CO2+ and N2O+, respectively. The analytical system can be operated automatically with sequential software programmed on a personal computer. The analytical precisions (the standard deviation of a single measurement) were better than 0.2‰ for δ15N of N2O and 0.3‰ for δ18O of N2O, in the case of more than 6.7 nmol N2O injection and better than 1.4‰ for δ15N of N2O and 2.6‰ for δ18O of N2O, in the case of more than 0.2 nmol N2O injection, respectively. Simultaneously, the analytical precisions were better than 0.07‰ for δ13C of CH4, in the case of more than 5.5 nmol CH4 infection and better than 2.1‰ for δ13C of CH4, when more than 0.024 nmol CH4 injection. In this manner, we can simultaneously determine stable isotopic compositions of a 120 mL water sample having

  14. An extractive removal step optimized for a high-throughput α-cellulose extraction method for δ13C and δ18O stable isotope ratio analysis in conifer tree rings.

    PubMed

    Lin, Wen; Noormets, Asko; King, John S; Sun, Ge; McNulty, Steve; Domec, Jean-Christophe

    2016-09-26

    Stable isotope ratios (δ(13)C and δ(18)O) of tree-ring α-cellulose are important tools in paleoclimatology, ecology, plant physiology and genetics. The Multiple Sample Isolation System for Solids (MSISS) was a major advance in the tree-ring α-cellulose extraction methods, offering greater throughput and reduced labor input compared to traditional alternatives. However, the usability of the method for resinous conifer species may be limited by the need to remove extractives from some conifer species in a separate pretreatment step. Here we test the necessity of pretreatment for α-cellulose extraction in loblolly pine (Pinus taeda L.), and the efficiency of a modified acetone-based ambient-temperature step for the removal of extractives (i) in loblolly pine from five geographic locations representing its natural range in the southeastern USA, and (ii) on five other common coniferous species (black spruce (Picea mariana Mill.), Fraser fir (Abies fraseri (Pursh) Poir.), Douglas fir (Pseudotsuga menziesii (Mirb.) Franco), Norway spruce (Picea abies (L.) Karst) and ponderosa pine (Pinus ponderosa D.)) with contrasting extractive profiles. The differences of δ(13)C values between the new and traditional pretreatment methods were within the precision of the isotope ratio mass spectrometry method used (±0.2‰), and the differences between δ(18)O values were not statistically significant. Although some unanticipated results were observed in Fraser fir, the new ambient-temperature technique was deemed as effective as the more labor-consuming and toxic traditional pretreatment protocol. The proposed technique requires a separate acetone-inert multiport system similar to MSISS, and the execution of both pretreatment and main extraction steps allows for simultaneous treatment of up to several hundred microsamples from resinous softwood, while the need of additional labor input remains minimal.

  15. Evolution of low-18O Icelandic crust

    NASA Astrophysics Data System (ADS)

    Pope, Emily C.; Bird, Dennis K.; Arnórsson, Stefán

    2013-07-01

    The Krafla central volcano in the neovolcanic zone of Iceland hosts a chemically diverse suite of magmas characterized by anomalously low δ18O values. A rhyolite magma intercepted by the Iceland Deep Drilling Project (IDDP) exploratory well at 2.1 km depth provided a unique opportunity to investigate the origins of an unerupted rhyolite melt in the primarily basaltic central volcano at Krafla. Here we compare whole rock hydrogen and oxygen isotopes of this melt to those of lavas within and near the caldera of the Krafla central volcano ranging from recent fissure eruptions to Plio--Pleistocene age (including analyses of 18 new samples, plus previously published values) in order to evaluate the petrogenesis of low-18O magmas within the neovolcanic zone of Iceland. Oxygen isotope values of the IDDP-1 melt (δ18O=+3.2±0.2‰) are within the range of Krafla eruptives that have a bimodal composition of olivine-tholeiite and rhyolite (δ18O=+1.6‰ to +4.5‰). Lavas show significantly more variability in hydrogen isotope values (δD=-161‰ to -92‰) than the IDDP-1 melt (-121±2‰), whose δD is comparable to local hydrothermal epidote (-127 to -108‰), and show significantly lower water contents than IDDP-1 (0.1-1.1 wt%, in contrast to ~1.8 wt%). Basaltic to dacitic lavas from the proximal Heidarspordur ridge volcanic zone have δ18O between +3.4‰ and +4.2‰ and δD between -105‰ and -99‰. Uniformity of oxygen isotopes in the Heidarspordur ridge lavas suggests that their magmatic compositional variations are a consequence of fractional crystallization. The δD of the glass sampled by IDDP-1 unequivocally identifies the source of silicic low-18O melts like those erupted from within the caldera of the Krafla volcano as anatexis of meteoric-hydrothermally altered basalts resulting from the intrusion of mantle-derived basaltic magma. Finally, mantle-derived basalts in both the Krafla central volcano and Heidarspordur ridge (MgO>5 wt%) have δ18O values lower

  16. NMR studies of two spliced leader RNAs using isotope labeling

    SciTech Connect

    Lapham, J.; Crothers, D.M.

    1994-12-01

    Spliced leader RNAs are a class of RNA molecules (<200 nts) involved in the trans splicing of messenger RNA found in trypanosomes, nematodes, and other lower eukaryotes. The spliced leader RNA from the trypanosome Leptomonas Collosoma exists in two alternate structural forms with similar thermal stabilities. The 54 nucleotides on the 5{prime} end of the SL molecule is structurally independent from the 3{prime} half of the RNA, and displays the two structural forms. Furthermore, the favored of the two structures was shown to contain anomalous nuclease sensitivity and thermal stability features, which suggests that there may be tertiary interactions between the splice site and other nucleotides in the 5{prime} end. Multidimensional NMR studies are underway to elucidate the structural elements present in the SL RNAs that give rise to their physical properties. Two spliced leader sequences have been studied. The first, the 54 nucleotides on the 5{prime} end of the L. Collosoma sequence, was selected because of earlier studies in our laboratory. The second sequence is the 5{prime} end of the trypanosome Crithidia Fasciculata, which was chosen because of its greater sequence homology to other SL sequences. Given the complexity of the NMR spectra for RNA molecules of this size, we have incorporated {sup 15}N/{sup 13}C-labeled nucleotides into the RNA. One of the techniques we have developed to simplify the spectra of these RNA molecules is isotope labeling of specific regions of the RNA. This has been especially helpful in assigning the secondary structure of molecules that may be able to adopt multiple conformations. Using this technique one can examine a part of the molecule without spectral interference from the unlabeled portion. We hope this approach will promote an avenue for studying the structure of larger RNAs in their native surroundings.

  17. Precise and accurate isotope fractionation factors (α17O, α18O and αD) for water and CaSO4·2H2O (gypsum)

    NASA Astrophysics Data System (ADS)

    Gázquez, Fernando; Evans, Nicholas P.; Hodell, David A.

    2017-02-01

    Gypsum (CaSO4·2H2O) is a hydrated mineral containing crystallization water, also known as gypsum hydration water (GHW). We determined isotope fractionation factors (α17O, α18O and αD) between GHW and free water of the mother solution in the temperature range from 3 °C to 55 °C at different salinities and precipitation rates. The hydrogen isotope fractionation factor (αDgypsum-water) increases by 0.0001 units per °C between 3 °C and 55 °C and salinities <150 g/L of NaCl. The αDgypsum-water is 0.9812 ± 0.0007 at 20 °C, which is in good agreement with previous estimates of 0.981 ± 0.001 at the same temperature. The α18Ogypsum-water slightly decreases with temperature by 0.00001 per °C, which is not significant over much of the temperature range considered for paleoclimate applications. Between 3 °C and 55 °C, α18Ogypsum-water averages 1.0035 ± 0.0002. This value is more precise than that reported previously (e.g. 1.0041 ± 0.0004 at 25 °C) and lower than the commonly accepted value of 1.004. We found that NaCl concentrations below 150 g/L do not significantly affect α18Ogypsum-water, but αDgypsum-water increases linearly with NaCl concentrations even at relatively low salinities, suggesting a salt correction is necessary for gypsum formed from brines. Unlike oxygen isotopes, the αDgypsum-water is affected by kinetic effects that increase with gypsum precipitation rate. As expected, the relationship of the fractionation factors for 17O and 18O follows the theoretical mass-dependent fractionation on Earth (θ = 0.529 ± 0.001). We provide specific examples of the importance of using the revised fractionation factors when calculating the isotopic composition of the fluids.

  18. Follow the Carbon: Isotopic Labeling Studies of Early Earth Aerosol

    NASA Astrophysics Data System (ADS)

    Hicks, Raea K.; Day, Douglas A.; Jimenez, Jose L.; Tolbert, Margaret A.

    2016-11-01

    Despite the faint young Sun, early Earth might have been kept warm by an atmosphere containing the greenhouse gases CH4 and CO2 in mixing ratios higher than those found on Earth today. Laboratory and modeling studies suggest that an atmosphere containing these trace gases could lead to the formation of organic aerosol haze due to UV photochemistry. Chemical mechanisms proposed to explain haze formation rely on CH4 as the source of carbon and treat CO2 as a source of oxygen only, but this has not previously been verified experimentally. In the present work, we use isotopically labeled precursor gases and unit-mass resolution (UMR) and high-resolution (HR) aerosol mass spectrometry to examine the sources of carbon and oxygen to photochemical aerosol formed in a CH4/CO2/N2 atmosphere. UMR results suggest that CH4 contributes 70-100% of carbon in the aerosol, while HR results constrain the value from 94% to 100%. We also confirm that CO2 contributes approximately 10% of the total mass to the aerosol as oxygen. These results have implications for the geochemical interpretations of inclusions found in Archean rocks on Earth and for the astrobiological potential of other planetary atmospheres.

  19. Follow the Carbon: Isotopic Labeling Studies of Early Earth Aerosol.

    PubMed

    Hicks, Raea K; Day, Douglas A; Jimenez, Jose L; Tolbert, Margaret A

    2016-11-01

    Despite the faint young Sun, early Earth might have been kept warm by an atmosphere containing the greenhouse gases CH4 and CO2 in mixing ratios higher than those found on Earth today. Laboratory and modeling studies suggest that an atmosphere containing these trace gases could lead to the formation of organic aerosol haze due to UV photochemistry. Chemical mechanisms proposed to explain haze formation rely on CH4 as the source of carbon and treat CO2 as a source of oxygen only, but this has not previously been verified experimentally. In the present work, we use isotopically labeled precursor gases and unit-mass resolution (UMR) and high-resolution (HR) aerosol mass spectrometry to examine the sources of carbon and oxygen to photochemical aerosol formed in a CH4/CO2/N2 atmosphere. UMR results suggest that CH4 contributes 70-100% of carbon in the aerosol, while HR results constrain the value from 94% to 100%. We also confirm that CO2 contributes approximately 10% of the total mass to the aerosol as oxygen. These results have implications for the geochemical interpretations of inclusions found in Archean rocks on Earth and for the astrobiological potential of other planetary atmospheres. Key Words: Atmosphere-Early Earth-Planetary atmospheres-Carbon dioxide-Methane. Astrobiology 16, 822-830.

  20. Multiplexed DNA sequencing and diagnostics by hybridization with enriched stable isotope labels

    SciTech Connect

    Arlinghaus, H.F.; Kwoka, M.N.; Guo, X.Q.; Jacobson, K.B.

    1997-04-15

    A new DNA diagnostic and sequencing system has been developed that uses time-of-flight resonance ionization mass spectrometry (TOF-RIMS) to provide a rapid method of analyzing stable isotope-labeled oligonucleotides in form 1 sequencing by hybridization (SBH). With form 1, the DNA is immobilized on a nylon membrane and enriched isotope-labeled individual oligonucleotide probes are free to seek out complementary DNAs during hybridization. The major advantage of this new approach is that multiple oligonucleotides can be labeled with different enriched isotopes and can all be simultaneously hybridized to the genosensor matrix. The probes can then be simultaneously detected with TOF-RIMS with high selectivity, sensitivity, and efficiency. By using isotopically enriched tin labels, up to 10 labeled oligonucleotides could be examined in a single hybridization to the DNA matrix. Greater numbers of labels are available if rare earth isotopes are employed. In the present study, matrices containing three different DNAs were prepared and simultaneously hybridized with two different probes under a variety of conditions. The results show that DNAs, immobilized on nylon surfaces, can be specifically hybridized to probes labeled with different enriched tin isotopes. Discrimination between complementary and noncomplementary sites of better than 100 was obtained in multiplexed samples. 34 refs., 5 figs.

  1. Preparation of soluble isotopically labeled human growth hormone produced in Escherichia coli.

    PubMed

    Lee, Jin-Hee; Jeong, Ji-Seon; Kim, Sook-Kyung; Song, Jimyeong; Lee, Ji Youn; Baek, Soyun; Choi, Jun-Hyuk

    2016-11-01

    Isotopically labeled proteins have been used as internal standards for mass spectrometry (MS)-based absolute protein quantification. Although this approach can provide highly accurate analyses of proteins of interest within a complex mixture, one of the major limitations of this method is the difficulty in preparing uniformly labeled standards. Human growth hormone (hGH) is one of the most important hormones that circulate throughout the body, and its measurement is primarily of interest in the diagnosis and treatment of growth disorders. In order to provide a useful internal standard for MS-based hGH measurement, we describe an efficient strategy to produce a potentially valuable, stable isotope-labeled hGH with high purity and yield. The strategy involves the following steps: solubilization of hGH under labeling conditions, detection of stable isotope incorporation, large-scale purification, analysis of the labeled protein, and assessment of the labeling efficiency. We show that the yield of soluble hGH under selective isotopic labeling conditions can be greatly increased by optimizing protein expression and extraction. Our efficient method for generating isotopically labeled hGH does not influence the structural integrity of hGH. Finally, we assessed the efficiency of stable isotope labeling at the intact protein level, and the result was further verified by amino acid analysis. These results clearly indicate that our labeling approach allows an almost complete incorporation of (13)C6(15)N4-arginine into the hGH expressed in E.coli without detectable isotope scrambling.

  2. Decadal patterns in δ18O of atmospheric CO2

    NASA Astrophysics Data System (ADS)

    Zakem, E.; White, J. W.

    2010-12-01

    The stable oxygen isotope 18O is unique to isotope ecology in that it links the hydrosphere to the carbon cycle. Since land biosphere fluxes are the dominant influences on 18O of atmospheric CO2, particularly on shorter times scales, analysis of atmospheric δ18O trends can provide useful insight into the terrestrial carbon cycle. The isotopic values imprinted by leaf water and soil water exchanges with CO2 out-compete those from ocean exchange, fossil fuel and biomass burning, and stratospheric reactions. The opposing isotopic imprints of photosynthesis and ecosystem respiration therefore control the majority of atmospheric 18O concentration. The resulting seasonal cycle in δ18O data of peaks during early summer, when photosynthesis dominates, and lows during early winter, when respiration dominates, has been clearly established. However, the reasons for the interannual variability of atmospheric 18O remain unknown. Studies have shown that the size and isotopic value of the “retrodiffusion” flux- the CO2 that enters and exits leaves without being fixed by photosynthesis- is a function of stomatal conductance, which is affected by the relative humidity in the surrounding atmosphere. We observe that data from numerous global sites shows a global decadal oscillation in δ18O, suggesting a climatological forcing. We compare decadal trends in δ18O with climate oscillations and the 11-year solar cycle, as well as relative humidity records, examining correlations and proposing associated mechanisms. Understanding the decadal patterns in atmospheric 18O of CO2 will shed light on global terrestrial carbon fluxes and the carbon-water interaction on decadal time scales, potentially helping to scale human versus natural impacts on this coupled system.

  3. Rapid 'de novo' peptide sequencing by a combination of nanoelectrospray, isotopic labeling and a quadrupole/time-of-flight mass spectrometer.

    PubMed

    Shevchenko, A; Chernushevich, I; Ens, W; Standing, K G; Thomson, B; Wilm, M; Mann, M

    1997-01-01

    Protein microanalysis usually involves the sequencing of gel-separated proteins available in very small amounts. While mass spectrometry has become the method of choice for identifying proteins in databases, in almost all laboratories 'de novo' protein sequencing is still performed by Edman degradation. Here we show that a combination of the nanoelectrospray ion source, isotopic end labeling of peptides and a quadrupole/ time-of-flight instrument allows facile read-out of the sequences of tryptic peptides. Isotopic labeling was performed by enzymatic digestion of proteins in 1:1 16O/18O water, eliminating the need for peptide derivatization. A quadrupole/time-of-flight mass spectrometer was constructed from a triple quadrupole and an electrospray time-of-flight instrument. Tandem mass spectra of peptides were obtained with better than 50 ppm mass accuracy and resolution routinely in excess of 5000. Unique and error tolerant identification of yeast proteins as well as the sequencing of a novel protein illustrate the potential of the approach. The high data quality in tandem mass spectra and the additional information provided by the isotopic end labeling of peptides enabled automated interpretation of the spectra via simple software algorithms. The technique demonstrated here removes one of the last obstacles to routine and high throughput protein sequencing by mass spectrometry.

  4. Multi-Proxy Palaeothermometry (δ18O, Mg/Ca, clumped isotopes) of Mid-Cretaceous Rudist Bivalves: Deciphering Stratigraphic and Seasonal Changes in Shallow-Marine Sea-Surface Temperature

    NASA Astrophysics Data System (ADS)

    Huck, S.; Steuber, T.; Bernasconi, S. M.; Heimhofer, U.

    2014-12-01

    The Cretaceous period is generally considered to have been a time of warm climate, but there is an on-going dispute about the existence of cool interludes - including the short-termed installation of polar ice caps. Mid-Cretaceous shoal-water ecosystems have been proven to show characteristic response modes (e.g., microencruster-blooms) to major climatic and environmental changes related to oceanic anoxic events (OAEs), times of widespread pelagic organic matter burial. Some biotic changes predate OAE1a by few 100kyrs, an observation that is in favour of gradual and pulsed volcanic CO2-outgassing as main trigger of this event. We aim at reconstructing the evolution of Barremian-Aptian sea-surface temperatures (SSTs) and SST seasonalities in the prelude, during and in the aftermath of OAE1a. The outer low-Mg calcite-shell layer of rudist bivalves (e.g., Toucasia), collected at carbonate platform settings in the (sub-)tropical Tethyan realm (France, Croatia, Spain), serves both as chemostratigraphic and palaeoclimatic archive. Sclerochronological variations in isotopic18O, δ13C, Mg/Ca, clumped isotopes) and geochemical composition (Sr, Fe, Mn, Ba, Ca/Mg) provide insights into seasonal and long-term palaeoclimatic and palaeoenvironmental changes. The outcome of this work will be of significance both for those studying the triggering factors of oceanic anoxic events and the palaeoecology of rudist bivalves.

  5. Assessment of temporal and spatial differences of source apportionment of nitrate in an urban river in China, using δ(15)N and δ(18)O values and an isotope mixing model.

    PubMed

    Zhang, Qianqian; Wang, Xiaoke; Sun, Feixiang; Sun, Jichao; Liu, Jingtao; Ouyang, Zhiyun

    2015-12-01

    Nitrate contamination in surface water has become an environmental problem widespread concern. In this study, environmental isotopes (δ(15)N-NO3 (-) and δ(18)O-NO3 (-)) and the chemical compositions of water samples from an urban river in Chongqing, China, were analyzed to evaluate the primary sources of nitrate pollution. A Bayesian isotope mixing model was applied to estimate the relative contributions of five potential NO3 (-) sources to river pollution (sewage/manure, soil N, NH4 (+) in fertilizer and precipitation, NO3 (-) fertilizer, and NO3 (-) in precipitation). The results show that the urban river was affected by NO3 (-) pollution from multiple sources. The major sources of NO3 (-) pollution in the dry season were sewage/manure (38-50 %) and soil N (22-26 %); in the wet season, the major sources of NO3 (-) pollution were sewage/manure (30-37 %), soil N (16-25 %), and precipitation (14-24 %). The higher contribution of N to the river water by precipitation indicates that atmospheric N deposition has become an important source of pollution in surface water in China. We conclude that domestic sewage is still the main contributor to NO3 (-) pollution in urban rivers in China. The discharge of domestic sewage into rivers should be prohibited as a priority measure to prevent NO3 (-) contamination.

  6. Water-Isotopes (2H, 3H, 18O) to trace the source and timing of recharge in a fractured granite aquifer in Western Kenya, Africa

    NASA Astrophysics Data System (ADS)

    Kralik, Martin; Whylidal, Stefan; Asunah, Francis; Sültenfuß, Jürgen

    2014-05-01

    The Vihiga District in West Kenya North-West of Lake Victoria is one of the most densely populated areas in Kenya with 1033 person per square kilometer. To find the most suitable location of an own well for a Primary School in this district, springs, school wells and creeks were sampled in the surroundings to get information about the hydrological cycle in the area. The Waluka Primary school (0.02134°N, 34.64311°E) is situated on the northern slope of the Maragoli Hills 20 km to the North-West of the Nyanzan provincial capital of Kisumu at the eastern shore of Lake Victoria. The hilly relief varies between 1535 - 1675m. The yearly precipitation is between 1200-1600 mm/a (23°C mean temperature) with biannual rainy seasons in which the long rains are generally from March to May as the Inter-Tropical Convergence Zone (ITCZ) moves northwards, and the short rains are typically from October to December as the ITCZ retreats southwards. A lateritic soil covers a thin alteration zone above the Precambrian Maragoli-Granite (Saggerson, 1952). Water circulates either in the thin alteration zone or in fault zones cutting through the Precambrian granite. From discharge measurements of two springs and a creek at the end of the dry season (February 2012) a minimum discharge of ca. 10-20 L/s km2 (300-600 mm) can be estimated. The water is of the alkaline sulfate-nitrate type with low mineralization (70-150 μ S/cm, 25°C) and a low pH of about 5 to 6. The delta oxygen-18 and deuterium value ranges between -2.84 to -1.98 oand -8.5 to - 3.9 o(VSMOW). The deuterium excess ranges from 11.7-14.2 oThe water of one spring and well close to the school have a tritium content of 1.42 - 1.62 TU. All groundwater has a low arsenium, fluorine and uranium content, which had only a short soil passage. The relatively elevated, but not problematic content in nitrate (10 - 16 mg/L) probaly reflects the intensive agricultural activities in this area. As the mean δ 18O values during the rainy

  7. Trypsin-catalyzed oxygen-18 labeling for quantitative proteomics

    SciTech Connect

    Qian, Weijun; Petritis, Brianne O.; Nicora, Carrie D.; Smith, Richard D.

    2011-07-01

    Stable isotope labeling based on relative peptide/protein abundance measurements is commonly applied for quantitative proteomics. Recently, trypsin-catalyzed oxygen-18 labeling has grown in popularity due to its simplicity, cost-effectiveness, and its ability to universally label peptides with high sample recovery. In (18)O labeling, both C-terminal carboxyl group atoms of tryptic peptides can be enzymatically exchanged with (18)O, thus providing the labeled peptide with a 4 Da mass shift from the (16)O-labeled sample. Peptide (18)O labeling is ideally suited for generating a labeled "universal" reference sample used for obtaining accurate and reproducible quantitative measurements across large number of samples in quantitative discovery proteomics.

  8. Strategies for Extending Metabolomics Studies with Stable Isotope Labelling and Fluxomics

    PubMed Central

    Srivastava, Anubhav; Kowalski, Greg M.; Callahan, Damien L.; Meikle, Peter J.; Creek, Darren J.

    2016-01-01

    This is a perspective from the peer session on stable isotope labelling and fluxomics at the Australian & New Zealand Metabolomics Conference (ANZMET) held from 30 March to 1 April 2016 at La Trobe University, Melbourne, Australia. This report summarizes the key points raised in the peer session which focused on the advantages of using stable isotopes in modern metabolomics and the challenges in conducting flux analyses. The session highlighted the utility of stable isotope labelling in generating reference standards for metabolite identification, absolute quantification, and in the measurement of the dynamic activity of metabolic pathways. The advantages and disadvantages of different approaches of fluxomics analyses including flux balance analysis, metabolic flux analysis and kinetic flux profiling were also discussed along with the use of stable isotope labelling in in vivo dynamic metabolomics. A number of crucial technical considerations for designing experiments and analyzing data with stable isotope labelling were discussed which included replication, instrumentation, methods of labelling, tracer dilution and data analysis. This report reflects the current viewpoint on the use of stable isotope labelling in metabolomics experiments, identifying it as a great tool with the potential to improve biological interpretation of metabolomics data in a number of ways. PMID:27706078

  9. Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis.

    PubMed

    Wang, Fangjun; Wei, Xiaoluan; Zhou, Hu; Liu, Jing; Figeys, Daniel; Zou, Hanfa

    2012-11-01

    Various enzyme reactors and online enzyme digestion strategies have been developed in recent years. These reactors greatly enhanced the detection sensitivity and proteome coverage in qualitative proteomics. However, these devices have higher rates of miscleavage in protein digestion. Therefore, we investigated the effect of online enzyme digestion on the quantification accuracy of quantitative proteomics using chemical or metabolic isotope labeling approaches. The incomplete digestion would introduce some unexpected variations in comparative quantification when the samples are digested and then chemically isotope labeled in different aliquots. Even when identical protein aliquots are processed on these devices using post-digestion chemical isotope labeling and the CVs of the ratios controlled to less than 50% in replicate analyses, about 10% of the quantified proteins have a ratio greater than two-fold, whereas in theory the ratio is 1:1. Interestingly, the incomplete digestion with enzyme reactor is not a problem when metabolic isotope labeling samples were processed because the proteins are isotopically labeled in vivo prior to their simultaneous digestion within the reactor. Our results also demonstrated that both high quantification accuracy and high proteome coverage can be achieved in comparative proteome quantification using online enzyme digestion even when a limited amount of metabolic isotope labeling samples is used (1683 proteins comparatively quantified from 10(5) Hela cells).

  10. Conifers, Angiosperm Trees, and Lianas: Growth, Whole-Plant Water and Nitrogen Use Efficiency, and Stable Isotope Composition (δ13C and δ18O) of Seedlings Grown in a Tropical Environment1[W][OA

    PubMed Central

    Cernusak, Lucas A.; Winter, Klaus; Aranda, Jorge; Turner, Benjamin L.

    2008-01-01

    Seedlings of several species of gymnosperm trees, angiosperm trees, and angiosperm lianas were grown under tropical field conditions in the Republic of Panama; physiological processes controlling plant C and water fluxes were assessed across this functionally diverse range of species. Relative growth rate, r, was primarily controlled by the ratio of leaf area to plant mass, of which specific leaf area was a key component. Instantaneous photosynthesis, when expressed on a leaf-mass basis, explained 69% of variation in r (P < 0.0001, n = 94). Mean r of angiosperms was significantly higher than that of the gymnosperms; within angiosperms, mean r of lianas was higher than that of trees. Whole-plant nitrogen use efficiency was also significantly higher in angiosperm than in gymnosperm species, and was primarily controlled by the rate of photosynthesis for a given amount of leaf nitrogen. Whole-plant water use efficiency, TEc, varied significantly among species, and was primarily controlled by ci/ca, the ratio of intercellular to ambient CO2 partial pressures during photosynthesis. Instantaneous measurements of ci/ca explained 51% of variation in TEc (P < 0.0001, n = 94). Whole-plant 13C discrimination also varied significantly as a function of ci/ca (R2 = 0.57, P < 0.0001, n = 94), and was, accordingly, a good predictor of TEc. The 18O enrichment of stem dry matter was primarily controlled by the predicted 18O enrichment of evaporative sites within leaves (R2 = 0.61, P < 0.0001, n = 94), with some residual variation explained by mean transpiration rate. Measurements of carbon and oxygen stable isotope ratios could provide a useful means of parameterizing physiological models of tropical forest trees. PMID:18599645

  11. Direct analysis of δ2H and δ18O in natural and enriched human urine using laser-based, Off-Axis Integrated Cavity Output Spectroscopy

    PubMed Central

    Berman, Elena S.F.; Fortsona, Susan L.; Snaith, Steven P.; Gupta, Manish; Baer, Douglas S.; Chery, Isabelle; Blanc, Stephane; Melanson, Edward L.; Thomson, Peter J; Speakman, John R.

    2012-01-01

    The stable isotopes of hydrogen (δ2H) and oxygen (δ18O) in human urine are measured during studies of total energy expenditure by the doubly labeled water method, measurement of total body water, and measurement of insulin resistance by glucose disposal among other applications. An ultrasensitive laser absorption spectrometer based on off-axis integrated cavity output spectroscopy was demonstrated for simple and inexpensive measurement of stable isotopes in natural isotopic abundance and isotopically enriched human urine. Preparation of urine for analysis was simple and rapid (approx. 25 samples per hour), requiring no decolorizing or distillation steps. Analysis schemes were demonstrated to address sample-to-sample memory while still allowing analysis of 45 natural or 30 enriched urine samples per day. The instrument was linear over a wide range of water isotopes (δ2H = −454 to +1702 ‰ and δ18O= −58.3 to +265 ‰). Measurements of human urine were precise to better than 0.65 ‰ 1σ for δ2H and 0.09 ‰ 1σ for δ18O for natural urines, 1.1 ‰ 1σ for δ2H and 0.13 ‰ 1σ for δ18O for low enriched urines, and 1.0 ‰ 1σ for δ2H and 0.08 ‰ 1σ for δ18O for high enriched urines. Furthermore, the accuracy of the isotope measurements of human urines was verified to better than ±0.81 ‰ in δ2H and ±0.13 ‰ in δ18O (average deviation) against three independent IRMS laboratories. The ability to immediately and inexpensively measure the stable isotopes of water in human urine is expected to increase the number and variety of experiments which can be undertaken. PMID:23075099

  12. Fully automated software solution for protein quantitation by global metabolic labeling with stable isotopes.

    PubMed

    Bindschedler, L V; Cramer, R

    2011-06-15

    Metabolic stable isotope labeling is increasingly employed for accurate protein (and metabolite) quantitation using mass spectrometry (MS). It provides sample-specific isotopologues that can be used to facilitate comparative analysis of two or more samples. Stable Isotope Labeling by Amino acids in Cell culture (SILAC) has been used for almost a decade in proteomic research and analytical software solutions have been established that provide an easy and integrated workflow for elucidating sample abundance ratios for most MS data formats. While SILAC is a discrete labeling method using specific amino acids, global metabolic stable isotope labeling using isotopes such as (15)N labels the entire element content of the sample, i.e. for (15)N the entire peptide backbone in addition to all nitrogen-containing side chains. Although global metabolic labeling can deliver advantages with regard to isotope incorporation and costs, the requirements for data analysis are more demanding because, for instance for polypeptides, the mass difference introduced by the label depends on the amino acid composition. Consequently, there has been less progress on the automation of the data processing and mining steps for this type of protein quantitation. Here, we present a new integrated software solution for the quantitative analysis of protein expression in differential samples and show the benefits of high-resolution MS data in quantitative proteomic analyses.

  13. Synthesis of isotopically labelled 2-isopropylthioxanthone from 2,2'-dithiosalicylic acid and deuterium cumene.

    PubMed

    Fang, Chao; Yang, Weicheng; Yang, Chao; Wang, Haoran; Sun, Kai; Luo, Yong

    2016-06-30

    Two efficient synthetic routes of stable deuterium labelled 2-isopropylthioxanthone were presented with 98.1% and 98.8% isotopic abundance in acceptable yields and excellent chemical purities. Their structures and the isotope-abundance were confirmed according to proton nuclear magnetic resonance and liquid chromatography-mass spectrometry.

  14. Coupling of continuous in situ ecosystem water vapor, precipitation, plant and soil water isotope (δ2H, δ18O and d-excess) measurements in Arctic Alaska to understand a changing water cycle

    NASA Astrophysics Data System (ADS)

    Welker, J. M.; Klein, E. S.; Leffler, J.; Cherry, J. E.; Young, J. M.

    2013-12-01

    A changing Arctic water cycle is focusing our efforts on the patterns and processes governing the exchange of water between the land surface and the atmosphere. We initiated a NSF EAGER study of the changing Arctic water cycle with a cross-scale water isotope (ecosystem-landscape and region) study employing tower, aircraft and satellite measurements in N Alaska. At the land surface, we measured in situ, continuous ecosystem δ2H and δ18O in water vapor isotopes along a vertical profile (0.1 to 3 m) in late winter, spring, and summer of 2013 at the Toolik Lake Field Station in Arctic Alaska. The continuous water vapor measurements are being combined with soil, plant and precipitation water isotope measurements, and species-level transpiration rates. Diurnal patterns of δ2H values in water vapor vary systematically from the soil surface, through the plant canopy and in the near boundary layer ranging between -200 ‰ at the surface to -240 ‰ at 3 m. These vertical patterns were also observed in d-excess ranging from 5 ‰ near the soil surface to -15 ‰ at 3 m. These patterns disappeared at night indicating their link with evapotranspiration. These data will serve as the foundation for a Bayesian model to articulate the sources, species, and processes governing the exchange of H2O with the lower boundary layer; and will also help to calibrate ecosystem, landscape (aircraft) and regional (satellite) measurements of the water vapor isotopes above different vegetation types (tussock tundra, riparian tundra, and recently burned tundra) and regions (Northern Foothills of the Brooks Range and the Arctic Coastal Plain). Continuous water vapor isotope traits in Arctic Tundra from the soil surface (0.02 m) to 3 m above the tundra for a window of time during the 2013 growing season. The diurnal patterns depict the evidence for transpiration and evaporation as the values above the soil (0.02 m) and in the canopy (0.2m) are enriched during the day and become depleted during

  15. Survey of groundwater isotopic composition (δ2H and δ18O) from the southwestern Edwards Aquifer and regionally associated aquifers

    NASA Astrophysics Data System (ADS)

    West, J. B.; Shallock, J. R.; Cooper, R.

    2009-12-01

    Aquifers are an important source of water for growing human populations, while they also support numerous critical ecosystem functions, including supplying water to springs and rivers necessary for terrestrial and aquatic ecosystems and the sometimes endangered species that are part of them. As human populations increase and the climate changes, demands on aquifer resources will continue to increase. To support these growing populations, urban areas in particular seek to develop additional water resources with, in some cases, not well-understood impacts on interconnected aquifers as well as the terrestrial and aquatic ecosystems that depend on spring-fed surface water. In addition, large-scale changes to terrestrial systems (e.g., removal of woody vegetation) can be motivated by a desire to enhance aquifer recharge, but an understanding of the effect of those efforts on the regional water cycle is hampered, in part, by a lack of detailed understanding of the interactions of various features below ground. It is critical therefore to understand the relationships between waters found in different formations, as well as among surface and ground waters to improve our understanding of the consequences of increasing human demands and the impacts of climate change. We have conducted a survey of wells in south central Texas that access major and minor aquifers, including the karst-type Edwards (BFZ) aquifer and several regional minor aquifers. The primary objective of this survey was to establish baseline information from which to develop further targeted research using event-based sampling and a range of additional data on precipitation, geology, and other spatially explicit information. We sampled multiple wells in an area west of San Antonio, TX that access the primary major and minor aquifers in the region. The isotopic compositions of water taken from these wells at various locations and depths were similar, suggesting that these aquifers experience significant mixing

  16. The evolution of 13C and 18O isotope composition of DIC in a calcite depositing film of water with isotope exchange between the DIC and a CO2 containing atmosphere, and simultaneous evaporation of the water. Implication to climate proxies from stalagmites: A theoretical model

    NASA Astrophysics Data System (ADS)

    Dreybrodt, Wolfgang; Romanov, Douchko

    2016-12-01

    The most widely applied climate proxies in speleothems are the isotope compositions of carbon and oxygen expressed by δ13C and δ18O values. However, mechanisms, which are not related to climate changes, overlay the climate signal. One is the temporal increase of both, δ13C and δ18O values by kinetic processes during precipitation of calcite. Isotope exchange between DIC in the water and the CO2 in the surrounding cave atmosphere can also change isotope composition. Here we present a theoretical model of the temporal isotope evolution of DIC in a thin water layer during precipitation of calcite and simultaneous isotope exchange with the cave atmosphere, and simultaneous evaporation of water. The exchange of oxygen isotopes in the DIC with those in the water is also considered. For drip times for Tdrip < 0.2τ, where τ is the precipitation time, we find for the change of the δ13C and δ18O values, respectively, after the time Tdrip ΔDIC(Tdrip) = ((λ + ɛ)Ceq/C0 - ɛ) Tdrip/τ + ((δeqatm - δ0) Tdrip/τinatm) + (δeqwater - δ0 - ɛw Tdrip/Tev) Tdrip/Twater The first term on the right hand side is the contribution from precipitation of calcite, the second stems from isotope exchange with the CO2 of the cave atmosphere, and the third results from isotope exchange between oxygen in the DIC and the oxygen in the water. λ, ε are kinetic parameters, τ is the time scale of precipitation, (δeqatm -δ0) and (δeqwater -δ0) are the differences between the corresponding initial δ-value δ0 and the value δeqatm,water if DIC were in isotope equilibrium with the atmosphere or in the case of oxygen with the water, respectively. τinatm and τwater are the time scales of approach to isotope equilibrium by the exchange reactions. Ceq is the concentration of DIC in chemical equilibrium with the CO2 in the cave atmosphere and C0 is the initial concentration, when the water drips to the stalagmite. Tev is the time needed for complete evaporation of the water layer. ε

  17. Stable isotope-labeling studies in metabolomics: new insights into structure and dynamics of metabolic networks

    PubMed Central

    Chokkathukalam, Achuthanunni; Kim, Dong-Hyun; Barrett, Michael P; Breitling, Rainer; Creek, Darren J

    2014-01-01

    The rapid emergence of metabolomics has enabled system-wide measurements of metabolites in various organisms. However, advances in the mechanistic understanding of metabolic networks remain limited, as most metabolomics studies cannot routinely provide accurate metabolite identification, absolute quantification and flux measurement. Stable isotope labeling offers opportunities to overcome these limitations. Here we describe some current approaches to stable isotope-labeled metabolomics and provide examples of the significant impact that these studies have had on our understanding of cellular metabolism. Furthermore, we discuss recently developed software solutions for the analysis of stable isotope-labeled metabolomics data and propose the bioinformatics solutions that will pave the way for the broader application and optimal interpretation of system-scale labeling studies in metabolomics. PMID:24568354

  18. Heavy Atom Labeled Nucleotides for Measurement of Kinetic Isotope Effects

    PubMed Central

    Weissman, Benjamin P.; Li, Nan-Sheng; York, Darrin; Harris, Michael; Piccirilli, Joseph A.

    2015-01-01

    Experimental analysis of kinetic isotope effects represents an extremely powerful approach for gaining information about the transition state structure of complex reactions not available through other methodologies. Implementation of this approach to the study of nucleic acid chemistry requires the synthesis of nucleobases and nucleotides enriched for heavy isotopes at specific positions. In this review we highlight current approaches to the synthesis of nucleic acids site-specifically enriched for heavy oxygen and nitrogen and their application in heavy atom isotope effect studies. PMID:25828952

  19. Noninvasive imaging of protein metabolic labeling in single human cells using stable isotopes and Raman microscopy.

    PubMed

    van Manen, Henk-Jan; Lenferink, Aufried; Otto, Cees

    2008-12-15

    We have combined nonresonant Raman microspectroscopy and spectral imaging with stable isotope labeling by amino acids in cell culture (SILAC) to selectively detect the incorporation of deuterium-labeled phenylalanine, tyrosine, and methionine into proteins in intact, single HeLa cells. The C-D stretching vibrational bands in these amino acids are observed in the 2100-2300 cm(-1) spectral region that is devoid of vibrational contributions from other, nondeuterated intracellular constituents. We found that incubation with deuterated amino acids for 8 h in cell culture already led to clearly detectable isotope-related signals in Raman spectra of HeLa cells. As expected, the level of isotope incorporation into proteins increased with incubation time, reaching 55% for deuterated phenylalanine after 28 h. Raman spectral imaging of HeLa cells incubated with deuterium-labeled amino acids showed similar spatial distributions for both isotope-labeled and unlabeled proteins, as evidenced by Raman ratio imaging. The SILAC-Raman methodology presented here combines the strengths of stable isotopic labeling of cells with the nondestructive and quantitative nature of Raman chemical imaging and is likely to become a powerful tool in both cell biology applications and research on tissues or whole organisms.

  20. Isotopic labeling of mammalian G protein-coupled receptors (GPCRs) heterologously expressed in Caenorhabditis elegans*

    PubMed Central

    Salom, David; Cao, Pengxiu; Yuan, Yiyuan; Miyagi, Masaru; Feng, Zhaoyang; Palczewski, Krzysztof

    2015-01-01

    High-resolution structural determination and dynamic characterization of membrane proteins by nuclear magnetic resonance (NMR) require their isotopic labeling. Although a number of labeled eukaryotic membrane proteins have been successfully expressed in bacteria, they lack posttranslational modifications and usually need to be refolded from inclusion bodies. This shortcoming of bacterial expression systems is particularly detrimental for the functional expression of G protein-coupled receptors (GPCRs), the largest family of drug targets, due to their inherent instability. In this work we show that proteins expressed by a eukaryotic organism can be isotopically labeled and produced with a quality and quantity suitable for NMR characterization. Using our previously described expression system in Caenorhabditis elegans, we showed the feasibility of labeling proteins produced by these worms with 15N,13C by providing them with isotopically labeled bacteria. 2H labeling also was achieved by growing C. elegans in presence of 70% heavy water. Bovine rhodopsin, simultaneously expressed in muscular and neuronal worm tissues, was employed as the ‘test’ GPCR to demonstrate the viability of this approach. Although the worms’ cell cycle was slightly affected by the presence of heavy isotopes, the final protein yield and quality was appropriate for NMR structural characterization. PMID:25461480

  1. 18O enrichment in phosphorus pools extracted from soybean leaves.

    PubMed

    Pfahler, Verena; Dürr-Auster, Thilo; Tamburini, Federica; Bernasconi, M Stefano; Frossard, Emmanuel

    2013-01-01

    The objective of this study was to investigate the isotopic composition of oxygen bound to phosphate (δ(18)O-PO(4)) in different phosphorus (P) pools in plant leaves. As a model plant we used soybean (Glycine max cv Toliman) grown in the presence of ample P in hydroponic cultures. The leaf blades were extracted with 0.3 M trichloroacetic acid (TCA) and with 10 M nitric acid. These extractions allowed measurement of the TCA-soluble reactive P (TCA P) that is rapidly cycled within the cell and the total leaf P. The difference between total leaf P and TCA P yielded the structural P which includes organic P compounds not extractable by TCA. P uptake and its translocation and transformation within the soybean plants lead to an (18)O enrichment of TCA P (δ(18)O-PO(4) between 16.9 and 27.5‰) and structural P (δ(18)O-PO(4) between 42.6 and 68.0 ‰) compared with 12.4‰ in the phosphate in the nutrient solution. δ(18)O values of phosphate extracted from soybean leaves grown under optimal conditions are greater than the δ(18)O-PO(4) values of the provided P source. Furthermore, the δ(18)O-PO(4) of TCA P seems to be controlled by the δ(18)O of leaf water and the activity of inorganic pyrophosphatase or other pyrophosphatases.

  2. Short-term natural δ13C and δ18O variations in pools and fluxes in a beech forest: the transfer of isotopic signal from recent photosynthates to soil respired CO2

    NASA Astrophysics Data System (ADS)

    Gavrichkova, O.; Proietti, S.; Moscatello, S.; Portarena, S.; Battistelli, A.; Matteucci, G.; Brugnoli, E.

    2011-10-01

    The fate of photosynthetic products within the plant-soil continuum determines how long the reduced carbon resides within the ecosystem and when it returns back to the atmosphere in the form of respiratory CO2. We have tested the possibility of measuring natural variation in δ13C and δ18O to disentangle the potential times needed to transfer carbohydrates produced by photosynthesis down to trunk, roots and, in general, to belowground up to its further release in the form of soil respiration into the atmosphere in a beech (Fagus sylvatica) forest. We have measured the variation in stable carbon and oxygen isotope compositions in plant material and in soil respired CO2 every three hours for three consecutive days. Possible steps and different signs of post-photosynthetic fractionation during carbon translocation were also identified. A 12 h-periodicity was observed for variation in δ13C in soluble sugars in the top crown leaves and it can be explained by starch day/night dynamics in synthesis and breakdown and by stomatal limitations under elevated vapour pressure deficits. Photosynthetic products were transported down the trunk and mixed with older carbon pools, therefore causing the dampening of the δ13C signal variation. The strongest periodicity of 24 h was found in δ13C in soil respiration indicating changes in root contribution to the total CO2 efflux. Other non-biological causes like diffusion fractionation and advection induced by gas withdrawn from the measurement chamber complicate data interpretation on this step of C transfer path. Nevertheless, it was possible to identify the speed of carbohydrates' translocation from the point of assimilation to the trunk breast height because leaf-imprinted enrichment of δ18O in soluble sugars was less modified along the downward transport and was well related to environmental parameters potentially linked to stomatal conductance. The speed of carbohydrates translocation from the site of assimilation to the trunk

  3. Glacial-interglacial shifts in global and regional precipitation δ18O

    NASA Astrophysics Data System (ADS)

    Jasechko, S.; Lechler, A.; Pausata, F. S. R.; Fawcett, P. J.; Gleeson, T.; Cendón, D. I.; Galewsky, J.; LeGrande, A. N.; Risi, C.; Sharp, Z. D.; Welker, J. M.; Werner, M.; Yoshimura, K.

    2015-03-01

    Previous analyses of past climate changes have often been based on site-specific isotope records from speleothems, ice cores, sediments and groundwaters. However, in most studies these dispersed records have not been integrated and synthesized in a comprehensive manner to explore the spatial patterns of precipitation isotope changes from the last ice age to more recent times. Here we synthesize 88 globally-distributed groundwater, cave calcite, and ice core isotope records spanning the last ice age to the late-Holocene. Our data-driven review shows that reconstructed precipitation δ18O changes from the last ice age to the late-Holocene range from -7.1‰ (ice age δ18O < late-Holocene δ18O) to +1.8‰ (ice age δ18O > late-Holocene δ18O) with wide regional variability. The majority (75%) of reconstructions have lower ice age δ18O values than late-Holocene δ18O values. High-magnitude, negative glacial-interglacial precipitation δ18O shifts (ice age δ18O < late-Holocene δ18O by more than 3‰) are common at high latitudes, high altitudes and continental interiors. Conversely, lower-magnitude, positive glacial-interglacial precipitation δ18O shifts (ice age δ18O > late-Holocene δ18O by less than 2‰) are most common along subtropical coasts. Broad, global patterns of glacial-interglacial precipitation δ18O shifts are consistent with stronger-than-modern isotopic distillation of air masses during the last ice age, likely impacted by larger global temperature differences between the tropics and the poles. Further, to complement our synthesis of proxy-record precipitation δ18O, we compiled isotope enabled general circulation model simulations of recent and last glacial maximum climate states. Simulated precipitation δ18O from five general circulation models show better inter-model and model-observation agreement in the sign of δ18O changes from the last ice age to present day in temperate and polar regions than in the tropics. Further model precipitation

  4. Quantifying Kinase-Specific Phosphorylation Stoichiometry Using Stable Isotope Labeling In a Reverse In-Gel Kinase Assay.

    PubMed

    Li, Xiang; Cox, Jonathan T; Huang, Weiliang; Kane, Maureen; Tang, Keqi; Bieberich, Charles J

    2016-12-06

    Despite recent advancements in large-scale phosphoproteomics, methods to quantify kinase-specific phosphorylation stoichiometry of protein substrates are lacking. We developed a method to quantify kinase-specific phosphorylation stoichiometry by combining the reverse in-gel kinase assay (RIKA) with high-resolution liquid chromatography-mass spectrometry (LC-MS). Beginning with predetermined ratios of phosphorylated to nonphosphorylated protein kinase CK2 (CK2) substrate molecules, we employed (18)O-labeled adenosine triphosphate ((18)O-ATP) as the phosphate donor in a RIKA, then quantified the ratio of (18)O- versus (16)O-labeled tryptic phosphopeptide using high mass accuracy mass spectrometry (MS). We demonstrate that the phosphorylation stoichiometry determined by this method across a broad percent phosphorylation range correlated extremely well with the predicted value (correlation coefficient = 0.99). This approach provides a quantitative alternative to antibody-based methods of determining the extent of phosphorylation of a substrate pool.

  5. Isotope Labeling for Solution and Solid-State NMR Spectroscopy of Membrane Proteins

    PubMed Central

    Verardi, Raffaello; Traaseth, Nathaniel J.; Masterson, Larry R.; Vostrikov, Vitaly V.; Veglia, Gianluigi

    2013-01-01

    In this chapter, we summarize the isotopic labeling strategies used to obtain high-quality solution and solid-state NMR spectra of biological samples, with emphasis on integral membrane proteins (IMPs). While solution NMR is used to study IMPs under fast tumbling conditions, such as in the presence of detergent micelles or isotropic bicelles, solid-state NMR is used to study the structure and orientation of IMPs in lipid vesicles and bilayers. In spite of the tremendous progress in biomolecular NMR spectroscopy, the homogeneity and overall quality of the sample is still a substantial obstacle to overcome. Isotopic labeling is a major avenue to simplify overlapped spectra by either diluting the NMR active nuclei or allowing the resonances to be separated in multiple dimensions. In the following we will discuss isotopic labeling approaches that have been successfully used in the study of IMPs by solution and solid-state NMR spectroscopy. PMID:23076578

  6. Open system sulphate reduction in a diagenetic environment - Isotopic analysis of barite (δ34S and δ18O) and pyrite (δ34S) from the Tom and Jason Late Devonian Zn-Pb-Ba deposits, Selwyn Basin, Canada

    NASA Astrophysics Data System (ADS)

    Magnall, J. M.; Gleeson, S. A.; Stern, R. A.; Newton, R. J.; Poulton, S. W.; Paradis, S.

    2016-05-01

    Highly positive δ34S values in sulphide minerals are a common feature of shale hosted massive sulphide deposits (SHMS). Often this is attributed to near quantitative consumption of seawater sulphate, and for Paleozoic strata of the Selwyn Basin (Canada), this is thought to occur during bacterial sulphate reduction (BSR) in a restricted, euxinic water column. In this study, we focus on drill-core samples of sulphide and barite mineralisation from two Late Devonian SHMS deposits (Tom and Jason, Macmillan Pass, Selwyn Basin), to evaluate this euxinic basin model. The paragenetic relationship between barite, pyrite and hydrothermal base metal sulphides has been determined using transmitted and reflected light microscopy, and backscatter electron imaging. This petrographic framework provides the context for in-situ isotopic microanalysis (secondary ion mass spectrometry; SIMS) of barite and pyrite. These data are supplemented by analyses of δ34S values for bulk rock pyrite (n = 37) from drill-core samples of un-mineralised (barren), siliceous mudstone, to provide a means by which to evaluate the mass balance of sulphur in the host rock. Three generations of barite have been identified, all of which pre-date hydrothermal input. Isotopically, the three generations of barite have overlapping distributions of δ34S and δ18O values (+22.5‰ to +33.0‰ and +16.4‰ to +18.3‰, respectively) and are consistent with an origin from modified Late Devonian seawater. Radiolarian tests, enriched in barium, are abundant within the siliceous mudstones, providing evidence that primary barium enrichment was associated with biologic activity. We therefore propose that barite formed following remobilisation of productivity-derived barium within the sediment, and precipitated within diagenetic pore fluids close to the sediment water interface. Two generations of pyrite are texturally associated with barite: framboidal pyrite (py-I), which has negative δ34S values (-23‰ to -28

  7. [Use of DNA-methylases as reagents for the production of isotopically labeled DNAs].

    PubMed

    Bur'ianov, Ia I; Bogdarina, I G; Nesterenko, V F; Baev, A A

    1982-04-01

    The method of incorporation of an isotopic label into DNA by means of DNA-methyltransferases (DNA-methylases) is proposed. DNA was no degraded and retained its biological activity in the DNA-methylase reaction. The specific activity of labelled DNA preparations can be increased, using the mixtures of different DNA-methylases in the enzymatic reaction. An isotopic label was incorporated into DNA, using DNA methylases M. .EcoRII, M.Eco dam and M.EcoMRE600 dcmI. An average activity of 1 microgram of labelled DNA preparations produced by S-adenosylmethionine (methyl-3H) with specific activity of 15 CU/mmol mas about 1 x 10(5) cpm.

  8. Structural determination of larger proteins using stable isotope labeling and NMR spectroscopy

    SciTech Connect

    Unkefer, C.; Hernandez, G.; Springer, P.; Trewhella, J.; Blumenthal, D.; Lidstrom, M.

    1996-04-01

    The project sought to employ stable isotope labeling and NMR spectroscopy to study protein structures and provide insight into important biochemical problems. A methylotrophic bacterial expression system has been developed for uniform deuterium and carbon-13 labeling of proteins for structural studies. These organisms grow using methanol as the sole source of carbon and energy. Because isotopically labeled methanol is relatively inexpensive, the methylotrophs are ideal for expressing proteins labeled uniformly with deuterium and/or carbon-13. This expression system has been employed to prepare deuterated troponin C. NMR spectroscopy measurements have been made on the inhibitory peptide from troponin I (residues 96--115), both as the free peptide and the peptide complexed with deuterated troponin C. Proton-NMR spectroscopy resonance-signal assignments have been made for the free peptide.

  9. Identification of RNA sequence isomer by isotope labeling and LC-MS/MS.

    PubMed

    Li, Siwei; Limbach, Patrick A

    2014-11-01

    Recently, we developed a method for modified ribonucleic acid (RNA) analysis based on the comparative analysis of RNA digests (CARD). Within this CARD approach, sequence or modification differences between two samples are identified through differential isotopic labeling of two samples. Components present in both samples will each be labeled, yielding doublets in the CARD mass spectrum. Components unique to only one sample should be detected as singlets. A limitation of the prior singlet identification strategy occurs when the two samples contain components of unique sequence but identical base composition. At the first stage of mass spectrometry, these sequence isomers cannot be differentiated and would appear as doublets rather than singlets. However, underlying sequence differences should be detectable by collision-induced dissociation tandem mass spectrometry (CID MS/MS), as y-type product ions will retain the original enzymatically incorporated isotope label. Here, we determine appropriate instrumental conditions that enable CID MS/MS of isotopically labeled ribonuclease T1 (RNase T1) digestion products such that the original isotope label is maintained in the product ion mass spectrum. Next, we demonstrate how y-type product ions can be used to differentiate singlets and doublets from isomer sequences. We were then able to extend the utility of this approach by using CID MS/MS for the confirmation of an expected RNase T1 digestion product within the CARD analysis of an Escherichia coli mutant strain even in the presence of interfering and overlapping digestion products from other transfer RNAs.

  10. Quantitative Microbial Ecology through Stable Isotope Probing

    PubMed Central

    Mau, Rebecca L.; Schwartz, Egbert; Caporaso, J. Gregory; Dijkstra, Paul; van Gestel, Natasja; Koch, Benjamin J.; Liu, Cindy M.; McHugh, Theresa A.; Marks, Jane C.; Morrissey, Ember M.; Price, Lance B.

    2015-01-01

    Bacteria grow and transform elements at different rates, and as yet, quantifying this variation in the environment is difficult. Determining isotope enrichment with fine taxonomic resolution after exposure to isotope tracers could help, but there are few suitable techniques. We propose a modification to stable isotope probing (SIP) that enables the isotopic composition of DNA from individual bacterial taxa after exposure to isotope tracers to be determined. In our modification, after isopycnic centrifugation, DNA is collected in multiple density fractions, and each fraction is sequenced separately. Taxon-specific density curves are produced for labeled and nonlabeled treatments, from which the shift in density for each individual taxon in response to isotope labeling is calculated. Expressing each taxon's density shift relative to that taxon's density measured without isotope enrichment accounts for the influence of nucleic acid composition on density and isolates the influence of isotope tracer assimilation. The shift in density translates quantitatively to isotopic enrichment. Because this revision to SIP allows quantitative measurements of isotope enrichment, we propose to call it quantitative stable isotope probing (qSIP). We demonstrated qSIP using soil incubations, in which soil bacteria exhibited strong taxonomic variations in 18O and 13C composition after exposure to [18O]water or [13C]glucose. The addition of glucose increased the assimilation of 18O into DNA from [18O]water. However, the increase in 18O assimilation was greater than expected based on utilization of glucose-derived carbon alone, because the addition of glucose indirectly stimulated bacteria to utilize other substrates for growth. This example illustrates the benefit of a quantitative approach to stable isotope probing. PMID:26296731

  11. Quantifying plant phenotypes with isotopic labeling and metabolic flux analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analyses of metabolic flux using stable isotopes in plants have traditionally been restricted to tissues with presumed homogeneous cell populations such as developing seeds, cell suspensions, or cultured roots and root tips. It is now possible to describe these and other more complex tissues such a...

  12. Stable isotope labeling of oligosaccharide cell surface antigens

    SciTech Connect

    Unkefer, C.J.; Silks, L.A. III; Martinez, R.A.

    1998-12-31

    The overall goal of this Laboratory Directed Research and Development (LDRD) project was to develop new methods for synthesis of {sup 13}C-labeled oligosaccharides that are required for nuclear magnetic resonance (NMR) studies of their solution conformation. Oligosaccharides are components of the cell`s outer surface and are involved in important processes such as cell-cell recognition and adhesion. Recently, Danishefsky and coworkers at Slone-Kettering Cancer Center developed a method for the solid-phase chemical synthesis of oligosaccharides. The specific goal of this LDRD project was to prepare uniform {sup 13}C-labeled aldohexose precursors required for the solid-phase synthesis of the Lewis blood-group antigenic determinants. We report the synthesis of {sup 13}C-labeled D-glucal, D-galactal and Fucosyl precursors. We have been collaborating with the Danishefsky group on the synthesis of the Lewis oligosaccharides and the NMR analysis of their solution conformation.

  13. 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.

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

    SciTech Connect

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

    1986-12-01

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

  15. A free-air system for long-term stable carbon isotope labeling of adult forest trees

    EPA Science Inventory

    Stable carbon (C) isotopes, in particular employed in labeling experiments, are an ideal tool to broaden our understanding of C dynamics in trees and forest ecosystems. Here, we present a free-air exposure system, named isoFACE, designed for long-term stable C isotope labeling in...

  16. Addressing the current bottlenecks of metabolomics: Isotopic Ratio Outlier Analysis™, an isotopic-labeling technique for accurate biochemical profiling.

    PubMed

    de Jong, Felice A; Beecher, Chris

    2012-09-01

    Metabolomics or biochemical profiling is a fast emerging science; however, there are still many associated bottlenecks to overcome before measurements will be considered robust. Advances in MS resolution and sensitivity, ultra pressure LC-MS, ESI, and isotopic approaches such as flux analysis and stable-isotope dilution, have made it easier to quantitate biochemicals. The digitization of mass spectrometers has simplified informatic aspects. However, issues of analytical variability, ion suppression and metabolite identification still plague metabolomics investigators. These hurdles need to be overcome for accurate metabolite quantitation not only for in vitro systems, but for complex matrices such as biofluids and tissues, before it is possible to routinely identify biomarkers that are associated with the early prediction and diagnosis of diseases. In this report, we describe a novel isotopic-labeling method that uses the creation of distinct biochemical signatures to eliminate current bottlenecks and enable accurate metabolic profiling.

  17. Isotopic labelling studies on far-infrared spectra of nickel-histamine complexes

    NASA Astrophysics Data System (ADS)

    Drożdżewski, Piotr; Kordon, Ewa

    2000-11-01

    Nickel-histamine (hm) complexes type [Ni(hm)Cl 2] and [Ni(hm) 3] X2 (Where X=Cl, Br, I, ClO 4) were investigated in the far-infrared region. Metal isotope labelling and deuteration effects were employed for observed band assignments. Metal-ligand vibrations were discussed and correlated with the structures of the complexes.

  18. Practical considerations in the use of stable isotope labelled compounds as tracers in clinical studies.

    PubMed

    Thompson, G N; Pacy, P J; Ford, G C; Halliday, D

    1989-05-01

    Increasingly widespread usage of stable isotope tracers to aid clinical diagnosis and support basic research has stemmed from both advances in mass spectrometry and the availability of competitively priced labelled compounds. Stable isotopes have been used generally to investigate normal and abnormal metabolic pathways, to estimate energy expenditure and body composition and to quantitate substrate flux and oxidation rates. Despite the fact that the underlying principles relating to the use of stable isotopes for in vivo studies are straightforward, careful consideration must be given to all aspects of human studies. This review highlights some of these, including choice of label and tracer molecule, mode of tracer administration and sampling site, analytical instrumentation, interpretation of data and ethical constraints.

  19. Stable-isotope-labeled carbohydrates and nucleosides: Synthesis and applications in chemistry and biology

    SciTech Connect

    Serianni, A.S.

    1994-12-01

    Carbohydrates play important roles in many key biochemical processes in living cells. For example, they are metabolized to produce energy, mediate cell-cell recognition, and play an indirect role (as constituents of DNA and RNA) in DNA replication, RNA transcription, and protein synthesis. These roles, and others of comparable biochemical significance, have been studied to varying extends with the use of stable isotopically labeled molecules, usually in conjunction with NMR spectroscopy and/or mass spectrometry. For example, carbohydrate metabolism has been monitored in vitro and in vivo with the use of isotopically labeled compounds. Molecular aspects of cell-cell recognition, mediated by cell-surface glycoproteins and glycolipids, have been probed through NMR studies of isotopically labeled oligosaccharides. More recently, the solution behavior of DNA and RNA has been examined through the use of labeled oligonucleotides. In all of these pursuits, the effort and expense to prepare labeled molecules, both of which can be substantial, are more than offset by the wealth of information derived from these studies. This information often cannot be accessed, or can be accessed only with great difficulty, using natural (unlabeled) compounds.

  20. Hydroponic isotope labeling of entire plants and high-performance mass spectrometry for quantitative plant proteomics.

    PubMed

    Bindschedler, Laurence V; Mills, Davinia J S; Cramer, Rainer

    2012-01-01

    Hydroponic isotope labeling of entire plants (HILEP) combines hydroponic plant cultivation and metabolic labeling with stable isotopes using (15)N-containing inorganic salts to label whole and mature plants. Employing (15)N salts as the sole nitrogen source for HILEP leads to the production of healthy-looking plants which contain (15)N proteins labeled to nearly 100%. Therefore, HILEP is suitable for quantitative plant proteomic analysis, where plants are grown in either (14)N- or (15)N-hydroponic media and pooled when the biological samples are collected for relative proteome quantitation. The pooled (14)N-/(15)N-protein extracts can be fractionated in any suitable way and digested with a protease for shotgun proteomics, using typically reverse phase liquid chromatography nanoelectrospray ionization tandem mass spectrometry (RPLC-nESI-MS/MS). Best results were obtained with a hybrid ion trap/FT-MS mass spectrometer, combining high mass accuracy and sensitivity for the MS data acquisition with speed and high-throughput MS/MS data acquisition, increasing the number of proteins identified and quantified and improving protein quantitation. Peak processing and picking from raw MS data files, protein identification, and quantitation were performed in a highly automated way using integrated MS data analysis software with minimum manual intervention, thus easing the analytical workflow. In this methodology paper, we describe how to grow Arabidopsis plants hydroponically for isotope labeling using (15)N salts and how to quantitate the resulting proteomes using a convenient workflow that does not require extensive bioinformatics skills.

  1. Investigation of spatio-temporal variability of water uptake in a groundwater-dependent ecosystem using a stable isotope approach (δ18O, δ2H): Pfyn Forest, Switzerland

    NASA Astrophysics Data System (ADS)

    Bertrand, G.; Masini, J.; Goldscheider, N.; Gobat, J. M.; Hunkeler, D.

    2012-04-01

    This work consists of an eco-hydrogeological study of the Pfyn Forest (46o17'35''N; 7o31'59''E, z = 550 m) which is a 6 km long alluvial zone in the upper Rhône valley, near Sierre (Wallis, Switzerland). From a hydrological point of view, the Rhône has a glacio-nival regime type in this area. Between low-flow and high-flow periods, groundwater levels strongly vary (about 8 m) near the main river-aquifer interaction zone in the most upgradient part of the site. In contrast, the downstream part of Pfyn is characterized by a low groundwater level fluctuation of about 1 or 2 m. From an ecological point of view, the riverine fringe at Pfyn presents a broadly recognized natural value but faces many threats due to human activities (derivation channel located upstream, gravel pits). Phytocoenosis vary from dry environments associations (with Scots pines, feather grass) upstream to active floodplain associations (with poplars, alders, willows) and likely dependent on groundwater, downstream. Between these two end-members, a transition mixed forest occurs. In the context of a potential hydrologic alteration due to global climatic change in a close future, this ecosystem should face modifications of the various water source (rainwater, groundwater) proportion and availability. In order to constrain the meteorological, hydrological, pedological and ecological factors governing water uptakes by trees, isotopic characterizations (δ18O, δ2H) of each water compartment (precipitations, groundwater, river, soil, xylem) coupled with the evaluation of the water balance, has been carried out. The investigation focused on 3 different sites located along a transect through the alluvial valley between April 2010 and February 2011, with a twice-monthly resolution. The data permit to obtain three major findings: - At first, an overview of both δ18O and δ2H data shows that rainwater, groundwater, soil water and plant water are usually located on the regional meteoric water line. For

  2. Escherichia coli cell-free protein synthesis and isotope labeling of mammalian proteins.

    PubMed

    Terada, Takaho; Yokoyama, Shigeyuki

    2015-01-01

    This chapter describes the cell-free protein synthesis method, using an Escherichia coli cell extract. This is a cost-effective method for milligram-scale protein production and is particularly useful for the production of mammalian proteins, protein complexes, and membrane proteins that are difficult to synthesize by recombinant expression methods, using E. coli and eukaryotic cells. By adjusting the conditions of the cell-free method, zinc-binding proteins, disulfide-bonded proteins, ligand-bound proteins, etc., may also be produced. Stable isotope labeling of proteins can be accomplished by the cell-free method, simply by using stable isotope-labeled amino acid(s) in the cell-free reaction. Moreover, the cell-free protein synthesis method facilitates the avoidance of stable isotope scrambling and dilution over the recombinant expression methods and is therefore advantageous for amino acid-selective stable isotope labeling. Site-specific stable isotope labeling is also possible with a tRNA molecule specific to the UAG codon. By the cell-free protein synthesis method, coupled transcription-translation is performed from a plasmid vector or a PCR-amplified DNA fragment encoding the protein. A milligram quantity of protein can be produced with a milliliter-scale reaction solution in the dialysis mode. More than a thousand solution structures have been determined by NMR spectroscopy for uniformly labeled samples of human and mouse functional domain proteins, produced by the cell-free method. Here, we describe the practical aspects of mammalian protein production by the cell-free method for NMR spectroscopy.

  3. δ18O anchoring to VPDB: calcite digestion with 18O-adjusted ortho-phosphoric acid.

    PubMed

    Wendeberg, Magnus; Richter, Jürgen M; Rothe, Michael; Brand, Willi A

    2011-04-15

    For anchoring CO(2) isotopic measurements on the δ(18)O(VPD-CO2) scale, the primary reference material (NBS 19 calcite) needs to be digested using concentrated ortho-phosphoric acid. During this procedure, great care must be taken to ensure that the isotopic composition of the liberated gas is accurate. Apart from controlling the reaction temperature to ±0.1 °C, the potential for oxygen isotope exchange between the produced CO(2) and water must be kept to a minimum. The water is usually assumed to reside on the walls in the headspace of the reaction vessel. We demonstrate here that a large fraction of the exchange may also occur with water inside the acid. Our results indicate that both exchange reactions have a significant impact on the results and may have largely been responsible for scale inconsistencies between laboratories in the past. The extent of CO(2)/H(2)O oxygen exchange depends on the concentration (amount of free water) in the acid. For acids with a nominal H(3)PO(4) mass fraction of less than 102%, oxygen isotope exchange can create a substantial isotopic bias during high-precision measurements with the degree of the alteration being proportional to the effective isotopic contrast between the acid and the CO(2) released from the calcite. Water evaporating from the acid at 25 °C has a δ(18)O value of -34.5‰ relative to the isotopic composition of the whole acid. This large fractionation is likely to occur in two steps; by exchange with phosphate, water inside the acid is decreased in oxygen-18 relative to the bulk acid by ∼ -22‰. This water is then fractionated further during evaporation. Oxygen exchange with both water inside the acid and water condensate in the headspace can contribute to the measured isotopic signature depending on the experimental parameters. The system employed for this study has been specifically designed to minimize oxygen exchange with water. However, the amount of altered CO(2) for a 95% H(3)PO(4) at 25 °C still

  4. Metabolic Flux Elucidation for Large-Scale Models Using 13C Labeled Isotopes

    PubMed Central

    Suthers, Patrick F.; Burgard, Anthony P.; Dasika, Madhukar S.; Nowroozi, Farnaz; Van Dien, Stephen; Keasling, Jay D.; Maranas, Costas D.

    2007-01-01

    A key consideration in metabolic engineering is the determination of fluxes of the metabolites within the cell. This determination provides an unambiguous description of metabolism before and/or after engineering interventions. Here, we present a computational framework that combines a constraint-based modeling framework with isotopic label tracing on a large-scale. When cells are fed a growth substrate with certain carbon positions labeled with 13C, the distribution of this label in the intracellular metabolites can be calculated based on the known biochemistry of the participating pathways. Most labeling studies focus on skeletal representations of central metabolism and ignore many flux routes that could contribute to the observed isotopic labeling patterns. In contrast, our approach investigates the importance of carrying out isotopic labeling studies using a more comprehensive reaction network consisting of 350 fluxes and 184 metabolites in Escherichia coli including global metabolite balances on cofactors such as ATP, NADH, and NADPH. The proposed procedure is demonstrated on an E. coli strain engineered to produce amorphadiene, a precursor to the anti-malarial drug artemisinin. The cells were grown in continuous culture on glucose containing 20% [U-13C]glucose; the measurements are made using GC-MS performed on 13 amino acids extracted from the cells. We identify flux distributions for which the calculated labeling patterns agree well with the measurements alluding to the accuracy of the network reconstruction. Furthermore, we explore the robustness of the flux calculations to variability in the experimental MS measurements, as well as highlight the key experimental measurements necessary for flux determination. Finally, we discuss the effect of reducing the model, as well as shed light onto the customization of the developed computational framework to other systems. PMID:17632026

  5. Absolute peptide quantification by lutetium labeling and nanoHPLC-ICPMS with isotope dilution analysis.

    PubMed

    Rappel, Christina; Schaumlöffel, Dirk

    2009-01-01

    The need of analytical methods for absolute quantitative protein analysis spurred research on new developments in recent years. In this work, a novel approach was developed for accurate absolute peptide quantification based on metal labeling with lutetium diethylenetriamine pentaacetic acid (Lu-DTPA) and nanoflow high-performance liquid chromatography-inductively coupled plasma isotope dilution mass spectrometry (nanoHPLC-ICP-IDMS). In a two-step procedure peptides were derivatized at amino groups with diethylenetriamine pentaacetic anhydride (DTPAA) followed by chelation of lutetium. Electrospray ionization mass spectrometry (ESI MS) of the reaction product demonstrated highly specific peptide labeling. Under optimized nanoHPLC conditions the labeled peptides were baseline-separated, and the excess labeling reagent did not interfere. A 176Lu-labeled spike was continuously added to the column effluent for quantification by ICP-IDMS. The recovery of a Lu-DTPA-labeled standard peptide was close to 100% indicating high labeling efficiency and accurate absolute quantification. The precision of the entire method was 4.9%. The detection limit for Lu-DTPA-tagged peptides was 179 amol demonstrating that lutetium-specific peptide quantification was by 4 orders of magnitude more sensitive than detection by natural sulfur atoms present in cysteine or methionine residues. Furthermore, the application to peptides in insulin tryptic digest allowed the identification of interfering reagents decreasing the labeling efficiency. An additional advantage of this novel approach is the analysis of peptides, which do not naturally feature ICPMS-detectable elements.

  6. Kinetic isotope effect of the {sup 16}O + {sup 36}O{sub 2} and {sup 18}O + {sup 32}O{sub 2} isotope exchange reactions: Dominant role of reactive resonances revealed by an accurate time-dependent quantum wavepacket study

    SciTech Connect

    Sun, Zhigang Yu, Dequan; Xie, Wenbo; Hou, Jiayi; Dawes, Richard; Guo, Hua

    2015-05-07

    The O + O{sub 2} isotope exchange reactions play an important role in determining the oxygen isotopic composition of a number of trace gases in the atmosphere, and their temperature dependence and kinetic isotope effects (KIEs) provide important constraints on our understanding of the origin and mechanism of these and other unusual oxygen KIEs important in the atmosphere. This work reports a quantum dynamics study of the title reactions on the newly constructed Dawes-Lolur-Li-Jiang-Guo (DLLJG) potential energy surface (PES). The thermal reaction rate coefficients of both the {sup 18}O + {sup 32}O{sub 2} and {sup 16}O + {sup 36}O{sub 2} reactions obtained using the DLLJG PES exhibit a clear negative temperature dependence, in sharp contrast with the positive temperature dependence obtained using the earlier modified Siebert-Schinke-Bittererova (mSSB) PES. In addition, the calculated KIE shows an improved agreement with the experiment. These results strongly support the absence of the “reef” structure in the entrance/exit channels of the DLLJG PES, which is present in the mSSB PES. The quantum dynamics results on both PESs attribute the marked KIE to strong near-threshold reactive resonances, presumably stemming from the mass differences and/or zero point energy difference between the diatomic reactant and product. The accurate characterization of the reactivity for these near-thermoneutral reactions immediately above the reaction threshold is important for correct characterization of the thermal reaction rate coefficients.

  7. REDOR NMR of stable-isotope-labeled protein binding sites

    SciTech Connect

    Schaefer, J.

    1994-12-01

    Rotational-echo, double resonance (REDOR) NMR, a new analytical spectroscopic technique for solids spinning at the magic angle, has been developed over the last 5 years. REDOR provides a direct measure of heteronuclear dipolar coupling between isolated pairs of labeled nuclei. In a solid with a {sup 13}C-{sup 15}N labeled pair, for example, the {sup 13}C rotational echoes that form each rotor period following a{sup 1}H-{sup 13}C cross-polarization transfer can be prevented from reaching full intensity by insertion of a {sup 15}N {pi} pulse each half rotor period. The REDOR difference (the difference between a {sup 13}C NMR spectrum obtained under these conditions and one obtained with no {sup 15}N {pi} pulses) has a strong dependence on the {sup 13}C-{sup 15}N dipolar coupling, and hence, the {sup 13}C-{sup 15}N internuclear distance. REDOR is described as double-resonance even though three radio frequencies (typically {sup 1}H, {sup 13}C, and {sup 15}N) are used because the protons are removed from the important evolution part of the experiment by resonant decoupling. The dephasing of magnetization in REDOR arises from a local dipolar {sup 13}C-{sup 15}N field gradient and involves no polarization transfer. REDOR has no dependence on {sup 13}C or {sup 15}N chemical-shift tensors and does not require resolution of a {sup 13}C-{sup 15}N coupling in the chemical-shift dimension.

  8. The role of soil processes in δ18O terrestrial climate proxies

    NASA Astrophysics Data System (ADS)

    Kanner, Lisa C.; Buenning, Nikolaus H.; Stott, Lowell D.; Timmermann, Axel; Noone, David

    2014-03-01

    A paleoclimate interpretation of a terrestrial hydrologic proxy such as the δ18O of tree cellulose or speleothem calcite may be biased or misinterpreted if the isotopic composition of the soil water from which the proxy originated undergoes isotopic exchange or fractionation. In this study, we use a global isotope-enabled land surface model to investigate how the δ18O of precipitation may be altered in a soil column due to evaporation and vertical moisture transport. In order to assess how precipitation and evaporation contribute to the soil water isotopic variability, we compare seasonal and interannual changes in simulated xylem water δ18O within a control simulation and in a suite of sensitivity experiments where the effects of precipitation δ18O, water vapor δ18O, and soil water evaporation are independently removed. The simulations, carried out for the period 1979 to 2004, reveal that in semiarid regions, such as the southwest United States, the seasonal cycle in xylem water δ18O is strongly affected by evaporative loss during the dry season and evaporation can also constitute as much as 50% of the interannual δ18O variance. Additional simulations, including soil water tagging experiments, indicate that upward fluxes of soil water occur during drier periods. For soil water δ18O profiles that are isotopically more depleted in 18O at depth, this imparts a low isotopic signature to xylem water δ18O during such dry intervals. Hence, without taking into account vertical moisture transport in the soils, low δ18O years could be misinterpreted as wet conditions (due to decreased evaporative enrichment) when instead drier conditions are equally as likely.

  9. Turnover of Leaf Waxes in Florida Slash Pine: Results of an Isotopic Labeling Experiment

    NASA Astrophysics Data System (ADS)

    Crumsey, J.; Conte, M. H.; Weber, J. C.; Mortazavi, B.; Smith, M.; Chanton, J.

    2006-12-01

    Isotopic discrimination of terrestrial photosynthesis, atmospheric CO2 concentration, and δ13CO2 are important parameters in global carbon models that are employed to estimate global carbon sources and sinks. Yet, terrestrial isotopic discrimination can be highly variable over space and time, yielding large uncertainties of terrestrial fluxes. The isotopic composition of plant wax aerosols in continental air masses can be used as an indirect measure of the spatial and temporal patterns of photosynthetic discrimination integrated over large (subcontinental) spatial scales. However, the temporal offset between wax biosynthesis and the wax aerosol isotopic signal of photosynthetic discrimination is not well constrained. To further our understanding of this temporal lag, this study sought to determine the turnover time of conifer leaf waxes by performing an isotopic labeling experiment. Four clonal pine saplings were placed in a tent and labeled with enriched 13CO2 for one year, while another four control saplings were grown under ambient CO2. At the end of the year long enrichment, the labeled saplings were removed from the tent and placed in ambient air, such that the wax turnover rate could be determined by analyzing the resultant isotopic and molecular changes. The results of this experiment indicated that after 80 days of sequestering ambient CO2, the wax (and soluble sugar) isotopic composition of the labeled saplings varied minimally. The molecular composition of the waxes, however, did change over time. From these results we concluded that waxes are turning over, but rather than being synthesized de novo from recently fixed carbon precursors they are synthesized using carbon from stored (labeled) carbon pools. Therefore, the δ13C of conifer leaf waxes in aerosols may not reflect recent photosynthetic discrimination, but instead represents photosynthetic discrimination integrated over a longer period of time. The implications of these findings are focused on

  10. Human neutrophil kinetics: modeling of stable isotope labeling data supports short blood neutrophil half-lives

    PubMed Central

    Lahoz-Beneytez, Julio; Elemans, Marjet; Zhang, Yan; Ahmed, Raya; Salam, Arafa; Block, Michael; Niederalt, Christoph; Macallan, Derek

    2016-01-01

    Human neutrophils have traditionally been thought to have a short half-life in blood; estimates vary from 4 to 18 hours. This dogma was recently challenged by stable isotope labeling studies with heavy water, which yielded estimates in excess of 3 days. To investigate this disparity, we generated new stable isotope labeling data in healthy adult subjects using both heavy water (n = 4) and deuterium-labeled glucose (n = 9), a compound with more rapid labeling kinetics. To interpret results, we developed a novel mechanistic model and applied it to previously published (n = 5) and newly generated data. We initially constrained the ratio of the blood neutrophil pool to the marrow precursor pool (ratio = 0.26; from published values). Analysis of heavy water data sets yielded turnover rates consistent with a short blood half-life, but parameters, particularly marrow transit time, were poorly defined. Analysis of glucose-labeling data yielded more precise estimates of half-life (0.79 ± 0.25 days; 19 hours) and marrow transit time (5.80 ± 0.42 days). Substitution of this marrow transit time in the heavy water analysis gave a better-defined blood half-life of 0.77 ± 0.14 days (18.5 hours), close to glucose-derived values. Allowing the ratio of blood neutrophils to mitotic neutrophil precursors (R) to vary yielded a best-fit value of 0.19. Reanalysis of the previously published model and data also revealed the origin of their long estimates for neutrophil half-life: an implicit assumption that R is very large, which is physiologically untenable. We conclude that stable isotope labeling in healthy humans is consistent with a blood neutrophil half-life of less than 1 day. PMID:27136946

  11. Discovery and validation of colonic tumor-associated proteins via metabolic labeling and stable isotopic dilution

    PubMed Central

    Huttlin, Edward L.; Chen, Xiaodi; Barrett-Wilt, Gregory A.; Hegeman, Adrian D.; Halberg, Richard B.; Harms, Amy C.; Newton, Michael A.; Dove, William F.; Sussman, Michael R.

    2009-01-01

    The unique biology of a neoplasm is reflected by its distinct molecular profile compared with normal tissue. To understand tumor development better, we have undertaken a quantitative proteomic search for abnormally expressed proteins in colonic tumors from ApcMin/+ (Min) mice. By raising pairs of Min and wild-type mice on diets derived from natural-abundance or 15N-labeled algae, we used metabolic labeling to compare protein levels in colonic tumor versus normal tissue. Because metabolic labeling allows internal control throughout sample preparation and analysis, technical error is minimized as compared with in vitro labeling. Several proteins displayed altered expression, and a subset was validated via stable isotopic dilution using synthetic peptide standards. We also compared gene and protein expression among tumor and nontumor tissue, revealing limited correlation. This divergence was especially pronounced for species showing biological change, highlighting the complementary perspectives provided by transcriptomics and proteomics. Our work demonstrates the power of metabolic labeling combined with stable isotopic dilution as an integrated strategy for the identification and validation of differentially expressed proteins using rodent models of human disease. PMID:19805096

  12. Three whole-wood isotopic reference materials, USGS54, USGS55, and USGS56, for δ2H, δ13C, δ15N, and δ18O measurements

    USGS Publications Warehouse

    Qi, Haiping; Coplen, Tyler B.; Jordan, James A.

    2016-01-01

    Comparative measurements of stable hydrogen and oxygen isotopes in wood are hampered by the lack of proper reference materials (RMs). The U.S. Geological Survey (USGS) has prepared three powdered, whole-wood RMs, USGS54 (Pinus contorta, Canadian lodgepole pine), USGS55 (Cordia cf. dodecandra, Mexican ziricote), and USGS56 (Berchemia cf. zeyheri, South African red ivorywood). The stable isotopes of hydrogen, oxygen, carbon, and nitrogen in these RMs span ranges as δ2HVSMOW from –150.4 to –28.2 mUr or ‰, as δ18OVSMOW from + 17.79 to + 27.23 mUr, as δ13CVPDB from –27.13 to –24.34 mUr, and as δ15N AIR-N2 from –2.42 to + 1.8 mUr. These RMs will enable users to normalize measurements of wood samples to isotope–delta scales, and they are intended primarily for the normalization of δ2H and δ18O measurements of unknown wood samples. However, they also are suitable for normalization of stable isotope measurements of carbon and nitrogen in wood samples. In addition, these RMs are suitable for inter-laboratory calibration for the dual-water suilibration procedure for the measurements of δ2HVSMOW values of non-exchangeable hydrogen. The isotopic compositions with 1-σ uncertainties, mass fractions of each element, and fractions of exchangeable hydrogen of these materials are:USGS54 (Pinus contorta, Canadian Lodgepole pine)δ2HVSMOW = –150.4 ± 1.1 mUr (n = 29), hydrogen mass fraction = 6.00 ± 0.04 % (n = 10)Fraction of exchangeable hydrogen = 5.4 ± 0.6 % (n = 29)δ18OVSMOW = + 17.79 ± 0.15 mUr (n = 18), oxygen mass fraction = 40.4 ± 0.2 % (n = 6)δ13CVPDB = –24.43 ± 0.02 mUr (n = 18), carbon mass fraction = 48.3 ± 0.4 % (n = 12)δ15NAIR-N2 = –2.42 ± 0.32 mUr (n = 17), nitrogen mass fraction = 0.05 % (n = 4)USGS55 (Cordia cf. dodecandra, Mexican ziricote)δ2HVSMOW = –28.2 ± 1.7 mUr (n = 30), hydrogen mass fraction = 5.65 ± 0.06 % (n = 10)Fraction of exchangeable

  13. Forward modeling of δ18O in Andean ice cores

    NASA Astrophysics Data System (ADS)

    Hurley, J. V.; Vuille, M.; Hardy, D. R.

    2016-08-01

    Tropical ice core archives are among the best dated and highest resolution from the tropics, but a thorough understanding of processes that shape their isotope signature as well as the simulation of observed variability remain incomplete. To address this, we develop a tropical Andean ice core isotope forward model from in situ hydrologic observations and satellite water vapor isotope measurements. A control simulation of snow δ18O captures the mean and seasonal trend but underestimates the observed intraseasonal variability. The simulation of observed variability is improved by including amount effects associated with South American cold air incursions, linking synoptic-scale disturbances and monsoon dynamics to tropical ice core δ18O. The forward model was calibrated with and run under present-day conditions but can also be driven with past climate forcings to reconstruct paleomonsoon variability. The model is transferable and may be used to render a (paleo)climatic context at other ice core locations.

  14. Analysis of mitochondrial metabolism in situ: Combining stable isotope labeling with selective permeabilization.

    PubMed

    Nonnenmacher, Yannic; Palorini, Roberta; d'Herouël, Aymeric Fouquier; Krämer, Lisa; Neumann-Schaal, Meina; Chiaradonna, Ferdinando; Skupin, Alexander; Wegner, Andre; Hiller, Karsten

    2016-12-15

    To date, it is well-established that mitochondrial dysfunction does not only play a vital role in cancer but also in other pathological conditions such as neurodegenerative diseases and inflammation. An important tool for the analysis of cellular metabolism is the application of stable isotope labeled substrates, which allow for the tracing of atoms throughout metabolic networks. While such analyses yield very detailed information about intracellular fluxes, the determination of compartment specific fluxes is far more challenging. Most approaches for the deconvolution of compartmented metabolism use computational models whereas experimental methods are rare. Here, we developed an experimental setup based on selective permeabilization of the cytosolic membrane that allows for the administration of stable isotope labeled substrates directly to mitochondria. We demonstrate how this approach can be used to infer metabolic changes in mitochondria induced by either chemical or genetic perturbations and give an outlook on its potential applications.

  15. Review of metabolic pathways activated in cancer cells as determined through isotopic labeling and network analysis.

    PubMed

    Dong, Wentao; Keibler, Mark A; Stephanopoulos, Gregory

    2017-02-10

    Cancer metabolism has emerged as an indispensable part of contemporary cancer research. During the past 10 years, the use of stable isotopic tracers and network analysis have unveiled a number of metabolic pathways activated in cancer cells. Here, we review such pathways along with the particular tracers and labeling observations that led to the discovery of their rewiring in cancer cells. The list of such pathways comprises the reductive metabolism of glutamine, altered glycolysis, serine and glycine metabolism, mutant isocitrate dehydrogenase (IDH) induced reprogramming and the onset of acetate metabolism. Additionally, we demonstrate the critical role of isotopic labeling and network analysis in identifying these pathways. The alterations described in this review do not constitute a complete list, and future research using these powerful tools is likely to discover other cancer-related pathways and new metabolic targets for cancer therapy.

  16. Chemical Ligation and Isotope Labeling to Locate Dynamic Effects during Catalysis by Dihydrofolate Reductase.

    PubMed

    Luk, Louis Y P; Ruiz-Pernía, J Javier; Adesina, Aduragbemi S; Loveridge, E Joel; Tuñón, Iñaki; Moliner, Vincent; Allemann, Rudolf K

    2015-07-27

    Chemical ligation has been used to alter motions in specific regions of dihydrofolate reductase from E. coli and to investigate the effects of localized motional changes on enzyme catalysis. Two isotopic hybrids were prepared; one with the mobile N-terminal segment containing heavy isotopes ((2) H, (13) C, (15) N) and the remainder of the protein with natural isotopic abundance, and the other one with only the C-terminal segment isotopically labeled. Kinetic investigations indicated that isotopic substitution of the N-terminal segment affected only a physical step of catalysis, whereas the enzyme chemistry was affected by protein motions from the C-terminal segment. QM/MM studies support the idea that dynamic effects on catalysis mostly originate from the C-terminal segment. The use of isotope hybrids provides insights into the microscopic mechanism of dynamic coupling, which is difficult to obtain with other studies, and helps define the dynamic networks of intramolecular interactions central to enzyme catalysis.

  17. Chemical Ligation and Isotope Labeling to Locate Dynamic Effects during Catalysis by Dihydrofolate Reductase†

    PubMed Central

    Luk, Louis Y. P.; Ruiz‐Pernía, J. Javier; Adesina, Aduragbemi S.; Loveridge, E. Joel

    2015-01-01

    Abstract Chemical ligation has been used to alter motions in specific regions of dihydrofolate reductase from E. coli and to investigate the effects of localized motional changes on enzyme catalysis. Two isotopic hybrids were prepared; one with the mobile N‐terminal segment containing heavy isotopes (2H, 13C, 15N) and the remainder of the protein with natural isotopic abundance, and the other one with only the C‐terminal segment isotopically labeled. Kinetic investigations indicated that isotopic substitution of the N‐terminal segment affected only a physical step of catalysis, whereas the enzyme chemistry was affected by protein motions from the C‐terminal segment. QM/MM studies support the idea that dynamic effects on catalysis mostly originate from the C‐terminal segment. The use of isotope hybrids provides insights into the microscopic mechanism of dynamic coupling, which is difficult to obtain with other studies, and helps define the dynamic networks of intramolecular interactions central to enzyme catalysis. PMID:26079622

  18. Implementation of a New Isotopic Tool (18O/16O in dissolved O2) for the Management of Oxygen in the Grand River, an Impacted Watershed in S. Ontario

    NASA Astrophysics Data System (ADS)

    Jamieson, T. S.; Schiff, S. L.; Taylor, W. D.

    2004-05-01

    Oxygen (O2) is essential for supporting life in aquatic ecosystems, and mediates the geochemistry of the aquatic environment. Recent analytical development now permits rapid analysis of the 18O/16O ratio of dissolved oxygen (DO) in water. The 18O/16O ratio provides insight into the main processes controlling DO concentrations in rivers: gas exchange, photosynthesis and respiration. Previously, this information was only available using indirect methods that involved laborious point scale measurements that are not amenable to routine sampling. The focus of this study is the Grand River, a Canadian Heritage River that drains into Lake Erie. The receives organic and nutrient inputs from sewage treatment plants, non-point agricultural sources, and serves as a source of drinking water and supports a significant recreational fishery. Dissolved oxygen concentrations in the Grand River can cycle from 2 mg/l to 14 mg/l in the summer during a 24 hour period. The concurrent shift in 18O/16O is greater than 10 permil on a diel basis. Rigorous sampling regimes are needed in dynamic systems that experience large fluxes in O2. Diurnal curves were analyzed for DO and 18O/16O in DO during different seasons. Gas exchange is an important constraint on the diurnal flux of DO. Currently, gas exchange coefficients for the Grand River are typically obtained using empirical estimates based on results from other rivers. Analysis of 18O/16O offers a new method to allow for the direct determination of the gas exchange coefficient for DO in rivers with diurnal cycles.

  19. RABA (Reductive Alkylation By Acetone): A novel stable isotope labeling approach for quantitative proteomics

    PubMed Central

    Zhai, Jianjun; Liu, Xiaoyan; Huang, Zhenyu; Zhu, Haining

    2009-01-01

    Quantitative proteomics is challenging and various stable isotope based approaches have been developed to meet the challenge.. Hereby we describe a simple, efficient, reliable and inexpensive method named RABA (reductive alkylation by acetone) to introduce stable isotopes to peptides for quantitative analysis. The RABA method leads to alkylation of N-terminal and lysine amino groups with isopropyl moiety. Using unlabeled (d0) and deuterium labeled (d6) acetone, a 6 Da mass split is introduced to each isopropyl modification between the light and heavy isotope labeled peptides, which is ideally suited for quantitative analysis. The reaction specificity, stoichoimetry, labeling efficiency and linear range of the RABA method has been thoroughly evaluated in this study using standard peptides, tryptic digest of proteins as well as human cell lysate. Reliable quantitative results have been consistently obtained in all experiments. We also applied the RABA method to quantitative analysis of proteins in spinal cords of transgenic mouse models of amyotrophic lateral sclerosis. Highly homologous proteins (transgenic human SOD1 and endogenous mouse SOD1) were distinguished and quantified using the method developed in this study. In addition, the quantitative results using the RABA approach were independently validated by Western blot. PMID:19419886

  20. Use of isotope-labeled aflatoxins for LC-MS/MS stable isotope dilution analysis of foods.

    PubMed

    Cervino, Christian; Asam, Stefan; Knopp, Dietmar; Rychlik, Michael; Niessner, Reinhard

    2008-03-26

    Aflatoxins are a group of very carcinogenic mycotoxins that can be found on a wide range of food commodities including nuts, cereals, and spices. In this study, the first LC-MS/MS stable isotope dilution assay (SIDA) for the determination of aflatoxins in foods was developed. The development of this method was enabled by easily accessible isotope-labeled (deuterated) aflatoxins B2 and G2, which were synthesized by catalytic deuteration of aflatoxin B1 and G1, purified, and well-characterized by NMR and MS. All four aflatoxins of interest (B1, B2, G1, and G2) were quantified in food samples by using these two labeled internal standards. The response factors (RF) of the linear calibrations were revealed to be matrix independent for labeled aflatoxin B2/aflatoxin B2 and labeled aflatoxin G2/aflatoxin G2. For labeled aflatoxin B 2/aflatoxin B 1 and labeled aflatoxin B2/aflatoxin G1 matrix-matched calibration was performed for the model matrices almonds and wheat flour, showing significant differences of the RFs. Limits of detection (LOD) were determined by applying a statistical approach in the presence of the two model matrices, yielding 0.31 microg/kg (aflatoxin B1), 0.09 microg/kg (aflatoxin B2), 0.38 microg/kg (aflatoxin G1), and 0.32 microg/kg (aflatoxin G2) for almonds (similar LODs were obtained for wheat flour). Recovery rates were between 90 and 105% for all analytes. Coefficients of variation (CV) of 12% (aflatoxin B1), 3.6% (aflatoxin B2), 14% (aflatoxin G1), and 4.8% (aflatoxin G2) were obtained from interassay studies. For further validation, a NIST standard reference food sample was analyzed for aflatoxins B1 and B2. The method was successfully applied to determine trace levels of aflatoxins in diverse food matrices such as peanuts, nuts, grains, and spices. Aflatoxin contents in these samples ranged from about 0.5 to 6 microg/kg.

  1. Late-Glacial to Late-holocene Shifts in Global Precipitation Delta(sup 18)O

    NASA Technical Reports Server (NTRS)

    Jasechko, S.; Lechler, A.; Pausata, F.S.R.; Fawcett, P.J.; Gleeson, T.; Cendon, D.I.; Galewsky, J.; LeGrande, A. N.; Risi, C.; Sharp, Z. D.; Welker, J. M.; Werner, M.; Yoshimura, K.

    2015-01-01

    Reconstructions of Quaternary climate are often based on the isotopic content of paleo-precipitation preserved in proxy records. While many paleo-precipitation isotope records are available, few studies have synthesized these dispersed records to explore spatial patterns of late-glacial precipitation delta(sup 18)O. Here we present a synthesis of 86 globally distributed groundwater (n 59), cave calcite (n 15) and ice core (n 12) isotope records spanning the late-glacial (defined as 50,000 to 20,000 years ago) to the late-Holocene (within the past 5000 years). We show that precipitation delta(sup 18)O changes from the late-glacial to the late-Holocene range from -7.1% (delta(sup 18)O(late-Holocene) > delta(sup 18)O(late-glacial) to +1.7% (delta(sup 18)O(late-glacial) > delta(sup 18)O(late-Holocene), with the majority (77) of records having lower late-glacial delta(sup 18)O than late-Holocene delta(sup 18)O values. High-magnitude, negative precipitation delta(sup 18)O shifts are common at high latitudes, high altitudes and continental interiors.

  2. UNiquant, a Program for Quantitative Proteomics Analysis Using Stable Isotope Labeling

    PubMed Central

    Huang, Xin; Tolmachev, Aleksey V.; Shen, Yulei; Liu, Miao; Huang, Lin; Zhang, Zhixin; Anderson, Gordon A.; Smith, Richard D.; Chan, Wing C.; Hinrichs, Steven H.; Fu, Kai; Ding, Shi-Jian

    2011-01-01

    Stable isotope labeling (SIL) methods coupled with nanoscale liquid chromatography and high resolution tandem mass spectrometry are increasingly useful for elucidation of the proteome-wide differences between multiple biological samples. Development of more effective programs for the sensitive identification of peptide pairs and accurate measurement of the relative peptide/protein abundance are essential for quantitative proteomic analysis. We developed and evaluated the performance of a new program, termed UNiquant, for analyzing quantitative proteomics data using stable isotope labeling. UNiquant was compared with two other programs, MaxQuant and Mascot Distiller, using SILAC-labeled complex proteome mixtures having either known or unknown heavy/light ratios. For the SILAC-labeled Jeko-1 cell proteome digests with known heavy/light ratios (H/L = 1:1, 1:5, and 1:10), UNiquant quantified a similar number of peptide pairs as MaxQuant for the H/L = 1:1 and 1:5 mixtures. In addition, UNiquant quantified significantly more peptides than MaxQuant and Mascot Distiller in the H/L = 1:10 mixtures. UNiquant accurately measured relative peptide/protein abundance without the need for post-measurement normalization of peptide ratios, which is required by the other programs. PMID:21158445

  3. Efficient segmental isotope labeling of multi-domain proteins using Sortase A.

    PubMed

    Freiburger, Lee; Sonntag, Miriam; Hennig, Janosch; Li, Jian; Zou, Peijian; Sattler, Michael

    2015-09-01

    NMR studies of multi-domain protein complexes provide unique insight into their molecular interactions and dynamics in solution. For large proteins domain-selective isotope labeling is desired to reduce signal overlap, but available methods require extensive optimization and often give poor ligation yields. We present an optimized strategy for segmental labeling of multi-domain proteins using the S. aureus transpeptidase Sortase A. Critical improvements compared to existing protocols are (1) the efficient removal of cleaved peptide fragments by centrifugal filtration and (2) a strategic design of cleavable and non-cleavable affinity tags for purification. Our approach enables routine production of milligram amounts of purified segmentally labeled protein for NMR and other biophysical studies.

  4. [Ligands of glutamate and dopamine receptors evenly labeled with hydrogen isotopes].

    PubMed

    Zolotarev, Iu A; Firstova, Iu Iu; Abaimov, D A; Dadaian, A K; Kosik, V S; Novikov, A V; Krasnov, N V; Vas'kovskiĭ, B V; Nazimov, I V; Kovalev, G I; Miasoedov, N F

    2009-01-01

    A reaction of high-temperature solid-phase catalytic isotope exchange (HSCIE) was studied for the preparation of tritium- and deuterium-labeled ligands of glutamate and dopamine receptors. Tritium-labeled (5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclopenten-5,10-imine ([G-(3)H]MK-801) and R(+)-7-hydroxy-N,N-di-n-propyl-2-aminotetraline ([G-(3)H]-7-OH-DPAT) were obtained with a specific activity of 210 and 120 Ci/mol, respectively. The isotopomeric distribution of deuterium-labeled ligands was studied using time-of-flight mass-spectrometer MX 5310 (ESI-o-TOF) with electrospray and orthogonal ion injection. Mean deuterium incorporation per ligand molecule was 11.09 and 3.21 atoms for [G-(2)H]MK-801 and [G-(2)H]-7-OH-DPAT, respectively. The isotope label was shown to be distributed all over the ligand molecule. The radioreceptor binding of tritium-labeled ligands [G-(3)H]MK-801 and [G-(3)H]-7-OH-DPAT was analyzed using the brain structure of Vistar rats. It was demonstrated that [G-(3)H]MK-801 specifically binds to hippocampus membranes with K(d) 8.3 +/- 1.4 nM, B(max) being 3345 +/- 300 fmol/mg protein. The [G-(3)H]-7-OH-DPAT ligand specifically binds to rat striatum membranes with K(d) 10.01 +/- 0.91 nM and B(max) 125 +/- 4.5 fmol/mg protein. It was concluded that the HSCIE reaction can be used for the preparation of highly tritium-labeled (+)-MK-801 and 7-OH-DPAT with retention of their physiological activities.

  5. A 240 ka terrestrial 18O record from a NE-Siberian loess-like permafrost paleosol-sequence based on a novel analytical 18O method

    NASA Astrophysics Data System (ADS)

    Tuthorn, M.; Zech, M.; Detsch, F.; Juchelka, D.; Kalbitz, K.; Mayr, C.; Werner, R.; Zech, R.; Zech, W.; Glaser, B.

    2012-04-01

    Recently, we developed a novel analytical tool for paleoclimate research based on compound-specific delta18O analyses of hemicellulose-derived monosaccharides using gas chromatography-pyrolysis-isotope ratio mass spectrometry (GC-Py-IRMS) (Zech and Glaser, 2009. Rapid Communications in Mass Spectrometry 23, 3522-3532). This method overcomes extraction, purification and hygroscopicity problems of so far applied cellulose methods based on TC/EA-IRMS delta18O analyses and allows establishing 18O records from sedimentary organic matter. Taking advantage of plant samples from a climate chamber experiment we can demonstrate that our novel method yields similar results like cellulose for plant material. Furthermore, we demonstrate using 18O-enriched water that the hydroxyl-groups of hemicelluloses are not prone to oxygen exchange reactions (Zech et al., 2012. Organic Geochemistry 42, 1470-1475). Ongoing methodological improvements will be shortly reported. By applying our novel 18O method to a loess-like permafrost paleosol-sequence we established a presumably 240 ka terrestrial 18O record for NE-Siberia. While the modern topsoil and the interglacial/-stadial paleosols reveal more positive delta18O values, the glacial paleosols reveal more negative delta18O values. The 18O variability is generally confirmed by a respective deltaD record which is based on sedimentary plant leafwax-derived n-alkanes. This finding suggests that our high-latitude 240 ka terrestrial 18O and D/H record from NE-Siberia reflects the temperature-dependent isotopic composition of precipitation and the increased isotopic enrichment of leaf-water during interglacials/-stadials.

  6. Recombinant isotope labeled and selenium quantified proteins for absolute protein quantification.

    PubMed

    Zinn, Nico; Winter, Dominic; Lehmann, Wolf D

    2010-03-15

    A novel, widely applicable method for the production of absolutely quantified proteins is described, which can be used as internal standards for quantitative proteomic studies based on mass spectrometry. These standards are recombinant proteins containing an isotope label and selenomethionine. For recombinant protein expression, assembly of expression vectors fitted to cell-free protein synthesis was conducted using the gateway technology which offers fast access to a variety of genes via open reading frame libraries and an easy shuttling of genes between vectors. The proteins are generated by cell-free expression in a medium in which methionine is exchanged against selenomethionine and at least one amino acid is exchanged by a highly stable isotope labeled analogue. After protein synthesis and purification, selenium is used for absolute quantification by element mass spectrometry, while the heavy amino acids in the protein serve as reference in subsequent analyses by LC-ESI-MS or MALDI-MS. Accordingly, these standards are denominated RISQ (for recombinant isotope labeled and selenium quantified) proteins. In this study, a protein was generated containing Lys+6 ([(13)C(6)]-lysine) and Arg+10 ([(13)C(6),(15)N(4)]-arginine) so that each standard tryptic peptide contains a labeled amino acid. Apolipoprotein A1 was synthesized as RISQ protein, and its use as internal standard led to quantification of a reference material within the specified value. Owing to their cell-free expression, RISQ proteins do not contain posttranslational modifications. Thus, correct quantitative data by ESI- or MALDI-MS are restricted to quantifications based on peptides derived from unmodified regions of the analyte protein. Therefore, besides serving as internal standards, RISQ proteins stand out as new tools for quantitative analysis of covalent protein modifications.

  7. Tracing bioavailability of ZnO nanoparticles using stable isotope labeling.

    PubMed

    Larner, Fiona; Dogra, Yuktee; Dybowska, Agnieszka; Fabrega, Julia; Stolpe, Björn; Bridgestock, Luke J; Goodhead, Rhys; Weiss, Dominik J; Moger, Julian; Lead, Jamie R; Valsami-Jones, Eugenia; Tyler, Charles R; Galloway, Tamara S; Rehkämper, Mark

    2012-11-06

    Zinc oxide nanoparticles (ZnO NPs) are widely used in commercial products and knowledge of their environmental fate is a priority for ecological protection. Here we synthesized model ZnO NPs that were made from and thus labeled with the stable isotope (68)Zn and this enables highly sensitive and selective detection of labeled components against high natural Zn background levels. We combine high precision stable isotope measurements and novel bioimaging techniques to characterize parallel water-borne exposures of the common mudshrimp Corophium volutator to (68)ZnO NPs, bulk (68)ZnO, and soluble (68)ZnCl(2) in the presence of sediment. C. volutator is an important component of coastal ecosystems where river-borne NPs will accumulate and is used on a routine basis for toxicity assessments. Our results demonstrate that ionic Zn from ZnO NPs is bioavailable to C. volutator and that Zn uptake is active. Bioavailability appears to be governed primarily by the dissolved Zn content of the water, whereby Zn uptake occurs via the aqueous phase and/or the ingestion of sediment particles with adsorbed Zn from dissolution of ZnO particles. The high sorption capacity of sediments for Zn thus enhances the potential for trophic transfer of Zn derived from readily soluble ZnO NPs. The uncertainties of our isotopic data are too large, however, to conclusively rule out any additional direct uptake route of ZnO NPs by C. volutator.

  8. Chemical imaging of biological materials by NanoSIMS using isotopic and elemental labels

    SciTech Connect

    Weber, P K; Fallon, S J; Pett-Ridge, J; Ghosal, S; Hutcheon, I D

    2006-04-10

    The NanoSIMS 50 combines unprecedented spatial resolution (as good as 50 nm) with ultra-high sensitivity (minimum detection limit of {approx}200 atoms). The NanoSIMS 50 incorporates an array of detectors, enabling simultaneous collection of 5 species originating from the same sputtered volume of a sample. The primary ion beam (Cs{sup +} or O{sup -}) can be scanned across the sample to produce quantitative secondary ion images. This capability for multiple isotope imaging with high spatial resolution provides a novel new approach to the study of biological materials. Studies can be made of sub-regions of tissues, mammalian cells, and bacteria. Major, minor and trace element distributions can be mapped on a submicron scale, growth and metabolism can be tracked using stable isotope labels, and biogenic origin can be determined based on composition. We have applied this technique extensively to mammalian and prokaryotic cells and bacterial spores. The NanoSIMS technology enables the researcher to interrogate the fate of molecules of interest within cells and organs through elemental and isotopic labeling. Biological applications at LLNL will be discussed.

  9. Transformation of ENSO-related rainwater to dripwater δ18O variability by vadose water mixing

    NASA Astrophysics Data System (ADS)

    Moerman, Jessica W.; Cobb, Kim M.; Partin, Judson W.; Meckler, A. Nele; Carolin, Stacy A.; Adkins, Jess F.; Lejau, Syria; Malang, Jenny; Clark, Brian; Tuen, Andrew A.

    2014-11-01

    Speleothem oxygen isotopes18O) are often used to reconstruct past rainfall δ18O variability, and thereby hydroclimate changes, in many regions of the world. However, poor constraints on the karst hydrological processes that transform rainfall signals into cave dripwater add significant uncertainty to interpretations of speleothem-based reconstructions. Here we present several 6.5 year, biweekly dripwater δ18O time series from northern Borneo and compare them to local rainfall δ18O variability. We demonstrate that vadose water mixing is the primary rainfall-to-dripwater transformation process at our site, where dripwater δ18O reflects amount-weighted rainfall δ18O integrated over the previous 3-10 months. We document large interannual dripwater δ18O variability related to the El Niño-Southern Oscillation (ENSO), with amplitudes inversely correlated to dripwater residence times. According to a simple stalagmite forward model, asymmetrical ENSO extremes produce significant offsets in stalagmite δ18O time series given different dripwater residence times. Our study highlights the utility of generating multiyear, paired time series of rainfall and dripwater δ18O to aid interpretations of stalagmite δ18O reconstructions.

  10. Safety-catch linker strategies for the production of radiopharmaceuticals labeled with positron-emitting isotopes.

    PubMed

    Maclean, Derek; Zhu, Jiang; Chen, Mingying; Hale, Ron; Satymurthy, Nagichettiar; Barrio, Jorge R

    2003-08-27

    A novel synthetic stratetegy for compounds labeled with the positron-emitting isotope carbon-11 is described. The use of precursors attached to a solid support via safety-catch linkers allows selective release of radiolabeled material, leaving unreacted precursor attached to the support. Two different linkers demonstrate the application to the preparation of radiolabeled N-alkyl tertiary amines and N-alkylsulfonamides. This technique is expected to lead to more widespread use of positron emission tomography for the in vivo analysis of compound behavior.

  11. Addressing Raman features of individual layers in isotopically labeled Bernal stacked bilayer graphene

    NASA Astrophysics Data System (ADS)

    Costa, Sara D.; Weis, Johan Ek; Frank, Otakar; Fridrichová, Michaela; Kalbac, Martin

    2016-06-01

    In this report important Raman modes for the evaluation of strain in graphene (the 2D and 2D‧) are analyzed. The isotope labeling is used to disentangle contribution of individual graphene layers of graphene bilayer to the studied Raman modes. It is shown that for Bernal-stacked bilayers, the 2D and the 2D‧ Raman modes have three distinct components that can be assigned to processes originating solely from the top graphene layer, bottom graphene layer, and from a combination of processes originating both from the top and bottom layers. The reported results thus enable addressing the properties of individual graphene layers in graphene bilayer by Raman spectroscopy.

  12. Metabolomics relative quantitation with mass spectrometry using chemical derivatization and isotope labeling

    DOE PAGES

    O'Maille, Grace; Go, Eden P.; Hoang, Linh; ...

    2008-01-01

    Comprehensive detection and quantitation of metabolites from a biological source constitute the major challenges of current metabolomics research. Two chemical derivatization methodologies, butylation and amination, were applied to human serum for ionization enhancement of a broad spectrum of metabolite classes, including steroids and amino acids. LC-ESI-MS analysis of the derivatized serum samples provided a significant signal elevation across the total ion chromatogram to over a 100-fold increase in ionization efficiency. It was also demonstrated that derivatization combined with isotopically labeled reagents facilitated the relative quantitation of derivatized metabolites from individual as well as pooled samples.

  13. 18O-Tracer Metabolomics Reveals Protein Turnover and CDP-Choline Cycle Activity in Differentiating 3T3-L1 Pre-Adipocytes

    PubMed Central

    Kirkwood, Jay S.; Miranda, Cristobal L.; Bobe, Gerd; Maier, Claudia S.; Stevens, Jan F.

    2016-01-01

    The differentiation of precursor cells into mature adipocytes (adipogenesis) has been an area of increased focus, spurred by a rise in obesity rates. Though our understanding of adipogenesis and its regulation at the cellular level is growing, many questions remain, especially regarding the regulation of the metabolome. The 3T3-L1 cell line is the most well characterized cellular model of adipogenesis. Using a time course metabolomics approach, we show that the 3T3-L1 preadipocyte metabolome is greatly altered during the first 48 hours of differentiation, where cells go through about two rounds of cell division, a process known as mitotic clonal expansion. Short-chain peptides were among several small molecules that were increased during mitotic clonal expansion. Additional indicators of protein turnover were also increased, including bilirubin, a degradation product of heme-containing proteins, and 3-methylhistidine, a post-translationally modified amino acid that is not reutilized for protein synthesis. To study the origin of the peptides, we treated differentiating preadipocytes with 18O labeled water and found that 18O was incorporated into the short chain peptides, confirming them, at least in part, as products of hydrolysis. Inhibitors of the proteasome or matrix metalloproteinases affected the peptide levels during differentiation, but inhibitors of autophagy or peptidases did not. 18O was also incorporated into several choline metabolites including cytidine 5'-diphosphocholine (CDP-choline), glycerophosphocholine, and several phosphatidylcholine species, indicative of phosphatidylcholine synthesis/degradation and of flux through the CDP-choline cycle, a hallmark of proliferating cells. 18O-Tracer metabolomics further showed metabolic labeling of glutamate, suggestive of glutaminolysis, also characteristic of proliferating cells. Together, these results highlight the utility of 18O isotope labeling in combination with metabolomics to uncover changes in

  14. Designer labels for plant metabolism: statistical design of isotope labeling experiments for improved quantification of flux in complex plant metabolic networks.

    PubMed

    Nargund, Shilpa; Sriram, Ganesh

    2013-01-27

    Metabolic fluxes are powerful indicators of cell physiology and can be estimated by isotope-assisted metabolic flux analysis (MFA). The complexity of the compartmented metabolic networks of plants has constrained the application of isotope-assisted MFA to them, principally because of poor identifiability of fluxes from the measured isotope labeling patterns. However, flux identifiability can be significantly improved by a priori design of isotope labeling experiments (ILEs). This computational design involves evaluating the effect of different isotope label and isotopomer measurement combinations on flux identifiability, and thereby identifying optimal labels and measurements toward evaluating the fluxes of interest with the highest confidence. This article reports ILE designs for two major, compartmented plant metabolic pathways - the pentose phosphate pathway (PPP) and γ-aminobutyric acid (GABA) shunt. Together, these pathways represent common motifs in plant metabolism including duplication of pathways in different subcellular compartments, reversible reactions and cyclic carbon flow. To compare various ILE designs, we employed statistical A- and D-optimality criteria. Our computations showed that 1,2-(13)C Glc is a powerful and robust label for the plant PPPs, given currently popular isotopomer measurement techniques (single quadrupole mass spectrometry [MS] and 2-D nuclear magnetic resonance [NMR]). Further analysis revealed that this label can estimate several PPP fluxes better than the popular label 1-(13)C Glc. Furthermore, the concurrent measurement of the isotopomers of hexose and pentose moieties synthesized exclusively in the cytosol or the plastid compartments (measurable through intracellular glucose or sucrose, starch, RNA ribose and histidine) considerably improves the identifiability of PPP fluxes in the individual compartments. Additionally, MS-derived isotopomer measurements outperform NMR-derived measurements in identifying PPP fluxes. The

  15. Comparisons of observed and modelled lake δ18O variability

    NASA Astrophysics Data System (ADS)

    Jones, Matthew D.; Cuthbert, Mark O.; Leng, Melanie J.; McGowan, Suzanne; Mariethoz, Gregoire; Arrowsmith, Carol; Sloane, Hilary J.; Humphrey, Kerenza K.; Cross, Iain

    2016-01-01

    With the substantial number of lake sediment δ18O records published in recent decades, a quantitative, process-based understanding of these systems can increase our understanding of past climate change. We test mass balance models of lake water δ18O variability against five years of monthly monitoring data from lakes with different hydrological characteristics, in the East-Midlands region of the UK, and the local isotope composition of precipitation. These mass balance models can explain up to 74% of the measured lake water isotope variability. We investigate the sensitivity of the model to differing calculations of evaporation amount, the amount of groundwater, and to different climatic variables. We show there is only a small range of values for groundwater exchange flux that can produce suitable lake water isotope compositions and that variations in evaporation and precipitation are both required to produce recorded isotope variability in lakes with substantial evaporative water losses. We then discuss the potential for this model to be used in a long-term, palaeo-scenario. This study demonstrates how long term monitoring of a lake system can lead to the development of robust models of lake water isotope compositions. Such systematics-based explanations allow us to move from conceptual, to more quantified reconstructions of past climates and environments.

  16. Using (18)O/(16)O exchange to probe an equilibrium space-charge layer at the surface of a crystalline oxide: method and application.

    PubMed

    De Souza, Roger A; Martin, Manfred

    2008-05-07

    The use of an (18)O/(16)O exchange experiment as a means for probing surface space-charge layers in oxides is examined theoretically and experimentally. On the basis of a theoretical treatment, isotope penetration profiles are calculated for (18)O/(16)O exchange across a gas-solid interface and subsequent diffusion of the labelled isotope through an equilibrium space-charge layer depleted of mobile oxygen vacancies and into a homogeneous bulk phase. Profiles calculated for a range of conditions all have a characteristic shape: a sharp drop in isotope fraction close to the surface followed by a normal bulk diffusion profile. Experimental (18)O profiles in an exchanged (001) oriented single crystal of Fe-doped SrTiO(3) were measured by time-of-flight secondary ion mass spectrometry (ToF-SIMS). By extracting the space-charge potential from such profiles, we demonstrate that this method allows the spatially resolved characterization of space-charge layers at the surfaces of crystalline oxides under thermodynamically well-defined conditions.

  17. Multiplexed Analysis of Cage and Cage Free Chicken Egg Fatty Acids Using Stable Isotope Labeling and Mass Spectrometry

    PubMed Central

    Torde, Richard G.; Therrien, Andrew J.; Shortreed, Michael R.; Smith, Lloyd M.; Lamos, Shane M.

    2014-01-01

    Binary stable isotope labeling couple with LC-ESI-MS has been used as a powerful non-targeted approach for the relative quantification of lipids, amino acids, and many other important metabolite classes. A multiplexed approach using three or more isotopic labeling reagents greatly reduces analytical run-time while maintaining excellent sensitivity and reproducibility. Three isotopic cholamine labeling reagents have been developed to take advantage of the pre-ionized character of cholamine, for ESI, and the ease by which stable isotopes can be incorporated into the cholamine structure. These three cholamine labeling reagents have been used to relatively quantify three fatty acid samples simultaneously. The quantification resulted in the observation of 12 fatty acids that had an average absolute error of 0.9% and an average coefficient of variation of 6.1%. Caged versus cage-free isotope labeling experiments showed that cage-free eggs have an increased level of omega-3 fatty acids as compared to caged eggs. This multiplexed fatty acid analysis provides an inexpensive and expedited tool for broad-based lipid profiling that will further aid discoveries in the mechanisms of fatty acid action in cells. PMID:24317525

  18. 18O/16O in CO2 evolved from goethite during some unusually rapid solid state α-FeOOH to α-Fe2O3 phase transitions: Test of an exchange model for possible use in oxygen isotope analyses of goethite

    NASA Astrophysics Data System (ADS)

    Yapp, Crayton J.

    2015-12-01

    The initial ∼60% of an isothermal vacuum dehydration of goethite can commonly be approximated by first order kinetics. Also, natural goethites contain small amounts of an Fe(CO3)OH component in apparent solid solution. The 18O/16O of CO2 evolved from the Fe(CO3)OH during isothermal vacuum dehydrations is related to the 18O/16O of the goethite by an apparent fractionation factor (αapp) that is, in turn, correlated with a first order rate constant, |m|. A kinetic exchange model predicts that αapp should decrease as |m| increases for a range of |m| that corresponds to relatively slow rates of dehydration. This pattern has been observed in published results. In contrast, for rapid rates of dehydration, αapp is predicted to increase with increasing |m|. Isothermal vacuum dehydrations of two natural goethites had unusually large values of |m| and provided serendipitous tests of this rapid-rate prediction. For these experiments, the measured values of αapp were consistent with patterns of variation predicted by the model. This allowed an estimate of the activation energy (E2) of a model parameter, K2, which is the rate constant for oxygen isotope exchange between CO2 and H2O during the solid-state goethite to hematite phase transition. The estimated value of E2 is only ∼9 kJ/mol. Heterogeneous catalysis tends to decrease the activation energies of gas reactions. Consequently, the inferred value of E2 suggests that goethite and/or hematite catalyze oxygen isotope exchange between CO2 and H2O during the solid-state phase change. Yield, δ13C, and δ18O values are routinely measured for increments of CO2 evolved from the Fe(CO3)OH component during isothermal vacuum dehydration of goethite. Model-predicted values of αapp can be combined with plateau δ18O values of the evolved CO2 to estimate the δ18O of the goethite with a less than optimal, but potentially useful, precision of about ±0.8‰. Therefore, a single analytical procedure (incremental dehydration

  19. Variable sea ice contributions to seawater δ18O on glacial-interglacial timescales

    NASA Astrophysics Data System (ADS)

    Brennan, C. E.; Weaver, A. J.; Eby, M.; Meissner, K. J.

    2011-12-01

    The oxygen isotope composition of seawater varies in time, mainly based on the amount of (depleted) ice stored on continents. Oxygen isotope records derived from ocean sediment cores serve as indicators of changes in both seawater temperature and continental ice volume. Seawater δ18O may contain a variable signature of sea ice production, especially at high latitudes. Sea ice growth produces isotopically enriched ice and depleted brine. Over glacial-interglacial cycles, changes in the sites and rates of sea ice production (and by extension sea ice meltwater and brine export) hold the potential to shift local to regional seawater isotopic chemistry. Neglecting variability in sea ice production may therefore superimpose error upon reconstructions employing high latitude δ18O records. We examine the effects of variability in sea ice production between glacial and interglacial climate states on seawater δ18O in the University of Victoria Earth System Climate Model. Oxygen isotopes are implemented in all components (ocean, atmosphere, land surface, and sea ice) of the coupled model. The role of glacial-interglacial sea ice variability is investigated in a set of model experiments. Here we isolate the seawater δ18O field due only to sea ice in the model. By contrasting the seawater δ18O fields due to sea ice resulting from the glacial and interglacial climates, we investigate the potential for variable sea ice formation to shift seawater δ18O.

  20. Parallel β-sheet vibrational couplings revealed by 2D IR spectroscopy of an isotopically labeled macrocycle: quantitative benchmark for the interpretation of amyloid and protein infrared spectra.

    PubMed

    Woys, Ann Marie; Almeida, Aaron M; Wang, Lu; Chiu, Chi-Cheng; McGovern, Michael; de Pablo, Juan J; Skinner, James L; Gellman, Samuel H; Zanni, Martin T

    2012-11-21

    Infrared spectroscopy is playing an important role in the elucidation of amyloid fiber formation, but the coupling models that link spectra to structure are not well tested for parallel β-sheets. Using a synthetic macrocycle that enforces a two stranded parallel β-sheet conformation, we measured the lifetimes and frequency for six combinations of doubly (13)C═(18)O labeled amide I modes using 2D IR spectroscopy. The average vibrational lifetime of the isotope labeled residues was 550 fs. The frequencies of the labels ranged from 1585 to 1595 cm(-1), with the largest frequency shift occurring for in-register amino acids. The 2D IR spectra of the coupled isotope labels were calculated from molecular dynamics simulations of a series of macrocycle structures generated from replica exchange dynamics to fully sample the conformational distribution. The models used to simulate the spectra include through-space coupling, through-bond coupling, and local frequency shifts caused by environment electrostatics and hydrogen bonding. The calculated spectra predict the line widths and frequencies nearly quantitatively. Historically, the characteristic features of β-sheet infrared spectra have been attributed to through-space couplings such as transition dipole coupling. We find that frequency shifts of the local carbonyl groups due to nearest neighbor couplings and environmental factors are more important, while the through-space couplings dictate the spectral intensities. As a result, the characteristic absorption spectra empirically used for decades to assign parallel β-sheet secondary structure arises because of a redistribution of oscillator strength, but the through-space couplings do not themselves dramatically alter the frequency distribution of eigenstates much more than already exists in random coil structures. Moreover, solvent exposed residues have amide I bands with >20 cm(-1) line width. Narrower line widths indicate that the amide I backbone is solvent

  1. Carbon allocation belowground in Pinus pinaster using stable carbon isotope pulse labeling technique

    NASA Astrophysics Data System (ADS)

    Dannoura, M.; Bosc, A.; Chipeaux, C.; Sartore, M.; Lambrot, C.; Trichet, P.; Bakker, M.; Loustau, D.; Epron, D.

    2010-12-01

    Carbon allocation belowground competes with aboveground growth and biomass production. In the other hand, it contributes to resource acquisition such as nutrient, water and carbon sequestration in soil. Thus, a better characterization of carbon flow from plant to soil and its residence time within each compartment is an important issue for understanding and modeling forest ecosystem carbon budget. 13C pulse labeling of whole crown was conducted at 4 seasons to study the fate of assimilated carbon by photosynthesis into the root on 12 year old Pinus pinaster planted in the INRA domain of Pierroton. Maritime pine is the most widely planted species in South-West Europe. Stem, root and soil CO2 effluxes and their isotope composition were measured continuously by tunable diode laser absorption spectroscopy with a trace gas analyzer (TGA 100A; Campbell Scientific) coupled to flow-through chambers. 13CO2 recovery and peak were observed in respiration of each compartment after labeling. It appeared sequentially from top of stem to bottom, and to coarse root. The maximum velocity of carbon transfer was calculated as the difference in time lag of recovery between two positions on the trunk or on the root. It ranged between 0.08-0.2 m h-1 in stem and between 0.04-0.12 m h-1 in coarse root. This velocity was higher in warmer season, and the difference between time lag of recovery and peak increased after first frost. Photosynthates arrived underground 1.5 to 5 days after labeling, at similar time in soil CO2 effluxes and coarse root respiration. 0.08-1.4 g of carbon was respired per tree during first 20 days following labeling. It presented 0.6 -10% of 13C used for labeling and it is strongly related to seasons. The isotope signal was detected in fine root organs and microbial biomass by periodical core sampling. The peak was observed 6 days after labeling in early summer while it was delayed more than 10 days in autumn and winter with less amount of carbon allocated

  2. Positional isotope exchange studies on enzyme using NMR spectroscopy

    SciTech Connect

    Matsunaga, T.O.

    1987-01-01

    The isotopically enriched compounds, /sup 18/O-..beta..,..gamma..-ATP and /sup 18/O bridge-labeled pyrophosphate, synthesized previously in this laboratory, were used to investigate and measure the exchange vs. turnover of substrates and products from their central complexes in four selected enzyme systems. Using hi-field /sup 31/P NMR, we were able to differentiate between /sup 18/O labeled in the bridge vs. the non-bridge positions by virtue of the isotope shift upon the phosphorus nuclei. The bridge to non-bridge scrambling of the label was quantitated and the exchange vs. turnover ratios under a variety of conditions was determined. Using the substrate inhibitor carboxycreatinine, PIX experiments with /sup 18/O-..beta..,..gamma..-ATP and creatine kinase were conducted. It was shown that carboxycreatinine and creatine kinase promoted exchange of the /sup 18/O label as determined by NMR. We have concluded that carboxycreatinine is either a substrate that catalyzes very slow turnover or it catalyzes exchange by a dissociative (SN/sub 1//sub P/) type of mechanism

  3. Multisubstrate Isotope Labeling and Metagenomic Analysis of Active Soil Bacterial Communities

    PubMed Central

    Verastegui, Y.; Cheng, J.; Engel, K.; Kolczynski, D.; Mortimer, S.; Lavigne, J.; Montalibet, J.; Romantsov, T.; Hall, M.; McConkey, B. J.; Rose, D. R.; Tomashek, J. J.; Scott, B. R.

    2014-01-01

    ABSTRACT Soil microbial diversity represents the largest global reservoir of novel microorganisms and enzymes. In this study, we coupled functional metagenomics and DNA stable-isotope probing (DNA-SIP) using multiple plant-derived carbon substrates and diverse soils to characterize active soil bacterial communities and their glycoside hydrolase genes, which have value for industrial applications. We incubated samples from three disparate Canadian soils (tundra, temperate rainforest, and agricultural) with five native carbon (12C) or stable-isotope-labeled (13C) carbohydrates (glucose, cellobiose, xylose, arabinose, and cellulose). Indicator species analysis revealed high specificity and fidelity for many uncultured and unclassified bacterial taxa in the heavy DNA for all soils and substrates. Among characterized taxa, Actinomycetales (Salinibacterium), Rhizobiales (Devosia), Rhodospirillales (Telmatospirillum), and Caulobacterales (Phenylobacterium and Asticcacaulis) were bacterial indicator species for the heavy substrates and soils tested. Both Actinomycetales and Caulobacterales (Phenylobacterium) were associated with metabolism of cellulose, and Alphaproteobacteria were associated with the metabolism of arabinose; members of the order Rhizobiales were strongly associated with the metabolism of xylose. Annotated metagenomic data suggested diverse glycoside hydrolase gene representation within the pooled heavy DNA. By screening 2,876 cloned fragments derived from the 13C-labeled DNA isolated from soils incubated with cellulose, we demonstrate the power of combining DNA-SIP, multiple-displacement amplification (MDA), and functional metagenomics by efficiently isolating multiple clones with activity on carboxymethyl cellulose and fluorogenic proxy substrates for carbohydrate-active enzymes. PMID:25028422

  4. The CW-CRDS spectra of the 16O/18O isotopologues of ozone between 5930 and 6340 cm-1—Part 3: 16O18O18O and 18O16O18O

    NASA Astrophysics Data System (ADS)

    De Backer, M.-R.; Barbe, A.; Starikova, E.; Tyuterev, Vl. G.; Mondelain, D.; Kassi, S.; Campargue, A.

    2013-09-01

    Our systematic investigation of the high sensitivity CW-Cavity Ring Down Spectra of 16O/18O ozone isotopologues at high vibrational excitation continues with the study of the 16O18O18O and 18O16O18O species. The first two papers of this series were devoted to the analysis of the same four bands of the 16O16O18O and 16O18O16O species in the 5930-6340 cm-1domain. Here, after a brief reminder of relevant experiment and theory, we report the analyses of two bands of 16O18O18O, vibrationally assigned as 2ν2+5ν3 and 2ν1+2ν2+3ν3_2 and three bands of 18O16O18O, assigned to 2ν1+2ν2+3ν3_1, 5ν1+ν3 and 3ν1+ν2+3ν3.They correspond to the highest vibration excitations observed so far for the 16O18O18O and 18O16O18O isotopologues. Altogether for the two new bands of 16O18O18O, 1214 rovibrational transitions were assigned up to Jmax=29 and for the three new bands of 18O16O18O, 948 rovibrational transitions were assigned up to Jmax=27. Observed line positions were fitted with root-mean squares deviations ranging from 0.005 to 0.011 cm-1, using effective Hamiltonian models accounting for dark state perturbations. The derived band centres and rotational constants are in good agreement with new theoretical calculations from the molecular potential function. The corresponding lists of 3365 lines are provided as Supplementary material.

  5. A 'hidden' 18O-enriched reservoir in the sub-arc mantle.

    PubMed

    Liu, Chuan-Zhou; Wu, Fu-Yuan; Chung, Sun-Lin; Li, Qiu-Li; Sun, Wei-Dong; Ji, Wei-Qiang

    2014-02-28

    Plate subduction continuously transports crustal materials with high-δ(18)O values down to the mantle wedge, where mantle peridotites are expected to achieve the high-δ(18)O features. Elevated δ(18)O values relative to the upper mantle value have been reported for magmas from some subduction zones. However, peridotites with δ(18)O values significantly higher than the well-defined upper mantle values have never been observed from modern subduction zones. Here we present in-situ oxygen isotope data of olivine crystals in Sailipu mantle xenoliths from South Tibet, which have been subjected to a long history of Tethyan subduction before the India-Asia collision. Our data identify for the first time a metasomatized mantle that, interpreted as the sub-arc lithospheric mantle, shows anomalously enriched oxygen isotopes (δ(18)O = +8.03 ± 0.28 ‰). Such a high-δ(18)O mantle commonly does not contribute significantly to typical island arc basalts. However, partial melting or contamination of such a high-δ(18)O mantle is feasible to account for the high-δ(18)O signatures in arc basalts.

  6. In vivo investigation of homocysteine metabolism to polyamines by high-resolution accurate mass spectrometry and stable isotope labeling.

    PubMed

    Ruseva, Silviya; Lozanov, Valentin; Markova, Petia; Girchev, Radoslav; Mitev, Vanio

    2014-07-15

    Polyamines are essential polycations, playing important roles in mammalian physiology. Theoretically, the involvement of homocysteine in polyamine synthesis via S-adenosylmethionine is possible; however, to our knowledge, it has not been established experimentally. Here, we propose an original approach for investigation of homocysteine metabolites in an animal model. The method is based on the combination of isotope-labeled homocysteine supplementation and high-resolution accurate mass spectrometry analysis. Structural identity of the isotope-labeled metabolites was confirmed by accurate mass measurements of molecular and fragment ions and comparison of the retention times and tandem mass spectrometry fragmentation patterns. Isotope-labeled methionine, spermidine, and spermine were detected in all investigated plasma and tissue samples. The induction of moderate hyperhomocysteinemia leads to an alteration in polyamine levels in a different manner. The involvement of homocysteine in polyamine synthesis and modulation of polyamine levels could contribute to a better understanding of the mechanisms connected with homocysteine toxicity.

  7. Chemical Ligation of Folded Recombinant Proteins: Segmental Isotopic Labeling of Domains for NMR Studies

    NASA Astrophysics Data System (ADS)

    Xu, Rong; Ayers, Brenda; Cowburn, David; Muir, Tom W.

    1999-01-01

    A convenient in vitro chemical ligation strategy has been developed that allows folded recombinant proteins to be joined together. This strategy permits segmental, selective isotopic labeling of the product. The src homology type 3 and 2 domains (SH3 and SH2) of Abelson protein tyrosine kinase, which constitute the regulatory apparatus of the protein, were individually prepared in reactive forms that can be ligated together under normal protein-folding conditions to form a normal peptide bond at the ligation junction. This strategy was used to prepare NMR sample quantities of the Abelson protein tyrosine kinase-SH(32) domain pair, in which only one of the domains was labeled with 15N Mass spectrometry and NMR analyses were used to confirm the structure of the ligated protein, which was also shown to have appropriate ligand-binding properties. The ability to prepare recombinant proteins with selectively labeled segments having a single-site mutation, by using a combination of expression of fusion proteins and chemical ligation in vitro, will increase the size limits for protein structural determination in solution with NMR methods. In vitro chemical ligation of expressed protein domains will also provide a combinatorial approach to the synthesis of linked protein domains.

  8. Quantitative Metabolome Analysis Based on Chromatographic Peak Reconstruction in Chemical Isotope Labeling Liquid Chromatography Mass Spectrometry.

    PubMed

    Huan, Tao; Li, Liang

    2015-07-21

    Generating precise and accurate quantitative information on metabolomic changes in comparative samples is important for metabolomics research where technical variations in the metabolomic data should be minimized in order to reveal biological changes. We report a method and software program, IsoMS-Quant, for extracting quantitative information from a metabolomic data set generated by chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS). Unlike previous work of relying on mass spectral peak ratio of the highest intensity peak pair to measure relative quantity difference of a differentially labeled metabolite, this new program reconstructs the chromatographic peaks of the light- and heavy-labeled metabolite pair and then calculates the ratio of their peak areas to represent the relative concentration difference in two comparative samples. Using chromatographic peaks to perform relative quantification is shown to be more precise and accurate. IsoMS-Quant is integrated with IsoMS for picking peak pairs and Zero-fill for retrieving missing peak pairs in the initial peak pairs table generated by IsoMS to form a complete tool for processing CIL LC-MS data. This program can be freely downloaded from the www.MyCompoundID.org web site for noncommercial use.

  9. Synthesis of stable isotopically labelled 3-methylfuran-2(5H)-one and the corresponding strigolactones.

    PubMed

    Cheng, Yun; Ding, Wen-hui; Long, Qin; Zhao, Min; Yang, Jun; Li, Xiao-qiang

    2015-07-01

    Conventional synthetic procedures of strigolactones (SLs) involve the independent synthesis of ring ABC and ring D, followed by a coupling of the two fragments. Here we prepared three kinds of stable, isotopically labelled D-ring analogues productively using a facile protocol. Then, a coupling of the D-rings to ring ABC produced three isotope-labelled SL derivatives. Moreover, (+)-D3-2'-epi-1A and (-)-ent-D3-2'-epi-1A with high enantiomeric purity were obtained via chiral resolution.

  10. The relationship between phytolith- and plant-water δ 18O values in grasses

    NASA Astrophysics Data System (ADS)

    Webb, Elizabeth A.; Longstaffe, Fred J.

    2003-04-01

    Information regarding climatic conditions during plant growth is preserved by the oxygen-isotope composition of biogenic silica (phytoliths) deposited in grasses. The O-isotope compositions of phytoliths and the plant water from which they precipitate are dependent on soil-water δ 18O values, relative humidity, evapotranspiration rates, and temperature. Plant water and phytoliths from two grass species, Ammophila breviligulata (C 3) and Calamovilfa longifolia (C 4) at Pinery Provincial Park in southwestern Ontario, Canada, were examined to determine the variability in their δ 18O values. Stem water was unfractionated from soil-water in oxygen isotopic composition and the δ 18O values of stem silica provide a good proxy for the soil water available to roots during the growing season. Greater spatial and temporal variation in the δ 18O values of water in the top 5 cm of the soil, and their enhanced sensitivity to evaporative 18O enrichment, are reflected in the generally higher δ 18O values of water in the shallow roots and rhizomes of these grasses. Water within the sheath and lower and upper leaf tissues experiences continual evaporation, becoming progressively enriched in 18O as it moves towards the tip of the leaf. However, the water from which leaf silica precipitates has not acquired the extreme 18O enrichment predicted using steady-state models, or measured for midday or average daily leaf water. Possible explanations for this behaviour include preferential deposition of silica at night; the existence of a secluded water fraction within the leaf, which experiences smaller diurnal variations in isotopic composition than leaf water at sites of evaporation; kinetic isotope effects during rapid precipitation of leaf silica; and incomplete exchange between the oxygen in the silicic acid and the leaf water.

  11. Multisubstrate isotope labeling and metagenomic analysis of active soil bacterial communities.

    PubMed

    Verastegui, Y; Cheng, J; Engel, K; Kolczynski, D; Mortimer, S; Lavigne, J; Montalibet, J; Romantsov, T; Hall, M; McConkey, B J; Rose, D R; Tomashek, J J; Scott, B R; Charles, T C; Neufeld, J D

    2014-07-15

    Soil microbial diversity represents the largest global reservoir of novel microorganisms and enzymes. In this study, we coupled functional metagenomics and DNA stable-isotope probing (DNA-SIP) using multiple plant-derived carbon substrates and diverse soils to characterize active soil bacterial communities and their glycoside hydrolase genes, which have value for industrial applications. We incubated samples from three disparate Canadian soils (tundra, temperate rainforest, and agricultural) with five native carbon ((12)C) or stable-isotope-labeled ((13)C) carbohydrates (glucose, cellobiose, xylose, arabinose, and cellulose). Indicator species analysis revealed high specificity and fidelity for many uncultured and unclassified bacterial taxa in the heavy DNA for all soils and substrates. Among characterized taxa, Actinomycetales (Salinibacterium), Rhizobiales (Devosia), Rhodospirillales (Telmatospirillum), and Caulobacterales (Phenylobacterium and Asticcacaulis) were bacterial indicator species for the heavy substrates and soils tested. Both Actinomycetales and Caulobacterales (Phenylobacterium) were associated with metabolism of cellulose, and Alphaproteobacteria were associated with the metabolism of arabinose; members of the order Rhizobiales were strongly associated with the metabolism of xylose. Annotated metagenomic data suggested diverse glycoside hydrolase gene representation within the pooled heavy DNA. By screening 2,876 cloned fragments derived from the (13)C-labeled DNA isolated from soils incubated with cellulose, we demonstrate the power of combining DNA-SIP, multiple-displacement amplification (MDA), and functional metagenomics by efficiently isolating multiple clones with activity on carboxymethyl cellulose and fluorogenic proxy substrates for carbohydrate-active enzymes. Importance: The ability to identify genes based on function, instead of sequence homology, allows the discovery of genes that would not be identified through sequence alone. This

  12. Effects of (18)O isotopic substitution on the rotational spectra and potential splitting in the OH-OH2 complex: improved measurements for (16)OH-(16)OH2 and (18)OH-(18)OH2, new measurements for the mixed isotopic forms, and ab initio calculations of the (2)A'-(2)A" energy separation.

    PubMed

    Brauer, Carolyn S; Sedo, Galen; Dahlke, Erin; Wu, Shenghai; Grumstrup, Erik M; Leopold, Kenneth R; Marshall, Mark D; Leung, Helen O; Truhlar, Donald G

    2008-09-14

    Rotational spectra have been observed for (16)OH-(16)OH(2), (16)OH-(18)OH(2), (18)OH-(16)OH(2), and (18)OH-(18)OH(2) with complete resolution of the nuclear magnetic hyperfine structure from the OH and water protons. Transition frequencies have been analyzed for each isotopic form using the model of Marshall and Lester [J. Chem. Phys. 121, 3019 (2004)], which accounts for partial quenching of the OH orbital angular momentum and the decoupling of the electronic spin from the OH molecular axis. The analysis accounts for both the ground ((2)A(')) and first electronically excited ((2)A(")) states of the system, which correspond roughly to occupancy by the odd electron in the p(y) and p(x) orbitals, respectively (where p(y) is in the mirror plane of the complex and p(x) is perpendicular to p(y) and the OH bond axis). The spectroscopic measurements yield a parameter, rho, which is equal to the vibrationally averaged (2)A(')-(2)A(") energy separation that would be obtained if spin-orbit coupling and rotation were absent. For the parent species, rho = -146.560 27(9) cm(-1). (18)O substitution on the water increases /rho/ by 0.105 29(10) cm(-1), while substitution on the OH decreases /rho/ by 0.068 64(11) cm(-1). In the OH-OH(2) complex, the observed value of rho implies an energy spacing between the rotationless levels of the (2)A(') and (2)A(") states of 203.76 cm(-1). Ab initio calculations have been performed with quadratic configuration interaction with single and double excitations (QCISD), as well as multireference configuration interaction (MRCI), both with and without the inclusion of spin-orbit coupling. The MRCI calculations with spin-orbit coupling perform the best, giving a value of 171 cm(-1) for the (2)A(')-(2)A(") energy spacing at the equilibrium geometry. Calculations along the large-amplitude bending coordinates of the OH and OH(2) moieties within the complex are presented and are shown to be consistent with a vibrational averaging effect as the main

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

    PubMed Central

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

    2009-01-01

    The stratospheric CO2 oxygen isotope budget is thought to be governed primarily by the O(1D)+CO2 isotope exchange reaction. However, there is increasing evidence that other important physical processes may be occurring that standard isotopic tools have been unable to identify. Measuring the distribution of the exceedingly rare CO2 isotopologue 16O13C18O, in concert with 18O and 17O abundances, provides sensitivities to these additional processes and, thus, is a valuable test of current models. We identify a large and unexpected meridional variation in stratospheric 16O13C18O, observed as proportions in the polar vortex that are higher than in any naturally derived CO2 sample to date. We show, through photochemical experiments, that lower 16O13C18O proportions observed in the midlatitudes are determined primarily by the O(1D)+CO2 isotope exchange reaction, which promotes a stochastic isotopologue distribution. In contrast, higher 16O13C18O proportions in the polar vortex show correlations with long-lived stratospheric tracer and bulk isotope abundances opposite to those observed at midlatitudes and, thus, opposite to those easily explained by O(1D)+CO2. We believe the most plausible explanation for this meridional variation is either an unrecognized isotopic fractionation associated with the mesospheric photochemistry of CO2 or temperature-dependent isotopic exchange on polar stratospheric clouds. Unraveling the ultimate source of stratospheric 16O13C18O enrichments may impose additional isotopic constraints on biosphere–atmosphere carbon exchange, biosphere productivity, and their respective responses to climate change. PMID:19564595

  14. Negative ion ESI-MS analysis of natural yellow dye flavonoids--An isotopic labelling study

    NASA Astrophysics Data System (ADS)

    McNab, Hamish; Ferreira, Ester S. B.; Hulme, Alison N.; Quye, Anita

    2009-07-01

    Flavonoids are amongst the most commonly used natural yellow colourants in paintings, as lakes, and in historical textiles as mordant dyes. In this paper, evidence from isotopically labelled substrates is used to propose negative ion electrospray collision induced decomposition mechanisms of flavones, flavonols and an isoflavone. These mechanisms include a retro-Diels-Alder fragmentation (observed for flavones and flavonols) and an M-122 fragmentation (characteristic of 3',4'-dihydroxyflavonols). In addition, the presence of a m/z 125 fragment ion is shown to be characteristic of 2'-hydroxyflavonols and an ion at m/z 149 is shown to be characteristic of 4'-hydroxyflavones. Applications of these methods are exemplified by the identification of a minor component of Dyer's camomile (Anthemis tinctoria L.) and the identification of the dye source in green threads sampled from an 18th Century Scottish tartan fragment.

  15. Isotope labelling to study molecular fragmentation during the dielectric barrier discharge wet reforming of methane

    NASA Astrophysics Data System (ADS)

    Montoro-Damas, Antonio M.; Gómez-Ramírez, Ana; Gonzalez-Elipe, Agustín R.; Cotrino, José

    2016-09-01

    Isotope labelling is used to study the wet plasma reforming of methane in a dielectric barrier discharge reactor using D2O and CH4 as reactants. Besides the formation of CO and hydrogen as main products, different partitions of H and D atoms are found in the hydrogen (i.e., H2, HD, D2), methane (i.e., CH4, CH3D and CH2D2) and water (D2O, DHO) molecules detected by mass spectrometry as outlet gases of the plasma process. The effect of operating parameters such as applied current, residence time and the addition of oxygen to the reaction mixture is correlated with the H/D distribution in these molecules, the overall reaction yield and the energetic efficiency of the process. The results prove the plasma formation of intermediate excited species that rendering water and methane instead of CO and hydrogen greatly contribute to decrease the overall energy efficiency of the reforming process.

  16. Immunoproteomics using polyclonal antibodies and stable isotope-labeled affinity-purified recombinant proteins.

    PubMed

    Edfors, Fredrik; Boström, Tove; Forsström, Björn; Zeiler, Marlis; Johansson, Henrik; Lundberg, Emma; Hober, Sophia; Lehtiö, Janne; Mann, Matthias; Uhlen, Mathias

    2014-06-01

    The combination of immuno-based methods and mass spectrometry detection has great potential in the field of quantitative proteomics. Here, we describe a new method (immuno-SILAC) for the absolute quantification of proteins in complex samples based on polyclonal antibodies and stable isotope-labeled recombinant protein fragments to allow affinity enrichment prior to mass spectrometry analysis and accurate quantification. We took advantage of the antibody resources publicly available from the Human Protein Atlas project covering more than 80% of all human protein-coding genes. Epitope mapping revealed that a majority of the polyclonal antibodies recognized multiple linear epitopes, and based on these results, a semi-automated method was developed for peptide enrichment using polyclonal antibodies immobilized on protein A-coated magnetic beads. A protocol based on the simultaneous multiplex capture of more than 40 protein targets showed that approximately half of the antibodies enriched at least one functional peptide detected in the subsequent mass spectrometry analysis. The approach was further developed to also generate quantitative data via the addition of heavy isotope-labeled recombinant protein fragment standards prior to trypsin digestion. Here, we show that we were able to use small amounts of antibodies (50 ng per target) in this manner for efficient multiplex analysis of quantitative levels of proteins in a human HeLa cell lysate. The results suggest that polyclonal antibodies generated via immunization of recombinant protein fragments could be used for the enrichment of target peptides to allow for rapid mass spectrometry analysis taking advantage of a substantial reduction in sample complexity. The possibility of building up a proteome-wide resource for immuno-SILAC assays based on publicly available antibody resources is discussed.

  17. Extrinsic labelling of staple food crops with isotopic iron does not consistently result in full equilibration: Revisiting the methodology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Extrinsic isotopic labeling of food Fe has been used for over 50 years to measure Fe absorption. This method is based on the assumption that complete equilibration occurs between the extrinsic and the intrinsic Fe prior to intestinal absorption. The present study tested this assumption via use of in...

  18. Highly efficient residue-selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain.

    PubMed

    Miyanoiri, Yohei; Ishida, Yojiro; Takeda, Mitsuhiro; Terauchi, Tsutomu; Inouye, Masayori; Kainosho, Masatsune

    2016-06-01

    We recently developed a practical protocol for preparing proteins bearing stereo-selectively (13)C-methyl labeled leucines and valines, instead of the commonly used (13)C-methyl labeled precursors for these amino acids, by E. coli cellular expression. Using this protocol, proteins with any combinations of isotope-labeled or unlabeled Leu and Val residues were prepared, including some that could not be prepared by the precursor methods. However, there is still room for improvement in the labeling efficiencies for Val residues, using the methods with labeled precursors or Val itself. This is due to the fact that the biosynthesis of Val could not be sufficiently suppressed, even by the addition of large amounts of Val or its precursors. In this study, we completely solved this problem by using a mutant strain derived from E. coli BL21(DE3), in which the metabolic pathways depending on two enzymes, dihydroxy acid dehydratase and β-isopropylmalate dehydrogenase, are completely aborted by deleting the ilvD and leuB genes, which respectively encode these enzymes. The ΔilvD E. coli mutant terminates the conversion from α,β-dihydroxyisovalerate to α-ketoisovalerate, and the conversion from α,β-dihydroxy-α-methylvalerate to α-keto-β-methylvalerate, which produce the preceding precursors for Val and Ile, respectively. By the further deletion of the leuB gene, the conversion from Val to Leu was also fully terminated. Taking advantage of the double-deletion mutant, ΔilvDΔleuB E. coli BL21(DE3), an efficient and residue-selective labeling method with various isotope-labeled Ile, Leu, and Val residues was established.

  19. Rapid biosynthesis of stable isotope-labeled peptides from a reconstituted in vitro translation system for targeted proteomics.

    PubMed

    Xian, Feng; Li, Shuwei; Liu, Siqi

    2015-01-01

    Stable isotope-labeled peptides are routinely used as internal standards (a.k.a. reference peptides) for absolute quantitation of proteins in targeted proteomics. These peptides can either be synthesized chemically on solid supports or expressed biologically by concatenating multiple peptides together to a large protein. Neither method, however, has required versatility, convenience, and economy for making a large number of reference peptides. Here, we describe the biosynthesis of stable isotope-labeled peptides from a reconstituted Escherichia coli in vitro translation system. We provide a detailed protocol on how to express these peptides with high purity and how to determine their concentrations with easiness. Our strategy offers a general, fast, and scalable approach for the easy preparation of labeled reference peptides, which will have broad application in both basic research and translational medicine.

  20. Assessing methane oxidation under landfill covers and its contribution to the above atmospheric CO{sub 2} levels: The added value of the isotope ({delta}{sup 13}C and {delta}{sup 18}O CO{sub 2}; {delta}{sup 13}C and {delta}D CH{sub 4}) approach

    SciTech Connect

    Widory, D.; Proust, E.; Bellenfant, G.; Bour, O.

    2012-09-15

    Highlights: Black-Right-Pointing-Pointer Comparison of the isotope and mass balance approaches to evaluate the level of methane oxidation within a landfill. Black-Right-Pointing-Pointer The level of methane oxidation is not homogenous under the landfill cover and is strongly correlated to the methane flux. Black-Right-Pointing-Pointer Isotope tracking of the contribution of the methane oxidation to the CO{sub 2} concentrations in the ambient air. - Abstract: We are presenting here a multi-isotope approach ({delta}{sup 13}C and {delta}{sup 18}O of CO{sub 2}; {delta}{sup 13}C and {delta}D of CH{sub 4}) to assess (i) the level(s) of methane oxidation during waste biodegradation and its migration through a landfill cover in Sonzay (France), and (ii) its contribution to the atmospheric CO{sub 2} levels above the surface. The isotope approach is compared to the more conventional mass balance approach. Results from the two techniques are comparable and show that the CH{sub 4} oxidation under the landfill cover is heterogenous, with low oxidation percentages in samples showing high biogas fluxes, which was expected in clay covers presenting fissures, through which CH{sub 4} is rapidly transported. At shallow depth, more immobile biogas pockets show a higher level of CH{sub 4} oxidation by the methanotrophic bacteria. {delta}{sup 13}C of CO{sub 2} samples taken at different heights (from below the cover up to 8 m above the ground level) were also used to identify and assess the relative contributions of its main sources both under the landfill cover and in the surrounding atmosphere.

  1. Evidence for a universal pathway of abscisic acid biosynthesis in higher plants from sup 18 O incorporation patterns

    SciTech Connect

    Zeevaart, J.A.D.; Heath, T.G.; Gage, D.A. )

    1989-12-01

    Previous labeling studies of abscisic acid (ABA) with {sup 18}O{sub 2} have been mainly conducted with water-stressed leaves. In this study, {sup 18}O incorporation into ABA of stressed leaves of various species was compared with {sup 18}O labeling of ABA of turgid leaves and of fruit tissue in different stages of ripening. In stressed leaves of all six species investigated, avocado (Persea americana), barley (Hordeum vulgare), bean (Phaseolus vulgaris), cocklebur (Xanthium strumarium), spinach (Spinacia oleracea), and tobacco (Nicotiana tabacum), {sup 18}O was most abundant in the carboxyl group, whereas incorporation of a second and third {sup 18}O in the oxygen atoms on the ring of ABA was much less prominent after 24 h in {sup 18}O{sub 2}. ABA from turgid bean leaves showed significant {sup 18}O incorporation, again with highest {sup 18}O enrichment in the carboxyl group. On the basis of {sup 18}O-labeling patterns observed in ABA from different tissues it is concluded that, despite variations in precusor pool sizes and intermediate turnover rates, there is a universal pathway of ABA biosynthesis in higher plants which involves cleavage of a larger precursor molecule, presumably an oxygenated carotenoid.

  2. Comparison of δ18O measurements in Nitrate by different combustion techniques

    USGS Publications Warehouse

    Revesz, Kinga; Böhlke, John Karl

    2002-01-01

    Three different KNO3 salts with δ18O values ranging from about −31 to +54‰ relative to VSMOW were used to compare three off-line, sealed glass tube combustion methods (widely used for isotope studies) with a more recently developed on-line carbon combustion technique. All methods yielded roughly similar isotope ratios for KNO3 samples with δ18O values in the midpoint of the δ18O scale near that of the nitrate reference material IAEA-NO-3 (around +21 to +25‰). This reference material has been used previously for one-point interlaboratory and intertechnique calibrations. However, the isotope ratio scale factors by all of the off-line combustion techniques are compressed such that they are between 0.3 and 0.7 times that of the on-line combustion technique. The contraction of the δ18O scale in the off-line preparations apparently is caused by O isotope exchange between the sample and the glass combustion tubes. These results reinforce the need for nitrate reference materials with δ18O values far from that of atmospheric O2, to improve interlaboratory comparability.

  3. A conceptual model for interpreting δ18O and δD biomarker records from terrestrial archives

    NASA Astrophysics Data System (ADS)

    Zech, Michael; Tuthorn, Mario; Detsch, Florian; Rozanski, Kazimierz; Zech, Roland; Zöller, Ludwig; Zech, Wolfgang; Glaser, Bruno

    2013-04-01

    The natural abundances of stable oxygen (18O/16O) and hydrogen isotopes (D/H) are valuable proxies for reconstructing paleoclimate history on global as well as on regional scale. While stable isotope analyses of sedimentary leaf wax-derived n-alkanes enables establishing δD biomarker records, we recently developed a method based on compound-specific δ18O analyses of hemicellulose sugars (Zech and Glaser, 2009), which now additionally allows establishing δ18O biomarker records from soil/sedimentary organic matter of terrestrial archives. Here we present a conceptual model for interpreting combined δ18O and δD biomarker records (Zech et al., submitted). Based on this model, we suggest that both δ18O and δD biomarker records primarily reflect the isotopic composition of paleoprecipitation modified by evaporative isotope enrichment of leaf water during transpiration. Considering biosynthetic fractionation factors allows reconstructing from combined δ18O and δD biomarker records the leaf water isotopic composition and the deuterium excess of the leaf water. The deuterium excess may serve as proxy for evaporative enrichment and allows reconstructing relative humidity using a Craig-Gordon model. Furthermore, the model allows calculating δ18O of the plant source water (δ18Osource water), which can be assumed to primarily reflect δ18O of paleoprecipitation. Hence, paleoclimatic conclusions in terms of temperature can be drawn in high latitude study areas and precipitation amount can be reconstructed in monsoon regions. Zech, M., Glaser, B., 2009. Compound-specific δ18O analyses of neutral sugars in soils using GC-Py-IRMS: problems, possible solutions and a first application. Rapid Commun. Mass Spectrom. 23, 3522-3532. Zech et al., 2013. A 220 ka terrestrial δ18O and deuterium excess biomarker record from an eolian permafrost paleosol sequence, NE-Siberia. Submitted to Chemical Geology.

  4. Isotope coded protein labeling coupled immunoprecipitation (ICPL-IP): a novel approach for quantitative protein complex analysis from native tissue.

    PubMed

    Vogt, Andreas; Fuerholzner, Bettina; Kinkl, Norbert; Boldt, Karsten; Ueffing, Marius

    2013-05-01

    High confidence definition of protein interactions is an important objective toward the understanding of biological systems. Isotope labeling in combination with affinity-based isolation of protein complexes has increased in accuracy and reproducibility, yet, larger organisms--including humans--are hardly accessible to metabolic labeling and thus, a major limitation has been its restriction to small animals, cell lines, and yeast. As composition as well as the stoichiometry of protein complexes can significantly differ in primary tissues, there is a great demand for methods capable to combine the selectivity of affinity-based isolation as well as the accuracy and reproducibility of isotope-based labeling with its application toward analysis of protein interactions from intact tissue. Toward this goal, we combined isotope coded protein labeling (ICPL)(1) with immunoprecipitation (IP) and quantitative mass spectrometry (MS). ICPL-IP allows sensitive and accurate analysis of protein interactions from primary tissue. We applied ICPL-IP to immuno-isolate protein complexes from bovine retinal tissue. Protein complexes of immunoprecipitated β-tubulin, a highly abundant protein with known interactors as well as the lowly expressed small GTPase RhoA were analyzed. The results of both analyses demonstrate sensitive and selective identification of known as well as new protein interactions by our method.

  5. Determining synthesis rates of individual proteins in zebrafish (Danio rerio) with low levels of a stable isotope labelled amino acid.

    PubMed

    Geary, Bethany; Magee, Kieran; Cash, Phillip; Young, Iain S; Whitfield, Phillip D; Doherty, Mary K

    2016-05-01

    The zebrafish is a powerful model organism for the analysis of human cardiovascular development and disease. Understanding these processes at the protein level not only requires changes in protein concentration to be determined but also the rate at which these changes occur on a protein-by-protein basis. The ability to measure protein synthesis and degradation rates on a proteome-wide scale, using stable isotope labelling in conjunction with mass spectrometry is now a well-established experimental approach. With the advent of more selective and sensitive mass spectrometers, it is possible to accurately measure lower levels of stable isotope incorporation, even when sample is limited. In order to challenge the sensitivity of this approach, we successfully determined the synthesis rates of over 600 proteins from the cardiac muscle of the zebrafish using a diet where either 30% or 50% of the L-leucine was replaced with a stable isotope labelled analogue ([(2) H7 ]L-leucine]. It was possible to extract sufficient protein from individual zebrafish hearts to determine the incorporation rate of the label into hundreds of proteins simultaneously, with the two labelling regimens showing a good correlation of synthesis rates.

  6. Evapotranspiration partitioning through in-situ oxygen isotope measurements in an oasis cropland

    NASA Astrophysics Data System (ADS)

    Wen, Xue-Fa

    2016-04-01

    The oxygen isotope compositions of ecosystem water pools and fluxes are useful tracers in the water cycle. As part of the Heihe Watershed Allied Telemetry Experimental Research (HiWATER) program, high-frequency and near-continuous in situ measurements of 18O composition of atmospheric vapor (δv) and of evapotranspiration (δET) were made with the flux-gradient method using a cavity ring-down spectroscopy water vapor isotope analyzer. At the sub-daily scale, we found, in conjunction with intensive isotopic measurements of other ecosystem water pools, that the differences between 18O composition of transpiration (δT) and of xylem water (δx) were negligible in early afternoon (13:00-15:00 Beijing time) when ET approached the daytime maximum, indicating isotopic steady state. At the daily scale, for the purpose of flux partitioning, δT was approximated by δx at early afternoon hours, and the 18O composition of soil evaporation (δE) was obtained from the Craig-Gordon model with a moisture-dependent soil resistance. The relative contribution of transpiration to evapotranspiration ranged from 0.71 to 0.96 with a mean of 0.87 ± 0.052 for the growing season according to the isotopic labeling, which was good agreement with soil lysimeter measurements showing a mean transpiration fraction of 0.86 ± 0.058. At the growing season scale, the predicted18O composition of runoff water was within the range of precipitation and irrigation water according to the isotopic mass conservation. The 18O mass conservation requires that the decreased δ18O of ET should be balanced by enhanced δ18O of runoff water. (Wen, XF*, Yang, B, Sun, XM, Lee, X. 2015. Evapotranspiration partitioning through in-situ oxygen isotope measurements in an oasis cropland. Agricultural and Forest Meteorology , doi:10.1016/j.agrformet.2015.12.003).

  7. Tracking the metabolic pulse of plant lipid production with isotopic labeling and flux analyses: Past, present and future.

    PubMed

    Allen, Doug K; Bates, Philip D; Tjellström, Henrik

    2015-04-01

    Metabolism is comprised of networks of chemical transformations, organized into integrated biochemical pathways that are the basis of cellular operation, and function to sustain life. Metabolism, and thus life, is not static. The rate of metabolites transitioning through biochemical pathways (i.e., flux) determines cellular phenotypes, and is constantly changing in response to genetic or environmental perturbations. Each change evokes a response in metabolic pathway flow, and the quantification of fluxes under varied conditions helps to elucidate major and minor routes, and regulatory aspects of metabolism. To measure fluxes requires experimental methods that assess the movements and transformations of metabolites without creating artifacts. Isotopic labeling fills this role and is a long-standing experimental approach to identify pathways and quantify their metabolic relevance in different tissues or under different conditions. The application of labeling techniques to plant science is however far from reaching it potential. In light of advances in genetics and molecular biology that provide a means to alter metabolism, and given recent improvements in instrumentation, computational tools and available isotopes, the use of isotopic labeling to probe metabolism is becoming more and more powerful. We review the principal analytical methods for isotopic labeling with a focus on seminal studies of pathways and fluxes in lipid metabolism and carbon partitioning through central metabolism. Central carbon metabolic steps are directly linked to lipid production by serving to generate the precursors for fatty acid biosynthesis and lipid assembly. Additionally some of the ideas for labeling techniques that may be most applicable for lipid metabolism in the future were originally developed to investigate other aspects of central metabolism. We conclude by describing recent advances that will play an important future role in quantifying flux and metabolic operation in plant

  8. ­Characterization of Reduced Magmatic C-O-H-N Volatiles By Isotopic Labeling

    NASA Astrophysics Data System (ADS)

    Falksen, E.; Armstrong, L. S.; Hirschmann, M. M.

    2014-12-01

    Characterization of COHN species in silicate melts [1-10] is required to understand the role of reduced volatiles in planetary and early Earth processes, including partitioning between planetary cores, mantles, and atmospheres during early differentiation. Vibrational spectroscopy has been used to examine volatile speciation, but for a number of absorptions there is uncertainty as to whether they relate to species containing N, C, or both [1,3]. In particular, an IR band at 3370 cm-1 is commonly attributed to N-H stretching [1,4,5,7], but associated Raman bands near 3280 cm-1 have also been attributed to alkyne (C-H) bonds [8-10]. The 3370 cm-1 IR band appears even in nominally N-free experiments owing to trapped air and is accompanied by a feature at 1615 cm-1 which could be caused by C=O or N-H bonds [1,3,8]. We sought to determine whether N and C were responsible for various IR bands by dissolving different isotopes of N and C in basaltic melts at high pressure and temperature and observing the shift in position of the resulting absorptions. Experiments were conducted at 1.2 GPa and 1400 oC and volatiles were added to a basaltic oxide mix in the form of unlabeled, 13C labeled, and 15N labeled urea [(NH2)2CO]. The resulting glasses were analyzed using FTIR and the theoretical band shifts were predicted based on a classical approximation of a diatomic molecule. Relative to isotopically normal glasses, bands at both 3370 cm-1 and 1615 cm-1 decrease by 4-8 wavenumbers for 15N and not at all for 13C, consistent with origination by N-H bonds in amines or metal-ammine complexes. [1] Stanley et al. (2014) GCA 129, 54-76. [2] Wetzel et al. (2013) PNAS 110, 8010-8013. [3] Armstrong et al. (in prep). [4] Kadik et al. (2011) Geochem. Int. 49, 429-438. [5] Kadik et al. (2013) PEPI 214, 14-24. [6]Mysen (2013) Chem. Geo. 346, 113-124. [7] Mysen et al. (2008) Am. Min. 93, 1760-1770. [8] Mysen et al. (2009) GCA 73, 1696-1710. [9] Dasgupta et al. (2013) GCA 102, 191-212. [10] Chi

  9. Application of SAIL phenylalanine and tyrosine with alternative isotope-labeling patterns for protein structure determination.

    PubMed

    Takeda, Mitsuhiro; Ono, Akira M; Terauchi, Tsutomu; Kainosho, Masatsune

    2010-01-01

    The extensive collection of NOE constraint data involving the aromatic ring signals is essential for accurate protein structure determination, although it is often hampered in practice by the pervasive signal overlapping and tight spin couplings for aromatic rings. We have prepared various types of stereo-array isotope labeled phenylalanines (epsilon- and zeta-SAIL Phe) and tyrosine (epsilon-SAIL Tyr) to overcome these problems (Torizawa et al. 2005), and proven that these SAIL amino acids provide dramatic spectral simplification and sensitivity enhancement for the aromatic ring NMR signals. In addition to these SAIL aromatic amino acids, we recently synthesized delta-SAIL Phe and delta-SAIL Tyr, which allow us to observe and assign delta-(13)C/(1)H signals very efficiently. Each of the various types of SAIL Phe and SAIL Tyr yields well-resolved resonances for the delta-, epsilon- or zeta-(13)C/(1)H signals, respectively, which can readily be assigned by simple and robust pulse sequences. Since the delta-, epsilon-, and zeta-proton signals of Phe/Tyr residues give rise to complementary NOE constraints, the concomitant use of various types of SAIL-Phe and SAIL-Tyr would generate more accurate protein structures, as compared to those obtained by using conventional uniformly (13)C, (15)N-double labeled proteins. We illustrated this with the case of an 18.2 kDa protein, Escherichia coli peptidyl-prolyl cis-trans isomerase b (EPPIb), and concluded that the combined use of zeta-SAIL Phe and epsilon-SAIL Tyr would be practically the best choice for protein structural determinations.

  10. 2H and 18O Freshwater Isoscapes of Scotland

    NASA Astrophysics Data System (ADS)

    Meier-Augenstein, Wolfram; Hoogewerff, Jurian; Kemp, Helen; Frew, Danny

    2013-04-01

    Scotland's freshwater lochs and reservoirs provide a vital resource for sustaining biodiversity, agriculture, food production as well as for human consumption. Regular monitoring of freshwaters by the Scottish Environment Protection Agency (SEPA) fulfils legislative requirements with regards to water quality but new scientific methods involving stable isotope analysis present an opportunity combining these mandatory monitoring schemes with fundamental research to inform and deliver on current and nascent government policies [1] through gaining a greater understanding of Scottish waters and their importance in the context of climate change, environmental sustainability and food security. For example, 2H and 18O isoscapes of Scottish freshwater could be used to underpin research and its applications in: • Climate change - Using longitudinal changes in the characteristic isotope composition of freshwater lochs and reservoirs as proxy, isoscapes will provide a means to assess if and how changes in temperature and weather patterns might impact on precipitation patterns and amount. • Scottish branding - Location specific stable isotope signatures of Scottish freshwater have the potential to be used as a tool for provenancing and thus protecting premium Scottish produce such as Scottish beef, Scottish soft fruit and Scottish Whisky. During 2011 and 2012, with the support of SEPA more than 110 samples from freshwater lochs and reservoirs were collected from 127 different locations across Scotland including the Highlands and Islands. Here we present the results of this sampling and analysis exercise isotope analyses in form of 2H and 18O isoscapes with an unprecedented grid resolution of 26.5 × 26.5 km (or 16.4 × 16.4 miles). [1] Adaptation Framework - Adapting Our Ways: Managing Scotland's Climate Risk (2009): Scotland's Biodiversity: It's in Your Hands - A strategy for the conservation and enhancement of biodiversity in Scotland (2005); Recipe For Success - Scotland

  11. Analysis of liposoluble carboxylic acids metabolome in human serum by stable isotope labeling coupled with liquid chromatography-mass spectrometry.

    PubMed

    Zhu, Quan-Fei; Zhang, Zheng; Liu, Ping; Zheng, Shu-Jian; Peng, Ke; Deng, Qian-Yun; Zheng, Fang; Yuan, Bi-Feng; Feng, Yu-Qi

    2016-08-19

    Fatty acids (FAs) are groups of liposoluble carboxylic acids (LCAs) and play important roles in various physiological processes. Abnormal contents or changes of FAs are associated with a series of diseases. Here we developed a strategy with stable isotope labeling combined with liquid chromatography-tandem mass spectrometry (IL-LC-MS) analysis for comprehensive profiling and relative quantitation of LCAs in human serum. In this strategy, a pair of isotope labeling reagents (2-dimethylaminoethylamine (DMED)) and d4-2-dimethylaminoethylamine (d4-DMED) were employed to selectively label carboxyl groups of LCAs. The DMED and d4-DMED labeled products can lose four characteristic neutral fragments of 45 and 49Da or 63 and 67Da in collision-induced dissociation. Therefore, quadruple neutral loss scan (QNLS) mode was established and used for non-targeted profiling of LCAs. The peak pairs of DMED and d4-DMED labeling with the same retention time, intensity and characteristic mass differences were extracted from the two NLS spectra respectively, and assigned as potential LCA candidates. Using this strategy, 241 LCA candidates were discovered in the human serum; 156 carboxylic acid compounds could be determined by searching HMDB and METLIN databases (FAs are over 90%) and 21 of these LCAs were successfully identified by standards. Subsequently, a modified pseudo-targeted method with multiple reaction monitoring (MRM) detection mode was developed and used for relative quantification of LCAs in human serum from type 2 diabetes mellitus (T2DM) patients and healthy controls. As a result, 81 LCAs were found to have significant difference between T2DM patients and healthy controls. Taken together, the isotope labeling combined with tandem mass spectrometry analysis demonstrated to be a powerful strategy for identification and quantification of LCA compounds in serum samples.

  12. Stable isotope N-phosphorylation labeling for Peptide de novo sequencing and protein quantification based on organic phosphorus chemistry.

    PubMed

    Gao, Xiang; Wu, Hanzhi; Lee, Kim-Chung; Liu, Hongxia; Zhao, Yufen; Cai, Zongwei; Jiang, Yuyang

    2012-12-04

    In this paper, we describe the development of a novel stable isotope N-phosphorylation labeling (SIPL) strategy for peptide de novo sequencing and protein quantification based on organic phosphorus chemistry. The labeling reaction could be performed easily and completed within 40 min in a one-pot reaction without additional cleanup procedures. It was found that N-phosphorylation labeling reagents were activated in situ to form labeling intermediates with high reactivity targeting on N-terminus and ε-amino groups of lysine under mild reaction conditions. The introduction of N-terminal-labeled phosphoryl group not only improved the ionization efficiency of peptides and increased the protein sequence coverage for peptide mass fingerprints but also greatly enhanced the intensities of b ions, suppressed the internal fragments, and reduced the complexity of the tandem mass spectrometry (MS/MS) fragmentation patterns of peptides. By using nano liquid chromatography chip/time-of-flight mass spectrometry (nano LC-chip/TOF MS) for the protein quantification, the obtained results showed excellent correlation of the measured ratios to theoretical ratios with relative errors ranging from 0.5% to 6.7% and relative standard deviation of less than 10.6%, indicating that the developed method was reproducible and precise. The isotope effect was negligible because of the deuterium atoms were placed adjacent to the neutral phosphoryl group with high electrophilicity and moderately small size. Moreover, the SIPL approach used inexpensive reagents and was amenable to samples from various sources, including cell culture, biological fluids, and tissues. The method development based on organic phosphorus chemistry offered a new approach for quantitative proteomics by using novel stable isotope labeling reagents.

  13. An efficient synthetic strategy for obtaining 4-methoxy carbon isotope labeled combretastatin A-4 phosphate and other Z-combretastatins.

    PubMed

    Pettit, George R; Minardi, Mathew D; Hogan, Fiona; Price, Pat M

    2010-03-26

    Human cancer and other clinical trials under development employing combretastatin A-4 phosphate (1b, CA4P) should benefit from the availability of a [(11)C]-labeled derivative for positron emission tomography (PET). In order to obtain a suitable precursor for addition of a [(11)C]methyl group at the penultimate step, several new synthetic pathways to CA4P were evaluated. Geometrical isomerization (Z to E) proved to be a challenge, but it was overcome by development of a new CA4P synthesis suitable for 4-methoxy isotope labeling.

  14. Amino acid-selective isotope labeling of proteins for nuclear magnetic resonance study: proteins secreted by Brevibacillus choshinensis.

    PubMed

    Tanio, Michikazu; Tanaka, Rikou; Tanaka, Takeshi; Kohno, Toshiyuki

    2009-03-15

    Here we report the first application of amino acid-type selective (AATS) isotope labeling of a recombinant protein secreted by Brevibacillus choshinensis for a nuclear magnetic resonance (NMR) study. To prepare the 15N-AATS-labeled protein, the transformed B. choshinensis was cultured in 15N-labeled amino acid-containing C.H.L. medium, which is commonly used in the Escherichia coli expression system. The analyses of the 1H-15N heteronuclear single quantum coherence (HSQC) spectra of the secreted proteins with a 15N-labeled amino acid demonstrated that alanine, arginine, asparagine, cysteine, glutamine, histidine, lysine, methionine, and valine are suitable for selective labeling, although acidic and aromatic amino acids are not suitable. The 15N labeling for glycine, isoleucine, leucine, serine, and threonine resulted in scrambling to specific amino acids. These results indicate that the B. choshinensis expression system is an alternative tool for AATS labeling of recombinant proteins, especially secretory proteins, for NMR analyses.

  15. Stable isotope applications in biomolecular structure and mechanisms. A meeting to bring together producers and users of stable-isotope-labeled compounds to assess current and future needs

    SciTech Connect

    Trewhella, J.; Cross, T.A.; Unkefer, C.J.

    1994-12-01

    Knowledge of biomolecular structure is a prerequisite for understanding biomolecular function, and stable isotopes play an increasingly important role in structure determination of biological molecules. The first Conference on Stable Isotope Applications in Biomolecular Structure and Mechanisms was held in Santa Fe, New Mexico, March 27--31, 1994. More than 120 participants from 8 countries and 44 institutions reviewed significant developments, discussed the most promising applications for stable isotopes, and addressed future needs and challenges. Participants focused on applications of stable isotopes for studies of the structure and function of proteins, peptides, RNA, and DNA. Recent advances in NMR techniques neutron scattering, EPR, and vibrational spectroscopy were highlighted in addition to the production and synthesis of labeled compounds. This volume includes invited speaker and poster presentations as well as a set of reports from discussion panels that focused on the needs of the scientific community and the potential roles of private industry, the National Stable Isotope Resource, and the National High Magnetic Field Laboratory in serving those needs. This is the leading abstract. Individual papers are processed separately for the database.

  16. A proteomic approach for quantitation of phosphorylation using stable isotope labeling in cell culture.

    PubMed

    Ibarrola, Nieves; Kalume, Dario E; Gronborg, Mads; Iwahori, Akiko; Pandey, Akhilesh

    2003-11-15

    Posttranslational modifications are major mechanisms of regulating protein activity and function in vertebrate cells. It is essential to obtain qualitative information about posttranslational modification patterns of proteins to understand signal transduction mechanisms in greater detail. However, it is equally important to measure the dynamics of posttranslational modifications such as phosphorylation to approach signaling networks from a systems biology perspective. Despite a number of advances, methods to quantitate posttranslational modifications remain difficult to implement due to a number of factors including lack of a generic method, elaborate chemical steps, and requirement for large amounts of sample. We have previously shown that stable isotope-containing amino acids in cell culture (SILAC) can be used to differentially label growing cell populations for quantitation of protein levels. In this report, we extend the use of SILAC as a novel proteomic approach for the relative quantitation of posttranslational modifications such as phosphorylation. We have used SILAC to quantitate the extent of known phosphorylation sites as well as to identify and quantitate novel phosphorylation sites.

  17. Tracking down sulphate-reducing microorganisms by molecular and isotope-labelling techniques

    NASA Astrophysics Data System (ADS)

    Loy, Alexander

    2010-05-01

    Sulphate-reducing microorganisms (SRM) are of great ecological importance for carbon compound degradation and sulphur cycling in many anoxic ecosystems, including marine sediments, peatlands, and oil reservoirs. However, the activity of SRM can result in oil souring and pipeline corrosion and thus is also an economic burden for the oil industry. Molecular diversity surveys based on rRNA genes and dsrAB, genes that encode major subunits of the dissimilatory sulfite reductase, indicate that our view of the natural diversity of SRM (as we know it from cultivation) is far from being complete. This enormous phylogenetic diversity complicates unbiased identification and quantification of SRM by molecular methods such as fluorescence in situ hybridization, real-time PCR or DNA microarrays. Combining these 16S rRNA and dsrAB-based molecular methods with substrate-mediated isotope labelling techniques is a potential solution for identification and functional characterization of yet uncultivated SRM. Using SRM in peatlands as an example, the problems and opportunities of these techniques for diagnosing and monitoring SRM in the environment will be discussed in this talk.

  18. LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

    NASA Technical Reports Server (NTRS)

    Zuwei, Wang; Boyd, Jason; Berens, Kurt L.; Putcha, Lakshmi

    2004-01-01

    Promethazine (PMZ) is taken by astronauts orally (PO), intramuscularly (IM) or rectally (PR) for space motion sickness. LC/MS method was developed with off-line solid phase extraction to measure plasma concentrations of PMZ given as stable isotope-labeled (SIL) formulations by the three different routes of administration simultaneously. Samples (0.5ml) were loaded on to Waters Oasis HLB co-polymer cartridges and eluted with 1.0 mL methanol. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 6 min. Acetonitrile/ ammonium acetate (30 mM) in water (3:2, v/v), pH 5.6 plus or minus 0.1, was used as the mobile phase for separation. Concentrations of PMZ, PMZ-d4 and PMZ-d7 and chlorpromazine (internal standard) were determined using a Micromass ZMD single quadrupole mass spectrometer with Electrospray Ionization (ESI). ESI mass spectra were acquired in positive ion mode with selected ion monitoring of [M+ H]dot plus. The method is rapid, reproducible and the assay specific parameters are listed in a table. A novel, sensitive and specific method for the measurement of PMZ and SIL PMZ in human plasma is reported.

  19. Isotope labeling-based quantitative proteomics of developing seeds of castor oil seed (Ricinus communis L.).

    PubMed

    Nogueira, Fábio C S; Palmisano, Giuseppe; Schwämmle, Veit; Soares, Emanuela L; Soares, Arlete A; Roepstorff, Peter; Domont, Gilberto B; Campos, Francisco A P

    2013-11-01

    In this study, we used a mass spectrometry-based quantification approach employing isotopic (ICPL) and isobaric (iTRAQ) labeling to investigate the pattern of protein deposition during castor oil seed (Ricinus communis L.) development, including that of proteins involved in fatty acid metabolism, seed-storage proteins (SSPs), toxins, and allergens. Additionally, we have used off-line hydrophilic interaction chromatography (HILIC) as a step of peptide fractionation preceding the reverse-phase nanoLC coupled to a LTQ Orbitrap. We were able to identify a total of 1875 proteins, and from these 1748 could be mapped to extant castor gene models, considerably expanding the number of proteins so far identified from developing castor seeds. Cluster validation and statistical analysis resulted in 975 protein trend patterns and the relative abundance of 618 proteins. The results presented in this work give important insights into certain aspects of the biology of castor oil seed development such as carbon flow, anabolism, and catabolism of fatty acid and the pattern of deposition of SSPs, toxins, and allergens such as ricin and 2S albumins. We also found, for the first time, some genes of SSP that are differentially expressed during seed development.

  20. Absolute Quantitation of Glycosylation Site Occupancy Using Isotopically Labeled Standards and LC-MS

    NASA Astrophysics Data System (ADS)

    Zhu, Zhikai; Go, Eden P.; Desaire, Heather

    2014-06-01

    N-linked glycans are required to maintain appropriate biological functions on proteins. Underglycosylation leads to many diseases in plants and animals; therefore, characterizing the extent of glycosylation on proteins is an important step in understanding, diagnosing, and treating diseases. To determine the glycosylation site occupancy, protein N-glycosidase F (PNGase F) is typically used to detach the glycan from the protein, during which the formerly glycosylated asparagine undergoes deamidation to become an aspartic acid. By comparing the abundance of the resulting peptide containing aspartic acid against the one containing non-glycosylated asparagine, the glycosylation site occupancy can be evaluated. However, this approach can give inaccurate results when spontaneous chemical deamidation of the non-glycosylated asparagine occurs. To overcome this limitation, we developed a new method to measure the glycosylation site occupancy that does not rely on converting glycosylated peptides to their deglycosylated forms. Specifically, the overall protein concentration and the non-glycosylated portion of the protein are quantified simultaneously by using heavy isotope-labeled internal standards coupled with LC-MS analysis, and the extent of site occupancy is accurately determined. The efficacy of the method was demonstrated by quantifying the occupancy of a glycosylation site on bovine fetuin. The developed method is the first work that measures the glycosylation site occupancy without using PNGase F, and it can be done in parallel with glycopeptide analysis because the glycan remains intact throughout the workflow.

  1. Novel diagnostics of metabolic dysfunction detected in breath and plasma by selective isotope-assisted labeling.

    PubMed

    Haviland, Julia A; Tonelli, Marco; Haughey, Dermot T; Porter, Warren P; Assadi-Porter, Fariba M

    2012-08-01

    Metabolomics is the study of a unique fingerprint of small molecules present in biological systems under healthy and disease conditions. One of the major challenges in metabolomics is validation of fingerprint molecules to identify specifically perturbed pathways in metabolic aberrations. This step is crucial to the understanding of budding metabolic pathologies and the ability to identify early indicators of common diseases such as obesity, type 2 diabetes mellitus, metabolic syndrome, polycystic ovary syndrome, and cancer. We present a novel approach to diagnosing aberrations in glucose utilization including metabolic pathway switching in a disease state. We used a well-defined prenatally exposed glucocorticoid mouse model that results in adult females with metabolic dysfunction. We applied the complementary technologies of nuclear magnetic resonance spectroscopy and cavity ring-down spectroscopy to analyze serial plasma samples and real-time breath measurements following selective (13)C-isotope-assisted labeling. These platforms allowed us to trace metabolic markers in whole animals and identify key metabolic pathway switching in prenatally glucocorticoid-treated animals. Total glucose flux is significantly proportionally increased through the major oxidative pathways of glycolysis and the pentose phosphate pathway in the prenatally glucocorticoid-treated animals relative to the control animals. This novel diagnostics approach is fast, noninvasive, and sensitive for determining specific pathway utilization, and provides a direct translational application in the health care field.

  2. Shape-Controlled Synthesis of Isotopic Yttrium-90-Labeled Rare Earth Fluoride Nanocrystals for Multimodal Imaging.

    PubMed

    Paik, Taejong; Chacko, Ann-Marie; Mikitsh, John L; Friedberg, Joseph S; Pryma, Daniel A; Murray, Christopher B

    2015-09-22

    Isotopically labeled nanomaterials have recently attracted much attention in biomedical research, environmental health studies, and clinical medicine because radioactive probes allow the elucidation of in vitro and in vivo cellular transport mechanisms, as well as the unambiguous distribution and localization of nanomaterials in vivo. In addition, nanocrystal-based inorganic materials have a unique capability of customizing size, shape, and composition; with the potential to be designed as multimodal imaging probes. Size and shape of nanocrystals can directly influence interactions with biological systems, hence it is important to develop synthetic methods to design radiolabeled nanocrystals with precise control of size and shape. Here, we report size- and shape-controlled synthesis of rare earth fluoride nanocrystals doped with the β-emitting radioisotope yttrium-90 ((90)Y). Size and shape of nanocrystals are tailored via tight control of reaction parameters and the type of rare earth hosts (e.g., Gd or Y) employed. Radiolabeled nanocrystals are synthesized in high radiochemical yield and purity as well as excellent radiolabel stability in the face of surface modification with different polymeric ligands. We demonstrate the Cerenkov radioluminescence imaging and magnetic resonance imaging capabilities of (90)Y-doped GdF3 nanoplates, which offer unique opportunities as a promising platform for multimodal imaging and targeted therapy.

  3. The influence of drinking water on the deltaD and delta18O values of house sparrow plasma, blood and feathers.

    PubMed

    Wolf, Nathan; Bowen, Gabriel J; Del Rio, Carlos Martinez

    2011-01-01

    We investigated the relationships between the δdeuterium (δD) and the δ(18)oxygen (δ(18)O) of drinking water and the δD and δ(18)O of blood plasma, red blood cells and feathers in house sparrows (Passer domesticus) fed on diets with identical hydrogen and oxygen isotopic compositions and five isotopically distinct drinking water treatments. We expected and, with only one exception ((18)O in blood plasma), found linear relationships between the δD and δ(18)O values of drinking water and those of bird tissues. The slopes of these relationships, which estimate the percentage contributions of drinking water to the tissue isotopic signatures, were lower than those of previous studies. We found significant differences in the δD and δ(18)O values of feathers, red blood cells and plasma solids. In feathers and red blood cells, δD and δ(18)O values were linearly correlated. Our results have three implications for isotopic field studies: (1) if the isotopic composition of drinking water differs from that of food, its effect on tissue isotope values can confound the assignment of animals to a site of origin; (2) comparisons of the δD and δ(18)O values of different tissues must account for inter-tissue discrimination factors; and (3) δD/δ(18)O linear relationships are probably as prevalent in animal systems as they are in geohydrological systems. These relationships may prove to be useful tools in animal isotopic ecology.

  4. Abstracts of the 24th international isotope society (UK group) symposium: synthesis and applications of labelled compounds 2015.

    PubMed

    Aigbirhio, F I; Allwein, S; Anwar, A; Atzrodt, J; Audisio, D; Badman, G; Bakale, R; Berthon, F; Bragg, R; Brindle, K M; Bushby, N; Campos, S; Cant, A A; Chan, M Y T; Colbon, P; Cornelissen, B; Czarny, B; Derdau, V; Dive, V; Dunscombe, M; Eggleston, I; Ellis-Sawyer, K; Elmore, C S; Engstrom, P; Ericsson, C; Fairlamb, I J S; Georgin, D; Godfrey, S P; He, L; Hickey, M J; Huscroft, I T; Kerr, W J; Lashford, A; Lenz, E; Lewinton, S; L'Hermite, M M; Lindelöf, Å; Little, G; Lockley, W J S; Loreau, O; Maddocks, S; Marguerit, M; Mirabello, V; Mudd, R J; Nilsson, G N; Owens, P K; Pascu, S I; Patriarche, G; Pimlott, S L; Pinault, M; Plastow, G; Racys, D T; Reif, J; Rossi, J; Ruan, J; Sarpaki, S; Sephton, S M; Simonsson, R; Speed, D J; Sumal, K; Sutherland, A; Taran, F; Thuleau, A; Wang, Y; Waring, M; Watters, W H; Wu, J; Xiao, J

    2016-04-01

    The 24th annual symposium of the International Isotope Society's United Kingdom Group took place at the Møller Centre, Churchill College, Cambridge, UK on Friday 6th November 2015. The meeting was attended by 77 delegates from academia and industry, the life sciences, chemical, radiochemical and scientific instrument suppliers. Delegates were welcomed by Dr Ken Lawrie (GlaxoSmithKline, UK, chair of the IIS UK group). The subsequent scientific programme consisted of oral presentations, short 'flash' presentations in association with particular posters and poster presentations. The scientific areas covered included isotopic synthesis, regulatory issues, applications of labelled compounds in imaging, isotopic separation and novel chemistry with potential implications for isotopic synthesis. Both short-lived and long-lived isotopes were represented, as were stable isotopes. The symposium was divided into a morning session chaired by Dr Rebekka Hueting (University of Oxford, UK) and afternoon sessions chaired by Dr Sofia Pascu (University of Bath, UK) and by Dr Alan Dowling (Syngenta, UK). The UK meeting concluded with remarks from Dr Ken Lawrie (GlaxoSmithKline, UK).

  5. The use of isotope effects to determine enzyme mechanisms.

    PubMed

    Cleland, W W

    2005-01-01

    Isotope effects are one of the most powerful kinetic tools for determining enzyme mechanisms. There are three methods of measurement. First, one can compare reciprocal plots with labeled and unlabeled substrates. The ratio of the slopes is the isotope effect on V/K, and the ratio of the vertical intercepts is the isotope effect on V(max). This is the only way to determine V(max) isotope effects, but is limited to isotope effects of 5% or greater. The second method is internal competition, where the labeled and unlabeled substrates are present at the same time and the change in their ratio in residual substrate or in product is used to calculate an isotope effect, which is that on V/K of the labeled reactant. This is the method used for tritium or (14)C, or with the natural abundances of (13)C, (15)N, or (18)O. The third method involves perturbations from equilibrium when a labeled substrate and corresponding unlabeled product are present at chemical equilibrium. This also gives just an isotope effect on V/K for the labeled reactant. The chemistry is typically not fully rate limiting, so that the isotope effect on V/K is given by: (x)(V/K)=((x)k+c(f)+c(r)(x)K(eq))/(1+c(f)+c(r)) where x defines the isotope (D, T, 13, 15, 18 for deuterium, tritium, (13)C, (15)N, or (18)O), and (x)(V/K), (x)k, and (x)K(eq) are the observed isotope effect, the intrinsic one on the chemical step, and the isotope effect on the equilibrium constant, respectively. The constants c(f) and c(r) are commitments in forward and reverse directions, and are the ratio of the rate constant for the chemical reaction and the net rate constant for release from the enzyme of the varied substrate (direct comparison) or labeled substrate (internal competition and equilibrium perturbation) for c(f), or the first product released or the one involved in the perturbation for c(r). The intrinsic isotope effect, (x)k, can be estimated by comparing deuterium and tritium isotope effects on V/K, or by comparing the

  6. Stereospecific Nickel-Catalyzed Cross-Coupling Reactions of Benzylic Ethers with Isotopically-Labeled Grignard Reagents

    PubMed Central

    2015-01-01

    In this manuscript we highlight the potential of stereospecific nickel-catalyzed cross-coupling reactions for applications in the pharmaceutical industry. Using an inexpensive and sustainable nickel catalyst, we report a gram-scale Kumada cross-coupling reaction. Reactions are highly stereospecific and proceed with inversion at the benzylic position. We also expand the scope of our reaction to incorporate isotopically labeled substituents. PMID:27458328

  7. Structural modeling of protein-RNA complexes using crosslinking of segmentally isotope-labeled RNA and MS/MS.

    PubMed

    Dorn, Georg; Leitner, Alexander; Boudet, Julien; Campagne, Sébastien; von Schroetter, Christine; Moursy, Ahmed; Aebersold, Ruedi; Allain, Frédéric H-T

    2017-03-27

    Ribonucleoproteins (RNPs) are key regulators of cellular function. We established an efficient approach, crosslinking of segmentally isotope-labeled RNA and tandem mass spectrometry (CLIR-MS/MS), to localize protein-RNA interactions simultaneously at amino acid and nucleotide resolution. The approach was tested on polypyrimidine tract binding protein 1 and U1 small nuclear RNP. Our method provides distance restraints to support integrative atomic-scale structural modeling and to gain mechanistic insights into RNP-regulated processes.

  8. Holocene tropical South American hydroclimate revealed from a decadally resolved lake sediment δ 18O record

    NASA Astrophysics Data System (ADS)

    Bird, Broxton W.; Abbott, Mark B.; Rodbell, Donald T.; Vuille, Mathias

    2011-10-01

    Oxygen isotope ratios of authigenic calcite (δ 18O cal) measured at annual to decadal resolution from Laguna Pumacocha document Andean precipitation variability during the last 11,200 years. Modern limnological data show that Pumacocha δ 18O cal reflects the average annual isotopic composition of the lake's surface waters (δ 18O lw), and that δ 18O lw tracks the isotopic composition of precipitation (δ 18O precip), which is largely controlled by the intensity of the South American summer monsoon (SASM). Based on these relationships we use down-core δ 18O cal measurements as a proxy for δ 18O precip that varies with the intensity of SASM precipitation. Pumacocha δ 18O cal increased rapidly between 11,200 and 10,300 yr B.P. from - 14.5‰ to - 10.5‰, reaching a maximum of - 10.3‰ by 9800 yr B.P. After 9800 yr B.P., δ 18O cal underwent a long-term decrease that tracked increasing Southern Hemisphere summer insolation, suggesting that enhanced SASM precipitation was linked to precessional forcing. Higher-frequency trends did not follow insolation and therefore represent other variability in the climate system. Millennial-scale trends from Pumacocha strongly resemble those from lower-resolution tropical Andean ice and lake core isotopic records, particularly the Huascaran ice core, and low elevation speleothems. These relationships suggest that tropical Andean isotopic records reflect variations in precipitation intensity related to precessional forcing rather than tropical temperatures. They also demonstrate a coherent pattern of SASM variability, although with differences between low elevation and Andean records during the late Glacial to Holocene transition and the late Holocene. Centennial and decadal SASM precipitation variability is also apparent. Reduced SASM rainfall occurred from 10,000-9200, 7000-5000, 1500-900 yr B.P. and during the last 100 years. Intensifications of the SASM occurred at 5000, 2200-1500, and 550-130 yr B.P. with the amplitude of

  9. Predicting animal δ18O: Accounting for diet and physiological adaptation

    NASA Astrophysics Data System (ADS)

    Kohn, Matthew J.

    1996-12-01

    Theoretical predictions and measured isotope variations indicate that diet and physiological adaptation have a significant impact on animals δ18O and cannot be ignored. A generalized model is therefore developed for the prediction of animal body water and phosphate δ18O to incorporate these factors quantitatively. Application of the model reproduces most published compositions and compositional trends for mammals and birds. A moderate dependence of animal δ18O on humidity is predicted for drought-tolerant animals, and the correlation between humidity and North American deer bone composition as corrected for local meteoric water is predicted within the scatter of the data. In contrast to an observed strong correlation between kangaroo δ18O and humidity (Δδ18O/Δh ∼ 2.5± 0.4‰/10%r.h.), the predicted humidity dependence is only 1.3 - 1.7‰/10% r.h., and it is inferred that drinking water in hot dry areas of Australia is enriched in 18O over rainwater. Differences in physiology and water turnover readily explain the observed differences in δ18O for several herbivore genera in East Africa, excepting antelopes. Antelope models are more sensitive to biological fractionations, and adjustments to the flux of transcutaneous water vapor within experimentally measured ranges allows their δ18O values to be matched. Models of the seasonal changes of forage composition for two regions with dissimilar climates show that significant seasonal variations in animal isotope composition are expected, and that animals with different physiologies and diets track climate differently. Analysis of different genera with disparate sensitivities to surface water and humidity will allow the most accurate quantification of past climate changes.

  10. A device for single leaf labelling with CO2 isotopes to study carbon allocation and partitioning in Arabidopsis thaliana

    PubMed Central

    2013-01-01

    Background Plant biomass consists primarily of carbohydrates derived from photosynthesis. Monitoring the assimilation of carbon via the Calvin-Benson cycle and its subsequent utilisation is fundamental to understanding plant growth. The use of stable and radioactive carbon isotopes, supplied to plants as CO2, allows the measurement of fluxes through the intermediates of primary photosynthetic metabolism, long-distance transport of sugars in the vasculature, and the synthesis of structural and storage components. Results Here we describe the design of a system for supplying isotopically labelled CO2 to single leaves of Arabidopsis thaliana. We demonstrate that the system works well using short pulses of 14CO2 and that it can be used to produce robust qualitative and quantitative data about carbon export from source leaves to the sink tissues, such as the developing leaves and the roots. Time course experiments show the dynamics of carbon partitioning between storage as starch, local production of biomass, and export of carbon to sink tissues. Conclusion This isotope labelling method is relatively simple to establish and inexpensive to perform. Our use of 14CO2 helps establish the temporal and spatial allocation of assimilated carbon during plant growth, delivering data complementary to those obtained in recent studies using 13CO2 and MS-based metabolomics techniques. However, we emphasise that this labelling device could also be used effectively in combination with 13CO2 and MS-based techniques. PMID:24252607

  11. 13C 18O clumping in speleothems: Observations from natural caves and precipitation experiments

    NASA Astrophysics Data System (ADS)

    Daëron, M.; Guo, W.; Eiler, J.; Genty, D.; Blamart, D.; Boch, R.; Drysdale, R.; Maire, R.; Wainer, K.; Zanchetta, G.

    2011-06-01

    The oxygen isotope composition of speleothems is an important proxy of continental paleoenvironments, because of its sensitivity to variations in cave temperature and drip water δ 18O. Interpreting speleothem δ 18O records in terms of absolute paleotemperatures and δ 18O values of paleo-precipitation requires quantitative separation of the effects of these two parameters, and correcting for possible kinetic isotope fractionation associated with precipitation of calcite out of thermodynamic equilibrium. Carbonate clumped-isotope thermometry, based on measurements of Δ47 (a geochemical variable reflecting the statistical overabundance of 13C 18O bonds in CO 2 evolved from phosphoric acid digestion of carbonate minerals), potentially provides a method for absolute speleothem paleotemperature reconstructions independent of drip water composition. Application of this new technique to karst records is currently limited by the scarcity of published clumped-isotope studies of modern speleothems. The only modern stalagmite reported so far in the literature yielded a lower Δ47 value than expected for equilibrium precipitation, possibly due to kinetic isotope fractionation. Here we report Δ47 values measured in natural speleothems from various cave settings, in carbonate produced by cave precipitation experiments, and in synthetic stalagmite analogs precipitated in controlled laboratory conditions designed to mimic natural cave processes. All samples yield lower Δ47 and heavier δ 18O values than predicted by experimental calibrations of thermodynamic equilibrium in inorganic calcite. The amplitudes of these isotopic disequilibria vary between samples, but there is clear correlation between the amount of Δ47 disequilibrium and that of δ 18O. Even pool carbonates believed to offer excellent conditions for equilibrium precipitation of calcite display out-of-equilibrium δ 18O and Δ47 values, probably inherited from prior degassing within the cave system. In addition

  12. Minimization of sample requirement for delta18O in benzoic acid.

    PubMed

    Hagopian, William M; Jahren, A Hope

    2010-09-15

    The measurement of the oxygen stable isotope content in organic compounds has applications in many fields, ranging from paleoclimate reconstruction to forensics. Conventional High-Temperature Conversion (HTC) techniques require >20 microg of O for a single delta(18)O measurement. Here we describe a system that converts the CO produced by HTC into CO(2) via reduction within a Ni-furnace. This CO(2) is then concentrated cryogenically, and 'focused' into the isotope ratio mass spectrometry (IRMS) source using a low-flow He carrier gas (6-8 mL/min). We report analyses of benzoic acid (C(7)H(6)O(2)) reference materials that yielded precise delta(18)O measurement down to 1.3 microg of O, suggesting that our system could be used to decrease sample requirement for delta(18)O by more than an order of magnitude.

  13. Teasing Cellulose Isotopic Signals Apart by Chemical Methods

    NASA Astrophysics Data System (ADS)

    Sternberg, L. D.; Anderson, W. T.; Morrison, K.

    2002-12-01

    The δ18O and δD values of precipitation water correlates with changes in atmospheric circulation patterns and temperature. This has been the basis of many attempts to use fossil tree ring cellulose as a proxy for paleoclimate. Ideally by measuring δ18O and δD values of tree ring cellulose one would infer isotopic composition of water available for plant uptake, which presumably is the least isotopically altered from precipitation. Subsequently, paleo-temperatures or atmospheric circulation patterns at the time of cellulose formation could then be inferred. However, this goal is confounded by isotopic exchange processes occurring in the leaf. Our current understanding of the physiological/biochemical mechanisms operating during the labeling of carbohydrates by water during tree ring cellulose synthesis indicates that the isotopic composition of tree ring cellulose is a mixture of isotopic signals coming from source (CA 35 to 45%) and leaf (CA 55 to 65%) water. The isotopic composition of the latter component is radically modified from that of the original source water by factors such as relative humidity and leaf properties. Here we present a chemical method of derivatizing cellulose to tease these two signals apart. We analyze the isotopic composition of cellulose and its derivative and calculate the δ18O value of the oxygen attached to the second carbon of the glucose moieties in cellulose (2C-OH). A one to one relationship between δ18O values of this oxygen and that of water available for cellulose synthesis in seeds germinated in the presence of water having different δ18O values was observed. Indicating that 2C-OH undergoes complete exchange with water during the synthesis of cellulose from sucrose. This technique can potentially be an analytical tool in paleo-climatic and ecological studies, once the analytical techniques are refined so as to increase precision.

  14. Intracellular Isotope Localization in Ammonia sp. (Foraminifera) of Oxygen-Depleted Environments: Results of Nitrate and Sulfate Labeling Experiments

    PubMed Central

    Nomaki, Hidetaka; Bernhard, Joan M.; Ishida, Akizumi; Tsuchiya, Masashi; Uematsu, Katsuyuki; Tame, Akihiro; Kitahashi, Tomo; Takahata, Naoto; Sano, Yuji; Toyofuku, Takashi

    2016-01-01

    Some benthic foraminiferal species are reportedly capable of nitrate storage and denitrification, however, little is known about nitrate incorporation and subsequent utilization of nitrate within their cell. In this study, we investigated where and how much 15N or 34S were assimilated into foraminiferal cells or possible endobionts after incubation with isotopically labeled nitrate and sulfate in dysoxic or anoxic conditions. After 2 weeks of incubation, foraminiferal specimens were fixed and prepared for Transmission Electron Microscopy (TEM) and correlative nanometer-scale secondary ion mass spectrometry (NanoSIMS) analyses. TEM observations revealed that there were characteristic ultrastructural features typically near the cell periphery in the youngest two or three chambers of the foraminifera exposed to anoxic conditions. These structures, which are electron dense and ~200–500 nm in diameter and co-occurred with possible endobionts, were labeled with 15N originated from 15N-labeled nitrate under anoxia and were labeled with both 15N and 34S under dysoxia. The labeling with 15N was more apparent in specimens from the dysoxic incubation, suggesting higher foraminiferal activity or increased availability of the label during exposure to oxygen depletion than to anoxia. Our results suggest that the electron dense bodies in Ammonia sp. play a significant role in nitrate incorporation and/or subsequent nitrogen assimilation during exposure to dysoxic to anoxic conditions. PMID:26925038

  15. Biodegradation and mineralization of isotopically labeled TNT and RDX in anaerobic marine sediments.

    PubMed

    Ariyarathna, Thivanka; Vlahos, Penny; Smith, Richard W; Fallis, Stephen; Groshens, Thomas; Tobias, Craig

    2016-10-28

    The lack of knowledge on the fate of explosive compounds 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), particularly in marine ecosystems, constrains the application of bioremediation techniques in explosive-contaminated coastal sites. The authors present a comparative study on anaerobic biodegradation and mineralization of (15) N-nitro group isotopically labeled TNT and RDX in organic carbon-rich, fine-grained marine sediment with native microbial assemblages. Separate sediment slurry experiments were carried out for TNT and RDX at 23°C for 16 d. Dissolved and sediment-sorbed fractions of parent and transformation products, isotopic compositions of sediment, and mineralization products of the dissolved inorganic N pool ((15) NH4(+) ,(15) NO3(-) ,(15) NO2(-) , and (15) N2 ) were measured. The rate of TNT removal from the aqueous phase was faster (0.75 h(-1) ) than that of RDX (0.37 h(-1) ), and (15) N accumulation in sediment was higher in the TNT (13%) than the RDX (2%) microcosms. Mono-amino-dinitrotoluenes were identified as intermediate biodegradation products of TNT. Two percent of the total spiked TNT-N is mineralized to dissolved inorganic N through 2 different pathways: denitration as well as deamination and formation of NH4(+) , facilitated by iron and sulfate reducing bacteria in the sediments. The majority of the spiked TNT-N (85%) is in unidentified pools by day 16. Hexahydro-1,3,5-trinitro-1,3,5-triazine (10%) biodegrades to nitroso derivatives, whereas 13% of RDX-N in nitro groups is mineralized to dissolved inorganic N anaerobically by the end of the experiment. The primary identified mineralization end product of RDX (40%) is NH4(+) , generated through either deamination or mono-denitration, followed by ring breakdown. A reasonable production of N2 gas (13%) was seen in the RDX system but not in the TNT system. Sixty-eight percent of the total spiked RDX-N is in an unidentified pool by day 16 and may include

  16. Patterns of d18O in fish tissues in two Oregon Coast range streams

    EPA Science Inventory

    We are using stable isotopes of C, N, O and S (H planned) to study the ecology of coho salmon in streams of the Oregon Coast Range. As part of this work we have examined changes in d18O in coho salmon juveniles (from eggs to smolting) and sculpin (from 0.5 to 20 gm.). For fish...

  17. Calibration of speleothem δ18O records against hydroclimate instrumental records in Central Brazil

    NASA Astrophysics Data System (ADS)

    Moquet, J. S.; Cruz, F. W.; Novello, V. F.; Stríkis, N. M.; Deininger, M.; Karmann, I.; Santos, R. Ventura; Millo, C.; Apaestegui, J.; Guyot, J.-L.; Siffedine, A.; Vuille, M.; Cheng, H.; Edwards, R. L.; Santini, W.

    2016-04-01

    δ18O in speleothems is a powerful proxy for reconstruction of precipitation patterns in tropical and sub-tropical regions. The aim of this study is to calibrate the δ18O record of speleothems against historical precipitation and river discharge data in central Brazil, a region directly influenced by the Southern Atlantic Convergence Zone (SACZ), a major feature of the South American Monsoon System (SAMS). The present work is based on a sub-annual resolution speleothem record covering the last 141 years (the period between the years 1870 and 2011) from a cave in central Brazil. The comparison of this record with instrumental hydroclimate records since 1921 allows defining a strong relationship between precipitation variability and stable oxygen isotope ratios from speleothems. The results from a monitoring program of climatic parameters and isotopic composition of rainfall and cave seepage waters performed in the same cave, show that the rain δ18O variability is dominated by the amount effect in this region, while δ18O drip water remains almost constant over the monitored period (1.5 years). The δ18O of modern calcite, on the other hand, shows clear seasonal variations, with more negative values observed during the rainy season, which implies that other factors also influence the isotopic composition of carbonate. However, the relationship between δ18O of carbonate deposits and rainwater is supported by the results from the comparison between speleothem δ18O records and historical hydroclimate records. A significant correlation between speleothem δ18O and monsoon rainfall variability is observed on sub-decadal time scales, especially for the monsoon period (DJFM and NDJFM), once the rainfall record have been smoothed with a 7-9 years running mean. This study confirms that speleothem δ18O is directly associated with monsoon rainfall variability in central Brazil. The relationship between speleothem δ18O records and hydroclimatic historical records allows

  18. Probing Protein 3D Structures and Conformational Changes Using Electrochemistry-Assisted Isotope Labeling Cross-Linking Mass Spectrometry.

    PubMed

    Zheng, Qiuling; Zhang, Hao; Wu, Shiyong; Chen, Hao

    2016-05-01

    This study presents a new chemical cross-linking mass spectrometry (MS) method in combination with electrochemistry and isotope labeling strategy for probing both protein three-dimensional (3D) structures and conformational changes. For the former purpose, the target protein/protein complex is cross-linked with equal mole of premixed light and heavy isotope labeled cross-linkers carrying electrochemically reducible disulfide bonds (i.e., DSP-d0 and DSP-d8 in this study, DSP = dithiobis[succinimidyl propionate]), digested and then electrochemically reduced followed with online MS analysis. Cross-links can be quickly identified because of their reduced intensities upon electrolysis and the presence of doublet isotopic peak characteristics. In addition, electroreduction converts cross-links into linear peptides, facilitating MS/MS analysis to gain increased information about their sequences and modification sites. For the latter purpose of probing protein conformational changes, an altered procedure is adopted, in which the protein in two different conformations is cross-linked using DSP-d0 and DSP-d8 separately, and then the two protein samples are mixed in 1:1 molar ratio. The merged sample is subjected to digestion and electrochemical mass spectrometric analysis. In such a comparative cross-linking experiment, cross-links could still be rapidly recognized based on their responses to electrolysis. More importantly, the ion intensity ratios of light and heavy isotope labeled cross-links reveal the conformational changes of the protein, as exemplified by examining the effect of Ca(2+) on calmodulin conformation alternation. This new cross-linking MS method is fast and would have high value in structural biology. Graphical Abstract ᅟ.

  19. Probing Protein 3D Structures and Conformational Changes Using Electrochemistry-Assisted Isotope Labeling Cross-Linking Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Zheng, Qiuling; Zhang, Hao; Wu, Shiyong; Chen, Hao

    2016-05-01

    This study presents a new chemical cross-linking mass spectrometry (MS) method in combination with electrochemistry and isotope labeling strategy for probing both protein three-dimensional (3D) structures and conformational changes. For the former purpose, the target protein/protein complex is cross-linked with equal mole of premixed light and heavy isotope labeled cross-linkers carrying electrochemically reducible disulfide bonds (i.e., DSP-d0 and DSP-d8 in this study, DSP = dithiobis[succinimidyl propionate]), digested and then electrochemically reduced followed with online MS analysis. Cross-links can be quickly identified because of their reduced intensities upon electrolysis and the presence of doublet isotopic peak characteristics. In addition, electroreduction converts cross-links into linear peptides, facilitating MS/MS analysis to gain increased information about their sequences and modification sites. For the latter purpose of probing protein conformational changes, an altered procedure is adopted, in which the protein in two different conformations is cross-linked using DSP-d0 and DSP-d8 separately, and then the two protein samples are mixed in 1:1 molar ratio. The merged sample is subjected to digestion and electrochemical mass spectrometric analysis. In such a comparative cross-linking experiment, cross-links could still be rapidly recognized based on their responses to electrolysis. More importantly, the ion intensity ratios of light and heavy isotope labeled cross-links reveal the conformational changes of the protein, as exemplified by examining the effect of Ca2+ on calmodulin conformation alternation. This new cross-linking MS method is fast and would have high value in structural biology.

  20. Leaf morphological effects predict effective path length and enrichment of 18O in leaf water of different Eucalyptus species

    NASA Astrophysics Data System (ADS)

    Kahmen, A.; Merchant, A.; Callister, A.; Dawson, T. E.; Arndt, S. K.

    2006-12-01

    Stable isotopes have been a valuable tool to study water or carbon fluxes of plants and ecosystems. In particular oxygen isotopes18O) in leaf water or plant organic material are now beginning to be established as a simple and integrative measure for plant - water relations. Current δ18O models, however, are still limited in their application to a broad range of different species and ecosystems. It remains for example unclear, if species-specific effects such as different leaf morphologies need to be included in the models for a precise understanding and prediction of δ18O signals. In a common garden experiment (Currency Creek Arboretum, South Australia), where over 900 different Eucalyptus species are cultivated in four replicates, we tested effects of leaf morphology and anatomy on δ18O signals in leaf water of 25 different species. In particular, we determined for all species enrichment in 18O of mean lamina leaf water above source water (Δ18O) as related to leaf physiology as well as leaf thickness, leaf area, specific leaf area and weight and selected anatomical properties. Our data revealed that diurnal Δ18O in leaf water at steady state was significantly different among the investigated species and with differences up to 10% at midday. Fitting factors (effective path length) of leaf water Δ18O models were also significantly different among the investigated species and were highly affected by species-specific morphological parameters. For example, leaf area explained a high percentage of the differences in effective path length observed among the investigated species. Our data suggest that leaf water δ18O can act as powerful tool to estimate plant - water relations in comparative studies but that additional leaf morphological parameters need to be considered in existing δ18O models for a better interpretation of the observed δ18O signals.

  1. Stable Isotope Labeled n-Alkanes to Assess Digesta Passage Kinetics through the Digestive Tract of Ruminants

    PubMed Central

    Warner, Daniel; Ferreira, Luis M. M.; Breuer, Michel J. H.; Dijkstra, Jan; Pellikaan, Wilbert F.

    2013-01-01

    We describe the use of carbon stable isotope (13C) labeled n-alkanes as a potential internal tracer to assess passage kinetics of ingested nutrients in ruminants. Plant cuticular n-alkanes originating from intrinsically 13C labeled ryegrass plants were pulse dosed intraruminally in four rumen-cannulated lactating dairy cows receiving four contrasting ryegrass silage treatments that differed in nitrogen fertilization level (45 or 90 kg nitrogen ha−1) and maturity (early or late). Passage kinetics through the gastrointestinal tract were derived from the δ13C (i.e. the ratio 13C:12C) in apparently undigested fecal material. Isotopic enrichment was observed in a wide range of long-chain n-alkanes (C27–C36) and passage kinetics were determined for the most abundant C29, C31 and C33 n-alkanes, for which a sufficiently high response signal was detected by combustion isotope ratio mass spectrometry. Basal diet treatment and carbon chain length of n-alkanes did not affect fractional passage rates from the rumen (K1) among individual n-alkanes (3.71–3.95%/h). Peak concentration time and transit time showed a quantitatively small, significant (p≤0.002) increase with carbon chain length. K1 estimates were comparable to those of the 13C labeled digestible dry matter fraction (3.38%/h; r = 0.61 to 0.71; p≤0.012). A literature review has shown that n-alkanes are not fermented by microorganisms in the rumen and affirms no preferential depletion of 13C versus 12C. Our results suggest that 13C labeled n-alkanes can be used as nutrient passage tracers and support the reliability of the δ13C signature of digestible feed nutrients as a tool to measure nutrient-specific passage kinetics. PMID:24124493

  2. Mechanistic investigations aided by isotopic labeling. 10. Investigations of novel furan-2,3-dione rearrangements by oxygen-17 labeling

    SciTech Connect

    Kollenz, G.; Sterk, H.; Hutter, G. )

    1991-01-04

    The oxa 1,3-diene moiety in 4-benzoyl-5-phenylfuran-2,3-dione (1) adds aryl isocyanides or heterocumulenes via formal (4 + 1) or (4 + 2) cycloaddition processes. The unstable primary adducts undergo novel furandione rearrangements to intermediates in which the two oxygen atoms of the lactone moiety in (1) are equivalent. This equivalence was confirmed by {sup 17}O-labeling experiments using {sup 17}O NMR spectroscopic and mass spectroscopic measurements. Comparison of the {sup 17}O chemical shifts in (1), labeled either at the benzoyl and ring oxygens (1a-{sup 17}O) or at both exocyclic ring-carbonyl oxygens (1b-{sup 17}O), with those in the products (2-4) confirmed the proposed pathways of these rearrangements. Reactions involving carbodiimides, isocyanates, and ketene imines were investigated.

  3. Holocene record of precipitation seasonality from lake calcite δ18O in the central Rocky Mountains, United States

    USGS Publications Warehouse

    Anderson, Lesleigh

    2011-01-01

    A context for recent hydroclimatic extremes and variability is provided by a ~10 k.y. sediment carbonate oxygen isotope (??18O) record at 5-100 yr resolution from Bison Lake, 3255 m above sea level, in northwestern Colorado (United States). Winter precipitation is the primary water source for the alpine headwater lake in the Upper Colorado River Basin and lake water ??18O measurements reflect seasonal variations in precipitation ??18O. Holocene lake water ??18O variations are inferred from endogenic sedimentary calcite ??18O based on comparisons with historic watershed discharge records and tree-ring reconstructions. Drought periods (i.e., drier winters and/or a more rain-dominated seasonal precipitation balance) generally correspond with higher calcite ??18O values, and vice-versa. Early to middle Holocene ??18O values are higher, implying a rain-dominated seasonal precipitation balance. Lower, more variable ??18O values after ca. 3500 yr ago indicate a snow-dominated but more seasonally variable precipitation balance. The middle to late Holocene ??18O record corresponds with records of El Ni??o Southern Oscillation intensification that supports a teleconnection between Rocky Mountain climate and North Pacific sea-surface temperatures at decade to century time scales. ?? 2011 Geological Society of America.

  4. A precise method for the analysis of d18O of dissolved inorganic phosphate in seawater

    USGS Publications Warehouse

    McLaughlin, K.; Silva, S.; Kendall, C.; Stuart-Williams, Hilary; Paytan, A.

    2004-01-01

    A method for preparation and analysis of the oxygen isotope composition (d18O) of dissolved inorganic phosphate (DIP) has been developed and preliminary results for water samples from various locations are reported. Phosphate is extracted from seawater samples by coprecipitation with magnesium hydroxide. Phosphate is further purified through a series of precipitations and resin separation and is ultimately converted to silver phosphate. Silver phosphate samples are pyrolitically decomposed to carbon monoxide and analyzed for d18O. Silver phosphate samples weighing 0.7 mg (3.5 mol oxygen) can be analyzed routinely with an average standard deviation of about 0.3. There is no isotope fractionation during extraction and blanks are negligible within analytical error. Reproducibility was determined for both laboratory standards and natural samples by multiple analyses. A comparison between filtered and unfiltered natural seawater samples was also conducted and no appreciable difference was observed for the samples tested. The d18O values of DIP in seawater determined using this method range from 18.6 to 22.3, suggesting small but detectable natural variability in seawater. For the San Francisco Bay estuary DIP d18O is more variable, ranging from 11.4 near the San Joaquin River to 20.1 near the Golden Gate Bridge, and was well correlated with salinity, phosphate concentration, and d18O of water.

  5. Development And Evaluation Of Stable Isotope And Fluorescent Labeling And Detection Methodologies For Tracking Injected Bacteria During In Situ Bioremediation

    SciTech Connect

    Mark E. Fuller; Tullis C. Onstott

    2003-12-17

    This report summarizes the results of a research project conducted to develop new methods to label bacterial cells so that they could be tracked and enumerated as they move in the subsurface after they are introduced into the groundwater (i.e., during bioaugmentation). Labeling methods based on stable isotopes of carbon (13C) and vital fluorescent stains were developed. Both approaches proved successful with regards to the ability to effectively label bacterial cells. Several methods for enumeration of fluorescently-labeled cells were developed and validated, including near-real time microplate spectrofluorometry that could be performed in the field. However, the development of a novel enumeration method for the 13C-enriched cells, chemical reaction interface/mass spectrometry (CRIMS), was not successful due to difficulties with the proposed instrumentation. Both labeling methodologies were successfully evaluated and validated during laboratory- and field-scale bacterial transport experiments. The methods developed during this research should be useful for future bacterial transport work as well as other microbial ecology research in a variety of environments. A full bibliography of research articles and meeting presentations related to this project is included (including web links to abstracts and full text reprints).

  6. Late-Pleistocene precipitation δ18O interpolated across the global landmass

    NASA Astrophysics Data System (ADS)

    Jasechko, Scott

    2016-08-01

    Global water cycles, ecosystem assemblages, and weathering rates were impacted by the ˜4°C of global warming that took place over the course of the last glacial termination. Fossil groundwaters can be useful indicators of late-Pleistocene precipitation isotope compositions, which, in turn, can help to test hypotheses about the drivers and impacts of glacial-interglacial climate changes. Here, a global catalog of 126 fossil groundwater records is used to interpolate late-Pleistocene precipitation δ18O across the global landmass. The interpolated data show that extratropical late-Pleistocene terrestrial precipitation was near uniformly depleted in 18O relative to the late Holocene. By contrast, tropical δ18O responses to deglacial warming diverged; late-Pleistocene δ18O was higher-than-modern across India and South China but lower-than-modern throughout much of northern and southern Africa. Groundwaters that recharged beneath large northern hemisphere ice sheets have different Holocene-Pleistocene δ18O relationships than paleowaters that recharged subaerially, potentially aiding reconstructions of englacial transport in paleo ice sheets. Global terrestrial late-Pleistocene precipitation δ18O maps may help to determine 3-D groundwater age distributions, constrain Pleistocene mammal movements, and better understand glacial climate dynamics.

  7. Influence of glacial meltwater on equilibrium process of two Tibetan lakes indicated by δ18O

    NASA Astrophysics Data System (ADS)

    Gao, J.

    2009-12-01

    δ18O measurements based on systematic sampling and isotopic model have been adopted to study the affects of glacial meltwater in two lake basins (Lakes Yamdrok-tso and Puma Yum-tso) at two different elevations on the southern Tibetan Plateau. Temporally, δ18O values in precipitation and lake water display a seasonal fluctuation in both lakes. Spatially, δ18O values in the two lake basins increase by 10‰ from the termini of glaciers to the lake shores, by about 1‰ from the lakeshores to the lake center, by 0.4‰ from the water surface to depth in these lakes. The obvious annual δ18O variations indicate that lake water mixes sufficient in a short time. Model results show that glacial meltwater is an important factor on lake water equilibrium process. Equilibrium δ18O values decrease 0.8‰ for Yamdrok-tso Lake and 0.6‰ for Puma Yum-tso Lake when contributions of glacial meltwater to these lakes shrink by 60%. δ18O increases rapidly during the initial stages and then it takes a long time to approach the equilibrium value. The modeled results also show that the surface lake water temperature has only a little impact on this process.

  8. Regional Scale High Resolution δ18O Prediction in Precipitation Using MODIS EVI

    PubMed Central

    Huang, Cho-Ying; Wang, Chung-Ho; Lin, Shou-De; Lo, Yi-Chen; Huang, Bo-Wen; Hatch, Kent A.; Shiu, Hau-Jie; You, Cheng-Feng; Chang, Yuan-Mou; Shen, Sheng-Feng

    2012-01-01

    The natural variation in stable water isotope ratio data, also known as water isoscape, is a spatiotemporal fingerprint and a powerful natural tracer that has been widely applied in disciplines as diverse as hydrology, paleoclimatology, ecology and forensic investigation. Although much effort has been devoted to developing a predictive water isoscape model, it remains a central challenge for scientists to generate high accuracy, fine scale spatiotemporal water isoscape prediction. Here we develop a novel approach of using the MODIS-EVI (the Moderate Resolution Imagining Spectroradiometer-Enhanced Vegetation Index), to predict δ18O in precipitation at the regional scale. Using a structural equation model, we show that the EVI and precipitated δ18O are highly correlated and thus the EVI is a good predictor of precipitated δ18O. We then test the predictability of our EVI-δ18O model and demonstrate that our approach can provide high accuracy with fine spatial (250×250 m) and temporal (16 days) scale δ18O predictions (annual and monthly predictabilities [r] are 0.96 and 0.80, respectively). We conclude the merging of the EVI and δ18O in precipitation can greatly extend the spatial and temporal data availability and thus enhance the applicability for both the EVI and water isoscape. PMID:23029053

  9. Regional scale high resolution δ18O prediction in precipitation using MODIS EVI.

    PubMed

    Chan, Wei-Ping; Yuan, Hsiao-Wei; Huang, Cho-Ying; Wang, Chung-Ho; Lin, Shou-De; Lo, Yi-Chen; Huang, Bo-Wen; Hatch, Kent A; Shiu, Hau-Jie; You, Cheng-Feng; Chang, Yuan-Mou; Shen, Sheng-Feng

    2012-01-01

    The natural variation in stable water isotope ratio data, also known as water isoscape, is a spatiotemporal fingerprint and a powerful natural tracer that has been widely applied in disciplines as diverse as hydrology, paleoclimatology, ecology and forensic investigation. Although much effort has been devoted to developing a predictive water isoscape model, it remains a central challenge for scientists to generate high accuracy, fine scale spatiotemporal water isoscape prediction. Here we develop a novel approach of using the MODIS-EVI (the Moderate Resolution Imagining Spectroradiometer-Enhanced Vegetation Index), to predict δ(18)O in precipitation at the regional scale. Using a structural equation model, we show that the EVI and precipitated δ(18)O are highly correlated and thus the EVI is a good predictor of precipitated δ(18)O. We then test the predictability of our EVI-δ(18)O model and demonstrate that our approach can provide high accuracy with fine spatial (250×250 m) and temporal (16 days) scale δ(18)O predictions (annual and monthly predictabilities [r] are 0.96 and 0.80, respectively). We conclude the merging of the EVI and δ(18)O in precipitation can greatly extend the spatial and temporal data availability and thus enhance the applicability for both the EVI and water isoscape.

  10. Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT): a novel glycan-relative quantification strategy.

    PubMed

    Walker, S Hunter; Taylor, Amber D; Muddiman, David C

    2013-09-01

    The Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT) strategy for the sample preparation, data analysis, and relative quantification of N-linked glycans is presented. Glycans are derivatized with either natural (L) or stable-isotope labeled (H) hydrazide reagents and analyzed using reversed phase liquid chromatography coupled online to a Q Exactive mass spectrometer. A simple glycan ladder, maltodextrin, is first used to demonstrate the relative quantification strategy in samples with negligible analytical and biological variability. It is shown that after a molecular weight correction attributable to isotopic overlap and a post-acquisition normalization of the data to account for any systematic bias, a plot of the experimental H:L ratio versus the calculated H:L ratio exhibits a correlation of unity for maltodextrin samples mixed in different ratios. We also demonstrate that the INLIGHT approach can quantify species over four orders of magnitude in ion abundance. The INLIGHT strategy is further demonstrated in pooled human plasma, where it is shown that the post-acquisition normalization is more effective than using a single spiked-in internal standard. Finally, changes in glycosylation are able to be detected in complex biological matrices, when spiked with a glycoprotein. The ability to spike in a glycoprotein and detect change at the glycan level validates both the sample preparation and data analysis strategy, making INLIGHT an invaluable relative quantification strategy for the field of glycomics.

  11. Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT): A Novel Glycan-Relative Quantification Strategy

    NASA Astrophysics Data System (ADS)

    Walker, S. Hunter; Taylor, Amber D.; Muddiman, David C.

    2013-09-01

    The Individuality Normalization when Labeling with Isotopic Glycan Hydrazide Tags (INLIGHT) strategy for the sample preparation, data analysis, and relative quantification of N-linked glycans is presented. Glycans are derivatized with either natural (L) or stable-isotope labeled (H) hydrazide reagents and analyzed using reversed phase liquid chromatography coupled online to a Q Exactive mass spectrometer. A simple glycan ladder, maltodextrin, is first used to demonstrate the relative quantification strategy in samples with negligible analytical and biological variability. It is shown that after a molecular weight correction attributable to isotopic overlap and a post-acquisition normalization of the data to account for any systematic bias, a plot of the experimental H:L ratio versus the calculated H:L ratio exhibits a correlation of unity for maltodextrin samples mixed in different ratios. We also demonstrate that the INLIGHT approach can quantify species over four orders of magnitude in ion abundance. The INLIGHT strategy is further demonstrated in pooled human plasma, where it is shown that the post-acquisition normalization is more effective than using a single spiked-in internal standard. Finally, changes in glycosylation are able to be detected in complex biological matrices, when spiked with a glycoprotein. The ability to spike in a glycoprotein and detect change at the glycan level validates both the sample preparation and data analysis strategy, making INLIGHT an invaluable relative quantification strategy for the field of glycomics.

  12. Cellulose δ18O is an index of leaf-to-air vapor pressure difference (VPD) in tropical plants

    PubMed Central

    Kahmen, Ansgar; Sachse, Dirk; Arndt, Stefan K.; Tu, Kevin P.; Farrington, Heraldo; Vitousek, Peter M.; Dawson, Todd E.

    2011-01-01

    Cellulose in plants contains oxygen that derives in most cases from precipitation. Because the stable oxygen isotope composition, δ18O, of precipitation is associated with environmental conditions, cellulose δ18O should be as well. However, plant physiological models using δ18O suggest that cellulose δ18O is influenced by a complex mix of both climatic and physiological drivers. This influence complicates the interpretation of cellulose δ18O values in a paleo-context. Here, we combined empirical data analyses with mechanistic model simulations to i) quantify the impacts that the primary climatic drivers humidity (ea) and air temperature (Tair) have on cellulose δ18O values in different tropical ecosystems and ii) determine which environmental signal is dominating cellulose δ18O values. Our results revealed that ea and Tair equally influence cellulose δ18O values and that distinguishing which of these factors dominates the δ18O values of cellulose cannot be accomplished in the absence of additional environmental information. However, the individual impacts of ea and Tair on the δ18O values of cellulose can be integrated into a single index of plant-experienced atmospheric vapor demand: the leaf-to-air vapor pressure difference (VPD). We found a robust relationship between VPD and cellulose δ18O values in both empirical and modeled data in all ecosystems that we investigated. Our analysis revealed therefore that δ18O values in plant cellulose can be used as a proxy for VPD in tropical ecosystems. As VPD is an essential variable that determines the biogeochemical dynamics of ecosystems, our study has applications in ecological-, climate-, or forensic-sciences. PMID:21245322

  13. Secondary isotope effects in liquid chromatography behaviour of 2H and 3H labelled solutes and solvents.

    PubMed

    Valleix, Alain; Carrat, Sandrine; Caussignac, Céline; Léonce, Estelle; Tchapla, Alain

    2006-05-26

    water in mobile phases of the same composition (%, w/w) is compared. Independent of the nature of the organic modifier (methanol, acetonitrile or ethanol), the effect of replacing H2O with 2H2O in the mobile phase, is an increase in the retention factors and an improvement in the chromatographic resolution of isotopologue pairs. This increase in the resolution is not accompanied by a change in the chromatographic selectivity. The measurement of liquid-liquid extraction coefficients proves that the effect is mainly due to the modification of the phase ratio. In general the effect of 2H-labelled solvents (2H2O and C2H3CN) as mobile phase components, compared to their isotopically non-modified isomers, can be rationalized on the basis of their lower polarisabilities. Overall the use of perdeuterated rather than isotopically non-modified solvents as mobile phase components leads to the most efficient separation systems.

  14. High-frequency observations of δ2H and δ18O in storm rainfall

    NASA Astrophysics Data System (ADS)

    Stoecker, F.; Klaus, J.; Pangle, L. A.; Garland, C.; McDonnell, J. J.

    2012-12-01

    Stable isotopes ratios of hydrogen (2H/1H) and oxygen (18O/16O) are indispensable tools for investigation of the hydrologic cycle. Recent technological advances with laser spectroscopy now enable high-frequency measurement of key water cycle components. While the controls on rainfall isotope composition have been known generally for some time, our understanding of the effect of inter- and intra-storm processes on fine scale rainfall isotope composition is poorly understood. Here we present a new approach to observe inter- and intra-storm isotope variability in precipitation in high-frequency. We investigate the temporal development of δ2H and δ18O within and between discrete rainstorm. δ2H and δ18O in precipitation was measured from November 2011 to February 2012 in Corvallis, OR using a flow-cell combined with a Liquid Water Isotope Analyzer (LWIA-24d, Los Gatos Research, Inc.). The average sample frequency was 15 samples per hour, resulting in more than 3100 samples during the observation period. 27 separate rainstorms were identified in the dataset based on minimum inter-event time, minimum precipitation depth, and minimum number of isotope measurements. Event meteoric water lines were developed for each event. We observed short-term isotopic patterns (e.g., V-shaped trends), high-rate changes (5.3‰/h) and large absolute changes in isotopic composition (20‰) on intra-event scale. V-shaped trends appeared to be related to individual storm fronts detected by air temperature, cloud heights (NEXRAD radar echo tops) and cloud trajectories (Hybrid Single Particle Lagrangian Integrated Trajectory Model (HYSPLIT)). Despite this, we could detect no linear correlation between event-based isotopic variables (slope, δ2H-intercept, δ2H, δ18O) and the event meteoric water line. Furthermore, the composite event meteoric water line (i.e. the local meteoric water line) showed a wider spread for heavy isotopes than for light isotopes, caused presumably by different

  15. δ18O analysis of individual carbohydrates - a new method for GC-pyrolysis-IRMS

    NASA Astrophysics Data System (ADS)

    Lehmann, Marco M.; Fischer, Maria; Zech, Michael; Siegwolf, Rolf T. W.; Saurer, Matthias

    2015-04-01

    Measuring the oxygen isotopic composition (δ18O) of various plant tissues is a widely used tool to investigate biochemical and physiological processes. While we have a good understanding about the hydrological cycle in plants with an evaporative enrichment in 18O in leaf water, we still lack knowledge about the biochemical link between the oxygen atoms in leaf water, leaf assimilates, and stem cellulose and associated isotope fractionations. Especially, the influence of different environmental factors on δ18O of individual carbohydrates (i.e. sugars) and thus on δ18O of cellulose is not fully resolved. A better understanding of these processes may improve climatic reconstructions of tree-ring studies about past environmental conditions. However, further progress in this topic is limited since a precise and reliable method to determine δ18O of individual sugars has not been available yet. With our new approach we attempt to overcome this issue by establishing a new methylation derivatization method suitable for GC-pyrolysis -IRMS. A methyl group (CH3) was thereby added to all hydroxyl groups of a sugar (e.g., glucose, fructose, and sucrose) during a catalyzed one-pot reaction overnight in acetonitrile with methyl iodide (CH3-I) and silver oxide, making them amenable for GC analysis. First results show a very good precision for δ18O of sucrose, but also δ18O of other high-abundant sugars such as glucose and fructose could be measured for the first time. We successfully analyzed a standard mix of all three sugars and determined various other carbohydrates not only related to plant sciences (e.g. mannitol, lactose), showing promising δ18O results. First tests with real plant samples were performed to make this method available for determining δ18O of individual carbohydrates of diverse plant tissues. In future, this new methylation derivatization method should allow us analyzing plant samples of different field sites and of lab experiments to investigate the

  16. A facile method for expression and purification of 15N isotope-labeled human Alzheimer's β-amyloid peptides from E. coli for NMR-based structural analysis

    PubMed Central

    Armand, Tara; Ball, K. Aurelia; Chen, Anna; Pelton, Jeffrey G.; Wemmer, David E.; Head-Gordon, Teresa

    2016-01-01

    Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting millions of people worldwide. AD is characterized by the presence of extracellular plaques composed of aggregated/oligomerized β-amyloid peptides with Aβ42 peptide representing a major isoform in the senile plaques. Given the pathological significance of Aβ42 in the progression of AD, there is considerable interest in understanding the structural ensembles for soluble monomer and oligomeric forms of Aβ42. This report describes an efficient method to express and purify high quality 15N isotope-labeled Aβ42 for structural studies by NMR. The protocol involves utilization of an auto induction system with 15N isotope labeled medium, for high-level expression of Aβ42 as a fusion with IFABP. After the over-expression of the 15N isotope-labeled IFABP-Aβ42 fusion protein in the inclusion bodies, pure 15N isotope-labeled Aβ42 peptide is obtained following a purification method that is streamlined and improved from the method originally developed for the isolation of unlabeled Aβ42 peptide (Garai et al., 2009). We obtain a final yield of ∼6 mg/L culture for 15N isotope-labeled Aβ42 peptide. Mass spectrometry and 1H–15N HSQC spectra of monomeric Aβ42 peptide validate the uniform incorporation of the isotopic label. The method described here is equally applicable for the uniform isotope labeling with 15N and 13C in Aβ42 peptide as well as its other variants including any Aβ42 peptide mutants. PMID:26231074

  17. Determination of protein conformation by isotopically labelled cross-linking and dedicated software

    NASA Astrophysics Data System (ADS)

    Nielsen, Tina; Thaysen-Andersen, Morten; Larsen, Nanna; Jørgensen, Flemming S.; Houen, Gunnar; Højrup, Peter

    2007-12-01

    Chemical cross-linking in conjunction with mass spectrometry (MS) can be used for sensitive and rapid investigation of the three-dimensional structure of proteins at low resolution. However, the resulting data are very complex, and on the bioinformatic side, there still exists an urgent need for improving computer software for (semi-) automated cross-linking data analysis. In this study, we have developed dedicated software for rapid and confident identification and validation of cross-linked species using an isotopic labelled cross-linker approach in combination with MS. Deuterated (+4 Da) and non-deuterated (+0 Da) bis(sulfosuccinimidyl)suberate, BS3, was used as homobifunctional cross-linker to tag the cross-linked regions. Peptides generated from proteolysis were separated using high performance liquid chromatography, and peptide mass fingerprinting was obtained for the individual fractions using matrix-assisted laser-desorption ionisation time-of-flight (MALDI TOF) MS. The resulting peptide mass lists were combined and transferred to the program, ProteinXXX, which generated the theoretical mass values of all combinations of cross-linked peptides and dead-end cross-links and compared this to the obtained mass lists. In addition, screening for 4 Da-separated signals aided the identification of potential cross-linked species. Sequence information of these candidates was then obtained using MALDI TOF TOF. The cross-linked peptides could then be validated based on the match of the fragmentation pattern and the theoretical values produced by ProteinXXX. This semi-automated interpretation provided a high analysis speed of cross-linking data, with efficient and confident identification of cross-linked species. Four experiments using different conditions showed a high degree of reproducibility as only 1 and 2 cross-links out of 36 identified was not observed in two experiments. The method was tested using human placenta calreticulin (CRT). Based on the identified cross

  18. Determining the Composition and Stability of Protein Complexes Using an Integrated Label-Free and Stable Isotope Labeling Strategy

    PubMed Central

    Greco, Todd M.; Guise, Amanda J.; Cristea, Ileana M.

    2016-01-01

    In biological systems, proteins catalyze the fundamental reactions that underlie all cellular functions, including metabolic processes and cell survival and death pathways. These biochemical reactions are rarely accomplished alone. Rather, they involve a concerted effect from many proteins that may operate in a directed signaling pathway and/or may physically associate in a complex to achieve a specific enzymatic activity. Therefore, defining the composition and regulation of protein complexes is critical for understanding cellular functions. In this chapter, we describe an approach that uses quantitative mass spectrometry (MS) to assess the specificity and the relative stability of protein interactions. Isolation of protein complexes from mammalian cells is performed by rapid immunoaffinity purification, and followed by in-solution digestion and high-resolution mass spectrometry analysis. We employ complementary quantitative MS workflows to assess the specificity of protein interactions using label-free MS and statistical analysis, and the relative stability of the interactions using a metabolic labeling technique. For each candidate protein interaction, scores from the two workflows can be correlated to minimize nonspecific background and profile protein complex composition and relative stability. PMID:26867737

  19. Stable isotope-labeled vitamin D, metabolites and chemical analogs: Synthesis and use in mass spectrometric studies

    SciTech Connect

    Coldwell, R.D.; Trafford, D.J.; Varley, M.J.; Kirk, D.N.; Makin, H.L. )

    1990-10-01

    Methods for the measurement of vitamin D and its metabolites using stable isotope-labeled internal standards and mass spectrometry are reviewed. The synthesis of both labeled and unlabeled standards is illustrated, and details of the synthesis of (26,26,27,27,27(-2)H5)-25,26-dihydroxyvitamin D3 and (28,28,28(-2)H3)-24,25-dihydroxyvitamin D2 are given. The use of in vitro biologic systems for the production of further metabolites of deuterated 25-hydroxyvitamin D3 is discussed. Use of deuterated 25-hydroxydihydrotachysterol3 as a substrate in the isolated perfused rat kidney has provided valuable data for the assignment of structure to a number of metabolites of 25-hydroxydihydrotachysterol3 formed in this system. 51 refs.

  20. Temporal and spatial distributions of δ18O and δ2H in precipitation in Romania

    NASA Astrophysics Data System (ADS)

    Nagavciuc, Viorica; Bădăluță, Carmen-Andreea; Perșoiu, Aurel

    2015-04-01

    Stable isotope ratios of meteoric water have an important role in climatic, paleoclimatic, hydrological and meteorological studies. While such data are available from most of Europe, so far, in Romania (East Central Europe), no systematic study of the stable isotopic composition of precipitation exists. In this context, the aim of this study is to analyze the isotopic composition of rainwater, its temporal and spatial distribution, the identification of the main factors influencing these variations and the creation of the first map of spatial distribution of stable isotopes in precipitation in Romania. Between March 2012 and March 2014 we have collected monthly samples from 22 stations in Romania, which were subsequently analyzed for their δ18O and δ2H at the Stable Isotopes Laboratory, Stefan cel Mare University, Suceava, Romania. Precipitation in W and NW Romania plot along the GMWL, while those in the East are slightly below it, on an evaporative trend. The LMWL for Romania is defined as δ2H=7,27*δ18O + 6,92. The W-E gradient in the distribution of δ18O and δ2H are less marked than the N-S ones, with local influences dominating in areas of strong evaporation (intramountain basins, rain-shadow areas etc). In SW, and especially in autumn and winter, Meditteranean cyclones carry moisture from the Eastern Mediterranean, the δ18O and δ2H values in precipitation in the area plotting between the GMWL and the Eastern Mediterranean Meteoric Water Line. The isotopic composition of rainwater in Romania correlates well with air temperature, and is influenced to a lesser extent by other factors such as the amount of precipitation, topography configuration, the effect of continentalism and season of the year.

  1. A Review of Climatic Controls on δ18o in Precipitation over the Tibetan Plateau: Observations and Simulations

    NASA Astrophysics Data System (ADS)

    Yao, T.; Masson-Delmotte, V.; Gao, J.; Risi, C. M.

    2014-12-01

    Located at the convergence of air masses between the westerlies and monsoon, the Tibetan Plateau (TP) undergoes complex water cycle processes, which need to be documented and understood through a combination of variant methodologies. The stable oxygen isotope ratio (δ18O) in precipitation is an integrated tracer of the atmospheric processes and has been used worldwide. Since the 1990s, an intensive effort has been dedicated to studying precipitation isotopic composition at more than 20 stations in the TP. Based on these observations, we establish a database of precipitation δ18O and use different models to evaluate the climatic drivers of present-day precipitation δ18O over the TP. The spatial and temporal patterns of precipitation δ18O and their relationships with temperature and precipitation reveal three distinct domains, respectively associated with the influence of the westerlies (Northern TP), Indian monsoon (Southern TP) and transition in between. The seasonal patterns of precipitation δ18O are diverse in different domains. High-resolution atmospheric models equipped with stable isotopes capture the spatial and temporal patterns of precipitation δ18O and their relationships with moisture transport from the westerlies and Indian monsoon. Only in the westerlies domain are atmospheric models able to represent qualitatively and quantitatively the relationships between climate and precipitation δ18O. More significant temperature effect exists when either the westerlies or Indian monsoon is the sole dominant atmospheric process. The observed and simulated altitude-δ18O relationships strongly depend on the season and the domain (monsoon or westerlies). Our results have crucial implications for the interpretation of the abundant stable isotope information derived from natural climatic archives over the TP such as ice cores, lake sediments or tree rings, and for the application of atmospheric simulations to quantifying paleo-climate and paleo

  2. Reconstructing relative humidity from plant δ18O and δD as deuterium deviations from the global meteoric water line.

    EPA Science Inventory

    Cellulose δ18O and δD in preserved plant material can provide insights on climates and hydrological cycling in the distant past. However, most studies of plant cellulose have used only one isotope, most commonly δ18O, resulting in difficulties partitioning variation between chang...

  3. O18O and C18O observations of ρ Ophiuchi A

    NASA Astrophysics Data System (ADS)

    Liseau, R.; Larsson, B.; Bergman, P.; Pagani, L.; Black, J. H.; Hjalmarson, Å.; Justtanont, K.

    2010-02-01

    Context. Contrary to theoretical expectation, surprisingly low concentrations of molecular oxygen, O2, have been found in the interstellar medium. Telluric absorption makes ground based O2 observations essentially impossible and observations had to be done from space. Millimetre-wave telescopes on space platforms were necessarily small, which resulted in large, several arcminutes wide, beam patterns. Observations of the (NJ = 11-10) ground state transition of O2 with the Odin satellite resulted in a ≳ 5σ detection toward the dense core ρ {Oph A}. At the frequency of the line, 119 GHz, the Odin telescope has a beam width of 10', larger than the size of the dense core. Aims: The precise nature of the emitting source and its exact location and extent are therefore unknown. The current investigation is intended to remedy this. Methods: Although the Earth's atmosphere is entirely opaque to low-lying O2 transitions, it allows ground based observations of the much rarer 16O18O in favourable conditions and at much higher angular resolution with larger telescopes. In addition, ρ {Oph A} exhibits both multiple radial velocity systems and considerable velocity gradients. Extensive mapping of the region in the proxy C18O (J = 3-2) line can be expected to help identify the O2 source on the basis of its line shape and Doppler velocity. Line opacities were determined from observations of optically thin 13C18O (J = 3-2). During several observing periods, two C18O intensity maxima in ρ {Oph A} were searched for O18O in the (21-01) line at 234 GHz with the 12 m APEX telescope. These positions are associated also with peaks in the mm-continuum emission from dust. Results: Our observations resulted in an upper limit on the integrated O18O intensity of int T*A d\\upsilon < 0.01 K km s-1 (3 σ) into the 26.6 arcsec beam. Together with the C18O data, this leads to a ratio of N(C18O)/N(O18O) > 16. Combining Odin's O2 with the present O18O observations we infer an O2 abundance 5

  4. Seasonal 18O variations and groundwater recharge for three landscape types in central Pennsylvania, USA

    NASA Astrophysics Data System (ADS)

    O'Driscoll, M. A.; DeWalle, D. R.; McGuire, K. J.; Gburek, W. J.

    2005-03-01

    Seasonal 18O variations in precipitation, soil water, snowmelt, spring flow and stream baseflow were analyzed to characterize seasonal dynamics of groundwater recharge in three central Pennsylvania catchments. The catchments represented three common landscape types: Valley and Ridge-shale (Mahantango Creek), Valley and Ridge-carbonate (Buffalo Run), and Appalachian Plateau-sandstone (Benner Run). Samples were collected on a biweekly basis from May 18, 1999 to May 9, 2000. Precipitation, soil water, and baseflow isotopic composition data indicated that a seasonal recharge bias existed for these catchments, most recharge occurred in the fall, winter, and spring months. An altitude effect of -0.16 to -0.32‰/100 m change in elevation was discernible in precipitation, soil water, and stream baseflow isotopic compositions. Soils effectively damped seasonal variations of recharge 18O composition after depths of 1.62-2.85 m. The greatest damping of the annual isotopic composition signal occurred in the shallow soil layers (0-15 cm). In these and similar landscapes with thick soils the annual isotopic composition signal may be completely damped prior to reaching the stream as baseflow. Isotopic variations measured in stream baseflow are more likely to be caused by the shallow flowpath water relatively close to the streams. Baseflow stable isotope variations found on the basins studied suggested that residence times for subsurface waters to reach channels were much longer than the annual seasonal cycle of 18O in precipitation. Damping depths were similar for the three different catchments but it is not certain how spatially variable damping depths were within each catchment. This information would be useful in determining areas within catchments that contribute to short term isotopic composition fluctuations within streams ('new water'). Predictive models that determine isotopic damping depth from meteorological, soil and vegetation/land-use data can help develop a better

  5. Characterization of surface and ground water δ18O seasonal variation and its use for estimating groundwater residence times

    USGS Publications Warehouse

    Reddy, Michael M.; Schuster, Paul; Kendall, Carol; Reddy, Micaela B.

    2006-01-01

    18O is an ideal tracer for characterizing hydrological processes because it can be reliably measured in several watershed hydrological compartments. Here, we present multiyear isotopic data, i.e. 18O variations (δ18O), for precipitation inputs, surface water and groundwater in the Shingobee River Headwaters Area (SRHA), a well-instrumented research catchment in north-central Minnesota. SRHA surface waters exhibit δ18O seasonal variations similar to those of groundwaters, and seasonal δ18O variations plotted versus time fit seasonal sine functions. These seasonal δ18O variations were interpreted to estimate surface water and groundwater mean residence times (MRTs) at sampling locations near topographically closed-basin lakes. MRT variations of about 1 to 16 years have been estimated over an area covering about 9 km2 from the basin boundary to the most downgradient well. Estimated MRT error (±0·3 to ±0·7 years) is small for short MRTs and is much larger (±10 years) for a well with an MRT (16 years) near the limit of the method. Groundwater transit time estimates based on Darcy's law, tritium content, and the seasonal δ18O amplitude approach appear to be consistent within the limits of each method. The results from this study suggest that use of the δ18O seasonal variation method to determine MRTs can help assess groundwater recharge areas in small headwaters catchments.

  6. Characterization of surface and ground water δ18O seasonal variation and its use for estimating groundwater residence times

    USGS Publications Warehouse

    Reddy, Michael M.; Schuster, Paul F.; Kendall, Carol; Reddy, Micaela B.

    2006-01-01

    18O is an ideal tracer for characterizing hydrological processes because it can be reliably measured in several watershed hydrological compartments. Here, we present multiyear isotopic data, i.e. 18O variations (δ18O), for precipitation inputs, surface water and groundwater in the Shingobee River Headwaters Area (SRHA), a well-instrumented research catchment in north-central Minnesota. SRHA surface waters exhibit δ18O seasonal variations similar to those of groundwaters, and seasonal δ18O variations plotted versus time fit seasonal sine functions. These seasonal δ18O variations were interpreted to estimate surface water and groundwater mean residence times (MRTs) at sampling locations near topographically closed-basin lakes. MRT variations of about 1 to 16 years have been estimated over an area covering about 9 km2 from the basin boundary to the most downgradient well. Estimated MRT error (±0·3 to ±0·7 years) is small for short MRTs and is much larger (±10 years) for a well with an MRT (16 years) near the limit of the method. Groundwater transit time estimates based on Darcy's law, tritium content, and the seasonal δ18O amplitude approach appear to be consistent within the limits of each method. The results from this study suggest that use of the δ18O seasonal variation method to determine MRTs can help assess groundwater recharge areas in small headwaters catchments.

  7. Latitudinal gradients in greenhouse seawater δ(18) O: evidence from Eocene sirenian tooth enamel.

    PubMed

    Clementz, Mark T; Sewall, Jacob O

    2011-04-22

    The Eocene greenhouse climate state has been linked to a more vigorous hydrologic cycle at mid- and high latitudes; similar information on precipitation levels at low latitudes is, however, limited. Oxygen isotopic fluxes track moisture fluxes and, thus, the δ(18)O values of ocean surface waters can provide insight into hydrologic cycle changes. The offset between tropical δ(18)O values from sampled Eocene sirenian tooth enamel and modern surface waters is greater than the expected 1.0 per mil increase due to increased continental ice volume. This increased offset could result from suppression of surface-water δ(18)O values by a tropical, annual moisture balance substantially wetter than that of today. Results from an atmospheric general circulation model support this interpretation and suggest that Eocene low latitudes were extremely wet.

  8. Evaluation of stereo-array isotope labeling (SAIL) patterns for automated structural analysis of proteins with CYANA.

    PubMed

    Ikeya, Teppei; Terauchi, Tsutomu; Güntert, Peter; Kainosho, Masatsune

    2006-07-01

    Recently we have developed the stereo-array isotope labeling (SAIL) technique to overcome the conventional molecular size limitation in NMR protein structure determination by employing complete stereo- and regiospecific patterns of stable isotopes. SAIL sharpens signals and simplifies spectra without the loss of requisite structural information, thus making large classes of proteins newly accessible to detailed solution structure determination. The automated structure calculation program CYANA can efficiently analyze SAIL-NOESY spectra and calculate structures without manual analysis. Nevertheless, the original SAIL method might not be capable of determining the structures of proteins larger than 50 kDa or membrane proteins, for which the spectra are characterized by many broadened and overlapped peaks. Here we have carried out simulations of new SAIL patterns optimized for minimal relaxation and overlap, to evaluate the combined use of SAIL and CYANA for solving the structures of larger proteins and membrane proteins. The modified approach reduces the number of peaks to nearly half of that observed with uniform labeling, while still yielding well-defined structures and is expected to enable NMR structure determinations of these challenging systems.

  9. A software toolkit and interface for performing stable isotope labeling and top3 quantification using Progenesis LC-MS.

    PubMed

    Qi, Da; Brownridge, Philip; Xia, Dong; Mackay, Katherine; Gonzalez-Galarza, Faviel F; Kenyani, Jenna; Harman, Victoria; Beynon, Robert J; Jones, Andrew R

    2012-09-01

    Numerous software packages exist to provide support for quantifying peptides and proteins from mass spectrometry (MS) data. However, many support only a subset of experimental methods or instrument types, meaning that laboratories often have to use multiple software packages. The Progenesis LC-MS software package from Nonlinear Dynamics is a software solution for label-free quantitation. However, many laboratories using Progenesis also wish to employ stable isotope-based methods that are not natively supported in Progenesis. We have developed a Java programming interface that can use the output files produced by Progenesis, allowing the basic MS features quantified across replicates to be used in a range of different experimental methods. We have developed post-processing software (the Progenesis Post-Processor) to embed Progenesis in the analysis of stable isotope labeling data and top3 pseudo-absolute quantitation. We have also created export ability to the new data standard, mzQuantML, produced by the Proteomics Standards Initiative to facilitate the development and standardization process. The software is provided to users with a simple graphical user interface for accessing the different features. The underlying programming interface may also be used by Java developers to develop other routines for analyzing data produced by Progenesis.

  10. Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay

    PubMed Central

    Bhate, Manasi P.; Wylie, Benjamin J.; Thompson, Ameer; Tian, Lin; Nimigean, Crina; McDermott, Ann E.

    2013-01-01

    We report the expression, purification, liposome reconstitution and functional validation of uniformly 13C and 15N isotope labeled KcsA, a bacterial potassium channel that has high homology with mammalian channels, for solid-state NMR studies. The expression and purification is optimized for an average yield of ~ 35–40 milligrams per liter of M9 media in a time-efficient way. The protein purity is confirmed by gel electrophoresis and the protein concentration is quantified by UV-Vis absorption spectroscopy. Protocols to efficiently reconstitute KcsA into liposomes are also presented. The presence of liposomes is confirmed by cryo-electron microscopy images and the effect of magic angle spinning on liposome packing is shown. High-resolution solid-state NMR spectra of uniformly isotope labeled KcsA in these liposomes reveal that our protocol yields to a very homogenous KcsA sample with high signal to noise and several well-resolved residues in NMR spectra. Electrophysiology of our samples before and after solid-state NMR show that channel function and selectivity remain intact after the solid-state NMR. PMID:23916531

  11. Discovery of histone modification crosstalk networks by stable isotope labeling of amino acids in cell culture mass spectrometry (SILAC MS).

    PubMed

    Guan, Xiaoyan; Rastogi, Neha; Parthun, Mark R; Freitas, Michael A

    2013-08-01

    In this paper we describe an approach that combines stable isotope labeling of amino acids in cells culture, high mass accuracy liquid chromatography tandem mass spectrometry and a novel data analysis approach to accurately determine relative peptide post-translational modification levels. This paper describes the application of this approach to the discovery of novel histone modification crosstalk networks in Saccharomyces cerevisiae. Yeast histone mutants were generated to mimic the presence/absence of 44 well-known modifications on core histones H2A, H2B, H3, and H4. In each mutant strain the relative change in H3 K79 methylation and H3 K56 acetylation were determined using stable isotope labeling of amino acids in cells culture. This approach showed relative changes in H3 K79 methylation and H3 K56 acetylation that are consistent with known histone crosstalk networks. More importantly, this study revealed additional histone modification sites that affect H3 K79 methylation and H3 K56 acetylation.

  12. French summer droughts since 1326 AD: a reconstruction based on tree ring cellulose δ18O

    NASA Astrophysics Data System (ADS)

    Labuhn, I.; Daux, V.; Girardclos, O.; Stievenard, M.; Pierre, M.; Masson-Delmotte, V.

    2015-11-01

    The reconstruction of droughts is essential for the understanding of past drought dynamics, and can help evaluate future drought scenarios in a changing climate. This article presents a reconstruction of summer droughts in France based on annually resolved, absolutely dated chronologies of oxygen isotope ratios (δ18O) in tree ring cellulose from Quercus spp. Samples were taken from living trees and timber wood from historic buildings at two sites: Fontainebleau (48° 23' N, 2° 40' E; 1326-2000 AD) and Angoulême (45° 44' N, 0° 18' E; 1360-2004 AD). Cellulose δ18O from these sites proved to be a good proxy of summer climate, as the trees were sensitive to temperature and moisture availability. However, offsets in average δ18O values between tree cohorts necessitated a correction before joining them to the final chronologies. Using the corrected δ18O chronologies, we developed models based on linear regression to reconstruct drought, expressed by the standardized precipitation evapotranspiration index (SPEI). The significant correlations between the SPEI and cellulose δ18O (r ≈ -0.70), as well as the verification of the models by independent data support the validity of these reconstructions. At both sites, recent decades are characterized by increasing drought. Fontainebleau displays dominantly wetter conditions during earlier centuries, whereas the current drought intensity is not unprecedented in the Angoulême record. While the δ18O chronologies at the two studied sites are highly correlated during the 19th and 20th century, there is a significant decrease in the correlation coefficient between 1550 and 1800 AD, which indicates either a weaker climate sensitivity of the tree ring proxies during this period, or a more heterogeneous climate in the north and the south of France. Future studies of tree ring isotope networks might reveal if the seasonality and spatial patterns of past droughts can explain this decoupling.

  13. Synthesis of isotopically labeled versions of L-MTP-PE (mifamurtide) and MDP.

    PubMed

    Li, Yuexian; Plesescu, Mihaela; Prakash, Shimoga R

    2013-01-01

    L-MTP-PE (1), an immunomodulator and its metabolite MDP (4) were synthesized from labeled l-alanine and its protected derivative, respectively. The key intermediate product for the labeled L-MTP-PE synthesis, [(13) C3 ,D4 ]-alanyl-cephalin (2A), was synthesized from [(13) C3 ,D4 ]-l-alanine (3A) in three steps. The key intermediate product for labeled MDP synthesis, amine 11, was prepared from [(13) C3 ,(15) N]-Boc-l-alanine (5A) in two steps.

  14. Discovery and microassay of a nitrite-dependent carbonic anhydrase activity by stable-isotope dilution gas chromatography-mass spectrometry.

    PubMed

    Zinke, Maximilian; Hanff, Erik; Böhmer, Anke; Supuran, Claudiu T; Tsikas, Dimitrios

    2016-01-01

    The intrinsic activity of carbonic anhydrase (CA) is the hydration of CO2 to carbonic acid and its dehydration to CO2. CA may also function as esterase and phosphatase. Recently, we demonstrated that renal CA is mainly responsible for the reabsorption of nitrite (NO2(-)) which is the most abundant reservoir of the biologically highly potent nitric oxide (NO). By means of a stable-isotope dilution GC-MS method, we discovered a novel CA activity which strictly depends upon nitrite. We found that bovine erythrocytic CAII (beCAII) catalyses the incorporation of (18)O from H2 (18)O into nitrite at pH 7.4. After derivatization with pentafluorobenzyl bromide, gas chromatographic separation and mass spectrometric analysis, we detected ions at m/z 48 for singly (18)O-labelled nitrite ((16)O=N-(18)O(-)/(18)O=N-(16)O(-)) and at m/z 50 for doubly (18)O-labelled nitrite ((18)O=N-(18)O(-)) in addition to m/z 46 for unlabelled nitrite. Using (15)N-labelled nitrite ((15)NO2 (-), m/z 47) as an internal standard and selected-ion monitoring of m/z 46, m/z 48, m/z 50 and m/z 47, we developed a GC-MS microassay for the quantitative determination of the nitrite-dependent beCAII activity. The CA inhibitors acetazolamide and FC5 207A did not alter beCAII-catalysed formation of singly and doubly (18)O-labelled nitrite. Cysteine and the experimental CA inhibitor DIDS (a diisothiocyanate) increased several fold the beCAII-catalysed formation of the (18)O-labelled nitrite species. Cysteine, acetazolamide, FC5 207A, and DIDS by themselves had no effect on the incorporation of (18)O from H2 (18)O into nitrite. We conclude that erythrocytic CA possesses a nitrite-dependent activity which can only be detected when nitrite is used as the substrate and the reaction is performed in buffers of neutral pH values prepared in H2 (18)O. This novel CA activity, i.e., the nitrous acid anhydrase activity, represents a bioactivation of nitrite and may have both beneficial (via S-nitrosylation and subsequent

  15. From position-specific isotope labeling towards soil fluxomics: a novel toolbox to assess the microbial impact on biogeochemical cycles

    NASA Astrophysics Data System (ADS)

    Apostel, C.; Dippold, M. A.; Kuzyakov, Y.

    2015-12-01

    Understanding the microbial impact on C and nutrient cycles is one of the most important challenges in terrestrial biogeochemistry. Transformation of low molecular weight organic substances (LMWOS) is a key step in all biogeochemical cycles because 1) all high molecular substances pass the LMWOS pool during their degradation and 2) only LMWOS can be taken up by microorganisms intact. Thus, the transformations of LMWOS are dominated by biochemical pathways of the soil microorganisms. Thus, understanding fluxes and transformations in soils requires a detailed knowledge on the microbial metabolic network and its control mechanism. Tracing C fate in soil by isotopes became on of the most applied and promising biogeochemistry tools but studies were nearly exclusively based on uniformly labeled substances. However, such tracers do not allow the differentiation of the intact use of the initial substances from its transformation to metabolites. The novel tool of position-specific labeling enables to trace molecule atoms separately and thus to determine the cleavage of molecules - a prerequisite for metabolic tracing. Position-specific labeling of basic metabolites and quantification of isotope incorporation in CO2 and bulk soil enabled following the basic metabolic pathways of microorganisms. However, the combination of position-specific 13C labeling with compound-specific isotope analysis of microbial biomarkers and metabolites like phospholipid fatty acids (PLFA) or amino sugars revealed new insights into the soil fluxome: First, it enables tracing specific anabolic pathways in diverse microbial communities in soils e.g. carbon starvation pathways versus pathways reflecting microbial growth. Second, it allows identification of specific pathways of individual functional microbial groups in soils in situ. Tracing metabolic pathways and understanding their regulating factors are crucial for soil C fluxomics i.e. the unravaling of the complex network of C transformations

  16. Production and application of high quality stable isotope-labeled human immunoglobulin G1 for mass spectrometry analysis.

    PubMed

    Phillip, Amsler; Thierry, Wolf; Christian, Lanshoeft; Anja, Bettighofer; Jochen, Eisfeld; Thomas, Moenius; Claudia, Probst; Coralie, Etter; Olivier, Heudi

    2016-12-12

    Here, we describe the production of stable isotope-labeled human immunoglobulin G1 ([(13) C]-hIgG1) using [(13) C]-L-lysine/arginine-labeled hIgG1. The fermentation process was run in shake flasks containing labeled arginine and lysinethat were incorporated into the produced recombinant hIgG1. The [(13) C]-hIgG1 was purified, and label incorporation was determined to be >99% at all lysine and arginine moieties. Sequence coverage was confirmed by peptide mapping. [(13) C]-hIgG1 was then used as an internal standard (IS) for the development of a liquid chromatography-tandem mass spectrometry method applicable to the quantitative analysis of all human types of hIgG1 in rat serum. Four conserved peptides, namely, GPSVFPLAPSSK, TTPPVLDSDGSFFLYSK, VVSVLTVLHQDWLNGK, and FNWYVDGVEVHNAK, originating from different parts of the fraction crystallizable region of hIgG1, were used for quantitation of hIgG1 in rat serum. The calibration curves with a coefficient of determination (r(2) ) between 0.9950 and 0.9962 resulting from the peak area ratio of each peptide to its respective labeled IS were reproducible. A mean bias within ±20.0% of the nominal values and a precision of ≤20.0 % were obtained for the calibration standards and quality control samples for each peptide. [(13) C]-hIgG1 was shown as a suitable IS for quantitative hIgG1 analysis in preclinical species by LC-MS/MS.

  17. Assessing modern climatic controls on southern Sierra Nevada precipitation and speleothem δ18O

    NASA Astrophysics Data System (ADS)

    McCabe-Glynn, S. E.; Johnson, K. R.; Berkelhammer, M. B.

    2012-12-01

    Precipitation in the southwestern United States (SW US) is highly seasonal and exhibits inter-annual to inter-decadal variability. A 1154-year δ18O time series obtained from a southwestern Sierra Nevada Mountain stalagmite from Crystal Cave, CRC-3, (36.58°N; 118.56°W; 1540 m) reveals substantial decadal to multi-decadal variability closely linked to the Pacific Decadal Oscillation (PDO), and more specifically, to sea surface temperatures (SSTs) in the Kuroshio Extension region, which impact the atmospheric trajectory and isotopic composition of moisture reaching the study site. The instrumental portion of the CRC-3 δ18O time series suggests that more negative precipitation δ18O values are delivered from higher latitudes during positive phases of the PDO and/or when SSTs in the Kuroshio Extension region are anomalously cool, such as during La Niña events. In order to improve our understanding of the controls on speleothem δ18O in this region, we have conducted a detailed modern study of the climate, hydrology, and stable isotopic composition of meteoric waters (precipitation and drip water) at the cave. Here we present Crystal Cave drip logger results from 2010 to 2012, the isotopic composition of North American Deposition Program precipitation samples collected from 2001 to 2012 from several locations near our site including Ash Mountain (ASM), Sequoia National Park-Giant Forest (Ca75), and Yosemite National Park (Ca99), and isotopic composition of cave drip water and glass plate calcite. We also compare the δ18O values in the precipitation to satellite imagery, NCAR/NCEP data, and NOAA Hysplit Model backward trajectories between the sites. Results indicate that this site is particularly sensitive to "Pineapple Express" type storms, a persistent flow of atmospheric moisture and heavy rainfall extending from near the Hawaiian Islands to the coast of North America, which average about twice as much precipitation as other storms in the Sierra Nevada during

  18. Synthesis of isotopically labeled R- or S-[.sup.13C, .sup.2H] glycerols

    DOEpatents

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

    2008-01-22

    The present invention is directed to asymmetric chiral labeled glycerols including at least one chiral atom, from one to two .sup.13C atoms and from zero to four deuterium atoms bonded directly to a carbon atom, e.g., (2S) [1,2-.sup.13C.sub.2]glycerol and (2R) [1,2-.sup.13C.sub.2]glycerol, and to the use of such chiral glycerols in the preparation of labeled amino acids.

  19. Counter-diffusion of isotopically labeled trichloroethylene in silica gel and geosorbent micropores: Model development

    SciTech Connect

    McMillan, S.A.; Werth, C.J.

    1999-07-01

    A new model was developed to determine if reduced uptake rates observed during isotope exchange experiments could plausibly be attributed to sterically hindered counter-diffusion in one-dimensional micropores. During exchange experiments, hydrogenated trichloroethylene ({sup 1}HTCE) was displaced with deuterated TCE (DTCE) in the slow-desorbing sites of a silica gel, a groundwater sediment, and a clay and silt fraction. To describe this process, the model accounts for co- and counter-diffusion of TCE isotopes in one-dimensional micropores, where each micropore type is defined by a single codiffusion rate constant and a single counter-diffusion rate constant. For silica gel, isotope exchange was simulated in a single micropore type. For geosorbents, isotope exchange was simulated in a distribution of micropore types characterized by a {gamma} distribution of diffusion rate constants. Simulation results indicate that (1) the proposed model accounts for the mechanisms controlling isotope exchange in the silica gel and the groundwater sediment and (2) the rate of counter-diffusion is up to 6 times slower than the rate of codiffusion. This suggests that steric hindrance between counter-diffusing sorbates can significantly affect mass transfer and, consequently, the transport of chemical mixtures in the subsurface.

  20. 13CO/C18O Gradients across the Disks of Nearby Spiral Galaxies

    NASA Astrophysics Data System (ADS)

    Jiménez-Donaire, María J.; Cormier, Diane; Bigiel, Frank; Leroy, Adam K.; Gallagher, Molly; Krumholz, Mark R.; Usero, Antonio; Hughes, Annie; Kramer, Carsten; Meier, David; Murphy, Eric; Pety, Jérôme; Schinnerer, Eva; Schruba, Andreas; Schuster, Karl; Sliwa, Kazimierz; Tomicic, Neven

    2017-02-01

    We use the IRAM Large Program EMPIRE and new high-resolution ALMA data to measure 13CO(1-0)/C18O(1-0) intensity ratios across nine nearby spiral galaxies. These isotopologues of 12CO are typically optically thin across most of the area in galaxy disks, and this ratio allows us to gauge their relative abundance due to chemistry or stellar nucleosynthesis effects. Resolved 13CO/C18O gradients across normal galaxies have been rare due to the faintness of these lines. We find a mean 13CO/C18O ratio of 6.0 ± 0.9 for the central regions of our galaxies. This agrees well with results in the Milky Way, but differs from results for starburst galaxies (3.4 ± 0.9) and ultraluminous infrared galaxies (1.1 ± 0.4). In our sample, the 13CO/C18O ratio consistently increases with increasing galactocentric radius and decreases with increasing star formation rate surface density. These trends could be explained if the isotopic abundances are altered by fractionation; the sense of the trends also agrees with those expected for carbon and oxygen isotopic abundance variations due to selective enrichment by massive stars.

  1. High resolution δ17O-δ18O as a single mineral thermometer

    NASA Astrophysics Data System (ADS)

    Sharp, Z. D.; Sengupta, S.; Pack, A.

    2014-12-01

    The equilibrium relationship α17O/16Oa-b = (α18O/16Oa-b)θ makes the analysis of δ17O redundant for most terrestrial applications. However the θ term varies with temperature, so that ultra-high precision δ17O data provide additional information not available from δ18O alone. If the δ18O and δ17O values of formation water covary in a known way (e.g., meteoric water, ocean water), then a unique solution for both temperature and the δ18O of the formation fluids can be obtained from the combined δ18O-δ17O mineral values. The paired δ18O-δ17O values are in essence a single mineral thermometer. Unlike clumped isotopes or combined δ18O-δD data, the δ18O and δ17O values of a mineral have identical 'diagenetic potential', and will only be altered with a high F/R ratio. We have made an empirical determination of the temperature dependence on θ = -710/T2 + 0.5305 using Pleistocene diatom data from ODP Leg 177, Site 1093 (δ18O = 39.610, δ17O = 20.536‰), which is almost identical to Pack and Herwartz (EPSL, 2014). Application to ancient cherts gives the following results: The δ18O-δ17O values of cherts vary systematically with age, from Archean to Proterozoic to Phanerozoic. The Archean cherts are incompatible with modern seawater under any temperature conditions. Instead they have equilibrated with water of δ18O= -10±3 (‰ vs SMOW) at 50 to 70°C. These data support a lighter ocean in the Archean by ~5‰. Proterozoic cherts equilibrated at 35-50°C with meteoric water of -8±3‰ and Phanerozoic cherts equilibrated with mixed meteoric water/ocean water at similar temperatures and higher δ18O values (-3±3‰). The δ18O values of lacustrine diatoms from the Valles Caldera, NM, vary by over 20‰ between glacial and interglacial times. The combined δ18O-δ17O values of interglacial diatoms give T= ~12°C, δ18Ometeoric water = -9‰. A glacial age diatom sample gives T=<10°C, δ18Ometeoric water = -20‰. These data could not be obtained from the

  2. Combining position-specific 13C labeling with compound-specific isotope analysis: first steps towards soil fluxomics

    NASA Astrophysics Data System (ADS)

    Dippold, Michaela; Kuzyakov, Yakov

    2015-04-01

    Understanding the soil organic matter (SOM) dynamics is one of the most important challenges in soil science. Transformation of low molecular weight organic substances (LMWOS) is a key step in biogeochemical cycles because 1) all high molecular substances pass this stage during their decomposition and 2) only LMWOS will be taken up by microorganisms. Previous studies on LMWOS were focused on determining net fluxes through the LMWOS pool, but they rarely identified transformations. As LMWOS are the preferred C and energy source for microorganisms, the transformations of LMWOS are dominated by biochemical pathways of the soil microorganisms. Thus, understanding fluxes and transformations in soils requires a detailed kno