WorldWideScience

Sample records for isotope labeling techniques

  1. Availability of phosphorus in cow slurry using isotopic labelling technique

    International Nuclear Information System (INIS)

    Pongsakul, P.; Bertelsen, F.; Gissel-Nielsen, G.

    1988-01-01

    A pot experiment was conducted to evaluate the influence of cow slurry on P uptake by corn and to estimate the readily available P in the slurry by using an isotopic labelling techique. Water-soluble P in soil was increased and isotopic equilibrium of available P was attained after labelled slurry was mixed thoroughly throughout the soil. Labelled slurry applied at planting increased the P uptake by corn, whereas the same amount applied one week before harvest did not affect the P uptake. It was estimated that 46-54% of the total P uptake in plants is derived from the slurry. The readily available P (the L-value) in the slurry was at least 26 mg/kg which equals 3.7% of the total P. (author)

  2. Stable isotopes labelled compounds

    International Nuclear Information System (INIS)

    1982-09-01

    The catalogue on stable isotopes labelled compounds offers deuterium, nitrogen-15, and multiply labelled compounds. It includes: (1) conditions of sale and delivery, (2) the application of stable isotopes, (3) technical information, (4) product specifications, and (5) the complete delivery programme

  3. Isotopically labelled benzodiazepines

    International Nuclear Information System (INIS)

    Liebman, A.A.

    1987-01-01

    This paper reports on the benzodiazepines which are a class of therapeutic agents. Improvements in the analytical methodology in the areas of biochemistry and pharmacology were significant, particularly in the application of chromatographic and spectroscopic techniques. In addition, the discovery and subsequent development of tritium and carbon-14 as an analytical tool in the biological sciences were essentially post-world war II phenomena. Thus, as these new chemical entities were found to be biologically active, they could be prepared in labeled form for metabolic study, biological half-life determination (pharmacokinetics), tissue distribution study, etc. This use of tracer methodology has been liberally applied to the benzodiazepines and also more recently to the study of receptor-ligand interactions, in which tritium, carbon-11 or fluorine-18 isotopes have been used. The history of benzodiazepines as medicinal agents is indeed an interesting one; an integral part of that history is their use in just about every conceivable labeled form

  4. Carbon allocation belowground in Pinus pinaster using stable carbon isotope pulse labeling technique

    Science.gov (United States)

    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

  5. Melatonin labeled with hydrogen isotopes

    International Nuclear Information System (INIS)

    Dmitrevskaya, L.I.; Smushkevich, Yu.I.; Kurkovskaya, L.N.; Ponomarenko, N.K.; Suvorov, N.N.

    1989-01-01

    A study has been made of isotope exchange between melatonin and deuterium (D 2 O) or tritium (HTO) oxide under different conditions. The ease of isotope exchange for the indole ring hydrogens of melatonin in an acidic medium decreases over the series H 4 > H 2 H 6 >> H 7 , enabling the authors to process a route for production of melatonin labeled with hydrogen isotopes at positions 4,6, and 2 of the indole ring. A method has been suggested for producing melatonin labeled with hydrogen isotopes at position 2 by desulfurization of 2-(2,4-dinitro-phenylsulfenyl)melatonin at Ni(Re) (D)

  6. Melatonin labelled by hydrogen isotopes

    International Nuclear Information System (INIS)

    Dmitrevskaya, L.I.; Smushkevich, Yu.I.; Kurkovskaya, L.N.; Ponomarenko, N.K.; Suvorov, N.N.

    1988-01-01

    Isotope exchange of melatonin with deuterium (D 2 O) and tritium (HTO) oxides under different conditions is studied. Simplicity of isotope exchange of hydrogens of the indole ring of melatonin in the acidic medium decreases in series H 4 >H 2 >H 6 >>H 7 , that permits to suggest the way of melatonin preparation labelled by hydrogen isotopes in positions 4,6 and 2 of the indole ring. The way of melatonin preparation labelled by hydrogen isotopes in position 2 according to the reaction of desulfation 2-(2,4-dinitrophenylsulphenyl) melatonin at catalyst Ni(Re)(D) is suggested

  7. Protein labelling with stable isotopes: strategies

    International Nuclear Information System (INIS)

    Lirsac, P.N.; Gilles, N.; Jamin, N.; Toma, F.; Gabrielsen, O.; Boulain, J.C.; Menez, A.

    1994-01-01

    A protein labelling technique with stable isotopes has been developed at the CEA: a labelled complete medium has been developed, performing as well as the Luria medium, but differing from it because it contains not only free aminated acids and peptides, but also sugars (96% of D-glucopyrannose) and labelled nucleosides. These precursors are produced from a labelled photosynthetic micro-organisms biomass, obtained with micro-algae having incorporated carbon 13, nitrogen 15 and deuterium during their culture. Labelling costs are reduced. 1 fig., 1 tab., 3 refs

  8. Isotopically labelled pyrimidines and purines

    International Nuclear Information System (INIS)

    Balaban, A.T.; Bally, I.

    1987-01-01

    Among the three diazines, pyrimidine is by far the most important one because its derivatives uracil, thymine and cytosine are constituents of the ubiquitous deoxynucleic acids (DNA) and ribonucleic acids (RNA). Other derivatives of pyrimidine without condensed rings include barbiturates, alloxan, orotic acid and thiamine or vitamin B 1 . From the polycyclic derivatives of pyrimidine such as pteridine, alloxazine, and purine, the latter, through its derivatives adenine and guanine complete the list of bases which occur in DNA and RNA: in addition, other purine derivatives such as hypoxanthine, xanthine, theobromine, theophylline, caffeine and uric acid are important natural products with biological activity. The paper presents methods for preparing isotopically labeled pyrimidines as well as purine derivatives. For convenience, the authors describe separately carbon-labeled with radioisotopes 11 C (T 1/2 = 20.3 min) and 14 C (T 1/2 = 5736 years) or the stable isotope 13 C (natural abundance 1.1%) and then hydrogen-labeled systems with the radioisotope 3 H ≡ T (T 1/2 = 12.346 years) or with the stable isotope 2 H ≡ D (natural abundance 0.015%). We do not separate stable from radioactive isotopes because the synthetic methods are identical for the same element; however, the introduction of hydrogen isotopes into organic molecules is often performed by reactions such as isotope exchange which cannot take place in the case of carbon isotopes

  9. Pharmaceuticals labelled with stable isotopes

    International Nuclear Information System (INIS)

    Krumbiegel, P.

    1986-11-01

    The relatively new field of pharmaceuticals labelled with stable isotopes is reviewed. Scientific, juridical, and ethical questions are discussed concerning the application of these pharmaceuticals in human medicine. 13 C, 15 N, and 2 H are the stable isotopes mainly utilized in metabolic function tests. Methodical contributions are given to the application of 2 H, 13 C, and 15 N pharmaceuticals showing new aspects and different states of development in the field under discussion. (author)

  10. NMR studies of isotopically labeled RNA

    Energy Technology Data Exchange (ETDEWEB)

    Pardi, A. [Univ. of Colorado, Boulder, CO (United States)

    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.

  11. Gluconeogenesis from labeled carbon: estimating isotope dilution

    International Nuclear Information System (INIS)

    Kelleher, J.K.

    1986-01-01

    To estimate the rate of gluconeogenesis from steady-state incorporation of labeled 3-carbon precursors into glucose, isotope dilution must be considered so that the rate of labeling of glucose can be quantitatively converted to the rate of gluconeogenesis. An expression for the value of this isotope dilution can be derived using mathematical techniques and a model of the tricarboxylic acid (TCA) cycle. The present investigation employs a more complex model than that used in previous studies. This model includes the following pathways that may affect the correction for isotope dilution: 1) flux of 3-carbon precursor to the oxaloacetate pool via acetyl-CoA and the TCA cycle; 2) flux of 4- or 5-carbon compounds into the TCA cycle; 3) reversible flux between oxaloacetate (OAA) and pyruvate and between OAA and fumarate; 4) incomplete equilibrium between OAA pools; and 5) isotope dilution of 3-carbon tracers between the experimentally measured pool and the precursor for the TCA-cycle OAA pool. Experimental tests are outlined which investigators can use to determine whether these pathways are significant in a specific steady-state system. The study indicated that flux through these five pathways can significantly affect the correction for isotope dilution. To correct for the effects of these pathways an alternative method for calculating isotope dilution is proposed using citrate to relate the specific activities of acetyl-CoA and OAA

  12. Phosphorus availability to ryegrass from urban sewage sludges assessed by isotopic labeling and dilution technique, effect of irradiation

    International Nuclear Information System (INIS)

    El-Motaium, R.A.; Morel, C.

    2007-01-01

    Urban sewage sludge are widely used as alternative P input because of their high P content. Irradiation of sewage sludge is developed in Egypt to make safe spreading of sludge on agricultural fields. Five sludges were sampled from representative urban wastewater treatment plants located in Cairo, Egypt. Sub samples were irradiated using gamma radiation at 6 KGy dose. A pot experiment was conducted under plastic greenhouse using sandy P-deficient soil. The soil was homogeneously labelled with a radioactive solution of carrier-free 32 PO 4 (about 3 MBq kg / soil) to determine the contribution of the different sources of P in plant nutrition. Sludges were applied at 50 mg P/ kg soil. In addition, a control treatment was also carried out to analyze sludge P availability in comparison to a water soluble mineral fertilizer, i.e. 50 mg P/kg soil was applied as commercial monocalcium phosphate (SSP). A reference treatment was included without any P application but the radioactive solution was added. One gram of ryegrass seeds (Lolium multiflorium) was sown in each pot. The aerial parts of the ryegrass were harvested four times, every 3 weeks. Analysis of all harvests included 32 P and total P content was used to determine the respective contribution of soil and sludge to plant P nutrition and the plant available P (L value) using the isotopic dilution principle. The ratio of radioactive P to non-radioactive P, i.e. isotopic composition, was calculated. The total P content in the different urban sewage sludge was ranged from 6.2 to 13.8 g P/ kg and affected by irradiation. Because the soil was extremely P deficient (L value=0.3 mg P/ kg soil), P derived from seeds represented 99% of the P taken up when no P was applied and 87% when applying 50 mg P-SSP/ kg. After correcting the contribution of ryegrass seeds, sludge P contributed by 98% to plant nutrition and 7.2% of the applied P was recovered. The sludge type and irradiation factors did not significantly affect sludge P

  13. Measurement of Hepatic Protein Fractional Synthetic Rate with Stable Isotope Labeling Technique in Thapsigargin Stressed HepG2 Cells

    Science.gov (United States)

    Song, Juquan; Zhang, Xiao-jun; Boehning, Darren; Brooks, Natasha C.; Herndon, David N.; Jeschke, Marc G.

    2012-01-01

    Severe burn-induced liver damage and dysfunction is associated with endoplasmic reticulum (ER) stress. ER stress has been shown to regulate global protein synthesis. In the current study, we induced ER stress in vitro and estimated the effect of ER stress on hepatic protein synthesis. The aim was two-fold: (1) to establish an in vitro model to isotopically measure hepatic protein synthesis and (2) to evaluate protein fractional synthetic rate (FSR) in response to ER stress. Human hepatocellular carcinoma cells (HepG2) were cultured in medium supplemented with stable isotopes 1,2-13C2-glycine and L-[ring-13C6]phenylalanine. ER stress was induced by exposing the cells to 100 nM of thapsigargin (TG). Cell content was collected from day 0 to 14. Alterations in cytosolic calcium were measured by calcium imaging and ER stress markers were confirmed by Western blotting. The precursor and product enrichments were detected by GC-MS analysis for FSR calculation. We found that the hepatic protein FSR were 0.97±0.02 and 0.99±0.05%/hr calculated from 1,2-13C2-glycine and L-[ring-13C6]phenylalanine, respectively. TG depleted ER calcium stores and induced ER stress by upregulating p-IRE-1 and Bip. FSR dramatically decreased to 0.68±0.03 and 0.60±0.06%/hr in the TG treatment group (pisotope tracer incorporation technique is a useful method for studying the effects of ER stress on hepatic protein synthesis. PMID:22298954

  14. SAIL--stereo-array isotope labeling.

    Science.gov (United States)

    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.

  15. Isotopic labelling with carbon-14 and tritium

    International Nuclear Information System (INIS)

    Evans, E.A.

    1980-01-01

    In this paper general methods of isotopic labelling with 14 C and with 3 H are briefly reviewed with special attention to examples of compounds likely to be of wide interest in biological research. (author)

  16. Isotope techniques for hydrology

    International Nuclear Information System (INIS)

    1964-01-01

    In the body of the Panel's report specific conclusions and recommendations are presented in the context of each subject. The general consensus of the Panel is as follows: by the study of this report, the 1961 Panel report, the Proceedings of the March 1963 Tokyo Symposium and other reports of research and technological advances, isotope-technique applications to hydrologic problems have provided some useful avenues for understanding the nature of the hydrologic cycle and in the solution of specific engineering problems. Some techniques are developed thoroughly enough for fairly routine application as tools for use in the solution of practical problems, but further research and development is needed on other concepts to determined whether or not they can be beneficially applied to either research or engineering problems. A concerted effort is required on the part of both hydrologists and isotope specialists working as teams to assure that proper synthesis of scientific advances in the respective fields and translation of these advances into practical technology is achieved

  17. Isotope techniques for hydrology

    Energy Technology Data Exchange (ETDEWEB)

    1964-01-01

    In the body of the Panel's report specific conclusions and recommendations are presented in the context of each subject. The general consensus of the Panel is as follows: by the study of this report, the 1961 Panel report, the Proceedings of the March 1963 Tokyo Symposium and other reports of research and technological advances, isotope-technique applications to hydrologic problems have provided some useful avenues for understanding the nature of the hydrologic cycle and in the solution of specific engineering problems. Some techniques are developed thoroughly enough for fairly routine application as tools for use in the solution of practical problems, but further research and development is needed on other concepts to determined whether or not they can be beneficially applied to either research or engineering problems. A concerted effort is required on the part of both hydrologists and isotope specialists working as teams to assure that proper synthesis of scientific advances in the respective fields and translation of these advances into practical technology is achieved.

  18. Techniques for preparing isotopic targets

    International Nuclear Information System (INIS)

    Xu Guoji; Guan Shouren; Luo Xinghua; Sun Shuhua

    1987-12-01

    The techniques of making isotopic targets for nuclear physics experiments are introduced. Vacuum evaporation, electroplating, centrifugal precipitation, rolling and focused heavy-ion beam sputtering used to prepare various isotopic targets at IAE are described. Reduction-distillation with active metals and electrolytic reduction for converting isotope oxides to metals are mentioned. The stripping processes of producing self-supporting isotopic targets are summarized. The store methods of metallic targets are given

  19. Synthesis of isotopically labelled salicylates

    International Nuclear Information System (INIS)

    Hawkins, D.R.; Pryor, R.W.

    1981-01-01

    [ 13 C-carboxyl]Salicylic acid has been prepared by carbonation of 2-benzyloxybromobenzene followed by reductive debenzylation. Deuterium and tritium labelled salicylic acid and 2 H 2 / 13 C-salicylic acid were prepared by reduction of the 3,5-dibromo derivatives using Raney Ni-Al. Deuterium labelled salicylic acid containing up to four deuterium atoms was prepared by catalytic exchange with Raney Ni-Al in 5% NaOD/D 2 O. (author)

  20. Performance of isobaric and isotopic labeling in quantitative plant proteomics

    DEFF Research Database (Denmark)

    Nogueira, Fábio C S; Palmisano, Giuseppe; Schwämmle, Veit

    2012-01-01

    , and quantitation. In the present work, we have used LC-MS to compare an isotopic (ICPL) and isobaric (iTRAQ) chemical labeling technique to quantify proteins in the endosperm of Ricinus communis seeds at three developmental stages (IV, VI, and X). Endosperm proteins of each stage were trypsin-digested in...

  1. Analysis of the differentially expressed low molecular weight peptides in human serum via an N-terminal isotope labeling technique combining nano-liquid chromatography/matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Leng, Jiapeng; Zhu, Dong; Wu, Duojiao; Zhu, Tongyu; Zhao, Ningwei; Guo, Yinlong

    2012-11-15

    Peptidomics analysis of human serum is challenging due to the low abundance of serum peptides and interference from the complex matrix. This study analyzed the differentially expressed (DE) low molecular weight peptides in human serum integrating a DMPITC-based N-terminal isotope labeling technique with nano-liquid chromatography and matrix-assisted laser desorption/ionization mass spectrometry (nano-LC/MALDI-MS). The workflow introduced a [d(6)]-4,6-dimethoxypyrimidine-2-isothiocyanate (DMPITC)-labeled mixture of aliquots from test samples as the internal standard. The spiked [d(0)]-DMPITC-labeled samples were separated by nano-LC then spotted on the MALDI target. Both quantitative and qualitative studies for serum peptides were achieved based on the isotope-labeled peaks. The DMPITC labeling technique combined with nano-LC/MALDI-MS not only minimized the errors in peptide quantitation, but also allowed convenient recognition of the labeled peptides due to the 6 Da mass difference. The data showed that the entire research procedure as well as the subsequent data analysis method were effective, reproducible, and sensitive for the analysis of DE serum peptides. This study successfully established a research model for DE serum peptides using DMPITC-based N-terminal isotope labeling and nano-LC/MALDI-MS. Application of the DMPITC-based N-terminal labeling technique is expected to provide a promising tool for the investigation of peptides in vivo, especially for the analysis of DE peptides under different biological conditions. Copyright © 2012 John Wiley & Sons, Ltd.

  2. Synthesis of isotopically labeled ketamine

    OpenAIRE

    Stuchlíková, Lucie

    2011-01-01

    In this work were synthesized ketamine isotopomers. Ketamine is used in human medicine and veterinary sectors. It has very broad spectrum of pharmacological effects: anesthetic, analgesic, hallucinogenic, bronchodilator, cardiovascular and antidepressive, which is currently in the research. At first was synthesized precursor of ketamine, N- desmethylketamine which was subsequently labeled the deuterium, tritium and carbon- 14. For the determination of purity and identity mass spectrometry and...

  3. Advances in stable isotope assisted labeling strategies with information science.

    Science.gov (United States)

    Kigawa, Takanori

    2017-08-15

    Stable-isotope (SI) labeling of proteins is an essential technique to investigate their structures, interactions or dynamics by nuclear magnetic resonance (NMR) spectroscopy. The assignment of the main-chain signals, which is the fundamental first step in these analyses, is usually achieved by a sequential assignment method based on triple resonance experiments. Independently of the triple resonance experiment-based sequential assignment, amino acid-selective SI labeling is beneficial for discriminating the amino acid type of each signal; therefore, it is especially useful for the signal assignment of difficult targets. Various combinatorial selective labeling schemes have been developed as more sophisticated labeling strategies. In these strategies, amino acids are represented by combinations of SI labeled samples, rather than simply assigning one amino acid to one SI labeled sample as in the case of conventional amino acid-selective labeling. These strategies have proven to be useful for NMR analyses of difficult proteins, such as those in large complex systems, in living cells, attached or integrated into membranes, or with poor solubility. In this review, recent advances in stable isotope assisted labeling strategies will be discussed. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Synthesis of 15N isotope labeled alanine

    International Nuclear Information System (INIS)

    Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Sant'Ana, Carlos Roberto; Tagliassachi, Romulo Barbieri; Maximo, Everaldo; Prestes, Clelber Vieira

    2005-01-01

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

  5. Isotopically labelled vitamin D derivatives and processes for preparing same

    International Nuclear Information System (INIS)

    Deluca, H.R.; Schnoes, H.K.; Napoli, J.L.; Fivizzani, M.A.

    1981-01-01

    This invention relates to 26,27-isotopically labelled vitamin D 3 compounds, including radiolabelled vitamin D 3 compounds of high specific activity, methods for their preparation, and intermediates obtained in their synthesis. The method involves reacting an ester of a 26,27-dinor-vitamin D-25-carboxylic acid with an isotopically labelled methyl Grignard reagent or methyl lithium reagent to obtain a 26,27-isotopically labelled compound in which at least some of the H atoms and/or C atoms are heavy isotopes. (author)

  6. Label and Label-Free Detection Techniques for Protein Microarrays

    Directory of Open Access Journals (Sweden)

    Amir Syahir

    2015-04-01

    Full Text Available Protein microarray technology has gone through numerous innovative developments in recent decades. In this review, we focus on the development of protein detection methods embedded in the technology. Early microarrays utilized useful chromophores and versatile biochemical techniques dominated by high-throughput illumination. Recently, the realization of label-free techniques has been greatly advanced by the combination of knowledge in material sciences, computational design and nanofabrication. These rapidly advancing techniques aim to provide data without the intervention of label molecules. Here, we present a brief overview of this remarkable innovation from the perspectives of label and label-free techniques in transducing nano‑biological events.

  7. Stable isotope dimethyl labelling for quantitative proteomics and beyond

    Science.gov (United States)

    Hsu, Jue-Liang; Chen, Shu-Hui

    2016-01-01

    Stable-isotope reductive dimethylation, a cost-effective, simple, robust, reliable and easy-to- multiplex labelling method, is widely applied to quantitative proteomics using liquid chromatography-mass spectrometry. This review focuses on biological applications of stable-isotope dimethyl labelling for a large-scale comparative analysis of protein expression and post-translational modifications based on its unique properties of the labelling chemistry. Some other applications of the labelling method for sample preparation and mass spectrometry-based protein identification and characterization are also summarized. This article is part of the themed issue ‘Quantitative mass spectrometry’. PMID:27644970

  8. Stable isotope labeling strategy based on coding theory

    Energy Technology Data Exchange (ETDEWEB)

    Kasai, Takuma; Koshiba, Seizo; Yokoyama, Jun; Kigawa, Takanori, E-mail: kigawa@riken.jp [RIKEN Quantitative Biology Center (QBiC), Laboratory for Biomolecular Structure and Dynamics (Japan)

    2015-10-15

    We describe a strategy for stable isotope-aided protein nuclear magnetic resonance (NMR) analysis, called stable isotope encoding. The basic idea of this strategy is that amino-acid selective labeling can be considered as “encoding and decoding” processes, in which the information of amino acid type is encoded by the stable isotope labeling ratio of the corresponding residue and it is decoded by analyzing NMR spectra. According to the idea, the strategy can diminish the required number of labelled samples by increasing information content per sample, enabling discrimination of 19 kinds of non-proline amino acids with only three labeled samples. The idea also enables this strategy to combine with information technologies, such as error detection by check digit, to improve the robustness of analyses with low quality data. Stable isotope encoding will facilitate NMR analyses of proteins under non-ideal conditions, such as those in large complex systems, with low-solubility, and in living cells.

  9. Stable isotope labeling strategy based on coding theory

    International Nuclear Information System (INIS)

    Kasai, Takuma; Koshiba, Seizo; Yokoyama, Jun; Kigawa, Takanori

    2015-01-01

    We describe a strategy for stable isotope-aided protein nuclear magnetic resonance (NMR) analysis, called stable isotope encoding. The basic idea of this strategy is that amino-acid selective labeling can be considered as “encoding and decoding” processes, in which the information of amino acid type is encoded by the stable isotope labeling ratio of the corresponding residue and it is decoded by analyzing NMR spectra. According to the idea, the strategy can diminish the required number of labelled samples by increasing information content per sample, enabling discrimination of 19 kinds of non-proline amino acids with only three labeled samples. The idea also enables this strategy to combine with information technologies, such as error detection by check digit, to improve the robustness of analyses with low quality data. Stable isotope encoding will facilitate NMR analyses of proteins under non-ideal conditions, such as those in large complex systems, with low-solubility, and in living cells

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

    Energy Technology Data Exchange (ETDEWEB)

    Unkefer, C.J.; Martinez, R.A.; Silks, L.A. III [Los Alamos National Laboratory, NM (United States); Lodwig, S.N. [Centralia College, WA (United States)

    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. Stereoselective synthesis of stable-isotope-labeled amino acids

    International Nuclear Information System (INIS)

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

    1994-01-01

    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 α-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

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

    Science.gov (United States)

    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.

  13. Custom synthesis of isotope-labelled Apis mellifera Pheromone

    International Nuclear Information System (INIS)

    Conanan, Aida P.; Cortes, Nicole Marie A.; Daguno, Cristel Lyn R.; Templonuevo, Jose Angelo A.; Sucgang, Raymond J.

    2012-01-01

    The object of this study is to determine the optimum conditions for the synthesis of the isotope-labelled isopentyl acetate. Isopentyl acetate is widely used as a raw material in industries, in syntheses, and is utilized as a sex attractant (pheromone) by the bee species, Apis mellifera. The isotope labelling of isopentyl acetate will allow tracking of the fate and movement of the isopentyl acetate in the environment, in chemical transformations, and in biological systems. Esterification by alcoholysis of acetic acid was optimized for the preparation of Carbon-14( 14 C)-labelled isopentyl acetate from 14 C-labelled acetic acid and isoamyl alcohol. The different conditions studied were: (1) The effects of acid catalysis and/or reflux on the incorporation and retention of the isotope label on the product. The efficiency of label incorporation and retention was determined through the beta radioactivity of Carbon 14 in each of the synthetic constructs. Determination of the beta radioactivity concentration of 14 C in the isopentyl acetate product was done using low level liquid scintillation spectrometry. Each of the synthetic products was mixed with Ultima Gold scintillation cocktail in a low potassium glass scintillation vial, and analysed in a low-level Wallac 1414 scintillation counter. The application of catalysis without reflux resulted in the highest yield (35%). The same condition also resulted in the highest abundance of carbon isotope label with 2.40 Bequerels per cubic centimetre, Bq/cc (measurement unit for radioactivity). (author)

  14. Vinasse labelling with sup(15)N: use in mineral plants fertilization studies and the potential of the isotopic technique in studies of fermentative nitrogen metabolism of wine

    International Nuclear Information System (INIS)

    Lara C, W.A.; Trivelin, P.C.O.; Basso, L.C.

    1991-01-01

    A methodology for vinasse sup(15)N enrichment was developed under laboratory conditions through a fermentative process. Direct addition of sup(15)N-(NH sub(4)) sub(2)SO sub(4) 90.39 atoms % to the vintage tub (FESA procedure) was compared to the use of a previously enrichment sup(15)N yeast (FELE procedure) by the addition of the label to the multiplicative medium. The mean metabolic recovery of the tracer from the vinasse after fermentation was 4.2 and 11.1% per cycle and accumulated recovery was 3.4 and 33.3%, respectively for the FESA and FELE procedures. The potential of the use of sup(15)N label in studies of fermentative nitrogen metabolism is illustrated by the quantification of sup(15)N distribution among recycled yeast and wine. (author)

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

    Science.gov (United States)

    Hooker, Jacob Matthew [Port Jefferson, NY; Schonberger, Matthias [Mains, DE; Schieferstein, Hanno [Aabergen, DE; Fowler, Joanna S [Bellport, NY

    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.

  16. Preparation of radioactive labelled compounds. Pt. 2. 82Br labelled organic bromine compounds by isotopic exchange

    International Nuclear Information System (INIS)

    Otto, R.

    1988-05-01

    Studies on isotopic exchange between organic bromine compounds and 82 Br labelled dioxane dibromide in the presence of AlCl 3 are described. The results obtained enable to develop a simple and quick preparation method for the labelling with 82 Br [fr

  17. Reductive methods for isotopic labeling of antibiotics

    International Nuclear Information System (INIS)

    Champney, W.S.

    1989-01-01

    Methods for the reductive methylation of the amino groups of eight different antibiotics using 3 HCOH or H 14 COH are presented. The reductive labeling of an additional seven antibiotics by NaB 3 H 4 is also described. The specific activity of the methyl-labeled drugs was determined by a phosphocellulose paper binding assay. Two quantitative assays for these compounds based on the reactivity of the antibiotic amino groups with fluorescamine and of the aldehyde and ketone groups with 2,4-dinitrophenylhydrazine are also presented. Data on the cellular uptake and ribosome binding of these labeled compounds are also presented

  18. Living labeling techniques of mesenchymal stem cells

    International Nuclear Information System (INIS)

    Dong Qingyu; Chen Li

    2007-01-01

    Mesenchymal stem cells (MSCs) are well known for their self-renew and multi- differentiation potentiality. With the transplantation of the MSCs which can promote the regeneration and repair of the injured tissue, a new route for the treatment of dieases is hopeful to be effective. To trace the distribution, migration, proliferation and differentiation of the implanted MSCs, there need effective labeling techniques, especially living labeling techniques. (authors)

  19. Copper absorption from foods labelled intrinsically and extrinsically with Cu-65 stable isotope.

    Science.gov (United States)

    Harvey, L J; Dainty, J R; Beattie, J H; Majsak-Newman, G; Wharf, S G; Reid, M D; Fairweather-Tait, S J

    2005-03-01

    To determine copper absorption from copper containing foods labelled either intrinsically or extrinsically with a highly enriched Cu-65 stable isotope label. A longitudinal cross-over study. The study was conducted at the Institute of Food Research, Human Nutrition Unit, Norwich, UK. Subjects were recruited locally via advertisements placed around the Norwich Research Park. A total of 10 volunteers (nine female, one male) took part in the study, but not all volunteers completed each of the test meals. A highly enriched Cu-65 stable isotope label was administered to volunteers in the form of a reference dose or in breakfast test meals consisting of red wine, soya beans, mushrooms or sunflower seeds. Faecal monitoring and mass spectrometry techniques were used to estimate the relative quantities of copper absorbed from the different test meals. True copper absorption from the reference dose (54%) was similar to extrinsically labelled red wine (49%) and intrinsically labelled sunflower seeds (52%), but significantly higher than extrinsically labelled mushrooms (35%), intrinsically (29%) and extrinsically (15%) labelled soya beans and extrinsically labelled sunflower seed (32%) test meals. The use of Cu-65 extrinsic labels in copper absorption studies requires validation according to the food being examined; intrinsic and extrinsic labelling produced significantly different results for sunflower seeds.

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

    Czech Academy of Sciences Publication Activity Database

    Ek Weis, Johan; da Costa, Sara; Frank, Otakar; Kalbáč, Martin

    2014-01-01

    Roč. 5, č. 3 (2014), s. 549-554 ISSN 1948-7185 R&D Projects: GA MŠk LL1301 Institutional support: RVO:61388955 Keywords : Bernal * graphene * isotopic labeling Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 7.458, year: 2014

  1. Determination of the distribution and reaction of polysaccharides in wood cell walls by the isotope tracer technique, 6: Selective radio-labeling of mannan in ginkgo (Ginkgo biloba)

    International Nuclear Information System (INIS)

    Imai, T.; Terashima, N.; Yasuda, S.

    1997-01-01

    D-Mannose-[2-H-3] and GDP (guanosine diphosphate)-D-mannose-[mannose-1-H-3] were administered to the shoots of ginkgo (Ginkgo biloba L.) tolabel mannan selectively in the cell walls. To suppress the incorporation of radioactivity into the lignin and cellulose, the precursors were administered in the presence of the inhibitor of phenylalanine ammonia-lyase (PAL): namely, L-alpha-aminooxy-beta-phenylpropionic acid (AOPP) and the inhibitor of glucan synthesis: namely, 2-deoxy-D-glucose (2-DG) and 2.6-dichlorobenzonitrile (2.6-DCB). When D-mannose-[2-H-3] was administered in the absence of the inhibitors, great radioactivities were found in the mannose and glucose obtained by sulfuric acid hydrolysis of the newly-formed xylem, and also in the vanillin obtained by nitrobenzene oxidation. These results indicate that the radioactivity was incorporated not only into mannan but also into cellulose and lignin. When D-mannose-[2-H-3] was administered in the presence of both AOPP and 2-DG, the radioactivities of vanillin and glucose were decreased but that of mannose was not decreased. These results indicate that the incorporations of radioactivities into lignin and cellulose were suppressed by the inhibitors, but the incorporation into mannan was not interfered with. The treatment with 2,6-DCB lessened the incorporations of radioactivity into vanillin, xylose, mannose, and glucose of the newly formed xylem considerably which indicated that 2,6-DCB disturbed the metabolic activities of the plant fatally. Consequently, the selective radiolabeling of mannan in ginkgo was achieved by the administration of D-mannose-[2-H-3], in the presence of both AOPP and 2-DG, toa growing stem. In the case of GDP-D-mannose-[mannose-1-H-3], the radioactivity incorporated into the newly-formed xylem was very little, and the selectivity in labeling and the effects of the inhibitors were not clear

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

    Energy Technology Data Exchange (ETDEWEB)

    Lapham, J.; Crothers, D.M. [Yale Univ., New Haven, CT (United States)

    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.

  3. Custom isotope-labelling of apis mellifera pheromone

    International Nuclear Information System (INIS)

    Conanan, Aida P.; Cortes, Nicole Marie A.; Daguno, Cristel Lyn R.; Templonuevo, Jose Angelo A.; Sucgang, Raymond J.

    2012-01-01

    The object of this study is to determine the optimum conditions for the synthesis of isotope-labelled isoamyl acetate. Esterification by alcoholysis of acetic acid was optimized for the preparation of Carbon - 14 ( 14 C)-labelled isopentyl acetate from 14 C-labelled acetic acid and isopentyl alcohol. The optimization procedure defined the effects of catalysis, reflux time, and temperature. The application of catalysis without reflux resulted to the highest yield (40%); the same condition also resulted to the highest abundance of carbon isotope label with 2.40 disintegrations per minute per cubic centimetre, DPM/cc (measurement unit for radioactivity). Determination of the beta radioactivity concentration of 14 C in the isopentyl acetate product was done using low level liquid scintillation spectrometry. Each of the synthetic constructs was mixed with Ultima Gold scintillation cocktail in a glass scintillation vial, and analysed in a low-level Wallac 1414 scintillation counter. Samples were counted for 2 hours in a chamber temperature maintained at 14 degree centegrade. The catalysed reaction without reflux was established to be the most efficient scheme for the radiolabelling. The radiolabelled isoamyl acetate can give way to the synthesis of more complex substances which can be then tracked when they are introduced to a system through the carbon isotope label. (author)

  4. Existing and emerging technologies for measuring stable isotope labelled retinol in biological samples: isotope dilution analysis of body retinol stores.

    Science.gov (United States)

    Preston, Tom

    2014-01-01

    This paper discusses some of the recent improvements in instrumentation used for stable isotope tracer measurements in the context of measuring retinol stores, in vivo. Tracer costs, together with concerns that larger tracer doses may perturb the parameter under study, demand that ever more sensitive mass spectrometric techniques are developed. GCMS is the most widely used technique. It has high sensitivity in terms of sample amount and uses high resolution GC, yet its ability to detect low isotope ratios is limited by background noise. LCMSMS may become more accessible for tracer studies. Its ability to measure low level stable isotope tracers may prove superior to GCMS, but it is isotope ratio MS (IRMS) that has been designed specifically for low level stable isotope analysis through accurate analysis of tracer:tracee ratios (the tracee being the unlabelled species). Compound-specific isotope analysis, where GC is interfaced to IRMS, is gaining popularity. Here, individual 13C-labelled compounds are separated by GC, combusted to CO2 and transferred on-line for ratiometric analysis by IRMS at the ppm level. However, commercially-available 13C-labelled retinol tracers are 2 - 4 times more expensive than deuterated tracers. For 2H-labelled compounds, GC-pyrolysis-IRMS has now become more generally available as an operating mode on the same IRMS instrument. Here, individual compounds are separated by GC and pyrolysed to H2 at high temperature for analysis by IRMS. It is predicted that GC-pyrolysis-IRMS will facilitate low level tracer procedures to measure body retinol stores, as has been accomplished in the case of fatty acids and amino acids. Sample size requirements for GC-P-IRMS may exceed those of GCMS, but this paper discusses sample preparation procedures and predicts improvements, particularly in the efficiency of sample introduction.

  5. Isotope labeling strategies for NMR studies of RNA

    International Nuclear Information System (INIS)

    Lu, Kun; Miyazaki, Yasuyuki; Summers, Michael F.

    2010-01-01

    The known biological functions of RNA have expanded in recent years and now include gene regulation, maintenance of sub-cellular structure, and catalysis, in addition to propagation of genetic information. As for proteins, RNA function is tightly correlated with structure. Unlike proteins, structural information for larger, biologically functional RNAs is relatively limited. NMR signal degeneracy, relaxation problems, and a paucity of long-range 1 H- 1 H dipolar contacts have limited the utility of traditional NMR approaches. Selective isotope labeling, including nucleotide-specific and segmental labeling strategies, may provide the best opportunities for obtaining structural information by NMR. Here we review methods that have been developed for preparing and purifying isotopically labeled RNAs, as well as NMR strategies that have been employed for signal assignment and structure determination.

  6. Isotope-labelled folic acid derivatives

    International Nuclear Information System (INIS)

    Lewin, N.; Wong, E.T.

    1976-01-01

    The suggestion deals with the production of folic acid derivatives suitable as indicators or tracers for analyses of serum folates. These folic acid derivatives contain folic acid which is bound by one or both carboxyl groups to the amino nitrogen of compounds such as, e.g., tyramine, glycyl tyrosine, tyrosine, or the methyl ester of tyrosine. The derivative obtained can be substituted by a gamma emitter, e.g. the iodine isotope I 125. The radioactive derivative is used in the method for the competitive protein bonding to determine endogenic folates in the serum. (UWI) [de

  7. Implementation of isotope correlation technique for safeguards

    International Nuclear Information System (INIS)

    Persiani, P.J.; Bucher, R.G.

    1989-01-01

    The isotopic correlation technique (ICT) is based on the fundamental physics principle that the isotopic compositions of nuclear material in the fuel cycle systems contain information regarding the design and history of nuclear material flow from fuel fabrication, reactor operation, and through input to the reprocessing plant. Isotopic Correlation in conjunction with the gravimetric (or Pu/U) method for mass determination can be developed to provide an independent in-field verification of the reprocessing input accountancy at the dissolver and/or accountancy stage of the reprocessing plant. The Argonne National Laboratory program in isotope correlation techniques is based on three-dimensional reactor physics calculations of characteristic geometries/composition in each reactor class. 10 refs., 1 fig., 3 tabs

  8. Evaluation of a radioisotope labelling technique for measuring bacterial adherence on fabrics

    International Nuclear Information System (INIS)

    Youlo Hsieh; Timm, Debra; Merry, Joanne

    1986-01-01

    A technique utilizing tritiated thymidine labelled bacteria to quantify bacteria on fabrics has been evaluated. Quenching or self-absorption of isotope solution and labelled bacteria suspension by some of the fabrics has been observed. The extents of self-absorption of both isotope and labelled bacteria solutions on various fabrics was found to be dependent upon the fiber contents, i.e. the chemical compositions, of the substrata. This observation confirms that reduction of scintillation efficiency or self-absorption does occur when radio-labelled substances in suspensions were measured with the presence of some fabrics. Cautions should be taken when radio-labelling techniques are applied to detect isotope-labelled micro-organisms or other substances which are in contact with fabrics in the form of solutions. However, when there is no excess and nonattached labelled bacteria in the aqueous surrounding of the fabric, scintillation counting efficiency of the labelled bacteria on all fabrics studied remained constant over a period of 8 h. This indicates that the application of the described isotope labelling procedure is appropriate for quantifying adherent bacteria on fibrous substrate. (author)

  9. Adaptation of a Commonly Used, Chemically Defined Medium for Human Embryonic Stem Cells to Stable Isotope Labeling with Amino Acids in Cell Culture

    DEFF Research Database (Denmark)

    Liberski, A. R.; Al-Noubi, M. N.; Rahman, Z. H.

    2013-01-01

    Metabolic labeling with stable isotopes is a prominent technique for comparative quantitative proteomics, and stable isotope labeling with amino acids in cell culture (SILAC) is the most commonly used approach. SILAC is, however, traditionally limited to simple tissue culture regimens and only ra...

  10. Syntheses with isotopically labelled carbon. Methyl iodide, formaldehyde and cyanide

    International Nuclear Information System (INIS)

    Finn, R.D.; Boothe, T.E.; Vora, M.M.; Hildner, J.C.; Emran, A.M.; Kothari, P.J.

    1984-01-01

    Many of the uniquely labelled synthetic precursors currently employed in the design of sophisticated radiolabelled compounds have their origins in the field of hot atom chemistry. Particularly, the development during the past few years of automated, on-line synthetic procedures which combine the nuclear reaction, hot atom and classical chemistry, and rapid purification methods has allowed the incorporation of useful radionuclides into suitable compounds of chemical and biochemical interest. The application of isotopically labelled methyl iodide, formaldehyde, and cyanide anion as synthetic intermediates in research involving human physiology and nuclear medicine, as well as their contributions to other scientific methodology, is reviewed. (author)

  11. Isotope measurement techniques for atmospheric methane

    International Nuclear Information System (INIS)

    Lowe, D.; White, J.; Levin, I.; Wahlen, M.; Miller, J.B.; Bergamaschi, P.

    2002-01-01

    Measurement techniques for the carbon isotopic composition of atmospheric methane (δ 13 C) are described in detail as applied in several leading institutions active in this field since many years. The standard techniques with offline sample preparation and subsequent measurement by dual inlet isotope ratio mass spectrometry (IRMS) are compared with continuous flow IRMS. The potential use of infrared absorption spectroscopy is briefly discussed. Details on quality control and calibration are provided. Basic analytical aspects for the measurement of other species, 2 H and 14 C, are also given. (author)

  12. Subcellular SIMS imaging of isotopically labeled amino acids in cryogenically prepared cells

    International Nuclear Information System (INIS)

    Chandra, Subhash

    2004-01-01

    Ion microscopy is a potentially powerful technique for localization of isotopically labeled molecules. In this study, L-arginine and phenylalanine amino acids labeled with stable isotopes 13 C and 15 N were localized in cultured cells with the ion microscope at 500 nm spatial resolution. Cells were exposed to the labeled amino acids and cryogenically prepared. SIMS analyses were made in fractured freeze-dried cells. A dynamic distribution was observed from labeled arginine-treated LLC-PK 1 kidney cells at mass 28 ( 13 C 15 N) in negative secondaries, revealing cell-to-cell heterogeneity and preferential accumulation of the amino acid (or its metabolite) in the nucleus and nucleolus of some cells. The smaller nucleolus inside the nucleus was clearly resolved in SIMS images and confirmed by correlative light microscopy. The distribution of labeled phenylalanine contrasted with arginine as it was rather homogeneously distributed in T98G human glioblastoma cells. Images of 39 K, 23 Na and 40 Ca were also recorded to confirm the reliability of sample preparation and authenticity of the observed amino acid distributions. These observations indicate that SIMS techniques can provide a valuable technology for subcellular localization of nitrogen-containing molecules in proteomics since nitrogen does not have a radionuclide tracer isotope. Amino acids labeled with stable isotopes can be used as tracers for studying their transport and metabolism in distinct subcellular compartments with SIMS. Further studies of phenylalanine uptake in human glioblastoma cells may have special significance in boron neutron capture therapy (BNCT) as a boron analogue of phenylalanine, boronophenylalanine is a clinically approved compound for the treatment of brain tumors

  13. Subcellular SIMS imaging of isotopically labeled amino acids in cryogenically prepared cells

    Energy Technology Data Exchange (ETDEWEB)

    Chandra, Subhash

    2004-06-15

    Ion microscopy is a potentially powerful technique for localization of isotopically labeled molecules. In this study, L-arginine and phenylalanine amino acids labeled with stable isotopes {sup 13}C and {sup 15}N were localized in cultured cells with the ion microscope at 500 nm spatial resolution. Cells were exposed to the labeled amino acids and cryogenically prepared. SIMS analyses were made in fractured freeze-dried cells. A dynamic distribution was observed from labeled arginine-treated LLC-PK{sub 1} kidney cells at mass 28 ({sup 13}C{sup 15}N) in negative secondaries, revealing cell-to-cell heterogeneity and preferential accumulation of the amino acid (or its metabolite) in the nucleus and nucleolus of some cells. The smaller nucleolus inside the nucleus was clearly resolved in SIMS images and confirmed by correlative light microscopy. The distribution of labeled phenylalanine contrasted with arginine as it was rather homogeneously distributed in T98G human glioblastoma cells. Images of {sup 39}K, {sup 23}Na and {sup 40}Ca were also recorded to confirm the reliability of sample preparation and authenticity of the observed amino acid distributions. These observations indicate that SIMS techniques can provide a valuable technology for subcellular localization of nitrogen-containing molecules in proteomics since nitrogen does not have a radionuclide tracer isotope. Amino acids labeled with stable isotopes can be used as tracers for studying their transport and metabolism in distinct subcellular compartments with SIMS. Further studies of phenylalanine uptake in human glioblastoma cells may have special significance in boron neutron capture therapy (BNCT) as a boron analogue of phenylalanine, boronophenylalanine is a clinically approved compound for the treatment of brain tumors.

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

    International Nuclear Information System (INIS)

    Wright, P.E.

    1994-01-01

    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 13 C, 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 13 C and 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

  15. Synthetic routes to some isotopically labelled intermediates for diterpenoid biosynthesis

    International Nuclear Information System (INIS)

    Dawson, R.M.; Godfrey, I.M.; Hogg, R.W.; Knox, J.R.

    1989-01-01

    The exo-15-hydrogen of ent-kaurene can be exchanged through a reversible ene reaction in a convenient and efficient procedure which has the potential for giving high specific activity 3 H-labelling. Copalol, the (Z)-double bond stereoisomer, and the allylic alcohol isomers ent-manool and ent-epimanool have been obtained through divergent synthetic pathways involving a 15,16-bisnor ketone intermediate. These pathways have also allowed the four compounds to be obtained with 14 C-labelling. A method, involving a Wittig reaction to form a vinyl bromide intermediate, has been developed for obtaining copalol, as the trityl ether derivative, with stereospecific isotopic labelling of one or the other of the hydrogens of the exocyclic methylene group. 27 refs., figs

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

    Energy Technology Data Exchange (ETDEWEB)

    Wright, P.E. [Scripps Research Institute, La Jolla, CA (United States)

    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.

  17. Tests of intestinal absorption using carbon-14-labeled isotopes

    International Nuclear Information System (INIS)

    Fromm, H.; Sarva, R.P.

    1983-01-01

    Beta radiation-emitting isotopes are being used increasingly in diagnostic gastroenterology for the study of absorption. The major reason for the popularity of radioisotopes is that their use is convenient for patient and physician alike. They often obviate naso- or orointestinal intubation and the collection, storage, and analysis of stool. The radioactivity used for the studies of digestive and absorptive processes is small and is not hazardous. In spite of the safety of the radiolabeled compounds, their use is restricted in children and pregnant women. Therefore, for most tests, promising alternative methods that make use of the stable isotope of carbon, /sup 13/C, instead of the radioactive /sup 14/C have been developed. The analysis of stable isotopes requires more sophisticated technology than that of radioactive compounds, however. Only a few centers presently are equipped and staffed to analyze stable isotopes on a routine basis. In contrast, the analysis of radioactive isotopes has become a routine procedure in almost ever major laboratory. The last decade has brought the development of several radioactive absorption tests. The clinically most useful tests relate to the study of bile acid, fat, lactose, and xylose absorption. All of these tests utilize the excretion rate of /sup 14/CO/sub 2/ in breath after ingestion of a /sup 14/C-labeled compound as a measure of the rate of its absorption or malabsorption

  18. Proceedings of the study of environmental change using isotope techniques

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2002-10-01

    Global warming is acknowledged as a major crisis facing society today, principally due to anticipated impacts on the environment, and availability and distribution of water resources. Scientific understanding of recent human-induced climate change, as well as evaluation of potential mitigation strategies, is progressively being developed through studies of atmospheric greenhouse gases and modern water energy carbon cycling processes. These efforts have been advanced through study of past global climate changes to understand mechanisms that play a role in determining natural climate fluctuations observed in ice cores, lake and sea sediments, corals, paleo-groundwater, cave deposits, tree rings, and other archives. Predictive models incorporating natural and human-induced climate change processes contribute to a better appreciation for the sensitivity of climate to specific anthropogenic perturbations. Increasingly, isotopes are being integrated in climate change studies. For example, isotope methodologies offer substantial improvements in the ability to label the origin and fate of greenhouse gases, and for studying the water and carbon cycle response to past climate changes, a high priority area for action identified by the Intergovernmental Panel on Climate Change (IPCC). Isotopes are also widely used as diagnostic variables for validation of models aimed at providing a prognosis of future environmental conditions. The International Atomic Energy Agency (IAEA) has long supported research and development of isotope applications for climate studies. The joint IAEA/WMO Global Network for Isotopes in Precipitation managed by the IAEA has for the last four decades provided the basic isotope data necessary for integrating stable oxygen and hydrogen isotopes in climate models. The IAEA has also sponsored co-ordinated research projects on Isotope Variations of Carbon Dioxide and other Trace Gases in the Atmosphere and Isotope-Aided Studies of Atmospheric Carbon Dioxide

  19. Proceedings of the study of environmental change using isotope techniques

    International Nuclear Information System (INIS)

    2002-01-01

    Global warming is acknowledged as a major crisis facing society today, principally due to anticipated impacts on the environment, and availability and distribution of water resources. Scientific understanding of recent human-induced climate change, as well as evaluation of potential mitigation strategies, is progressively being developed through studies of atmospheric greenhouse gases and modern water energy carbon cycling processes. These efforts have been advanced through study of past global climate changes to understand mechanisms that play a role in determining natural climate fluctuations observed in ice cores, lake and sea sediments, corals, paleo-groundwater, cave deposits, tree rings, and other archives. Predictive models incorporating natural and human-induced climate change processes contribute to a better appreciation for the sensitivity of climate to specific anthropogenic perturbations. Increasingly, isotopes are being integrated in climate change studies. For example, isotope methodologies offer substantial improvements in the ability to label the origin and fate of greenhouse gases, and for studying the water and carbon cycle response to past climate changes, a high priority area for action identified by the Intergovernmental Panel on Climate Change (IPCC). Isotopes are also widely used as diagnostic variables for validation of models aimed at providing a prognosis of future environmental conditions. The International Atomic Energy Agency (IAEA) has long supported research and development of isotope applications for climate studies. The joint IAEA/WMO Global Network for Isotopes in Precipitation managed by the IAEA has for the last four decades provided the basic isotope data necessary for integrating stable oxygen and hydrogen isotopes in climate models. The IAEA has also sponsored co-ordinated research projects on Isotope Variations of Carbon Dioxide and other Trace Gases in the Atmosphere and Isotope-Aided Studies of Atmospheric Carbon Dioxide

  20. Characterization and relative quantification of phospholipids based on methylation and stable isotopic labeling[S

    Science.gov (United States)

    Cai, Tanxi; Shu, Qingbo; Liu, Peibin; Niu, Lili; Guo, Xiaojing; Ding, Xiang; Xue, Peng; Xie, Zhensheng; Wang, Jifeng; Zhu, Nali; Wu, Peng; Niu, Lili; Yang, Fuquan

    2016-01-01

    Phospholipids (PLs), one of the lipid categories, are not only the primary building blocks of cellular membranes, but also can be split to produce products that function as second messengers in signal transduction and play a pivotal role in numerous cellular processes, including cell growth, survival, and motility. Here, we present an integrated novel method that combines a fast and robust TMS-diazomethane-based phosphate derivatization and isotopic labeling strategy, which enables simultaneous profiling and relative quantification of PLs from biological samples. Our results showed that phosphate methylation allows fast and sensitive identification of the six major PL classes, including their lysophospholipid counterparts, under positive ionization mode. The isotopic labeling of endogenous PLs was achieved by deuterated diazomethane, which was generated through acid-catalyzed hydrogen/deuterium (H/D) exchange and methanolysis of TMS-diazomethane during the process of phosphate derivatization. The measured H/D ratios of unlabeled and labeled PLs, which were mixed in known proportions, indicated that the isotopic labeling strategy is capable of providing relative quantitation with adequate accuracy, reproducibility, and a coefficient of variation of 9.1%, on average. This novel method offers unique advantages over existing approaches and presents a powerful tool for research of PL metabolism and signaling. PMID:26733148

  1. Isotope techniques in water resources development 1991

    International Nuclear Information System (INIS)

    1992-01-01

    Water resources are scarce in many parts of the world. Often, the only water resource is groundwater. Overuse usually invites a rapid decline in groundwater resources which are recharged insufficiently, or not at all, by prevailing climatic conditions. These and other problems currently encountered in hydrology and associated environmental fields have prompted an increasing demand for the utilization of isotope methods. Such methods have been recognized as being indispensable for solving problems such as the identification of pollution sources, characterization of palaeowater resources, evaluation of recharge and evaporative discharge under arid and semi-arid conditions, reconstruction of past climates, study of the interrelationships between surface and groundwater, dating of groundwater and validation of contaminant transport models. Moreover, in combination with other hydrogeological and geochemical methods, isotope techniques can provide useful hydrological information, such as data on the origin, replenishment and dynamics of groundwater. It was against this background that the International Atomic Energy Agency, in co-operation with the United Nations Educational, Scientific and Cultural Organization and the International Association of Hydrological Sciences, organized this symposium on the Use of Isotope Techniques in Water Resources Development, which took place in Vienna from 11 to 15 March 1991. The main themes of the symposium were the use of isotope techniques in solving practical problems of water resources assessment and development, particularly with respect to groundwater protection, and in studying environmental problems related to water, including palaeohydrological and palaeoclimatological problems. A substantial part of the oral presentations was concerned with the present state and trends in groundwater dating, and with some methodological aspects. These proceedings contain the papers of 37 oral and the extended synopses of 47 poster

  2. Stable isotope labeling of glycoprotein expressed in silkworms using immunoglobulin G as a test molecule

    Energy Technology Data Exchange (ETDEWEB)

    Yagi, Hirokazu [Nagoya City University, Faculty and Graduate School of Pharmaceutical Sciences (Japan); Nakamura, Masatoshi [National Institute of Agrobiological Sciences, Genetic Resources Conservation Research Unit, Genetic Resources Center (Japan); Yokoyama, Jun [Taiyo Nippon Sanso Corporation, Tsukuba Laboratories (Japan); Zhang, Ying; Yamaguchi, Takumi [National Institutes of Natural Sciences, Institute for Molecular Science and Okazaki Institute for Integrative Bioscience (Japan); Kondo, Sachiko [Nagoya City University, Faculty and Graduate School of Pharmaceutical Sciences (Japan); Kobayashi, Jun [Yamaguchi University, Department of Biological and Environmental Sciences, Faculty of Agriculture (Japan); Kato, Tatsuya; Park, Enoch Y. [Shizuoka University, Laboratory of Biotechnology, Research Institute of Green Science and Technology (Japan); Nakazawa, Shiori [Nagoya University, Sugashima Marine Biological Laboratory, Graduate School of Science (Japan); Hashii, Noritaka; Kawasaki, Nana [National Institute of Health Sciences, Division of Biological Chemistry and Biologicals (Japan); Kato, Koichi, E-mail: kkato@phar.nagoya-cu.ac.jp [Nagoya City University, Faculty and Graduate School of Pharmaceutical Sciences (Japan)

    2015-06-15

    Silkworms serve as promising bioreactors for the production of recombinant proteins, including glycoproteins and membrane proteins, for structural and functional protein analyses. However, lack of methodology for stable isotope labeling has been a major deterrent to using this expression system for nuclear magnetic resonance (NMR) structural biology. Here we developed a metabolic isotope labeling technique using commercially available silkworm larvae. The fifth instar larvae were infected with baculoviruses for co-expression of recombinant human immunoglobulin G (IgG) as a test molecule, with calnexin as a chaperone. They were subsequently reared on an artificial diet containing {sup 15}N-labeled yeast crude protein extract. We harvested 0.1 mg of IgG from larva with a {sup 15}N-enrichment ratio of approximately 80 %. This allowed us to compare NMR spectral data of the Fc fragment cleaved from the silkworm-produced IgG with those of an authentic Fc glycoprotein derived from mammalian cells. Therefore, we successfully demonstrated that our method enables production of isotopically labeled glycoproteins for NMR studies.

  3. Stable isotopic labeling-based quantitative targeted glycomics (i-QTaG).

    Science.gov (United States)

    Kim, Kyoung-Jin; Kim, Yoon-Woo; Kim, Yun-Gon; Park, Hae-Min; Jin, Jang Mi; Hwan Kim, Young; Yang, Yung-Hun; Kyu Lee, Jun; Chung, Junho; Lee, Sun-Gu; Saghatelian, Alan

    2015-01-01

    Mass spectrometry (MS) analysis combined with stable isotopic labeling is a promising method for the relative quantification of aberrant glycosylation in diseases and disorders. We developed a stable isotopic labeling-based quantitative targeted glycomics (i-QTaG) technique for the comparative and quantitative analysis of total N-glycans using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We established the analytical procedure with the chemical derivatizations (i.e., sialic acid neutralization and stable isotopic labeling) of N-glycans using a model glycoprotein (bovine fetuin). Moreover, the i-QTaG using MALDI-TOF MS was evaluated with various molar ratios (1:1, 1:2, 1:5) of (13) C6 /(12) C6 -2-aminobenzoic acid-labeled glycans from normal human serum. Finally, this method was applied to direct comparison of the total N-glycan profiles between normal human sera (n = 8) and prostate cancer patient sera (n = 17). The intensities of the N-glycan peaks from i-QTaG method showed a good linearity (R(2) > 0.99) with the amount of the bovine fetuin glycoproteins. The ratios of relative intensity between the isotopically 2-AA labeled N-glycans were close to the theoretical molar ratios (1:1, 1:2, 1:5). We also demonstrated that the up-regulation of the Lewis antigen (~82%) in sera from prostate cancer patients. In this proof-of-concept study, we demonstrated that the i-QTaG method, which enables to achieve a reliable comparative quantitation of total N-glycans via MALDI-TOF MS analysis, has the potential to diagnose and monitor alterations in glycosylation associated with disease states or biotherapeutics. © 2015 American Institute of Chemical Engineers.

  4. Raman spectroscopy of isotopically labeled two-layer graphene

    Czech Academy of Sciences Publication Activity Database

    Kalbáč, Martin; Kong, J.; Kavan, Ladislav; Dresselhaus, M. S.

    2012-01-01

    Roč. 249, č. 12 (2012), s. 2500-2502 ISSN 0370-1972 R&D Projects: GA AV ČR IAA400400911; GA AV ČR IAA400400804; GA AV ČR KAN200100801; GA MŠk ME09060; GA ČR GAP204/10/1677; GA ČR GBP208/12/G016; GA ČR(CZ) GAP208/12/1062 Institutional support: RVO:61388955 Keywords : electrochemical doping * isotope labeling * graphene Subject RIV: CG - Electrochemistry Impact factor: 1.489, year: 2012

  5. Isotope techniques in hydrology and sedimentology

    International Nuclear Information System (INIS)

    Bomtempo, Virgilio Lopardi

    1999-01-01

    Water is the foundation of all life on Earth. Although two thirds of the terrestrial surface are covered by water (estimated volume is 1.5 billion cubic meters), just 2% of this total are fresh water, most of it locked in glaciers, ice caps and in deep groundwater reservoirs. Only 2,000 cubic kilometers are estimated to be available for consumption. Water resources have become more and more scarce, and the utilization becomes increasingly costly, due to the impact caused by over-exploitation and by diversified fronts of pollution. Specialists have been working in the development and in the application of several techniques to face this problem and to produce sustainable solutions. Isotope techniques represent a group of widespread tools that have been used along many years, and have become outstanding in hydrological investigation. This paper introduces a comprehensive review of the isotope techniques, taking into account environmental isotopes (stable and radioactive), artificial tracers and the use of radioactive sealed sources. Potentialities and limitations, future perspectives, as well as risks and benefits are also discussed. (author)

  6. Development of versatile isotopic labeling reagents for profiling the amine submetabolome by liquid chromatography–mass spectrometry

    International Nuclear Information System (INIS)

    Zhou, Ruokun; Huan, Tao; Li, Liang

    2015-01-01

    Highlights: • Two new reagents were developed for chemical isotope labeling mass spectrometry (MS). • They could be used to label amine-containing metabolites in a metabolomic sample. • The labeled metabolites could be detected with much improved sensitivity in MS. • One of the reagents could also help generate useful MS/MS spectra for structural analysis. • These reagents should be useful for quantitative metabolomics. - Abstract: Metabolomic profiling involves relative quantification of metabolites in comparative samples and identification of the significant metabolites that differentiate different groups (e.g., diseased vs. controls). Chemical isotope labeling (CIL) liquid chromatography–mass spectrometry (LC–MS) is an enabling technique that can provide improved metabolome coverage and metabolite quantification. However, chemical identification of labeled metabolites can still be a challenge. In this work, a new set of isotopic labeling reagents offering versatile properties to enhance both detection and identification are described. They were prepared by a glycine molecule (or its isotopic counterpart) and an aromatic acid with varying structures through a simple three-step synthesis route. In addition to relatively low costs of synthesizing the reagents, this reaction route allows adjusting reagent property in accordance with the desired application objective. To date, two isotopic reagents, 4-dimethylaminobenzoylamido acetic acid N-hydroxylsuccinimide ester (DBAA-NHS) and 4-methoxybenzoylamido acetic acid N-hydroxylsuccinimide ester (MBAA-NHS), for labeling the amine-containing metabolites (i.e., amine submetabolome) have been synthesized. The labeling conditions and the related LC–MS method have been optimized. We demonstrate that DBAA labeling can increase the metabolite detectability because of the presence of an electrospray ionization (ESI)-active dimethylaminobenzoyl group. On the other hand, MBAA labeled metabolites can be fragmented

  7. Development of versatile isotopic labeling reagents for profiling the amine submetabolome by liquid chromatography–mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Ruokun; Huan, Tao; Li, Liang, E-mail: Liang.Li@ualberta.ca

    2015-06-30

    Highlights: • Two new reagents were developed for chemical isotope labeling mass spectrometry (MS). • They could be used to label amine-containing metabolites in a metabolomic sample. • The labeled metabolites could be detected with much improved sensitivity in MS. • One of the reagents could also help generate useful MS/MS spectra for structural analysis. • These reagents should be useful for quantitative metabolomics. - Abstract: Metabolomic profiling involves relative quantification of metabolites in comparative samples and identification of the significant metabolites that differentiate different groups (e.g., diseased vs. controls). Chemical isotope labeling (CIL) liquid chromatography–mass spectrometry (LC–MS) is an enabling technique that can provide improved metabolome coverage and metabolite quantification. However, chemical identification of labeled metabolites can still be a challenge. In this work, a new set of isotopic labeling reagents offering versatile properties to enhance both detection and identification are described. They were prepared by a glycine molecule (or its isotopic counterpart) and an aromatic acid with varying structures through a simple three-step synthesis route. In addition to relatively low costs of synthesizing the reagents, this reaction route allows adjusting reagent property in accordance with the desired application objective. To date, two isotopic reagents, 4-dimethylaminobenzoylamido acetic acid N-hydroxylsuccinimide ester (DBAA-NHS) and 4-methoxybenzoylamido acetic acid N-hydroxylsuccinimide ester (MBAA-NHS), for labeling the amine-containing metabolites (i.e., amine submetabolome) have been synthesized. The labeling conditions and the related LC–MS method have been optimized. We demonstrate that DBAA labeling can increase the metabolite detectability because of the presence of an electrospray ionization (ESI)-active dimethylaminobenzoyl group. On the other hand, MBAA labeled metabolites can be fragmented

  8. Isotope techniques in a water survey

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1959-10-15

    The circulation of water is one of the most interesting of natural phenomena. Exact knowledge of fluctuations in precipitation and other factors in water circulation is extremely important for areas which have a very limited water supply. The information about the circulation of water is also important for the disposal of radioactive wastes on land and in the sea. Before satisfactory methods of disposal can be devised, it is essential to know precisely whether and to what extent the wastes can be transferred from one place to another as a result of the circulation of water. One of the most efficient ways of gathering such information is to study the isotopic ratios of hydrogen and oxygen in water in different areas. Tritium can serve a s a tracer in the study of water circulation. A variety of information can be obtained by measurements of isotopic composition of water, e.g. the average age of the water molecule in a lake or age, size, storage time and flow rate of a groundwater body. The modern tools of hydrological research cannot be employed by every country, because measurements of the isotopic composition of water require great technical skill and scientific knowledge. Besides, interpretation of isotope data in terms of hydrology and climatology requires the knowledge of certain basic data for the whole world or at least for large areas. A more complete knowledge of the worldwide variations in the isotopic composition of water would greatly facilitate the interpretation of local conditions. Guided by these considerations, the International Atomic Energy Agency has decided to initiate a study to determine the world-wide distribution of hydrogen and oxygen isotopes in water. On the basis of this study, it will be possible to make available basic data for the use of any country that wishes to apply isotope techniques for hydrological and climatological research. Under this project, it is proposed to collect samples of rain, river and ocean water in different

  9. Labelled antibody techniques in glycoprotein estimation

    International Nuclear Information System (INIS)

    Hazra, D.K.; Ekins, R.P.; Edwards, R.; Williams, E.S.

    1977-01-01

    The problems in the radioimmunoassay of the glycoprotein hormones (pituitary LH, FSH and TSH and human chlorionic gonadotrophin HGG) are reviewed viz: limited specificity and sensitivity in the clinical context, interpretation of disparity between bioassay and radioimmunoassay, and interlaboratory variability. The advantages and limitations of the labelled antibody techniques - classical immonoradiometric methods and 2-site or 125 I-anti-IgG indirect labelling modifications are reviewed in general, and their theoretical potential in glycoprotein assays examined in the light of previous work. Preliminary experiments in the development of coated tube 2-site assay for glycoproteins using 125 I anti-IgG labelling are described, including conditions for maximizing solid phase extraction of the antigen, iodination of anti-IgG, and assay conditions such as effects of temperature of incubation with antigen 'hormonefree serum', heterologous serum and detergent washing. Experiments with extraction and antigen-specific antisera raised in the same or different species are described as exemplified by LH and TSH assay systems, the latter apparently promising greater sensitivity than radioimmunoassay. Proposed experimental and mathematical optimisation and validation of the method as an assay system is outlined, and the areas for further work delineated. (orig.) [de

  10. Synthesis of 14C-labeled and stable isotope-labeled CGS 16617

    International Nuclear Information System (INIS)

    Chaudhuri, N.K.; Markus, B.; Sung Mingsang

    1988-01-01

    The synthesis of a 14 C-labeled and two stable isotope-labeled analogs of CGS 16617 is described. The synthetic method involved the preparation of tetrahydro-3-bromo-1-benzazepin-2-one, labeled with a 14 C or four deuterium atoms, followed by introduction of two side chains at 1- and 3-positions. The labeled bromobenzazepinones were prepared by Beckmann rearrangement of bromo-oximes of α-tetralones, obtained by cyclization of labeled benzenebutanoic acids. The 14 C-labeled acid was prepared by hydrolysis of the nitrile, prepared by reaction of 3-bromopropylbenzene and K 14 CN. The tetradeutero acid was prepared from ethyl phenylpropynoate by catalytic reduction of the triple bond with deuterium gas, followed by reduction of the deuterated ester with lithium aluminium hydride and conversion of the resulting alcohol into the carboxylic acid. The acetic acid side chain was introduced by N-alkylation with ethyl bromoacetate or ethyl bromoacetate-1, 2- 13 C 2 followed by hydrolysis, and the L-lysine side chain, by reaction with L-(-)-3-amino-ε-caprolactam followed by hydrolysis of the caprolactam ring. (author)

  11. Management of ground water using isotope techniques

    International Nuclear Information System (INIS)

    Romani, Saleem

    2004-01-01

    Ground water play a major role in national economy and sustenance of life and environment. Prevalent water crisis in India includes falling water table, water quality deterioration, water logging and salinity. Keeping in view the increasing thrust on groundwater resources and the present scenario of availability vis-a vis demand there is a need to reorient our approach to ground water management. The various ground water management options require proper understanding of ground water flow system. Isotopes are increasingly being applied in hydrogeological investigations as a supplementary tool for assessment of aquifer flow and transport characteristics. Isotope techniques coupled with conventional hydrogeological and hydrochemical methods can bring in greater accuracy in the conceptualization of hydrogeological control mechanism. The use of isotope techniques in following areas can certainly be of immense help in implementing various ground water management options in an efficient manner. viz.Interaction between the surface water - groundwater systems to plan conjunctive use of surface and ground water. Establishing hydraulic interconnections between the aquifers in a multi aquifer system. Depth of circulation of water and dating of ground water. Demarcating ground water recharge and discharge areas. Plan ground water development in coastal aquifers to avoid sea water ingress. Development of flood plain aquifer. (author)

  12. The synthesis of a tritium, carbon-14, and stable isotope-labeled cathepsin C inhibitors.

    Science.gov (United States)

    Allen, Paul; Bragg, Ryan A; Caffrey, Moya; Ericsson, Cecilia; Hickey, Michael J; Kingston, Lee P; Elmore, Charles S

    2017-02-01

    As part of a medicinal chemistry program aimed at developing a highly potent and selective cathepsin C inhibitor, tritium, carbon-14, and stable isotope-labeled materials were required. The synthesis of tritium-labeled methanesulfonate 5 was achieved via catalytic tritiolysis of a chloro precursor, albeit at a low radiochemical purity of 67%. Tritium-labeled AZD5248 was prepared via a 3-stage synthesis, utilizing amide-directed hydrogen isotope exchange. Carbon-14 and stable isotope-labeled AZD5248 were successfully prepared through modifications of the medicinal chemistry synthetic route, enabling the use of available labeled intermediates. Copyright © 2016 John Wiley & Sons, Ltd.

  13. Graceful, harmonious and magic type labelings relations and techniques

    CERN Document Server

    López, Susana C

    2017-01-01

    Aimed toward upper undergraduate and graduate students in mathematics, this book examines the foremost forms of graph labelings including magic, harmonious, and graceful labelings. An overview of basic graph theory concepts and notation is provided along with the origins of graph labeling. Common methods and techniques are presented introducing readers to links between graph labels. A variety of useful techniques are presented to analyze and understand properties of graph labelings. The classical results integrated with new techniques, complete proofs, numerous exercises, and a variety of open problems, will provide readers with a solid understanding of graph labelings.

  14. Use of stable isotope techniques in soil organic matter studies

    International Nuclear Information System (INIS)

    Gerzabek, M.H.

    1998-01-01

    Plants differ distinctly in their C-isotopic composition. The largest differences occur between plant species with different photosynthetic pathways. C 3 - and C 4 -plants are differentiated by approximately 1.4% on the δ-scale (approx. -2.7% 13 C versus -1.3% 13 C). Modern elemental analyser - mass spectrometer combinations reach accuracies of at least 0.01% δ 13 C. Therefore, the difference between C 3 and C 4 plants is sufficient to be used for tracer studies. Several investigations of soil organic mater (SOM) turnover under field conditions were undertaken using the fact that the vegetation cover changed between C 3 and C 4 plants. The discrimination between SOM originating from indigenous vegetation (forest, C 3 ) and sugar cane (C 4 ) after 50 years of cropping introducing two SOM compartments of different stability was described. Another example is the change from prairie vegetation (C 4 ) to different C 3 -crops and the evaluation of the carbon origin at or near equilibrium. More recent studies use 15 N-labelled C 4 -plant residues or 13 C-labelled C 3 -plants to elucidate the fate of carbon and nitrogen in soils developed under C 3 -vegetation. Both in situ experiments and laboratory incubations were used to evaluate carbon and nitrogen fluxes from crop residues. Physical fractionation of bulk soil into particle sizes proved to be of advantage to follow short and long-term dynamics of crop residues within SOM. Changes in the natural abundance of 13 C and 15 N within soil profiles can elucidate leaching or mineralization of humic substances. Changes in the natural abundance of stable isotopes are also possible due to the application of organic manures, quantification, however is not easy because of the small isotopic differences between soil and manure carbon and nitrogen. 15 N labelling of soil nitrogen has been widely used in the last two decades to quantify biological nitrogen fixation. Considerable progress has been made due to the isotope dilution

  15. Closing the gap between T-cell life span estimates from stable isotope-labeling studies in mice and humans

    NARCIS (Netherlands)

    Westera, Liset; Drylewicz, Julia; den Braber, Ineke; Mugwagwa, Tendai; van der Maas, Iris; Kwast, Lydia; Volman, Thomas; van de Weg-Schrijver, Elise H. R.; Bartha, István; Spierenburg, Gerrit; Gaiser, Koos; Ackermans, Mariëtte T.; Asquith, Becca; de Boer, Rob J.; Tesselaar, Kiki; Borghans, José A. M.

    2013-01-01

    Quantitative knowledge of the turnover of different leukocyte populations is a key to our understanding of immune function in health and disease. Much progress has been made thanks to the introduction of stable isotope labeling, the state-of-the-art technique for in vivo quantification of cellular

  16. Isotope correlation and mass spectrometry techniques for irradiated fuel assay

    International Nuclear Information System (INIS)

    Deron, S.

    1985-01-01

    This paper outlines the methods used to account for fissionable materials in irradiated nuclear fuel elements entering reprocessing plants. Verification is accomplished at three mass balance stations in the plant. Techniques employed fall into two categories: isotopic and isotope dilution analyses by mass spectometry and isotope correlation techniques. These methods are discussed in some detail

  17. Evaluation of leucocytes labelling technique with 99m Tc - HMPAO

    International Nuclear Information System (INIS)

    Moraes, D.W.; Marinho, N.S.; Rezende, M.E.X.; Buchpiguel, C.A.; Rockmann, T.M.B.

    1992-01-01

    An evaluation of the leukocytes labelling technique with HMPAO- 99m Tc, using low volume of blood and keeping the labelling efficiency and cellular viability in compatible levels with those preconized by the literature is presented. (C.G.C.)

  18. The Helikon technique for isotope enrichment

    International Nuclear Information System (INIS)

    Haarhoff, P.C.

    1976-11-01

    The separating element employed in the UCOR process for uranium enrichment has an enriched stream which is much smaller than the depleted stream. To deal with this small cut and to exploit the full potential of the process, a new cascade technique has been developed, the so-called helikon technique. It is based on the principle that an axial flow compressor can simultaneously compress a number of streams of different isotopic composition, which flow through it in parallel, without any significant mixing between them. The technique makes it possible to achieve the desired enrichment with a relatively small number of separating units, by making the best use of the high enrichment factor available. A further feature of the helikon technique is that a module yields an enrichment factor which is not constant, but can vary. In this way a cascade can be built up from modules of a fixed size, which is a great advantage when compared to conventional cascade arrangements where several unit sizes are required. A general theoretical treatment of the helikon technique is given and the similarity between helikon and conventional cascades is pointed out. Practical helikon cascades are subsequently discussed on the basis of the UCOR process

  19. Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones.

    Science.gov (United States)

    Liokatis, Stamatios

    2017-03-26

    Asymmetrically modified nucleosomes contain the two copies of a histone (sister histones) decorated with distinct sets of Post-translational Modifications (PTMs). They are newly identified species with unknown means of establishment and functional implications. Current analytical methods are inadequate to detect the copy-specific occurrence of PTMs on the nucleosomal sister histones. This protocol presents a biochemical method for the in vitro reconstitution of nucleosomes containing differentially isotope-labeled sister histones. The generated complex can be also asymmetrically modified, after including a premodified histone pool during refolding of histone subcomplexes. These asymmetric nucleosome preparations can be readily reacted with histone-modifying enzymes to study modification cross-talk mechanisms imposed by the asymmetrically pre-incorporated PTM using nuclear magnetic resonance (NMR) spectroscopy. Particularly, the modification reactions in real-time can be mapped independently on the two sister histones by performing different types of NMR correlation experiments, tailored for the respective isotope type. This methodology provides the means to study crosstalk mechanisms that contribute to the formation and propagation of asymmetric PTM patterns on nucleosomal complexes.

  20. Use of the doubly labeled water technique in humans during heavy sustained exercise

    International Nuclear Information System (INIS)

    Westerterp, K.R.; Saris, W.H.; van Es, M.; ten Hoor, F.

    1986-01-01

    We measured energy expenditure with the doubly labeled water technique during heavy sustained exercise in the Tour de France, a bicycle race lasting more than 3 wk. Four subjects were observed for consecutive intervals of 7, 8, and 7 days. Each interval started with an oral isotope dose to reach an excess isotope level of 200 ppm 18O and 130 ppm 2H. The biological half-lives of the isotopes were between 2.25 and 3.80 days. Energy expenditure was compared with simultaneous measurements of energy intake, and body mass and body composition did not change significantly. The doubly labeled water technique gave higher values for energy expenditure than the food record technique. The discrepancy showed a systematic increment from the first to the third interval, being 12.9 +/- 7.9, 21.4 +/- 9.8, and 35.3 +/- 4.4% of the energy expenditure calculated from dietary intake, respectively. Possible explanations for the discrepancy are discussed. The subjects reached an average daily metabolic rate of 3.4-3.9 or 4.3-5.3 times basal metabolic rate based on the food record technique and the doubly labeled water technique, respectively. Thus, when measured with the same technique, the energetic ceiling for performance in humans is comparable with that of animals like birds

  1. Pyrolysis of Cigarette Ingredients Labelled with Stable Isotopes

    Directory of Open Access Journals (Sweden)

    Stotesbury S

    2014-12-01

    Full Text Available It is important to know how tobacco additives behave when cigarettes are smoked, whether they transfer intact to the smoke or whether there is any decomposition during smoking. Pyrolysis-GC-MS is a technique that can be focussed upon the effects of combustion from a single material free from interference from the complex mixture of different components present in the smoke. However, because pyrolysis is a model technique, the results need to be validated by comparison with cigarette smoke chemistry. In a previous paper we presented such a method for modelling the smoke chemistry from a burning cigarette using pyrolysis-GC-MS. The transfer and the extent of degradation of anisole, p-anisaldehyde, benzaldehyde, isoamylisovalerate, methyl trans-cinnamate and vanillin within a burning cigarette were estimated using this pyrolysis method. When these data were compared with results from smoke studies from 14C-analogues of the materials, the high levels of transfer predicted by pyrolysis were found to be generally consistent with the smoke chemistry data. However, there were still two outstanding issues. Firstly, there was some ambiguity in the labelled study about whether vanillin actually transferred without degradation or not. Furthermore, the results from the 14C-labelled study showed a greater extent of degradation for p-anisaldehyde than that indicated from the pyrolysis experiments. The purpose of the current study was to present some new information obtained to address these questions by better understanding the effect upon the smoke chemistry from adding vanillin and p-anisaldehyde, and the relationship between the smoke chemistry and the pyrolysis results. Components were identified in the smoke from cigarettes loaded with p-anisaldehyde and vanillin labelled with 18O and 13C. The extent of degradation from each additive was estimated by identifying labelled degradation products in the smoke. Because there was a clear distinction between the

  2. 13C metabolic flux analysis: optimal design of isotopic labeling experiments.

    Science.gov (United States)

    Antoniewicz, Maciek R

    2013-12-01

    Measuring fluxes by 13C metabolic flux analysis (13C-MFA) has become a key activity in chemical and pharmaceutical biotechnology. Optimal design of isotopic labeling experiments is of central importance to 13C-MFA as it determines the precision with which fluxes can be estimated. Traditional methods for selecting isotopic tracers and labeling measurements did not fully utilize the power of 13C-MFA. Recently, new approaches were developed for optimal design of isotopic labeling experiments based on parallel labeling experiments and algorithms for rational selection of tracers. In addition, advanced isotopic labeling measurements were developed based on tandem mass spectrometry. Combined, these approaches can dramatically improve the quality of 13C-MFA results with important applications in metabolic engineering and biotechnology. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Extensions of the techniques for the accelerated unspecific isotopic labelling of organic compounds; Applications nouvelles des techniques accelererees du marquage non specifique de composes organiques; Rasprostranenie metodov uskoreniya mecheniya nespetsifichnymi izotopami organicheskikh slozhnykh soedinenij; Nuevas aplicaciones de las tecnicas de marcacion inespecifica acelerada de compuestos organicos

    Energy Technology Data Exchange (ETDEWEB)

    Ghanem, N A [National Research Centre, El-Dokki, Cairo (Egypt); Westermark, T [Division of Physical Chemistry, Royal Institute of Technology, Stockholm 70 (Sweden)

    1962-03-15

    The paper aims at reviewing new means of obtaining acceleration in the reaction of a radioactive gas with a target material to be labelled and examples of successful cases. The techniques include the use of microwave and radiowave discharges, glow discharges affected through DC or AC sources, and simply ultraviolet radiation. Specific activities of more than 100 mc/g in purified significant quantities of products were obtained. Periods of exposure to discharge were of the order of one minute and decomposition was limited in most cases. Calculations of utilization of the energy absorbed during the labelling process were compared with the energy from ionizing radiation used by Wilzbach and others, and G values for isotope replacement are given. In polystyrene labelling, evidence is found that the mechanisms is governed by an atom-molecule reaction. (author) [French] Les auteurs passent en revue des methodes recentes visant a accelerer la reaction d'un gaz radioactif avec des matieres a marquer; ils citent des cas de reussite. Les techniques etudiees comportent l'emploi de decharges de micro-ondes et d'ondes hertziennes, de decharges luminescentes par des sources de courant continu ou alternatif et de simples rayons ultraviolets. On a obtenu des activites specifiques superieures a 100 mc/g dans des quantites importantes de produits purifies. Les temps d'exposition a la decharge etaient de l'ordre d'une minute; dans la plupart des cas, la decomposition etait limitee. Les auteurs comparent la quantite d'energie absorbee au cours du marquage a celle qui est necessaire lorsqu'on emploie des rayonnements ionisants, comme l'ont fait Wilzbach et autres; ils donnent les valeurs G de remplacement des isotopes. Dans le marquage du polystyrene, il semble bien que le processus soit regi par une reaction atome-molecule. (author) [Spanish] El objeto de la memoria es pasar revista a nuevos metodos para acelerar la reaccion de un gas radiactivo con el material de blanco que se desea

  4. Analysis of [U-13C6]glucose in human plasma using liquid chromatography/isotope ratio mass spectrometry compared with two other mass spectrometry techniques

    NARCIS (Netherlands)

    Schierbeek, H.; Moerdijk-Poortvliet, T.C.W.; van den Akker, C.H.P.; te Braake, F.W.J.; Boschker, H.T.S.; van Goudoever, J.B.

    2009-01-01

    The use of stable isotope labelled glucose provides insight into glucose metabolism. The 13C-isotopic enrichment of glucose is usually measured by gas chromatography/mass spectrometry (GC/MS) or gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). However, in both techniques

  5. Review of advanced techniques for waste canister labeling

    International Nuclear Information System (INIS)

    Culbreth, W.G.; Bhagi, B.K.; Kanjerla, A.

    1994-01-01

    Technology has produced several new labeling techniques that may meet the needs of a nuclear waste repository. New methods must be capable of providing permanent labels to clearly identify the contents of each package containing high-level spent nuclear fuel. Several new techniques, along with their benefits and problems, are discussed

  6. SIMSISH technique does not alter the apparent isotopic composition of bacterial cells.

    Directory of Open Access Journals (Sweden)

    Olivier Chapleur

    Full Text Available In order to identify the function of uncultured microorganisms in their environment, the SIMSISH method, combining in situ hybridization (ISH and nanoscale secondary ion mass spectrometry (nanoSIMS imaging, has been proposed to determine the quantitative uptake of specific labelled substrates by uncultured microbes at the single cell level. This technique requires the hybridization of rRNA targeted halogenated DNA probes on fixed and permeabilized microorganisms. Exogenous atoms are introduced into cells and endogenous atoms removed during the experimental procedures. Consequently differences between the original and the apparent isotopic composition of cells may occur. In the present study, the influence of the experimental procedures of SIMSISH on the isotopic composition of carbon in E. coli cells was evaluated with nanoSIMS and compared to elemental analyser-isotopic ratio mass spectrometer (EA-IRMS measurements. Our results show that fixation and hybridization have a very limited, reproducible and homogeneous influence on the isotopic composition of cells. Thereby, the SIMSISH procedure minimizes the contamination of the sample by exogenous atoms, thus providing a means to detect the phylogenetic identity and to measure precisely the carbon isotopic composition at the single cell level. This technique was successfully applied to a complex sample with double bromine - iodine labelling targeting a large group of bacteria and a specific archaea to evaluate their specific (13C uptake during labelled methanol anaerobic degradation.

  7. SIMSISH Technique Does Not Alter the Apparent Isotopic Composition of Bacterial Cells

    Science.gov (United States)

    Chapleur, Olivier; Wu, Ting-Di; Guerquin-Kern, Jean-Luc; Mazéas, Laurent; Bouchez, Théodore

    2013-01-01

    In order to identify the function of uncultured microorganisms in their environment, the SIMSISH method, combining in situ hybridization (ISH) and nanoscale secondary ion mass spectrometry (nanoSIMS) imaging, has been proposed to determine the quantitative uptake of specific labelled substrates by uncultured microbes at the single cell level. This technique requires the hybridization of rRNA targeted halogenated DNA probes on fixed and permeabilized microorganisms. Exogenous atoms are introduced into cells and endogenous atoms removed during the experimental procedures. Consequently differences between the original and the apparent isotopic composition of cells may occur. In the present study, the influence of the experimental procedures of SIMSISH on the isotopic composition of carbon in E. coli cells was evaluated with nanoSIMS and compared to elemental analyser-isotopic ratio mass spectrometer (EA-IRMS) measurements. Our results show that fixation and hybridization have a very limited, reproducible and homogeneous influence on the isotopic composition of cells. Thereby, the SIMSISH procedure minimizes the contamination of the sample by exogenous atoms, thus providing a means to detect the phylogenetic identity and to measure precisely the carbon isotopic composition at the single cell level. This technique was successfully applied to a complex sample with double bromine – iodine labelling targeting a large group of bacteria and a specific archaea to evaluate their specific 13C uptake during labelled methanol anaerobic degradation. PMID:24204855

  8. Isotopic labeling affects 1,25-dihydroxyvitamin D metabolism

    International Nuclear Information System (INIS)

    Halloran, B.P.; Bikle, D.D.; Castro, M.E.; Gee, E.

    1989-01-01

    Isotope substitution can change the biochemical properties of vitamin D. To determine the effect of substituting 3H for 1H on the metabolism of 1,25(OH)2D3, we measured the metabolic clearance rate and renal metabolism of unlabeled and 3H-labeled 1,25(OH)2D3. Substitution of 3H for 1H on carbons 26 and 27 [1,25(OH)2[26,27(n)-3H]D3] or on carbons 23 and 24 [1,25(OH)2[23,24(n)-3H]D3] reduced the in vivo metabolic clearance rate of 1,25(OH)2D3 by 36% and 37%, respectively, and reduced the in vitro renal catabolism of 1,25(OH)2D3 by 11% and 54%, respectively. Substitutions of 3H for 1H on carbons 23 and 24 as opposed to carbons 26 and 27 reduced conversion of [3H]1,25(OH)2D3 to [3H]1,24,25(OH)2D3 by 25% and to putative 24-oxo-1,23,25-dihydroxyvitamin D3 by 1600%. These results indicate that substitution of 3H for 1H on carbons 26 and 27 or on carbons 23 and 24 can reduce the metabolic clearance rate and in vitro metabolism of 1,25(OH)2D3 and quantitatively alter the pattern of metabolic products produced

  9. Improved segmental isotope labeling of proteins and application to a larger protein

    International Nuclear Information System (INIS)

    Otomo, Takanori; Teruya, Kenta; Uegaki, Koichi; Yamazaki, Toshio; Kyogoku, Yoshimasa

    1999-01-01

    A new isotope labeling technique for peptide segments in a protein sample was recently established using the protein splicing element intein [Yamazaki et al. (1998) J. Am. Chem. Soc., 120, 5591-5592]. This method makes it possible to observe signals of a selected amino (N-) or carboxyl (C-) terminal region along a peptide chain. However, there is a problem with the yield of the segmentally labeled protein. In this paper, we report an increase in the yield of the protein that enables the production of sufficient amounts of segmentally 13 C/ 15 N-labeled protein samples. This was achieved by improvement of the expression level of the N-terminal fragment in cells and the efficiency of refolding into the active splicing conformation. The N-terminal fragment was expressed as a fused protein with the cellulose binding domain at its N-terminus, which was expressed as an insoluble peptide in cells and the expression level was increased. Incubation with 2.5 M urea and 50% glycerol increased the efficiency of the refolding greatly, thereby raising the final yields of the ligated proteins. The feasibility of application of the method to a high-molecular-weight protein was demonstrated by the results for a maltose binding protein consisting of 370 amino acids. All four examined joints in the maltose binding protein were successfully ligated to produce segmentally labeled protein samples

  10. Isotopes in medical diagnosis. New techniques reviewed at Vienna symposium

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1959-04-15

    The medical uses of radioisotopes cover diagnosis and therapy as well as clinical research. There is specialized equipment not only detects the radiations from a radioactive substance inside the body, but also produces a two-dimensional visual image of the size and functional condition of tissue that absorbs this substance. This is known as medical radioisotope scanning, aimed at determining the distribution of a radioisotope within an organ which specifically concentrates a radioisotope. Methods of scanning are exceedingly complex, and although a variety of equipment is now available for the automatic determination of the distribution of a radioisotope in an organ, experience of their use is still limited. To facilitate an adequate exchange of views and a pooling of experience in this field, the International Atomic Energy Agency and the World Health Organization recently arranged a seminar to discuss the techniques and results of scanning. Topics discussed included problems of collimation, sensitivity patterns of various collimating systems, optimum isotope concentrations, essential requirements for obtaining satisfactory visualization by a scanner, basic principles of scintillation counting, a the scintillation camera, isotope scanning for studies on liver disorders and liver tumours, radiogold used for liver scanning, scanning the liver and pancreas with the help of positron-emitting isotopes, measuring the distribution of radioisotopes in deep lying tissues by detection of 'bremsstrahlen', 'profile counting', brain tumour scanning with the usage of radioarsenic or radioiodine-labelled albumen, different aspects of thyroid scanning, value and methods of scanning in thyroid cancer as well as scanning procedures in non-cancerous thyroid disease. During the discussions Dr. Keroe (IAEA) demonstrated a new electronic device, the use of which results in a substantial increase of contrast and resolution of the recordings of a scanning machine, and which is the first

  11. Testing isotopic labeling with [¹³C₆]glucose as a method of advanced glycation sites identification.

    Science.gov (United States)

    Kielmas, Martyna; Kijewska, Monika; Stefanowicz, Piotr; Szewczuk, Zbigniew

    2012-12-01

    The Maillard reaction occurring between reducing sugars and reactive amino groups of biomolecules leads to the formation of a heterogeneous mixture of compounds: early, intermediate, and advanced glycation end products (AGEs). These compounds could be markers of certain diseases and of the premature aging process. Detection of Amadori products can be performed by various methods, including MS/MS techniques and affinity chromatography on immobilized boronic acid. However, the diversity of the structures of AGEs makes detection of these compounds more difficult. The aim of this study was to test a new method of AGE identification based on isotope (13)C labeling. The model protein (hen egg lysozyme) was modified with an equimolar mixture of [(12)C(6)]glucose and [(13)C(6)]glucose and then subjected to reduction of the disulfide bridges followed by tryptic hydrolysis. The digest obtained was analyzed by LC-MS. The glycation products were identified on the basis of characteristic isotopic patterns resulting from the use of isotopically labeled glucose. This method allowed identification of 38 early Maillard reaction products and five different structures of the end glycation products. This isotopic labeling technique combined with LC-MS is a sensitive method for identification of advanced glycation end products even if their chemical structure is unknown. Copyright © 2012 Elsevier Inc. All rights reserved.

  12. Radioisotope labeling technique for vapor density measurements of volatile inorganic species

    International Nuclear Information System (INIS)

    Peterson, E.J.; Caird, J.A.; Hessler, J.P.; Hoekstra, H.R.; Williams, C.W.

    1979-01-01

    A new method for complexed metal ion vapor density measurement involving labeling the metal ions of interest with a radioactive isotope is described. The isotope chosen in the present work is unstable and leads to emission of a characteristic γ ray. Thus the γ-counting rate was related to the number density of complexed metal ions in the vapor phase. This technique is applicable to the study of any volatile inorganic species, but in the present study has been used to measure vapor densities of complex species in the TbCl 3 -AlCl 3 system by using tracer 160 Tb. 4 figures, 2 tables

  13. Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells

    International Nuclear Information System (INIS)

    Takahashi, Hideo; Shimada, Ichio

    2010-01-01

    The preparation of stable isotope-labeled proteins is necessary for the application of a wide variety of NMR methods, to study the structures and dynamics of proteins and protein complexes. The E. coli expression system is generally used for the production of isotope-labeled proteins, because of the advantages of ease of handling, rapid growth, high-level protein production, and low cost for isotope-labeling. However, many eukaryotic proteins are not functionally expressed in E. coli, due to problems related to disulfide bond formation, post-translational modifications, and folding. In such cases, other expression systems are required for producing proteins for biomolecular NMR analyses. In this paper, we review the recent advances in expression systems for isotopically labeled heterologous proteins, utilizing non-E. coli prokaryotic and eukaryotic cells.

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

    NARCIS (Netherlands)

    van Manen, H.J.; Lenferink, Aufrid T.M.; Otto, Cornelis

    2008-01-01

    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

  15. Gaseous isotope correlation technique for safeguards at reprocessing facilities

    International Nuclear Information System (INIS)

    Ohkubo, Michiaki.

    1988-03-01

    The isotope correlation technique based on gaseous stable fission products can be used as a means of verifying the input measurement to fuel reprocessing plants. This paper reviews the theoretical background of the gaseous fission product isotope correlation technique. The correlations considered are those between burnup and various isotopic ratios of Kr and Xe nuclides. The feasibility of gaseous ICT application to Pu input accountancy of reprocessing facilities is also discussed. The technique offers the possibility of in situ measurement verification by the inspector. (author). 16 refs, 7 figs

  16. Nuclear material safeguards surveillance and accountancy by isotope correlation techniques

    International Nuclear Information System (INIS)

    Persiani, P.J.; Goleb, J.A.

    1981-01-01

    This paper presents the initial phase of the US study program and involves the computation of isotopic correlations for the LMFBR fuel cycle. The LWR fuel cycle phase of the study is currently in progress. The selection of the more safeguards effective functionals will depend not only on the level of reliability of isotope correlation technique (ICT) for verification, but also on the capability and difficulty of developing measurement methods. Performance characteristics of existing and proposed isotopic measurement techniques cover the general areas including assay and isotopic wet chemistry and NDA: (1) simultaneous multicomponent analysis techniques, (2) mass spectrometry, (3) x-ray fluorescence or densitometry with high flux monochromatic x-ray sources and high dispersion spectrometers, (4) passive and active neutron interrogation, (5) high level gamma-ray spectroscopy, (6) coulometry, and (7) potentiometry. The measurement capabilities and inherent limitations of these systems are to be evaluated in terms of total systems, operational mode, sample preparation requirements and consequent effect on dissolver solution representation, accuracy and precision estimates (if available), development status of the technique, and development requirements. The isotopic correlation technique shows considerable promise for use in verifying the initial isotopic composition and burnup of discharged assemblies based on the measured ratios of several key isotopes, obtained probably at the dissolver stage in reprocessing. This technique should, for example, easily be capable of indicating the exchange of a blanket assembly for a fuel assembly

  17. Isotopic labelling studies for a gold-catalysed skeletal rearrangement of alkynyl aziridines

    Directory of Open Access Journals (Sweden)

    Neil Spencer

    2011-06-01

    Full Text Available Isotopic labelling studies were performed to probe a proposed 1,2-aryl shift in the gold-catalysed cycloisomerisation of alkynyl aziridines into 2,4-disubstituted pyrroles. Two isotopomers of the expected skeletal rearrangement product were identified using 13C-labelling and led to a revised mechanism featuring two distinct skeletal rearrangements. The mechanistic proposal has been rationalised against the reaction of a range of 13C- and deuterium-labelled substrates.

  18. Effects of must concentration techniques on wine isotopic parameters.

    Science.gov (United States)

    Guyon, Francois; Douet, Christine; Colas, Sebastien; Salagoïty, Marie-Hélène; Medina, Bernard

    2006-12-27

    Despite the robustness of isotopic methods applied in the field of wine control, isotopic values can be slightly influenced by enological practices. For this reason, must concentration technique effects on wine isotopic parameters were studied. The two studied concentration techniques were reverse osmosis (RO) and high-vacuum evaporation (HVE). Samples (must and extracted water) have been collected in various French vineyards. Musts were microfermented at the laboratory, and isotope parameters were determined on the obtained wine. Deuterium and carbon-13 isotope ratios were studied on distilled ethanol by nuclear magnetic resonance (NMR) and isotope ratio mass spectrometry (IRMS), respectively. The oxygen-18 ratio was determined on extracted and wine water using IRMS apparatus. The study showed that the RO technique has a very low effect on isotopic parameters, indicating that this concentration technique does not create any isotopic fractionation, neither at sugar level nor at water level. The effect is notable for must submitted to HVE concentration: water evaporation leads to a modification of the oxygen-18 ratio of the must and, as a consequence, ethanol deuterium concentration is also modified.

  19. Technique of leukocyte harvesting and labeling: problems and perspectives

    International Nuclear Information System (INIS)

    McAfee, J.G.; Subramanian, G.; Gagne, G.

    1984-01-01

    Mixed leukocyte suspensions obtained after gravity sedimentation of red cells and labeled with 111 In lipophilic chelates are now widely used clinically for abscess localization at many medical centers. So far, labeling with 111 In-oxine or tropolone has been more successful than any 99 mTc method. More sophisticated approaches are available for isolation and labeling of specific leukocyte cell types, to study their migration in vivo. The most significant advances in cell harvesting include newer density gradients for isopyknic centrifugation, centrifugal elutriation, and flow cytometry. Unlike current radioactive agents which label many cell types indiscriminately, more selective ligands are being developed which bind to specific cell surface receptors. These will label certain leukocyte populations or subtypes while not reacting with others, thereby avoiding laborious separation techniques. Monoclonal antibodies against leukocyte cell-surface antigens appear particularly promising as agents for selective cell labeling

  20. Lymphocyte labelling technique for the exploration of kidney transplants

    International Nuclear Information System (INIS)

    Guey, A.; Touraine, J.L.; Collard, M.; Claveyrolas, P.; Bouteiller, O. de; Traeger, J.

    The labelling technique is developed with a precise clinical exploration in view and has to take into account the following rules or conditions: - the blood sample must be smaller than 20 ml; - the manipulation must not last more than 3 hours; - the immunological properties of the labelled lymphocytes must be kept intact; - the solution reinjected into the patient must contain no aggregates, be absolutely sterile and possess a radioactivity above 1mCi. The technique of extraction and labelling from a sample of about 15ml is described. The main factors responsible for the quality of the labelling are analysed, together with the labelling and irradiation dose effects on certain properties of the lymphocytes (viability, rosette E formation, proliferative response to mitogens) [fr

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

    Energy Technology Data Exchange (ETDEWEB)

    Schaefer, J. [Washington Univ., St. Louis, MO (United States)

    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.

  2. Evaluation of the ''Sabana de Bogota'' aquifer using isotopes techniques

    International Nuclear Information System (INIS)

    Torres, E.; Jimenez, G.; Obando, E.; Sanchez, L.; Alayon, E.

    1987-01-01

    Groundwater from the Sabana de Bogota area was studied by means of isotopic techniques such as isotope analysis, injection of tracers for determination of hydrogeological parameters, velocity measurements using tracers. These results combined with the ones obtained by conventional techniques were used to determine recharge area, origin, equipotencial lines and direction of groundwater. The results show that a combined application of classical and nuclear methods may establish rather reasonable hydrogeological concepts without investing much time and money

  3. Fatty acids labelled in the. omega. -position with iodine isotopes

    Energy Technology Data Exchange (ETDEWEB)

    Mathieu, J.P.; Busquet, G.; Comet, M. (Universite Scientifique et Medicale de Grenoble, 38 - La Tronche (France)); Riche, F.; Vidal, M. (Laboratoire d' Etudes Dynamiques et Structurales de la Selectivite, 38 - Grenoble (France)); Coornaert, S.; Bardy, A. (CEA, Centre de Saclay, 91 - Gif-sur-Yvette (France)); Godart, J. (Institut des Sciences Nucleaires, 38 - Grenoble (France))

    1982-01-01

    The synthesis of saturated acetylenic and olefinic (Z or E) ..omega..-iodinated fatty acids has been carried out and their labelling with iodine-131 or 123 by exchange I/sup -/, *I/sup -/ has been studied. The influence of several parameters -water and fatty acid concentrations, specific activity, labelling solution acidity, iodine carrier presence- on this exchange reaction has been noted, enabling experimental conditions to be defined that produce labelling yields of greater than 95%. These results should lead to widespread clinical use of iodine labelled fatty acids.

  4. Investigation into reaction of heterogenous isotopic exchange with gaseoUs tritium in solution for preparation labelled lipid compounds

    International Nuclear Information System (INIS)

    Shevchenko, V.P.; Myasoedov, N.F.

    1983-01-01

    The applicability of the method of heterogeneous catalytic isotopic exchange with gaseous tritium in the solution for the production of labelled lipide preparations is studied. Labelled saturated and unsaturated aliphatic acids, prostaglandins, phospholipides and sphingolipides are prepared

  5. Synthesis of 13C and 2H labelled retinals: spectroscopic investigations on isotopically labelled rhodopsin and bacteriorhodopsin

    International Nuclear Information System (INIS)

    Pardoen, J.A.

    1986-01-01

    In order to develop probes of the structure of chromophores, the author introduces isotopic modifications at specific chromophoric positions as structural probes. To obtain bacteriorhodopsin, rhodopsin and their photoproducts labelled in the chromophore at selected positions, bacterioopsin and opsin were reacted with the appropriate labelled a11-trans and 11-cis retinals. The author describes the synthesis of a11-trans retinal selectively 13 C labelled at different positions. The characterization of these labelled a11-trans retinals by mass spectrometry, 300 MHz 1 H NMR and 75 MHz 13 C NMR spectroscopy is given. The photochemical preparation and isolation of the pure 9-, 11- and 13-cis forms is described in the experimental part. (Auth.)

  6. 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

    International Nuclear Information System (INIS)

    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

  7. 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

    Energy Technology Data Exchange (ETDEWEB)

    Trewhella, J.; Cross, T.A.; Unkefer, C.J. [eds.

    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.

  8. Lung transit of /sup 111/Indium-labelled granulocytes. Relationship to labelling techniques

    Energy Technology Data Exchange (ETDEWEB)

    Saverymuttu, S.H.; Peters, A.M.; Danpure, H.J.; Reavy, H.J.; Osman, S.; Lavender, J.P. (Hammersmith Hospital, London, England)

    1983-01-01

    The early in vivo distribution of /sup 111/Indium-labelled granulocytes, recorded by dynamic imaging using a gamma camera and computer, varied according to the separation and labelling technique. Following i.v. bolus injection, 4 kinetic patterns could be identified: (A) rapid transit through the pulmonary vasculature, (B) delayed transit through the lung with clearance by about 30 min, (C) complete retention by the lung, for up to 10 min, followed by slow release over a period of 1 to 2 h, (D) delayed transit through the lung with a similar time course to (B) but with subsequent heavy liver uptake. Granulocytes labelled with /sup 111/In-tropolonate and maintained in plasma throughout the labelling procedure, whether injected as a 'pure' (separated by plasma-enriched density gradient centrifugation) or 'crude' (seprated by differential centrifugation) preparation, displayed type A kinetics, thought to most closely represent the normal behaviour of granulocytes. 'Crude' cells labelled in saline with /sup 111/In-acetylacetonate displayed type B kinetics. 'Pure' cells isolated on Percoll-saline and labelled in saline with /sup 111/In-acetylacetonate displayed type C kinetics, thought to represent granulocyte 'stimulation' and/or damage, or type D kientics, thought to represent severe damage. The importance is stressed of labelling granulocytes for kinetic studies with a technique that results in minimal alteration of cell behaviour.

  9. Cell-free expression and stable isotope labelling strategies for membrane proteins

    International Nuclear Information System (INIS)

    Sobhanifar, Solmaz; Reckel, Sina; Junge, Friederike; Schwarz, Daniel; Kai, Lei; Karbyshev, Mikhail; Loehr, Frank; Bernhard, Frank; Doetsch, Volker

    2010-01-01

    Membrane proteins are highly underrepresented in the structural data-base and remain one of the most challenging targets for functional and structural elucidation. Their roles in transport and cellular communication, furthermore, often make over-expression toxic to their host, and their hydrophobicity and structural complexity make isolation and reconstitution a complicated task, especially in cases where proteins are targeted to inclusion bodies. The development of cell-free expression systems provides a very interesting alternative to cell-based systems, since it circumvents many problems such as toxicity or necessity for the transportation of the synthesized protein to the membrane, and constitutes the only system that allows for direct production of membrane proteins in membrane-mimetic environments which may be suitable for liquid state NMR measurements. The unique advantages of the cell-free expression system, including strong expression yields as well as the direct incorporation of almost any combination of amino acids with very little metabolic scrambling, has allowed for the development of a wide-array of isotope labelling techniques which facilitate structural investigations of proteins whose spectral congestion and broad line-widths may have earlier rendered them beyond the scope of NMR. Here we explore various labelling strategies in conjunction with cell-free developments, with a particular focus on α-helical transmembrane proteins which benefit most from such methods.

  10. Dynamics of N2O production pathways analyzed by 15N18O isotope labeling

    DEFF Research Database (Denmark)

    Jensen, Marlene Mark; Ma, Chun; Lavik, Gaute

    Nitrous oxide production associated with biological nitrogen transformations can contribute substantially to the CO2 footprint of both man-made and natural systems, but the pathways and regulation of N2O production are poorly understood. We developed a 15N/18O dual isotope labelling technique...

  11. Kinetic isotope effects in reaction of ferment oxidation of tritium-labelled D-galactosamine

    International Nuclear Information System (INIS)

    Akulov, G.P.; Korsakova, N.A.

    1992-01-01

    Primary, secondary and intramolecular kinetic isotopic effects in reaction of ferment oxidation of D-galactosamine labelled by tritium in position 6, were measured. When comparing values of the effects with previously obtained results for similar reaction D-[6- 3 H]galactose, it was ascertained that the presence of aminogroup in galactopyranosyl mainly affects kinetics of substrate-ferment complex formation stage. The possibility to use kinetic isotope effects for increase in molar activity of D-galactosamine, labelled by tritium in position 6, is shown

  12. Advisory group meeting on stable isotope labelled compounds in biomedical studies

    International Nuclear Information System (INIS)

    Vera Ruiz, H.; Parr, R.M.

    1985-11-01

    The programme of the meeting was restricted to topics involving applications of stable isotopes of the lighter elements (H, C, N, O). The current status of stable isotope techniques and applications in nutritional and biomedical studies, the applicability of these techniques in developing countries and the IAEA's future programmes on this topic were discussed

  13. Radiation and isotopic techniques in the study and control of piroplasms of cattle: a review

    International Nuclear Information System (INIS)

    Irvin, A.D.; Brocklesby, D.W.; Purnell, R.E.

    1979-01-01

    Radiation and isotopic techniques have been used in a number of ways to study Babesia and Theileria parasites (piroplasms), and in attempts to vaccinate animals against diseases induced by these agents. Work involving the piroplasm infections of cattle is reviewed in this paper. Some success has been achieved by using radiation to modify tick or bovine stages of parasites, for use as potential vaccines. Radiation has also been used to induce immuno-suppression in laboratory animals, in an effort to render them susceptible to bovine piroplasms, particularly Theileria. Isotope tracer techniques have been used for labelling parasites, and for studying metabolic pathways. Encouraging results, particularly with regard to vaccine development, indicate the importance of radiation techniques in the study and control of piroplasms of cattle. (Auth.)

  14. The choice of label and measurement technique in tracer studies of body protein metabolism in man

    International Nuclear Information System (INIS)

    James, W.P.T.; Sender, P.M.; Garlick, P.J.; Waterlow, J.C.

    1975-01-01

    The turnover of non-serum proteins in man has had limited study despite the physiological importance of maintaining the balance between synthesis and breakdown of body proteins. Body protein is usually considered as a single pool and breakdown rates are often measured by monitoring excreted label at intervals after pulse labelling with radioactive or 15 N amino acids. No label has yet been used for measuring tissue protein breakdown in man which is free from the major problem of label re-utilization. All measurements of breakdown rates, eg. with 75 Se-selenomethionine, 15 N- or 14 C-glycine, give rate constants which are too low. The heterogeneity of body proteins also means that an estimate of the weighted average breakdown rate can only be obtained after following the excretion of isotope for a long period, perhaps of the order of 3-4 half-lives which, for man, would be 100 days after labelling. We therefore use infusions with either 14 C- or 15 N-labelled amino acids to measure breakdown and synthesis rates: these values are less affected by problems of protein heterogeneity. Single injection techniques are subject to more error than constant infusions of label because of the difficulty of defining the precursor activity. 15 N labelling need not be confined to essential amino acids if total protein rather than amino acid turnover is studied: the latter involves measurements of the labelled amino acid itself which is difficult with 15 N because of the small amounts of free amino acid nitrogen available. Carbon labelling of non-essential amino acids is unsuitable for studies of protein turnover and the choice of the position of the label on the molecule is important when labelled essential amino acids are employed. Short-term changes in protein metabolism are evaluated better with amino acids with a small pool size; the equilibration time in the excretory bicarbonate pool is also shorter than in the urea pool so that 15 N is less useful than carbon labelling. We

  15. A new method for the labelling of proteins with radioactive arsenic isotopes

    Energy Technology Data Exchange (ETDEWEB)

    Jennewein, M. [Institute of Nuclear Chemistry, Johannes Gutenberg University of Mainz, Fritz-Strassmann-Weg 2, 55128 Mainz (Germany); Hermanne, A. [VUB Cyclotron, University of Brussels, Laarbeeklaan 103, 1090 Brussels (Belgium); Mason, R.P. [Department of Radiology, Advanced Radiological Sciences, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas (United States); Thorpe, P.E. [Department of Pharmacology and Simmons and Hamon Cancer Centers, University of Texas Southwestern Medical Center at Dallas, Dallas, TX (United States); Roesch, F. [Institute of Nuclear Chemistry, Johannes Gutenberg University of Mainz, Fritz-Strassmann-Weg 2, 55128 Mainz (Germany)]. E-mail: frank.roesch@uni-mainz.de

    2006-12-20

    Radioarsenic labelled radiopharmaceuticals could be a valuable asset to positron emission tomography. In particular, the long half-lives of {sup 72}As (T{sub 1/2}=26h) and {sup 74}As (T{sub 1/2}=17.8d) allow to investigate slow physiological or metabolical processes, like the enrichment and distribution of monoclonal antibodies (mab) in tumour tissue. In this work, a new method for the labelling of proteins with various radioactive arsenic isotopes was developed. For this purpose, two proteins, namely a chimeric IgG{sub 3} monoclonal antibody, ch3G4, directed against anionic phospholipids, and Rituxan (Rituximab), were labelled as a proof of principle with no-carrier-added radioarsenic isotopes ({sup 74}As and {sup 77}As). The developed labelling chemistry gives high yields (>99.9%), is reliable and could easily be transferred to automated labelling systems in a clinical environment. At least for the mab used in this work, this route of radioarsenic labelling does not affect the immunoreactivity of the product. The arsenic label stays stable for up to 72h at the molecular mass of the monoclonal antibody, which is in particular relevant to follow the pharmacology and pharmacokinetics of the labelled mab for several days.

  16. Direct isotope determination of isotopically labelled lipids by field desorption mass spectrometry

    International Nuclear Information System (INIS)

    Lehmann, W.D.; Kessler, M.

    1982-01-01

    Lipids labelled with deuterium or carbon-14 have been investigated by field desorption mass spectrometry for determination of their degree of labelling. This application is demonstrated for free fatty acids, cholesterol, cholesteryl esters, triglycerides, and L-α-phosphatidylcholines. Comparison of the molecular ion groups of the non-labelled and of the labelled compounds enables a fast and reliable determination of the degree of labelling. For multiply labelled compounds the label distribution is also obtained from the molecular ion group. In addition, for cholesteryl esters and for phosphatidylcholines structurally significant fragment ions provide information about the position of the label. Several hundred nanograms of the compound are typically required for a single analysis with a relative standard error of 0.5-2% in the value calculated for atom% hydrogen-2 or for the specific carbon-14 activity. (orig.) [de

  17. Application of isotope-labelled compounds in the study of the chemical stability of pesticides

    International Nuclear Information System (INIS)

    Roesseler, M.; Luther, D.; Abendroth, H.C.; Koch, H.

    1980-01-01

    The user of pesticides requires specific biological modes of action from the corresponding commercial products. Impurities and degradation products may cause uncontrollable toxicological reactions. Profound knowledge of the chemical stability of the effective substance in question and its formulations under storage conditions as well as under those of analytical sample preparation and detection is required. Radioisotope labelled effective substances dimethoate and 1-butyl-amino-cyclohexane-phosphonic acid dibutyl ester are used to study storage stability of the pure effective substance and its formulations; effects of selected impurities, such as technical by-products, moisture or water content, binding or carrier materials, organic solvents, chemical stabilizers and other formulation components on storage properties; temperature dependence of storage stability; selection of suitable analytical techniques for quantitative determination of the effective substance without interference effects from any by-product; reduction of the necessary analytical expense; disclosure of sources of error in the application of usual analytical techniques; improvement of possibilities of an immediate and clearer discrimination between types and amounts of compounds in a chemical system consisting of one pesticide and its degradation or reaction products at the beginning and at the end of an experimental or reaction period. Radiochemical analytical techniques, such as radio thin-layer chromatography (also combined with liquid scintillation counting), radio gas chromatography, autoradiography and isotope dilution analysis were used. Results are discussed, especially of experiments on dimethoate and its technical by-products

  18. Affordable uniform isotope labeling with 2H, 13C and 15N in insect cells

    International Nuclear Information System (INIS)

    Sitarska, Agnieszka; Skora, Lukasz; Klopp, Julia; Roest, Susan; Fernández, César; Shrestha, Binesh; Gossert, Alvar D.

    2015-01-01

    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

  19. Animal science: Isotopes and nuclear techniques

    International Nuclear Information System (INIS)

    Vercoe, J.E.

    1978-01-01

    In many regions of the world, animal production is limited by poor growth, reproductive performance and milk output of livestock, thus limiting the availability of animal products such as meat, milk, leather, etc., for use by man. Reduced animal production results from inadequate or unbalanced nutrition, lack of adaptation to climatic conditions, and parasitic and other diseases The production from animals can be improved on the one hand by improving genotypes (the breeds and strains within a given species) to make them better suited to the environment to which they are exposed, and on the other hand by modifying the components of the environment which are limiting the production. In practice, a combination of these two strategies is usually desirable but the relative importance of each is determined by short- and long-term economic considerations and the likely environmental consequences. Isotopes and radiation play an important part in identifying and alleviating the genetic and environmental limitations to animal production

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

    International Nuclear Information System (INIS)

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

    1986-01-01

    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 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 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 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 2 production

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

    Science.gov (United States)

    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...

  2. Soil organic matter (SOM) dynamics determined by stable isotope techniques

    International Nuclear Information System (INIS)

    Gerzabek, M. H.

    1998-09-01

    Being aware of limitations and possible bias the 13 C natural abundance technique using the different 13 C enrichments in plants with differing photosynthetic pathways in a powerful tool to quantify turnover processes, both in long-term field studies and short-term laboratory experiments. Special care is needed in choosing reference plots and the proper number of replicate samples. The combination of 13 C and 14 C measurements has a high potential for a further improvement of isotope techniques in SOM studies. Natural abundance of 15 N is less powerful with respect to quantification of SOM processes than the isotope dilution technique. However its usefulness could be distinctly improved by introducing other stable isotopes into the studies.(author)

  3. Stable isotope measurement techniques for atmospheric greenhouse gases

    International Nuclear Information System (INIS)

    2002-01-01

    The technical requirements to perform useful measurements of atmospheric greenhouse gas concentrations and of their isotope ratios are of direct relevance for all laboratories engaged in this field. A meaningful interpretation of isotopes in global models on sources and sinks of CO 2 and other greenhouse gases depends on strict laboratory protocols and data quality control measures ensuring comparable data in time and space. Only with this precondition met, the isotope techniques can serve as a potentially powerful method for reducing uncertainties in the global CO 2 budgets and for tracing pathways and interaction of terrestrial, oceanic, and atmospheric pools of carbon. This publication provides four contributions describing methods for the determination of the isotopic composition of trace gases in atmospheric air and in ice cores. These contributions have been indexed separately

  4. Expeditious syntheses of stable and radioactive isotope-labeled anticonvulsant agent, JNJ-26990990, and its metabolites.

    Science.gov (United States)

    Lin, Ronghui; Weaner, Larry E; Hoerr, David C; Salter, Rhys; Gong, Yong

    2013-01-01

    Syntheses of stable and radioactive isotope-labeled anticonvulsant agent, JNJ-26990990, that is, N-(benzo[b]thien-3-ylmethyl)-sulfamide and its metabolites are described. [(13)C(15)N]Benzo[b]thiophene-3-carbonitrile was first prepared by coupling of 3-bromo-benzo[b]thiophene with [(13)C(15)N]-copper cyanide. The resultant [(13)C(15)N]benzo[b]thiophene-3-carbonitrile was reduced with lithium aluminum deuteride to give [(13)CD2(15)N]benzo[b]thiophen-3-yl-methylamine; which was then coupled with sulfamide to afford [(13)CD2(15)N]-N-(benzo[b]thien-3-ylmethyl)-sulfamide, the stable isotope-labeled compound with four stable isotope atoms. Direct oxidation of [(13)CD2(15)N]-N-(benzo[b]thien-3-ylmethyl)-sulfamide with hydrogen peroxide and peracetic acid gave the stable isotope-labeled sulfoxide and sulfone metabolites. On the other hand, radioactive (14)C-labeled N-(benzo[b]thien-3-ylmethyl)-sulfamide was prepared conveniently by sequential coupling of 3-bromo-benzo[b]thiophene with [(14)C]-copper cyanide, reduction of the carbonitrile to carboxaldehyde, and reductive amination with sulfamide. Copyright © 2013 John Wiley & Sons, Ltd.

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

    Czech Academy of Sciences Publication Activity Database

    da Costa, Sara; Ek Weis, Johan; Frank, Otakar; Fridrichová, Michaela; Kalbáč, Martin

    2016-01-01

    Roč. 3, č. 2 (2016), 025022 ISSN 2053-1583 R&D Projects: GA MŠk LL1301 Institutional support: RVO:61388955 Keywords : graphene bilayer * Raman spectroscopy * isotope labeling Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 6.937, year: 2016

  6. Mass spectrometric studies of stable isotope-labelled carboxylic acid derivatives

    International Nuclear Information System (INIS)

    Andersson, B.Aa.; Dinger, F.; Dinh-Nguyen, N.

    1975-01-01

    Low resolution mass spectra of deuterium and carbon-13 labelled fatty acid pyrrolidides are discussed. The simple fragmentation pattern of pyrrolidides makes them superior to other derivatives, regarding location of isotopes. Deuteriation of ethylenic fatty acid pyrrolidides therefore seems to be an improved method to locate carbon-carbon double bonds by mass spectrometry. (author)

  7. Identification of miRNA targets with stable isotope labeling by amino acids in cell culture

    DEFF Research Database (Denmark)

    Vinther, Jeppe; Hedegaard, Mads Marquardt; Gardner, Paul Phillip

    2006-01-01

    miRNAs are small noncoding RNAs that regulate gene expression. We have used stable isotope labeling by amino acids in cell culture (SILAC) to investigate the effect of miRNA-1 on the HeLa cell proteome. Expression of 12 out of 504 investigated proteins was repressed by miRNA-1 transfection...

  8. Temperature-induced strain and doping in monolayer and bilayer isotopically labeled graphene

    Czech Academy of Sciences Publication Activity Database

    Verhagen, Timotheus; Drogowska, Karolina; Kalbáč, Martin; Vejpravová, Jana

    2015-01-01

    Roč. 92, č. 12 (2015), "125437-1"-"125437-9" ISSN 1098-0121 R&D Projects: GA ČR GA15-02196S; GA MŠk LL1301 Institutional support: RVO:68378271 ; RVO:61388955 Keywords : isotopically labeled graphene * temperature dependence * Raman spectroscopy * phonons Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 3.736, year: 2014

  9. Isotope effect study of κ-(BEDT-TTF)2Cu(NCS)2: Labeling in the anion

    International Nuclear Information System (INIS)

    Kini, A.M.; Wang, H.H.; Schlueter, J.A.

    1995-01-01

    Since the initial discovery of organic superconductivity in 1979, a large number of organic superconductors have now been synthesized. However, the mechanism of electron-pairing in these novel superconductors has remained largely unresolved. Isotope effect studies constitute an important experimental tool for the investigation of whether or not the electron-pairing mechanism in organic superconductors is phonon-mediated, as in conventional superconductors. Recent isotope effect studies in the authors' laboratory, involving seven different isotopically labeled BEDT-TTF (or ET) derivatives, have demonstrated the following: (1) intramolecular phonon modes involving C double-bond C and Csingle bondS stretching vibrations in the ET donor molecule are not the dominant mediators of electron-pairing, and (2) in κ-(ET) 2 Cu(NCS) 2 , there exist two competing isotope effects--a normal mass effect, i.e., lowering of T c upon isotopic labeling, when the ET molecular mass is increased by concurrent 13 C and 34 S labeling, in addition to an inverse isotope effect upon deuterium labeling in ET. It is of great interest to investigate if there is an isotope effect when the charge-compensating anions, which are also located within the non-conducting layer in the superconducting cation-radical salts, are isotopically labeled. The existence of an isotope effect when the anions are labeled would be indicative of electron-pairing with the mediation of vibrational frequencies associated with the anions. In this paper, the authors present the results of the first isotope effect study in which isotopic labeling in the anion portion of κ-(ET) 2 Cu(NCS) 2 is carried out. The authors find no isotope effect when the carbon and nitrogen atoms of the thiocyanate groups in the anion are replaced with 13 C and 15 N isotopes

  10. Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation.

    Science.gov (United States)

    Pabst, Martin; Benešová, Iva; Fagerer, Stephan R; Jacobsen, Mathias; Eyer, Klaus; Schmidt, Gregor; Steinhoff, Robert; Krismer, Jasmin; Wahl, Fabian; Preisler, Jan; Zenobi, Renato

    2016-01-04

    We introduce a stable isotope labeling approach for glycopeptides that allows a specific glycosylation site in a protein to be quantitatively evaluated using mass spectrometry. Succinic anhydride is used to specifically label primary amino groups of the peptide portion of the glycopeptides. The heavy form (D4(13)C4) provides an 8 Da mass increment over the light natural form (H4(12)C4), allowing simultaneous analysis and direct comparison of two glycopeptide profiles in a single MS scan. We have optimized a protocol for an in-solution trypsin digestion, a one-pot labeling procedure, and a post-labeling solid-phase extraction to obtain purified and labeled glycopeptides. We provide the first demonstration of this approach by comparing IgG1 Fc glycopeptides from polyclonal IgG samples with respect to their galactosylation and sialylation patterns using MALDI MS and LC-ESI-MS.

  11. Assessing of distribution, mobility and bioavailability of exogenous Pb in agricultural soils using isotopic labeling method coupled with BCR approach

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Zhi-Yong, E-mail: zhyhuang@jmu.edu.cn [College of Bioengineering, Jimei University, Xiamen 361021 (China); Xie, Hong [College of Bioengineering, Jimei University, Xiamen 361021 (China); Shandong Vocational Animal Science and Veterinary College, Weifang 261061 (China); Cao, Ying-Lan [College of Bioengineering, Jimei University, Xiamen 361021 (China); Cai, Chao [Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021 (China); Zhang, Zhi [College of Bioengineering, Jimei University, Xiamen 361021 (China)

    2014-02-15

    Highlights: • Large amounts of exogenous Pb were found to distribute in reducible fractions. • Very few of exogenous Pb were found to distribute in acid-extractable fractions. • More than 60% of exogenous Pb in rhizosphere soils lost after planting. • Isotopic labeling method and SEP enable to explore Pb bioavailability in soil. -- Abstract: The contamination of Pb in agricultural soils is one of the most important ecological problems, which potentially results in serious health risk on human health through food chain. Hence, the fate of exogenous Pb contaminated in agricultural soils is needed to be deeply explored. By spiking soils with the stable enriched isotopes of {sup 206}Pb, the contamination of exogenous Pb{sup 2+} ions in three agricultural soils sampled from the estuary areas of Jiulong River, China was simulated in the present study, and the distribution, mobility and bioavailability of exogenous Pb in the soils were investigated using the isotopic labeling method coupled with a four-stage BCR (European Community Bureau of Reference) sequential extraction procedure. Results showed that about 60–85% of exogenous Pb was found to distribute in reducible fractions, while the exogenous Pb in acid-extractable fractions was less than 1.0%. After planting, the amounts of exogenous Pb presenting in acid-extractable, reducible and oxidizable fractions in rhizospheric soils decreased by 60–66%, in which partial exogenous Pb was assimilated by plants while most of the metal might transfer downward due to daily watering and applying fertilizer. The results show that the isotopic labeling technique coupled with sequential extraction procedures enables us to explore the distribution, mobility and bioavailability of exogenous Pb contaminated in soils, which may be useful for the further soil remediation.

  12. Assessing of distribution, mobility and bioavailability of exogenous Pb in agricultural soils using isotopic labeling method coupled with BCR approach

    International Nuclear Information System (INIS)

    Huang, Zhi-Yong; Xie, Hong; Cao, Ying-Lan; Cai, Chao; Zhang, Zhi

    2014-01-01

    Highlights: • Large amounts of exogenous Pb were found to distribute in reducible fractions. • Very few of exogenous Pb were found to distribute in acid-extractable fractions. • More than 60% of exogenous Pb in rhizosphere soils lost after planting. • Isotopic labeling method and SEP enable to explore Pb bioavailability in soil. -- Abstract: The contamination of Pb in agricultural soils is one of the most important ecological problems, which potentially results in serious health risk on human health through food chain. Hence, the fate of exogenous Pb contaminated in agricultural soils is needed to be deeply explored. By spiking soils with the stable enriched isotopes of 206 Pb, the contamination of exogenous Pb 2+ ions in three agricultural soils sampled from the estuary areas of Jiulong River, China was simulated in the present study, and the distribution, mobility and bioavailability of exogenous Pb in the soils were investigated using the isotopic labeling method coupled with a four-stage BCR (European Community Bureau of Reference) sequential extraction procedure. Results showed that about 60–85% of exogenous Pb was found to distribute in reducible fractions, while the exogenous Pb in acid-extractable fractions was less than 1.0%. After planting, the amounts of exogenous Pb presenting in acid-extractable, reducible and oxidizable fractions in rhizospheric soils decreased by 60–66%, in which partial exogenous Pb was assimilated by plants while most of the metal might transfer downward due to daily watering and applying fertilizer. The results show that the isotopic labeling technique coupled with sequential extraction procedures enables us to explore the distribution, mobility and bioavailability of exogenous Pb contaminated in soils, which may be useful for the further soil remediation

  13. Sensing site-specific structural characteristics and chirality using vibrational circular dichroism of isotope labeled peptides.

    Science.gov (United States)

    Keiderling, Timothy A

    2017-12-01

    Isotope labeling has a long history in chemistry as a tool for probing structure, offering enhanced sensitivity, or enabling site selection with a wide range of spectroscopic tools. Chirality sensitive methods such as electronic circular dichroism are global structural tools and have intrinsically low resolution. Consequently, they are generally insensitive to modifications to enhance site selectivity. The use of isotope labeling to modify vibrational spectra with unique resolvable frequency shifts can provide useful site-specific sensitivity, and these methods have been recently more widely expanded in biopolymer studies. While the spectral shifts resulting from changes in isotopic mass can provide resolution of modes from specific parts of the molecule and can allow detection of local change in structure with perturbation, these shifts alone do not directly indicate structure or chirality. With vibrational circular dichroism (VCD), the shifted bands and their resultant sign patterns can be used to indicate local conformations in labeled biopolymers, particularly if multiple labels are used and if their coupling is theoretically modeled. This mini-review discusses selected examples of the use of labeling specific amides in peptides to develop local structural insight with VCD spectra. © 2017 Wiley Periodicals, Inc.

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

    International Nuclear Information System (INIS)

    Serianni, A.S.

    1994-01-01

    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

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

    Energy Technology Data Exchange (ETDEWEB)

    Serianni, A.S. [Univ. of Notre Dame, IN (United States)

    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.

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

    Science.gov (United States)

    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.

  17. Process efficiency in isotope techniques by microelectronics

    International Nuclear Information System (INIS)

    Ziegler, J.; Boelke, L.; Nagli, G.; Schaelicke, W.

    1987-01-01

    The wide application of tracer techniques as a diagnostic tool in nuclear medicine requires the dosage of radioactive tracer solutions in a volume range of 0.5 to 10 ml with a high degree of precision. Two types of computer controlled filling and closing machines for injection bottles are described. Safe handling of radioactive solutions is taken in special consideration. (author)

  18. Site-specific Orientation of an α-helical Peptide Ovispirin-1 from Isotope Labeled SFG Spectroscopy

    Science.gov (United States)

    Ding, Bei; Laaser, Jennifer E.; Liu, Yuwei; Wang, Pengrui; Zanni, Martin T.; Chen, Zhan

    2013-01-01

    Sum-frequency generation (SFG) vibrational spectroscopy is often used to probe the backbone structures and orientations of polypeptides at surfaces. Using the ovispirin-1 polypeptide at the solid/liquid interface of polystyrene, we demonstrate for the first time that SFG can probe the polarization response of a single isotope labeled residue. To interpret the spectral intensities, we simulated the spectra using an excitonic Hamiltonian approach. We show that the polarization dependence of either the label or the unlabeled amide I band alone does not provide sufficient structural constraints to obtain both the tilt and the twist of the ovispirin helix at a solid/liquid interface, but that both can be determined from the polarization dependence of the complete spectrum. For ovispirin, the detailed analysis of the polarized SFG experimental data shows that the helix axis is tilted at roughly 138 degrees from the surface normal, and the transition dipole of the isotope labeled C=O group is tilted at 23 degrees from the surface normal, with the hydrophobic region facing the polystyrene surface. We further demonstrated that the Hamiltonian approach is able to address the coupling effect and the structural disorder. For comparison, we also collected the FTIR spectrum of ovispirin under similar conditions, which reveals the enhanced sensitivity of SFG for structural studies of single monolayer peptide surfaces. Our study provides insight into how structural and environmental effects appear in SFG spectra of the amide I band and establishes that SFG of isotope labeled peptides will be a powerful technique for elucidating secondary structures with residue-by-residue resolution. PMID:24228619

  19. Site-specific orientation of an α-helical peptide ovispirin-1 from isotope-labeled SFG spectroscopy.

    Science.gov (United States)

    Ding, Bei; Laaser, Jennifer E; Liu, Yuwei; Wang, Pengrui; Zanni, Martin T; Chen, Zhan

    2013-11-27

    Sum-frequency generation (SFG) vibrational spectroscopy is often used to probe the backbone structures and orientations of polypeptides at surfaces. Using the ovispirin-1 polypeptide at the solid/liquid interface of polystyrene, we demonstrate for the first time that SFG can probe the polarization response of a single-isotope-labeled residue. To interpret the spectral intensities, we simulated the spectra using an excitonic Hamiltonian approach. We show that the polarization dependence of either the label or the unlabeled amide I band alone does not provide sufficient structural constraints to obtain both the tilt and the twist of the ovispirin helix at a solid/liquid interface, but that both can be determined from the polarization dependence of the complete spectrum. For ovispirin, the detailed analysis of the polarized SFG experimental data shows that the helix axis is tilted at roughly 138° from the surface normal, and the transition dipole of the isotope-labeled C═O group is tilted at 23° from the surface normal, with the hydrophobic region facing the polystyrene surface. We further demonstrate that the Hamiltonian approach is able to address the coupling effect and the structural disorder. For comparison, we also collected the FTIR spectrum of ovispirin under similar conditions, which reveals the enhanced sensitivity of SFG for structural studies of single monolayer peptide surfaces. Our study provides insight into how structural and environmental effects appear in SFG spectra of the amide I band and establishes that SFG of isotope-labeled peptides will be a powerful technique for elucidating secondary structures with residue-by-residue resolution.

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

    Science.gov (United States)

    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. © 2015 Elsevier Inc. All rights reserved.

  1. Use of hydrological modelling and isotope techniques in Guvenc basin

    International Nuclear Information System (INIS)

    Altinbilek, D.

    1991-07-01

    The study covers the work performed under Project No. 335-RC-TUR-5145 entitled ''Use of Hydrologic Modelling and Isotope Techniques in Guvenc Basin'' and is an initial part of a program for estimating runoff from Central Anatolia Watersheds. The study presented herein consists of mainly three parts: 1) the acquisition of a library of rainfall excess, direct runoff and isotope data for Guvenc basin; 2) the modification of SCS model to be applied to Guvenc basin first and then to other basins of Central Anatolia for predicting the surface runoff from gaged and ungaged watersheds; and 3) the use of environmental isotope technique in order to define the basin components of streamflow of Guvenc basin. 31 refs, figs and tabs

  2. Isotope techniques in water resources development and management. Proceedings

    International Nuclear Information System (INIS)

    1999-01-01

    The 10th International Symposium on Isotope Techniques in Water Resources Development and Management was organized by the International Atomic Energy Agency in co-operation with UNESCO, WMO and International Association of Hydrological Sciences and was held at IAEA Headquarters, Vienna, during 10-14 May 1999. The symposium provided an international forum for assessing the status and recent advances in isotope applications to water resources and an exchange of information on the following main themes: processes at the interface between the atmosphere and hydrosphere; investigations in surface waters and groundwaters: their origin, dynamics, interrelations; problems and techniques for investigating sedimentation; water resources issues: pollution, source and transport of contaminants, salinization, water-rock interaction and processes in geothermal systems; isotope data interpretation and evaluation methodologies: modelling approaches. The proceedings contain the 46 papers presented and extended synopses of poster presentations; each of them was indexed individually

  3. Development of computational technique for labeling magnetic flux-surfaces

    International Nuclear Information System (INIS)

    Nunami, Masanori; Kanno, Ryutaro; Satake, Shinsuke; Hayashi, Takaya; Takamaru, Hisanori

    2006-03-01

    In recent Large Helical Device (LHD) experiments, radial profiles of ion temperature, electric field, etc. are measured in the m/n=1/1 magnetic island produced by island control coils, where m is the poloidal mode number and n the toroidal mode number. When the transport of the plasma in the radial profiles is numerically analyzed, an average over a magnetic flux-surface in the island is a very useful concept to understand the transport. On averaging, a proper labeling of the flux-surfaces is necessary. In general, it is not easy to label the flux-surfaces in the magnetic field with the island, compared with the case of a magnetic field configuration having nested flux-surfaces. In the present paper, we have developed a new computational technique to label the magnetic flux-surfaces. This technique is constructed by using an optimization algorithm, which is known as an optimization method called the simulated annealing method. The flux-surfaces are discerned by using two labels: one is classification of the magnetic field structure, i.e., core, island, ergodic, and outside regions, and the other is a value of the toroidal magnetic flux. We have applied the technique to an LHD configuration with the m/n=1/1 island, and successfully obtained the discrimination of the magnetic field structure. (author)

  4. Validation of radioisotopic labelling techniques in gastric emptying studies

    International Nuclear Information System (INIS)

    Corinaldesi, R.; Stanghellini, V.; Raiti, C.; Calamelli, R.; Salgemini, R.; Barbara, L.; Zarabini, G.E.

    1987-01-01

    Several techniques are currently employed to label solid and liquid foods with gamma-emitting radioisotopes in order to carry out gamma-camera gastric emptying studies. The present study describes an in vitro technique for evaluating the labelling stability of some of the most commonly employed radiomarkers of both the solid and liquid phases. Technetium-99m-sulphur colloid ( 99m Tc-SC) in vivo and in vitro labelled liver of chickens and other animal species appears to be almost ideal marker of the solid phase (97% of radioactivity still bound to the solid phase after incubation in gastric juice for 90 minutes). On the contrary, 51 CrCl 3 -beef ground meat (81%) and 99m Tc-SC egg white (69%) are unsatisfactory markers of the solid phase. Likewise, 99m Tc-DTPA and 111 In-DTPA cannot be considered satisfactory fluid-phase agents, because of the high proportion of radioactivity that leaves the liquid phase to become bound to the solid phase (respectively 76% and 49% after 90 minutes of incubation). This validation technique appears to be simple, feasible and reprodicible, and can be applied in any Nuclear Medicine Department to evaluate the validity of the labelling procedures, in order to improve the accuracy of the results of radioisotopic gastric emptying studies

  5. Assessing of distribution, mobility and bioavailability of exogenous Pb in agricultural soils using isotopic labeling method coupled with BCR approach.

    Science.gov (United States)

    Huang, Zhi-Yong; Xie, Hong; Cao, Ying-Lan; Cai, Chao; Zhang, Zhi

    2014-02-15

    The contamination of Pb in agricultural soils is one of the most important ecological problems, which potentially results in serious health risk on human health through food chain. Hence, the fate of exogenous Pb contaminated in agricultural soils is needed to be deeply explored. By spiking soils with the stable enriched isotopes of (206)Pb, the contamination of exogenous Pb(2+) ions in three agricultural soils sampled from the estuary areas of Jiulong River, China was simulated in the present study, and the distribution, mobility and bioavailability of exogenous Pb in the soils were investigated using the isotopic labeling method coupled with a four-stage BCR (European Community Bureau of Reference) sequential extraction procedure. Results showed that about 60-85% of exogenous Pb was found to distribute in reducible fractions, while the exogenous Pb in acid-extractable fractions was less than 1.0%. After planting, the amounts of exogenous Pb presenting in acid-extractable, reducible and oxidizable fractions in rhizospheric soils decreased by 60-66%, in which partial exogenous Pb was assimilated by plants while most of the metal might transfer downward due to daily watering and applying fertilizer. The results show that the isotopic labeling technique coupled with sequential extraction procedures enables us to explore the distribution, mobility and bioavailability of exogenous Pb contaminated in soils, which may be useful for the further soil remediation. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Development of techniques for furthering the use of isotope radiography

    International Nuclear Information System (INIS)

    Wamorkar, R.R.; Singh, G.; Kalurkar, A.R.; Jagasia, N.S.; Malhotra, H.K.

    1977-01-01

    Isotope radiography techniques in the non-conventional applications such as : (a) hot radiography - a method for examination of radioactive materials, and (b) thin section radiography for inspection of biological specimen and non-metallic specimen are described. Also, a process used for the reproduction of gamma-graphs of welds and castings has been discussed in detail. (author)

  7. Isotope techniques in the study of environmental change

    International Nuclear Information System (INIS)

    1998-01-01

    The International Atomic Energy Agency, among its efforts to promote the wider use of isotope techniques in hydrology and related environmental disciplines, organized an International Symposium on Applications of Isotope Techniques in Studying Past and Current Environmental Changes in the Hydrosphere and the Atmosphere, held in 1993 in Vienna. The broad scientific interest and abundant participation in the symposium, as well as the rapid progress seen in this field since then, encouraged the IAEA to organize a second symposium of this kind. It was held from 14 to 18 April 1997 in Vienna and brought together 177 scientists representing 46 Member States, UNESCO, FAO, WMO, the World Climate Research Programme (WCRP) and the International Geosphere-Biosphere Programme (IGBP). The symposium was structured in five sessions of oral presentations, a poster session and a round table discussion that focused on trends and future requirements in isotope hydrology and on applications in climate and environmental research. The major themes covered by the presentations included the use of isotopic tracers in studies of atmospheric and hydrospheric changes and of the human impact on water and the environment. Special emphasis was placed on isotopic archives of climatic and environmental change. Some contributions addressed new technical approaches

  8. Fuel reprocessing data validation using the isotope correlation technique

    International Nuclear Information System (INIS)

    Persiani, P.J.; Bucher, R.G.; Pond, R.B.; Cornella, R.J.

    1990-01-01

    The Isotope Correlation Technique (ICT), in conjunction with the gravimetric (Pu/U ratio) method for mass determination, provides an independent verification of the input accountancy at the dissolver or accountancy stage of the reprocessing plant. The Isotope Correlation Technique has been applied to many classes of domestic and international reactor systems (light-water, heavy-water, and graphite reactors) operating in a variety of modes (power, research, and production reactors), and for a variety of reprocessing fuel cycle management strategies. Analysis of reprocessing operations data based on isotopic correlations derived for assemblies in a PWR environment and fuel management scheme, yielded differences between the measurement-derived and ICT-derived plutonium mass determinations of (- 0.02 ± 0.23)% for the measured U-235 and (+ 0.50 ± 0.31)% for the measured Pu-239, for a core campaign. The ICT analyses has been implemented for the plutonium isotopics in a depleted uranium assembly in a heavy-water, enriched uranium system and for the uranium isotopes in the fuel assemblies in light-water, highly-enriched systems

  9. Isotopic Abundance and Chemical Purity Analysis of Stable Isotope Deuterium Labeled Sudan I

    Directory of Open Access Journals (Sweden)

    CAI Yin-ping;LEI Wen;ZHENG Bo;DU Xiao-ning

    2014-02-01

    Full Text Available It is important that to analysis of the isotopic abundance and chemical purity of Sudan I-D5, which is the internal standard of isotope dilution mass spectrometry. The isotopic abundance of Sudan I-D5 is detected by “mass cluster” classification method and LC-MS. The repeatability and reproducibility experiments were carried out by using different mass spectrometers and different operators. The RSD was less than 0.1%, so the repeatability and reproducibility were satisfactory. The accuracy and precision of the isotopic abundance analysis method was good with the results of F test and t test. The high performance liquid chromatography (HPLC had been used for detecting the chemical purity of Sudan I-D5 as external standard method.

  10. Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics

    International Nuclear Information System (INIS)

    Shen, Weifeng; Han, Wei; Li, Yunong; Meng, Zhiqi; Cai, Leiming; Li, Liang

    2016-01-01

    Silkworm (Bombyx mori) is a very useful target insect for evaluation of endocrine disruptor chemicals (EDCs) due to mature breeding techniques, complete endocrine system and broad basic knowledge on developmental biology. Comparative metabolomics of silkworms with and without EDC exposure offers another dimension of studying EDCs. In this work, we report a workflow on metabolomic profiling of silkworm hemolymph based on high-performance chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) and demonstrate its application in studying the metabolic changes associated with the pesticide dichlorodiphenyltrichloroethane (DDT) exposure in silkworm. Hemolymph samples were taken from mature silkworms after growing on diet that contained DDT at four different concentrations (1, 0.1, 0.01, 0.001 ppm) as well as on diet without DDT as controls. They were subjected to differential "1"2C-/"1"3C-dansyl labeling of the amine/phenol submetabolome, LC-UV quantification of the total amount of labeled metabolites for sample normalization, and LC-MS detection and relative quantification of individual metabolites in comparative samples. The total concentration of labeled metabolites did not show any significant change between four DDT-treatment groups and one control group. Multivariate statistical analysis of the metabolome data set showed that there was a distinct metabolomic separation between the five groups. Out of the 2044 detected peak pairs, 338 and 1471 metabolites have been putatively identified against the HMDB database and the EML library, respectively. 65 metabolites were identified by the dansyl library searching based on the accurate mass and retention time. Among the 65 identified metabolites, 33 positive metabolites had changes of greater than 1.20-fold or less than 0.83-fold in one or more groups with p-value of smaller than 0.05. Several useful biomarkers including serine, methionine, tryptophan, asymmetric dimethylarginine, N

  11. Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics

    Energy Technology Data Exchange (ETDEWEB)

    Shen, Weifeng [Key Laboratory of Detection for Pesticide Residues, Ministry of Agriculture (China); Sericultural Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Han, Wei; Li, Yunong [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada); Meng, Zhiqi [Sericultural Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Cai, Leiming, E-mail: cailm@mail.zaas.ac.cn [Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou (China); Li, Liang, E-mail: Liang.Li@ualberta.ca [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada)

    2016-10-26

    Silkworm (Bombyx mori) is a very useful target insect for evaluation of endocrine disruptor chemicals (EDCs) due to mature breeding techniques, complete endocrine system and broad basic knowledge on developmental biology. Comparative metabolomics of silkworms with and without EDC exposure offers another dimension of studying EDCs. In this work, we report a workflow on metabolomic profiling of silkworm hemolymph based on high-performance chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) and demonstrate its application in studying the metabolic changes associated with the pesticide dichlorodiphenyltrichloroethane (DDT) exposure in silkworm. Hemolymph samples were taken from mature silkworms after growing on diet that contained DDT at four different concentrations (1, 0.1, 0.01, 0.001 ppm) as well as on diet without DDT as controls. They were subjected to differential {sup 12}C-/{sup 13}C-dansyl labeling of the amine/phenol submetabolome, LC-UV quantification of the total amount of labeled metabolites for sample normalization, and LC-MS detection and relative quantification of individual metabolites in comparative samples. The total concentration of labeled metabolites did not show any significant change between four DDT-treatment groups and one control group. Multivariate statistical analysis of the metabolome data set showed that there was a distinct metabolomic separation between the five groups. Out of the 2044 detected peak pairs, 338 and 1471 metabolites have been putatively identified against the HMDB database and the EML library, respectively. 65 metabolites were identified by the dansyl library searching based on the accurate mass and retention time. Among the 65 identified metabolites, 33 positive metabolites had changes of greater than 1.20-fold or less than 0.83-fold in one or more groups with p-value of smaller than 0.05. Several useful biomarkers including serine, methionine, tryptophan, asymmetric dimethylarginine, N

  12. Mechanistic Insights into Catalytic Ethanol Steam Reforming Using Isotope-Labeled Reactants.

    Science.gov (United States)

    Crowley, Stephen; Castaldi, Marco J

    2016-08-26

    The low-temperature ethanol steam reforming (ESR) reaction mechanism over a supported Rh/Pt catalyst has been investigated using isotope-labeled EtOH and H2 O. Through strategic isotope labeling, all nonhydrogen atoms were distinct from one another, and allowed an unprecedented level of understanding of the dominant reaction pathways. All combinations of isotope- and non-isotope-labeled atoms were detected in the products, thus there are multiple pathways involved in H2 , CO, CO2 , CH4 , C2 H4 , and C2 H6 product formation. Both the recombination of C species on the surface of the catalyst and preservation of the C-C bond within ethanol are responsible for C2 product formation. Ethylene is not detected until conversion drops below 100 % at t=1.25 h. Also, quantitatively, 57 % of the observed ethylene is formed directly through ethanol dehydration. Finally there is clear evidence to show that oxygen in the SiO2 -ZrO2 support constitutes 10 % of the CO formed during the reaction. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. On structural identifiability analysis of the cascaded linear dynamic systems in isotopically non-stationary 13C labelling experiments.

    Science.gov (United States)

    Lin, Weilu; Wang, Zejian; Huang, Mingzhi; Zhuang, Yingping; Zhang, Siliang

    2018-06-01

    The isotopically non-stationary 13C labelling experiments, as an emerging experimental technique, can estimate the intracellular fluxes of the cell culture under an isotopic transient period. However, to the best of our knowledge, the issue of the structural identifiability analysis of non-stationary isotope experiments is not well addressed in the literature. In this work, the local structural identifiability analysis for non-stationary cumomer balance equations is conducted based on the Taylor series approach. The numerical rank of the Jacobian matrices of the finite extended time derivatives of the measured fractions with respect to the free parameters is taken as the criterion. It turns out that only one single time point is necessary to achieve the structural identifiability analysis of the cascaded linear dynamic system of non-stationary isotope experiments. The equivalence between the local structural identifiability of the cascaded linear dynamic systems and the local optimum condition of the nonlinear least squares problem is elucidated in the work. Optimal measurements sets can then be determined for the metabolic network. Two simulated metabolic networks are adopted to demonstrate the utility of the proposed method. Copyright © 2018 Elsevier Inc. All rights reserved.

  14. A general procedure for isotopic (deuterium) labelling of non-steroidal antiinflammatory 2-arylpropionic acids

    International Nuclear Information System (INIS)

    Castell, J.V.; Martinez, L.A.; Universidad Politecnica de Valencia; Miranda, M.A.; Tarrega, Pilar

    1994-01-01

    Alkaline treatment of nonsteroidal antiinflammatory 2-arylpropionic acids in deuterium oxide led in all cases to isotopic exchange of the proton located at the α-position of the side chain. Monodeuteration was observed in the case of carprofen, ibuprofen, ketoprofen, fenoprofen, flurbiprofen and naproxen. Additional exchange of one or two protons of the heterocyclic ring occurred in indoprofen, suprofen and tiaprofenic acid. The isotopic labelling survived under the conditions required to perform in vitro photoallergic studies (photolysis in non-deuterated aqueous media). (Author)

  15. A general procedure for isotopic (deuterium) labelling of non-steroidal antiinflammatory 2-arylpropionic acids

    Energy Technology Data Exchange (ETDEWEB)

    Castell, J.V. (Valencia Univ. Hospital (Spain). Centro de Investigacion); Martinez, L.A. (Valencia Univ. Hospital (Spain). Centro de Investigacion Universidad Politecnica de Valencia (Spain). Dept. de Quimica); Miranda, M.A.; Tarrega, Pilar (Universidad Politecnica de Valencia (Spain). Dept. de Quimica)

    1994-01-01

    Alkaline treatment of nonsteroidal antiinflammatory 2-arylpropionic acids in deuterium oxide led in all cases to isotopic exchange of the proton located at the [alpha]-position of the side chain. Monodeuteration was observed in the case of carprofen, ibuprofen, ketoprofen, fenoprofen, flurbiprofen and naproxen. Additional exchange of one or two protons of the heterocyclic ring occurred in indoprofen, suprofen and tiaprofenic acid. The isotopic labelling survived under the conditions required to perform in vitro photoallergic studies (photolysis in non-deuterated aqueous media). (Author).

  16. Quantification of isotope-labelled and unlabelled folates in plasma, ileostomy and food samples.

    Science.gov (United States)

    Büttner, Barbara E; Öhrvik, Veronica E; Witthöft, Cornelia M; Rychlik, Michael

    2011-01-01

    New stable isotope dilution assays were developed for the simultaneous quantitation of [(13)C(5)]-labelled and unlabelled 5-methyltetrahydrofolic acid, 5-formyltetrahydrofolic acid, folic acid along with unlabelled tetrahydrofolic acid and 10-formylfolic acid in clinical samples deriving from human bioavailability studies, i.e. plasma, ileostomy samples, and food. The methods were based on clean-up by strong anion exchange followed by LC-MS/MS detection. Deuterated analogues of the folates were applied as the internal standards in the stable isotope dilution assays. Assay sensitivity was sufficient to detect all relevant folates in the respective samples as their limits of detection were below 0.62 nmol/L in plasma and below 0.73 μg/100 g in food or ileostomy samples. Quantification of the [(13)C(5)]-label in clinical samples offers the possibility to differentiate between folate from endogenous body pools and the administered dose when executing bioavailability trials.

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

    Science.gov (United States)

    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. © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

  18. Systematic NMR Analysis of Stable Isotope Labeled Metabolite Mixtures in Plant and Animal Systems: Coarse Grained Views of Metabolic Pathways

    Science.gov (United States)

    Chikayama, Eisuke; Suto, Michitaka; Nishihara, Takashi; Shinozaki, Kazuo; Hirayama, Takashi; Kikuchi, Jun

    2008-01-01

    Background Metabolic phenotyping has become an important ‘bird's-eye-view’ technology which can be applied to higher organisms, such as model plant and animal systems in the post-genomics and proteomics era. Although genotyping technology has expanded greatly over the past decade, metabolic phenotyping has languished due to the difficulty of ‘top-down’ chemical analyses. Here, we describe a systematic NMR methodology for stable isotope-labeling and analysis of metabolite mixtures in plant and animal systems. Methodology/Principal Findings The analysis method includes a stable isotope labeling technique for use in living organisms; a systematic method for simultaneously identifying a large number of metabolites by using a newly developed HSQC-based metabolite chemical shift database combined with heteronuclear multidimensional NMR spectroscopy; Principal Components Analysis; and a visualization method using a coarse-grained overview of the metabolic system. The database contains more than 1000 1H and 13C chemical shifts corresponding to 142 metabolites measured under identical physicochemical conditions. Using the stable isotope labeling technique in Arabidopsis T87 cultured cells and Bombyx mori, we systematically detected >450 HSQC peaks in each 13C-HSQC spectrum derived from model plant, Arabidopsis T87 cultured cells and the invertebrate animal model Bombyx mori. Furthermore, for the first time, efficient 13C labeling has allowed reliable signal assignment using analytical separation techniques such as 3D HCCH-COSY spectra in higher organism extracts. Conclusions/Significance Overall physiological changes could be detected and categorized in relation to a critical developmental phase change in B. mori by coarse-grained representations in which the organization of metabolic pathways related to a specific developmental phase was visualized on the basis of constituent changes of 56 identified metabolites. Based on the observed intensities of 13C atoms of

  19. Studies of phosphorus-containing fertilizer uptake in soils by 32P isotope labelling

    International Nuclear Information System (INIS)

    Fueleky, Gyoergy; Osztoics, Andrasne; Papne Kranitz, Erzsebet

    1983-01-01

    Breeding experiments were carried out with rye-grass (Lolium perenne L.) on two soil types to determine the plant uptake of phosphorus from naturally occuring element and from that added to the soil by superphosphate fertilizers. 32 P isotope labelling and radiometric measuring method were applied. In addition to the determination of phosphorus uptake, the phosphorus contents of the soil from its natural stock and from the fertilizer for both soil types can be determined by this method. (A.L.)

  20. The synthesis of tritium, carbon-14 and stable isotope labelled selective estrogen receptor degraders.

    Science.gov (United States)

    Bragg, Ryan A; Bushby, Nick; Ericsson, Cecilia; Kingston, Lee P; Ji, Hailong; Elmore, Charles S

    2016-09-01

    As part of a Medicinal Chemistry program aimed at developing an orally bioavailable selective estrogen receptor degrader, a number of tritium, carbon-14, and stable isotope labelled (E)-3-[4-(2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indol-1-yl)phenyl]prop-2-enoic acids were required. This paper discusses 5 synthetic approaches to this compound class. Copyright © 2016 John Wiley & Sons, Ltd.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-03-04

    We present UNiquant, a new software program for analyzing stable isotope labeling (SIL) based quantitative proteomics data. UNiquant surpassed the performance of two other platforms, MaxQuant and Mascot Distiller, using complex proteome mixtures having either known or unknown heavy/light ratios. UNiquant is compatible with a broad spectrum of search engines and SIL methods, providing outstanding peptide pair identification and accurate measurement of the relative peptide/protein abundance.

  2. Architecture and dynamics of isotopically labelled macromolecules by nuclear magnetic resonance spectroscopy

    International Nuclear Information System (INIS)

    Matwiyoff, N.A.

    1979-01-01

    The use of 13 C is considered using NMR spectra of cell suspensions. Biochemical reaction kinetics are still unclear in the study of environmental and structural perturbations of amino acids and peptides; thus needs still exist for this labelling technique

  3. A new technique of ion beam tritium labelling

    International Nuclear Information System (INIS)

    Zhang Nianbao; Sheng Shugang; Yao Fuzeng

    1990-01-01

    In this paper a new technique is reported for tritium labelling of proteins, peptides and other nonvolatile organic compounds. A tritium ion beam is accelerated to bombard solid sample target for producing tritium exchange with hydrogen. The tritium labelling method has been applied to tritiated soybean trypsin inhibitor, ribonuclease A, elastin, pachyman and others totalled 11. After purifying by dialysis, ion exchange chromatography and gel filtration, the tritiated proteins and polysaccharide were obtained with specific activity over 37 GBq/mmol, without decomposition and with biological activity well preserved. By amino acid analysis of tritiated protein it was shown that the relative specific radioactivities for His., Tyr. and Phe. residues were higher while those for Val., Ile. and Ser. residues were lower

  4. A new technique for ion beam tritium labelling

    International Nuclear Information System (INIS)

    Zhang Nianbao; Sheng Shugang; Yao Fuzeng

    1990-06-01

    An advanced technique, the ion beam tritium labelling method (IBTL), used for labelling proteins, peptides and other nonvolatile organic compounds is introduced. In this method the excited tritium ion beam is accelerated and then bombs a solid sample target in which tritium exchanging for hydrogen is taken place. The IBTL has been used for preparation of tritiated soybean trypsin inhibitor, ribonuclease A, elastin and pachyman etc. After purifing by dialysis, ion exchange chromatography and gel filtration, the tritiated proteins and polysaccharide were obtained with the specific activity over 37 GBq/mmol, the function of tritiated decomposition products was not found. The product was shown to have native biological activity. Amino acid analysis of tritiated protein showed that the relative specific radioactivities were higher for His., Tyr. and Phe. but lower for Val., Ile. and Ser

  5. Geothermal investigations with isotope and geochemical techniques in Latin America

    International Nuclear Information System (INIS)

    1992-03-01

    The IAEA Co-ordinated Research Programme (CRP) for Latin America on the Use of Isotope and Geochemical Techniques in Geothermal Exploration started in 1984. The first activity carried out was a Seminar on isotope and geochemical techniques in geothermal exploration, which took place in June 1984 in Morelia, Mexico. During the seminar, which was attended by representatives of the institutions which later took part in the programme, the objectives, main research lines, and geothermal fields to be studied during the CRP were discussed. The first research contracts were awarded towards the end of 1984. The field work started in 1985 and continued through 1990. During the implementation of the CRP a considerable number of geothermal fields were studied in the nine participating countries. The investigations carried out were geochemically quite comprehensive in most cases, but in some others they were still in a reconnaissance stage when the CRP ended: the latter studies are not reported in these proceedings, but the data obtained are in principle available from the relevant national institutions. While investigations with conventional geochemical techniques had already started in several fields before 1985, isotope methods were applied for the first time in all cases during this CRP. Due to the remoteness and high elevation of many of the fields studied and the adverse meteorological conditions during long periods of the year, the investigations could not proceed rapidly: this is the main reason for the unusually long duration of the CRP, which could be concluded only after more than five years after its inception

  6. Nuclear material safeguards surveillance and accountancy by isotope correlation techniques

    International Nuclear Information System (INIS)

    Persiani, P.J.; Goleb, J.A.; Kroc, T.K.

    1981-11-01

    The purpose of this study is to investigate the applicability of isotope correlation techniques (ICT) to the Light Water Reactor (LWR) and the Liquid Metal Fast Breeder Reactor (LMFBR) fuel cycles for nuclear material accountancy and safeguards surveillance. The isotopic measurement of the inventory input to the reprocessing phase of the fuel cycle is the primary direct determination that an anomaly may exist in the fuel management of nuclear material. The nuclear materials accountancy gap which exists between the fabrication plant output and the input to the reprocessing plant can be minimized by using ICT at the dissolver stage of the reprocessing plant. The ICT allows a level of verification of the fabricator's fuel content specifications, the irradiation history, the fuel and blanket assemblies management and scheduling within the reactor, and the subsequent spent fuel assembly flows to the reprocessing plant. The investigation indicates that there exist relationships between isotopic concentration which have predictable, functional behavior over a range of burnup. Several cross-correlations serve to establish the initial core assembly-averaged composition. The selection of the more effective functionals will depend not only on the level of reliability of ICT for verification, but also on the capability, accuracy and difficulty of developing measurement methods. The propagation of measurement errors on the correlation functions and respective sensitivities to isotopic compositional changes have been examined and found to be consistent with current measurement methods

  7. Isotope-based immunological techniques. Their use in assessment of immune competence and the study of immune responses to pathogens

    International Nuclear Information System (INIS)

    Duffus, W.P.H.

    1984-01-01

    The influence of isotope-based techniques on both assessment of immune competence and immune response to pathogens is discussed. Immunodeficiencies acquired as a result of factors like malnutrition and concomitant disease can severely affect not only attempts to intensify and improve production but also successful immune response against important vaccines such as rinderpest and foot-and-mouth disease. Isotope-based techniques, with their accuracy, speed and small sample volume, are ideally suited for assessing immunocompetence. One of the main drawbacks remains antigen purity, an area where research should now be concentrated. Lymphocyte transformation is widely used to assess cell-mediated immuno-competence but techniques to assess biological functions such as phagocytosis and cell-mediated cytotoxicity could more usefully reflect immune status. These latter techniques utilize isotopes such as 3 H, 14 C, 32 P and 125 I. Investigation of specific cell-mediated immune response often requires a labelled target. Suitable isotopes such as 51 Cr, 99 Tcsup(m), 75 Se and 3 H are compared for their capacity to label both mammalian and parasite targets. Suggestions are made on a number of areas of research that might usefully be encouraged and supported in order to improve applied veterinary immunology in tropical countries. (author)

  8. Sulfur Isotope Exchange between S-35 Labeled Inorganic Sulfur-Compounds in Anoxic Marine-Sediments

    DEFF Research Database (Denmark)

    FOSSING, H.; THODEANDERSEN, S.; JØRGENSEN, BB

    1992-01-01

    Isotope exchange reactions between S-35-labeled sulfur compounds were studied in anoxic estuarine sediment slurries at 21-degrees-C and pH 7.4-7.7. Two experiments labeled with radioactive elemental sulfur (S-35-degrees) and one labeled with radioactive sulfate ((SO42-)-S-35) were performed as time......% of the total S-35 was recovered in the SIGMA-HS- pool in less than 1.5 h. With no detectable SIGMA-HS- (less than 1-mu-M) in the slurry, 58% of the total S-35 was observed in the pyrite pool within 1.5 h. The FeS pool received up to 31% of all S-35 added. The rapid S-35 incorporation from S-35-degrees...... into SIGMA-HS- and FeS pools was explained by isotope exchange reactions. In contrast, there was evidence that the radioactivity observed in the 'pyrite pool' was caused by adhesion of the added S-35-degrees to the FeS2 grains. In all S-35-degrees-labeled experiments we also observed oxidation...

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

    Energy Technology Data Exchange (ETDEWEB)

    Freiburger, Lee, E-mail: lee.freiburger@tum.de; Sonntag, Miriam, E-mail: miriam.sonntag@mytum.de; Hennig, Janosch, E-mail: janosch.hennig@helmholtz-muenchen.de [Helmholtz Zentrum München, Institute of Structural Biology (Germany); Li, Jian, E-mail: lijianzhongbei@163.com [Chinese Academy of Sciences, Tianjin Institute of Industrial Biotechnology (China); Zou, Peijian, E-mail: peijian.zou@helmholtz-muenchen.de; Sattler, Michael, E-mail: sattler@helmholtz-muenchen.de [Helmholtz Zentrum München, Institute of Structural Biology (Germany)

    2015-09-15

    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.

  10. Reliable LC-MS quantitative glycomics using iGlycoMab stable isotope labeled glycans as internal standards.

    Science.gov (United States)

    Zhou, Shiyue; Tello, Nadia; Harvey, Alex; Boyes, Barry; Orlando, Ron; Mechref, Yehia

    2016-06-01

    Glycans have numerous functions in various biological processes and participate in the progress of diseases. Reliable quantitative glycomic profiling techniques could contribute to the understanding of the biological functions of glycans, and lead to the discovery of potential glycan biomarkers for diseases. Although LC-MS is a powerful analytical tool for quantitative glycomics, the variation of ionization efficiency and MS intensity bias are influencing quantitation reliability. Internal standards can be utilized for glycomic quantitation by MS-based methods to reduce variability. In this study, we used stable isotope labeled IgG2b monoclonal antibody, iGlycoMab, as an internal standard to reduce potential for errors and to reduce variabililty due to sample digestion, derivatization, and fluctuation of nanoESI efficiency in the LC-MS analysis of permethylated N-glycans released from model glycoproteins, human blood serum, and breast cancer cell line. We observed an unanticipated degradation of isotope labeled glycans, tracked a source of such degradation, and optimized a sample preparation protocol to minimize degradation of the internal standard glycans. All results indicated the effectiveness of using iGlycoMab to minimize errors originating from sample handling and instruments. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Cooperation of CMEA member states in the field of the manufacture and use of stable isotopes and compounds thus labelled

    International Nuclear Information System (INIS)

    Ertel, G.; Ewald, G.

    1977-01-01

    The contribution presents a survey of scientific-technical cooperation of CMEA member states in the field of stable isotopes, it deals with the specialization of stable isotope production and compounds thus labelled, and gives the prospects for further development of this cooperation. (HK) [de

  12. Application of isotope techniques to investigate groundwater pollution

    International Nuclear Information System (INIS)

    1998-10-01

    This publication is a compilation of scientific results from the Co-ordinated Research Project (CRP) on the Application of Isotope Techniques to Investigate Groundwater Pollution which was implemented from 1995 to 1997. The conclusions of the CRP were presented by scientists from the following participating Member States: Austria, Brazil, China, Czech Republic, France, Hungary, India, Israel, Italy, New Zealand, Pakistan, Poland, Senegal and the United Kingdom. The CRP was implemented in recognition of the importance of protecting groundwater resources, and promoting the role of isotope techniques when integrated to classical hydrological methods to identify the sources and mechanisms of by which pollution takes place. The results of the CRP are expected to find practical applications in tackling hydrological problems encountered in technical co-operation projects of the IAEA. This publication could also provide a contribution toward the continuing efforts of various sectors to investigate, mitigate and control the threat of groundwater pollution. This publication includes the results of 16 investigations dealing with isotopes of hydrogen, carbon, nitrogen, oxygen and sulfur integrated to some extent with the classical hydrological tools of investigation. Each document in this compilation is provided with abstract and index

  13. Measurement of apolipoprotein E and amyloid β clearance rates in the mouse brain using bolus stable isotope labeling

    Science.gov (United States)

    2012-01-01

    Background Abnormal proteostasis due to alterations in protein turnover has been postulated to play a central role in several neurodegenerative diseases. Therefore, the development of techniques to quantify protein turnover in the brain is critical for understanding the pathogenic mechanisms of these diseases. We have developed a bolus stable isotope-labeling kinetics (SILK) technique coupled with multiple reaction monitoring mass spectrometry to measure the clearance of proteins in the mouse brain. Results Cohorts of mice were pulse labeled with 13 C6-leucine and the brains were isolated after pre-determined time points. The extent of label incorporation was measured over time using mass spectrometry to measure the ratio of labeled to unlabeled apolipoprotein E (apoE) and amyloid β (Aβ). The fractional clearance rate (FCR) was then calculated by analyzing the time course of disappearance for the labeled protein species. To validate the technique, apoE clearance was measured in mice that overexpress the low-density lipoprotein receptor (LDLR). The FCR in these mice was 2.7-fold faster than wild-type mice. To demonstrate the potential of this technique for understanding the pathogenesis of neurodegenerative disease, we applied our SILK technique to determine the effect of ATP binding cassette A1 (ABCA1) on both apoE and Aβ clearance. ABCA1 had previously been shown to regulate both the amount of apoE in the brain, along with the extent of Aβ deposition, and represents a potential molecular target for lowering brain amyloid levels in Alzheimer's disease patients. The FCR of apoE was increased by 1.9- and 1.5-fold in mice that either lacked or overexpressed ABCA1, respectively. However, ABCA1 had no effect on the FCR of Aβ, suggesting that ABCA1 does not regulate Aβ metabolism in the brain. Conclusions Our SILK strategy represents a straightforward, cost-effective, and efficient method to measure the clearance of proteins in the mouse brain. We expect that

  14. Stable isotope labeled n-alkanes to assess digesta passage kinetics through the digestive tract of ruminants

    NARCIS (Netherlands)

    Warner, D.; Ferreira, L.M.M.; Breuer, M.J.H.; Dijkstra, J.; Pellikaan, W.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

  15. Recycling of an amino acid label with prolonged isotope infusion: Implications for kinetic studies

    International Nuclear Information System (INIS)

    Schwenk, W.F.; Tsalikian, E.; Beaufrere, B.; Haymond, M.W.

    1985-01-01

    To investigate whether recycling of a labeled amino acid would occur after 24 h of infusion, two groups of normal volunteers were infused with [ 3 H]leucine and alpha-[ 14 C]-ketoisocaproate for 4 h and [ 2 H 3 ]leucine for either 4 or 24 h (groups I and II, respectively). Entry of [ 2 H 3 ]leucine at steady state into the plasma space was indistinguishable from its infusion rate for group I but 30% higher (P less than 0.001) than this rate for group II, demonstrating significant recycling of label. After discontinuation of the infusions, isotope disappearance from the plasma space was followed for 2 h. The 3 H and 14 C decay data for both groups suggest that plasma leucine and alpha- ketoisocaproate are derived from a single intracellular pool in the postabsorptive state. In group I, the 3 H and 2 H labels decayed identically; whereas, in group II, the decay of [ 2 H 3 ]-leucine and alpha- [ 2 H 3 ]ketoisocaproate was slower (P less than 0.01) than the decay of [ 3 H]leucine and alpha-[ 3 H]ketoisocaproate, confirming re-entry of label after a 24-h infusion. Therefore kinetic values calculated from models assuming no recycling of labeled amino acids are most likely not quantitative and must be interpreted with care when flux does not change or decreases

  16. Selectively dispersed isotope labeling for protein structure determination by magic angle spinning NMR

    Energy Technology Data Exchange (ETDEWEB)

    Eddy, Matthew T. [Massachusetts Institute of Technology, Department of Chemistry (United States); Belenky, Marina [Brandeis University, Department of Chemistry (United States); Sivertsen, Astrid C. [Massachusetts Institute of Technology, Francis Bitter Magnet Laboratory (United States); Griffin, Robert G. [Massachusetts Institute of Technology, Department of Chemistry (United States); Herzfeld, Judith, E-mail: herzfeld@brandeis.edu [Brandeis University, Department of Chemistry (United States)

    2013-10-15

    The power of nuclear magnetic resonance spectroscopy derives from its site-specific access to chemical, structural and dynamic information. However, the corresponding multiplicity of interactions can be difficult to tease apart. Complimentary approaches involve spectral editing on the one hand and selective isotope substitution on the other. Here we present a new 'redox' approach to the latter: acetate is chosen as the sole carbon source for the extreme oxidation numbers of its two carbons. Consistent with conventional anabolic pathways for the amino acids, [1-{sup 13}C] acetate does not label {alpha} carbons, labels other aliphatic carbons and the aromatic carbons very selectively, and labels the carboxyl carbons heavily. The benefits of this labeling scheme are exemplified by magic angle spinning spectra of microcrystalline immunoglobulin binding protein G (GB1): the elimination of most J-couplings and one- and two-bond dipolar couplings provides narrow signals and long-range, intra- and inter-residue, recoupling essential for distance constraints. Inverse redox labeling, from [2-{sup 13}C] acetate, is also expected to be useful: although it retains one-bond couplings in the sidechains, the removal of CA-CO coupling in the backbone should improve the resolution of NCACX spectra.

  17. Automatic isotope gas analysis of tritium labelled organic materials Pt. 1

    International Nuclear Information System (INIS)

    Gacs, I.; Mlinko, S.

    1978-01-01

    A new automatic procedure developed to convert tritium in HTO hydrogen for subsequent on-line gas counting is described. The water containing tritium is introduced into a column prepared from molecular sieve-5A and heated to 550 deg C. The tritium is transferred by isotopic exchange into hydrogen flowing through the column. The radioactive gas is led into an internal detector for radioactivity measurement. The procedure is free of memory effects, provides quantitative recovery with analytical reproducibility better than 0.5% rel. at a preset number of counts. The experimental and analytical results indicate that isotopic exchange between HTO and hydrogen over a column prepared from alumina or molecular sieve-5A can be successfully applied for the quantitative transfer of tritium from HTO into hydrogen for on-line gas countinq. This provides an analytical procedure for the automatic determination of tritium in water with an analytical reproducibility better than 0.5% rel. The exchange process will also be suitable for rapid tritium transfer from water formed during the decomposition of tritium-labelled organic compounds or biological materials. The application of the procedure in automatic isotope gas analysis of organic materials labelled with tritium will be described in subsequent papers (Parts II and III). (T.G.)

  18. Production of large quantities of isotopically labeled protein in Pichia pastoris by fermentation

    International Nuclear Information System (INIS)

    Wood, Matthew J.; Komives, Elizabeth A.

    1999-01-01

    Heterologous expression in Pichia pastoris has many of the advantages of eukaryotic expression, proper folding and disulfide bond formation, glycosylation, and secretion. Contrary to other eukaryotic systems, protein production from P.pastoris occurs in simple minimal defined media making this system attractive for production of labeled proteins for NMR analysis. P.pastoris is therefore the expression system of choice for NMR of proteins that cannot be refolded from inclusion bodies or that require post-translational modifications for proper folding or function. The yield of expressed proteins from P.pastoris depends critically on growth conditions, and attainment of high cell densities by fermentation has been shown to improve protein yields by 10-100-fold. Unfortunately, the cost of the isotopically enriched fermentation media components, particularly 15NH4OH, is prohibitively high. We report fermentation methods that allow for both 15N- labeling from (15NH4)2SO4 and 13C-labeling from 13C-glucose or 13C-glycerol of proteins produced in Pichia pastoris. Expression of an 83 amino acid fragment of thrombomodulin with two N-linked glycosylation sites shows that fermentation is more cost effective than shake flask growth for isotopic enrichment

  19. International symposium on isotope techniques in water resources development and management. Book of extended synopses

    International Nuclear Information System (INIS)

    1999-05-01

    The papers presented at the symposium dealt with environmental isotope measurements and tracer techniques in studies on sedimentation, groundwater recharge, dynamics and pollution; discussed isotope data interpretation; and presented recent advances in analytical techniques for isotope hydrology. This document contains 139 extended synopses; each of them was indexed individually

  20. International symposium on isotope techniques in water resources development and management. Book of extended synopses

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-05-01

    The papers presented at the symposium dealt with environmental isotope measurements and tracer techniques in studies on sedimentation, groundwater recharge, dynamics and pollution; discussed isotope data interpretation; and presented recent advances in analytical techniques for isotope hydrology. This document contains 139 extended synopses; each of them was indexed individually Refs, figs, tabs

  1. Isotope technique in JPS dam surveillance: its potential

    International Nuclear Information System (INIS)

    Sabri Hassan

    2006-01-01

    Controlling seepage is one of the most important requirements for safe dams. Any leakage at an earth embankment may be potentially dangerous since rapid internal erosion may quickly enlarge an initially minor defect. Thus dam owners need to have thorough surveillance programs that can forewarn of impending problems from seepage or other factors influencing the safety of dams. In carrying out dam surveillance works, all possible efforts should be considered and foreseeing the potential of isotope technique, JPS (Department of Irrigation and Drainage, Malaysia) and MINT (Malaysian Institute for Nuclear Technology Research) participated actively in the UNDP/RCA/IAEA program under RAS/8/093 project sponsored by the International Atomic Energy Agency (IAEA). Through these activities, it was noted that the technique demonstrated very promising potentials such as in assisting dam site selections, site investigations, watershed studies, dam and reservoir design, leakage investigations and sediments related issues, the two latter ones being relatively critical during the operational life of the dam. Establishment of baseline isotopic characteristics (or fingerprint), hydrochemistry, electrical conductivity and temperature profiles is underway for all JPS dams to be later utilized in diagnosing seepage related issues it is suggested that application of this technique be extended to other dam owners nationwide. (Author)

  2. Determination of zinc nutrient in the soil using isotope technique

    International Nuclear Information System (INIS)

    Suwadji, E.

    1975-01-01

    In this experiment the availability of soil Zn nutrient in various soil conditions (dry and submerged), and the efficiency of the application of Zn fertilizer in rice nutrition were measured in glasshouse using isotope dilution technique. The amount of soil Zn nutrient available to plants can be expressed in 'E' and 'L' values. Submerged conditions generally showed an increase in the 'E' and 'L' value compared to dry conditions. Mixed treatment with ZnSO 4 fertilizer is more efficient for Zn absorption than surface treatment. (author)

  3. Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis

    DEFF Research Database (Denmark)

    Harkness, Linda; Prokhorova, Tatyana A; Kassem, Moustapha

    2012-01-01

    The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass...... spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we...

  4. Dansylation isotope labeling liquid chromatography mass spectrometry for parallel profiling of human urinary and fecal submetabolomes

    Energy Technology Data Exchange (ETDEWEB)

    Su, Xiaoling [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Wang, Nan [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2 (Canada); Chen, Deying [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Li, Yunong [Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2 (Canada); Lu, Yingfeng [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Huan, Tao [Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2 (Canada); Xu, Wei [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Li, Liang, E-mail: Liang.Li@ualberta.ca [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China); Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2 (Canada); Li, Lanjuan, E-mail: ljli@zju.edu.cn [State Key Laboratory and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003 (China)

    2016-01-15

    Human urine and feces can be non-invasively collected for metabolomics-based disease biomarker discovery research. Because urinary and fecal metabolomes are thought to be different, analysis of both biospecimens may generate a more comprehensive metabolomic profile that can be better related to the health state of an individual. Herein we describe a method of using differential chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) for parallel metabolomic profiling of urine and feces. Dansylation labeling was used to quantify the amine/phenol submetabolome changes among different samples based on {sup 12}C-labeling of individual samples and {sup 13}C-labeling of a pooled urine or pooled feces and subsequent analysis of the {sup 13}C-/{sup 12}C-labeled mixture by LC-MS. The pooled urine and pooled feces are further differentially labeled, mixed and then analyzed by LC-MS in order to relate the metabolite concentrations of the common metabolites found in both biospecimens. This method offers a means of direct comparison of urinary and fecal submetabolomes. We evaluated the analytical performance and demonstrated the utility of this method in the analysis of urine and feces collected daily from three healthy individuals for 7 days. On average, 2534 ± 113 (n = 126) peak pairs or metabolites could be detected from a urine sample, while 2507 ± 77 (n = 63) peak pairs were detected from a fecal sample. In total, 5372 unique peak pairs were detected from all the samples combined; 3089 and 3012 pairs were found in urine and feces, respectively. These results reveal that the urine and fecal metabolomes are very different, thereby justifying the consideration of using both biospecimens to increase the probability of finding specific biomarkers of diseases. Furthermore, the CIL LC-MS method described can be used to perform parallel quantitative analysis of urine and feces, resulting in more complete coverage of the human metabolome

  5. Dansylation isotope labeling liquid chromatography mass spectrometry for parallel profiling of human urinary and fecal submetabolomes

    International Nuclear Information System (INIS)

    Su, Xiaoling; Wang, Nan; Chen, Deying; Li, Yunong; Lu, Yingfeng; Huan, Tao; Xu, Wei; Li, Liang; Li, Lanjuan

    2016-01-01

    Human urine and feces can be non-invasively collected for metabolomics-based disease biomarker discovery research. Because urinary and fecal metabolomes are thought to be different, analysis of both biospecimens may generate a more comprehensive metabolomic profile that can be better related to the health state of an individual. Herein we describe a method of using differential chemical isotope labeling (CIL) liquid chromatography mass spectrometry (LC-MS) for parallel metabolomic profiling of urine and feces. Dansylation labeling was used to quantify the amine/phenol submetabolome changes among different samples based on "1"2C-labeling of individual samples and "1"3C-labeling of a pooled urine or pooled feces and subsequent analysis of the "1"3C-/"1"2C-labeled mixture by LC-MS. The pooled urine and pooled feces are further differentially labeled, mixed and then analyzed by LC-MS in order to relate the metabolite concentrations of the common metabolites found in both biospecimens. This method offers a means of direct comparison of urinary and fecal submetabolomes. We evaluated the analytical performance and demonstrated the utility of this method in the analysis of urine and feces collected daily from three healthy individuals for 7 days. On average, 2534 ± 113 (n = 126) peak pairs or metabolites could be detected from a urine sample, while 2507 ± 77 (n = 63) peak pairs were detected from a fecal sample. In total, 5372 unique peak pairs were detected from all the samples combined; 3089 and 3012 pairs were found in urine and feces, respectively. These results reveal that the urine and fecal metabolomes are very different, thereby justifying the consideration of using both biospecimens to increase the probability of finding specific biomarkers of diseases. Furthermore, the CIL LC-MS method described can be used to perform parallel quantitative analysis of urine and feces, resulting in more complete coverage of the human metabolome. - Highlights: • A

  6. Solid state NMR of isotope labelled murine fur: a powerful tool to study atomic level keratin structure and treatment effects

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Wai Ching Veronica; Narkevicius, Aurimas; Chow, Wing Ying; Reid, David G.; Rajan, Rakesh [University of Cambridge, Department of Chemistry (United Kingdom); Brooks, Roger A. [University of Cambridge, Department of Trauma and Orthopaedic Surgery, Addenbrooke’s Hospital (United Kingdom); Green, Maggie [University of Cambridge, Central Biomedical Resources, School of Clinical Medicine (United Kingdom); Duer, Melinda J., E-mail: mjd13@cam.ac.uk [University of Cambridge, Department of Chemistry (United Kingdom)

    2016-10-15

    We have prepared mouse fur extensively {sup 13}C,{sup 15}N-labelled in all amino acid types enabling application of 2D solid state NMR techniques which establish covalent and spatial proximities within, and in favorable cases between, residues. {sup 13}C double quantum–single quantum correlation and proton driven spin diffusion techniques are particularly useful for resolving certain amino acid types. Unlike 1D experiments on isotopically normal material, the 2D methods allow the chemical shifts of entire spin systems of numerous residue types to be determined, particularly those with one or more distinctively shifted atoms such as Gly, Ser, Thr, Tyr, Phe, Val, Leu, Ile and Pro. Also the partial resolution of the amide signals into two signal envelopes comprising of α-helical, and β-sheet/random coil components, enables resolution of otherwise overlapped α-carbon signals into two distinct cross peak families corresponding to these respective secondary structural regions. The increase in resolution conferred by extensive labelling offers new opportunities to study the chemical fate and structural environments of specific atom and amino acid types under the influence of commercial processes, and therapeutic or cosmetic treatments.

  7. Quantification of Labile Soil Mercury by Stable Isotope Dilution Techniques

    Science.gov (United States)

    Shetaya, Waleed; Huang, Jen-How; Osterwalder, Stefan; Alewell, Christine

    2016-04-01

    Mercury (Hg) is a toxic element that can cause severe health problems to humans. Mercury is emitted to the atmosphere from both natural and anthropogenic sources and can be transported over long distances before it is deposited to aquatic and terrestrial environments. Aside from accumulation in soil solid phases, Hg deposited in soils may migrate to surface- and ground-water or enter the food chain, depending on its lability. There are many operationally-defined extraction methods proposed to quantify soil labile metals. However, these methods are by definition prone to inaccuracies such as non-selectivity, underestimation or overestimation of the labile metal pool. The isotopic dilution technique (ID) is currently the most promising method for discrimination between labile and non-labile metal fractions in soil with a minimum disturbance to soil-solid phases. ID assesses the reactive metal pool in soil by defining the fraction of metal both in solid and solution phases that is isotopically-exchangeable known as the 'E-value'. The 'E-value' represents the metal fraction in a dynamic equilibrium with the solution phase and is potentially accessible to plants. This is carried out by addition of an enriched metal isotope to soil suspensions and quantifying the fraction of metal that is able to freely exchange with the added isotope by measuring the equilibrium isotopic ratio by ICP-MS. E-value (mg kg-1) is then calculated as follows: E-Value = (Msoil/ W) (CspikeVspike/ Mspike) (Iso1IAspike -Iso2IAspikeRss / Iso2IAsoil Rss - Iso1IAsoil) where M is the average atomic mass of the metal in the soil or the spike, W is the mass of soil (kg), Cspike is the concentration of the metal in the spike (mg L-1), Vspike is the volume of spike (L), IA is isotopic abundance, and Rss is the equilibrium ratio of isotopic abundances (Iso1:Iso2). Isotopic dilution has been successfully applied to determine E-values for several elements. However, to our knowledge, this method has not yet

  8. Root activity evaluation in tree crops using isotopic techniques

    International Nuclear Information System (INIS)

    Calvache, Marcelo

    1991-01-01

    This paper discusses the methdology used to evalute root activity of the crops utilizing the technique of soil injection with solutions marked with isotopes. Some of the experimental data obtained with coffee, citrus and oil palm are also presented. Ovel all, these tree crops present a higher root activity in soil layers close to the surface (0-20 cm) and to a distance from the trunk which varies with age, season and variety. The most important conclusions are: 1. The isotope injection technique using 3 2 P , 1 5 N , or 8 5 R b, allow direct and reliable determination of root activity in these tree crops. 2. Root activity of three crops depends on age of the tree, variety, moisture content of the soil and soil type. 3. Soil moisture is the most influencial factor affecting root activity. This is turn depends on the irrigation method employed. 4. From the practical view point, the best distance from the trunk to apply fertilizer in the one wich has highest root activity closest to the soil surface

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2005-07-01

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

  10. Nuclear and isotopic techniques for addressing nutritional problems, with special reference to current applications in developing countries.

    Science.gov (United States)

    Iyengar, Venkatesh

    2002-03-01

    Nuclear and isotopic techniques are valuable tools in human nutritional research studies. Isotopes, both radioactive and nonradioactive, enable detailed evaluations of nutrient intake, body composition, energy expenditure, status of micronutrients, and nutrient bioavailability. In recent times, isotopic methods have been widely used in a number of coordinated research projects and technical cooperation projects of the International Atomic Energy Agency's Nutrition Programme. The doubly labeled water technique combines the use of the stable isotopes oxygen-18 and hydrogen-2 (deuterium) to measure total energy expenditure in free-living human subjects, and to investigate the magnitude and causes of both undernutrition and the emergence of obesity in developing countries. The deuterium dilution technique is a reliable tool to measure breastmilk intake and thereby infant growth and development. In collaboration with the World Health Organization's Growth Monitoring Program, this technique is being used to generate new data on growth standards for children in developing countries. This technique is also used in the measurement of body composition by the estimation of lean body mass and fat mass in individuals. Stable isotopes of iron and zinc have been successfully used to assess the nutritional impact of several nationwide food supplementation-programs conducted on pregnant and lactating women and children in both industrialized and developing countries. Isotopic techniques are especially suitable for monitoring changes in body composition, energy metabolism, and mineral status (with particular reference to osteoporosis) in the elderly. Nuclear methods have also served to develop models for a physiological reference man in Asia in support of radiological health and safety issues, for establishing elemental composition of foods, and for measurement of pollutants in the environment.

  11. Identifying and quantifying proteolytic events and the natural N terminome by terminal amine isotopic labeling of substrates.

    Science.gov (United States)

    Kleifeld, Oded; Doucet, Alain; Prudova, Anna; auf dem Keller, Ulrich; Gioia, Magda; Kizhakkedathu, Jayachandran N; Overall, Christopher M

    2011-09-22

    Analysis of the sequence and nature of protein N termini has many applications. Defining the termini of proteins for proteome annotation in the Human Proteome Project is of increasing importance. Terminomics analysis of protease cleavage sites in degradomics for substrate discovery is a key new application. Here we describe the step-by-step procedures for performing terminal amine isotopic labeling of substrates (TAILS), a 2- to 3-d (depending on method of labeling) high-throughput method to identify and distinguish protease-generated neo-N termini from mature protein N termini with all natural modifications with high confidence. TAILS uses negative selection to enrich for all N-terminal peptides and uses primary amine labeling-based quantification as the discriminating factor. Labeling is versatile and suited to many applications, including biochemical and cell culture analyses in vitro; in vivo analyses using tissue samples from animal and human sources can also be readily performed. At the protein level, N-terminal and lysine amines are blocked by dimethylation (formaldehyde/sodium cyanoborohydride) and isotopically labeled by incorporating heavy and light dimethylation reagents or stable isotope labeling with amino acids in cell culture labels. Alternatively, easy multiplex sample analysis can be achieved using amine blocking and labeling with isobaric tags for relative and absolute quantification, also known as iTRAQ. After tryptic digestion, N-terminal peptide separation is achieved using a high-molecular-weight dendritic polyglycerol aldehyde polymer that binds internal tryptic and C-terminal peptides that now have N-terminal alpha amines. The unbound naturally blocked (acetylation, cyclization, methylation and so on) or labeled mature N-terminal and neo-N-terminal peptides are recovered by ultrafiltration and analyzed by tandem mass spectrometry (MS/MS). Hierarchical substrate winnowing discriminates substrates from the background proteolysis products and

  12. The use of environmental isotope techniques in hydrology

    International Nuclear Information System (INIS)

    Qatan, Z.

    1990-01-01

    It is a very short report about the second coordinating meeting and workshop on the Regional Arab Middle-East Technical Cooperation Project, entitled 'use of environmental isotope techniques in hydrogeological investigations' was organized by the IAEA and the Syrian Atomic Energy Commission and was held in Damascus, Syria in the period 10-14 December 1989. About 27 participants, involved in the implementation of this cooperation project, from Jordan, United Arab Emirates, Saudi Arabia and Syria together with 3 technical experts from IAEA and the Earth Science Institute of Hanover University (F.R.G) have attended this meeting. The main objective of the meeting was to discuss the overall review of hydrogeological activities and development in these four countries, after the first sampling results already obtained, and to give a new plan of the activities to be implemented in the future. The meeting had also included some lectures, given by the experts, on the evaluations and the interpretations of isotope results, obtained from arid and semi-arid zones in some Middle-East countries. (author)

  13. A guide through the computational analysis of isotope-labeled mass spectrometry-based quantitative proteomics data: an application study

    Directory of Open Access Journals (Sweden)

    Haußmann Ute

    2011-06-01

    Full Text Available Abstract Background Mass spectrometry-based proteomics has reached a stage where it is possible to comprehensively analyze the whole proteome of a cell in one experiment. Here, the employment of stable isotopes has become a standard technique to yield relative abundance values of proteins. In recent times, more and more experiments are conducted that depict not only a static image of the up- or down-regulated proteins at a distinct time point but instead compare developmental stages of an organism or varying experimental conditions. Results Although the scientific questions behind these experiments are of course manifold, there are, nevertheless, two questions that commonly arise: 1 which proteins are differentially regulated regarding the selected experimental conditions, and 2 are there groups of proteins that show similar abundance ratios, indicating that they have a similar turnover? We give advice on how these two questions can be answered and comprehensively compare a variety of commonly applied computational methods and their outcomes. Conclusions This work provides guidance through the jungle of computational methods to analyze mass spectrometry-based isotope-labeled datasets and recommends an effective and easy-to-use evaluation strategy. We demonstrate our approach with three recently published datasets on Bacillus subtilis 12 and Corynebacterium glutamicum 3. Special focus is placed on the application and validation of cluster analysis methods. All applied methods were implemented within the rich internet application QuPE 4. Results can be found at http://qupe.cebitec.uni-bielefeld.de.

  14. Isotopic exchange between CO2 and H2O and labelling kinetics of photosynthetic oxygen

    International Nuclear Information System (INIS)

    Gerster, Richard

    1971-01-01

    The reaction of carbon dioxide with water has been studied by measuring the rate of oxygen exchange between C 18 O 2 and H 2 16 O. The mathematical treatment of the kinetics allows to determine with accuracy the diffusion flow between the gas and the liquid phase, in the same way as the CO 2 hydration rate. The velocity constant of this last process, whose value gives the in situ enzymatic activity of carbonic anhydrase, has been established in the case of chloroplast and Euglena suspensions and of aerial leaves. The study of the isotopic exchange between C 18 O 2 and a vegetable submitted to alternations of dark and light has allowed to calculate the isotopic abundance of the metabolized CO 2 whose value has been compared to that of the intracellular water and that of photosynthetic oxygen. In addition, a new method using 13 C 18 O 2 gives the means to measure with accuracy eventual isotopic effects. The labelling kinetics of the oxygen evolved by Euglena suspensions whose water has been enriched with 18 O have been established at different temperatures. (author) [fr

  15. Sectoral programming mission isotope techniques for geothermal development. Philippines. UNDP sectoral support

    International Nuclear Information System (INIS)

    Froehlich, K.; Sun, Y.

    1995-10-01

    This report discusses the accomplishments of IAEA Technical Cooperation project PHI/8/016 ''Isotope Techniques in Geothermal Hydrology''. It is intended to help Philippine National Oil Company's Energy Development Corporation (PNOC-EDC) in use of isotope techniques for geothermal development. This report discusses outcomes of the mission, conclusions and recommendations on applications of isotopes techniques in geothermal agro-industrial plants and geothermal hydrology

  16. Teams explore water supplies in Latin America: Interest raised in isotope techniques

    International Nuclear Information System (INIS)

    Gonfiantini, R.

    1984-01-01

    Latin American countries are particularly interested in applying isotope techniques in hydrology and other fields of earth sciences. Ten research contracts already have been awarded under the IAEA's Co-ordinated Research Programme on the Application of Isotope Techniques in Hydrology in the Latin American Region. In July 1984, IAEA and UNESCO sponsored a regional seminar for Latin America on the use of isotope techniques in water resources management in Buenos Aires

  17. International conference on the study of environmental change using isotope techniques. Book of extended synopses

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2001-04-01

    This publication contains extended synopses of the oral and poster presentations delivered at the meeting. The main topics of the meeting included: Isotopes in the atmosphere and the hydrosphere; Interaction between the atmosphere and the hydrosphere; Isotope indicators of past climatic and environmental changes; and Advances in isotope and other analytical techniques. The individual papers have been indexed separately.

  18. International conference on the study of environmental change using isotope techniques. Book of extended synopses

    International Nuclear Information System (INIS)

    2001-04-01

    This publication contains extended synopses of the oral and poster presentations delivered at the meeting. The main topics of the meeting included: Isotopes in the atmosphere and the hydrosphere; Interaction between the atmosphere and the hydrosphere; Isotope indicators of past climatic and environmental changes; and Advances in isotope and other analytical techniques. The individual papers have been indexed separately

  19. Formation of Hydroxymethyl DNA Adducts in Rats Orally Exposed to Stable Isotope Labeled Methanol

    Science.gov (United States)

    Lu, Kun; Gul, Husamettin; Upton, Patricia B.; Moeller, Benjamin C.; Swenberg, James A.

    2012-01-01

    Methanol is a large volume industrial chemical and widely used solvent and fuel additive. Methanol’s well known toxicity and use in a wide spectrum of applications has raised long-standing environmental issues over its safety, including its carcinogenicity. Methanol has not been listed as a carcinogen by any regulatory agency; however, there are debates about its carcinogenic potential. Formaldehyde, a metabolite of methanol, has been proposed to be responsible for the carcinogenesis of methanol. Formaldehyde is a known carcinogen and actively targets DNA and protein, causing diverse DNA and protein damage. However, formaldehyde-induced DNA adducts arising from the metabolism of methanol have not been reported previously, largely due to the absence of suitable DNA biomarkers and the inability to differentiate what was due to methanol compared with the substantial background of endogenous formaldehyde. Recently, we developed a unique approach combining highly sensitive liquid chromatography-mass spectrometry methods and exposure to stable isotope labeled chemicals to simultaneously quantify formaldehyde-specific endogenous and exogenous DNA adducts. In this study, rats were exposed daily to 500 or 2000 mg/kg [13CD4]-methanol by gavage for 5 days. Our data demonstrate that labeled formaldehyde arising from [13CD4]-methanol induced hydroxymethyl DNA adducts in multiple tissues in a dose-dependent manner. The results also demonstrated that the number of exogenous DNA adducts was lower than the number of endogenous hydroxymethyl DNA adducts in all tissues of rats administered 500 mg/kg per day for 5 days, a lethal dose to humans, even after incorporating an average factor of 4 for reduced metabolism due to isotope effects of deuterium-labeled methanol into account. PMID:22157354

  20. Isotopic techniques to study phosphorus cycling in soils

    International Nuclear Information System (INIS)

    Manjaiah, K.M.; Sreenivasa Chari, M.; Sachdev, P.; Sachdev, M.S.

    2008-01-01

    A sound understanding of phosphorus cycling in soil system is essential in order to manage this system in a sustainable manner. Phosphorus transformations are characterized by physico-chemical (sorption-desorption) and biological processes . The transformation rates need to be taken into account while developing nutrient management strategies for economical and sustainable production. One of the important tools and the method gaining popularity for determining the gross transformation rates of nutrients in the soil is the isotopic dilution technique. The major processes in the soil-plant system which determine the distribution and bioavailability of phosphorus in various inorganic and organic soil components consist of: (1) the dissolution of soil mineral phosphates, (2) retention of phosphorus by inorganic soil constituents, (3) decomposition of organic phosphorus contained in plant, animal and microbial detritus and (4) Immobilization of phosphorus via the soil microbial biomass and plan uptake

  1. Isotopic evaluations of dynamic and plant uptake of N in soil amended with 15N-labelled sewage sludge

    International Nuclear Information System (INIS)

    Kchaou, R.; Khelil, M. N.; Rejeb, S.; Gharbi, F.; Henchi, B.; Hernandez, T.; Destain, J. P.

    2010-01-01

    Field experiments were conducted to evaluate the use of a novel 15N isotope technique for comparing the dynamics of N derived from sewage sludge applied to sorghum to the dynamics of N derived from the commercial fertilizer, urea. The treatments included a control, sludge applied at three rates (3, 6 and 9 t/ha, or 113, 226 and 338 kg N/ha) and N-urea applied at three rates (150, 250 and 350 kg N/ha). Recovery of 15N -labelled sludge was similar for the different nitrogen rates applied , with a mean value of 27%. However, the recovery of 15N -urea decreased as the rate of N application increased (from 38% to 27%). Approximately 22% and 19% of the 15N from sludge and urea, respectively, remained in the 0-60 cm layer of soil, most of which was present in the 0-20 cm layer. Furthermore, losses of 15N -labelled fertilizer were not affected by the N fertilization source, and the greatest losses, which were measured in response to the highest N application rate, were 59%. (authors)

  2. Developing isotope techniques for water exploration in the Sahel

    International Nuclear Information System (INIS)

    Aranyossy, J.F.; Louvat, D.; Maksoudi, M.

    1991-01-01

    The IAEA in the early 1980s started its first activities on hydrogeology in Mali, Niger, and Senegal through its technical co-operation programme. These activities included, first of all, local aquifer studies with environmental isotopes. The studies were found to be particularly useful for obtaining specific information on, for example, relationships between aquifers and between surface water and groundwater, and the existence or absence of recent recharge. Moreover, the IAEA established the basic infrastructures needed for the application of artificial tracer techniques and the use of nuclear instrumentation in hydrology and hydrosedimentology. Several factors underscored the need for co-ordinating the scientific activities already being carried out under existing bilateral projects. Firstly, the ''horizontal'' flow of information was very poor in spite of the similarity between the hydrological and hydrogeological problems encountered in the three countries. Secondly, political frontiers do not generally follow the boundaries of the hydrological basins, so that the same aquifer can be shared between several countries. Thirdly, the costly infrastructure established by the IAEA in certain countries deserved to be utilized at the regional level. To meet these concerns, an African regional project was launched 4 years ago on the development of isotope and nuclear techniques in hydrology in the Sahelian countries. Its main objectives were to further broaden ongoing studies in the participating countries; to strengthen training; to promote exchange of information on various previous studies; to foster co-operation between the different institutes involved in these studies; and to reinforce the infrastructures established by the IAEA and their utilization in the region

  3. Efficient production of isotopically labeled proteins by cell-free synthesis: A practical protocol

    Energy Technology Data Exchange (ETDEWEB)

    Torizawa, Takuya; Shimizu, Masato [Crest, Jst (Japan); Taoka, Masato [Tokyo Metropolitan University, Graduate School of Science (Japan); Miyano, Hiroshi [Ajinomoto Co., Inc. Institute of Life Sciences (Japan); Kainosho, Masatsune [Crest, Jst (Japan)], E-mail: kainosho@nmr.chem.metro-u.ac.jp

    2004-11-15

    We provide detailed descriptions of our refined protocols for the cell-free production of labeled protein samples for NMR spectroscopy. These methods are efficient and overcome two critical problems associated with the use of conventional Escherichia coli extract systems. Endogenous amino acids normally present in E. coli S30 extracts dilute the added labeled amino acids and degrade the quality of NMR spectra of the target protein. This problem was solved by altering the protocol used in preparing the S30 extract so as to minimize the content of endogenous amino acids. The second problem encountered in conventional E. coli cell-free protein production is non-uniformity in the N-terminus of the target protein, which can complicate the NMR spectra. This problem was solved by adding a DNA sequence to the construct that codes for a cleavable N-terminal peptide tag. Addition of the tag serves to increase the yield of the protein as well as to ensure a homogeneous protein product following tag cleavage. We illustrate the method by describing its stepwise application to the production of calmodulin samples with different stable isotope labeling patterns for NMR analysis.

  4. The evaluation of new and isotopically labeled isoindoline nitroxides and an azaphenalene nitroxide for EPR oximetry

    Science.gov (United States)

    Khan, Nadeem; Blinco, James P.; Bottle, Steven E.; Hosokawa, Kazuyuki; Swartz, Harold M.; Micallef, Aaron S.

    2011-01-01

    Isoindoline nitroxides are potentially useful probes for viable biological systems, exhibiting low cytotoxicity, moderate rates of biological reduction and favorable Electron Paramagnetic Resonance (EPR) characteristics. We have evaluated the anionic (5-carboxy-1,1,3,3-tetramethylisoindolin-2-yloxyl; CTMIO), cationic (5-(N,N,N-trimethylammonio)-1,1,3,3-tetramethylisoindolin-2-yloxyl iodide, QATMIO) and neutral (1,1,3,3-tetramethylisoindolin-2-yloxyl; TMIO) nitroxides and their isotopically labeled analogues (2H12- and/or 2H12-15N-labeled) as potential EPR oximetry probes. An active ester analogue of CTMIO, designed to localize intracellularly, and the azaphenalene nitroxide 1,1,3,3-tetramethyl-2,3-dihydro-2-azaphenalen-2-yloxyl (TMAO) were also studied. While the EPR spectra of the unlabeled nitroxides exhibit high sensitivity to O2 concentration, deuteration resulted in a loss of superhyperfine features and a subsequent reduction in O2 sensitivity. Labeling the nitroxides with 15N increased the signal intensity and this may be useful in decreasing the detection limits for in vivo measurements. The active ester nitroxide showed approximately 6% intracellular localization and low cytotoxicity. The EPR spectra of TMAO nitroxide indicated an increased rigidity in the nitroxide ring, due to dibenzo-annulation. PMID:21665499

  5. Labeling pharmaceuticals with radioactive isotopes. Technical progress report, December 1, 1975--November 30, 1976

    International Nuclear Information System (INIS)

    Blau, M.; Bender, M.A.

    1976-01-01

    The purpose of this research is to prepare iodo- and bromo-aliphatic amino acid analogs labeled with γ-emitting isotopes ( 131 I, 123 I and 77 Br) for possible use as pancreas localizing agents. Studies on the halogen exchange reaction (I- for Cl-) for the synthesis of β-iodo-α-aminobutyric acid (a valine analog) have suggested that the iodo compound was formed initially. However, the desired compound cannot be isolated because of its chemical instability. Distribution studies in rats with the crude halogen exchange reaction mixture confirmed this finding. Studies on the addition of hydrogen iodine to allylglycine under various conditions for the synthesis of γ-iodo-α-aminopentanoic acid (a leucine analog) suffered the same obstacle; the chemical instability of the desired iodo compound precludes isolation and characterization. Convinced that the iodo analogs were too unstable for use as practical localizing agents, we turned to the possible use of Br for CH 3 substituted amino acids. The 14 C labeled β-bromo-α-aminobutyric acid methyl ester was synthesized. This methyl ester will be hydrolyzed and the distribution of free amino acid will be studied. Labeled with 77 Br this compound might be useful for pancreas localization

  6. Efficient production of isotopically labeled proteins by cell-free synthesis: A practical protocol

    International Nuclear Information System (INIS)

    Torizawa, Takuya; Shimizu, Masato; Taoka, Masato; Miyano, Hiroshi; Kainosho, Masatsune

    2004-01-01

    We provide detailed descriptions of our refined protocols for the cell-free production of labeled protein samples for NMR spectroscopy. These methods are efficient and overcome two critical problems associated with the use of conventional Escherichia coli extract systems. Endogenous amino acids normally present in E. coli S30 extracts dilute the added labeled amino acids and degrade the quality of NMR spectra of the target protein. This problem was solved by altering the protocol used in preparing the S30 extract so as to minimize the content of endogenous amino acids. The second problem encountered in conventional E. coli cell-free protein production is non-uniformity in the N-terminus of the target protein, which can complicate the NMR spectra. This problem was solved by adding a DNA sequence to the construct that codes for a cleavable N-terminal peptide tag. Addition of the tag serves to increase the yield of the protein as well as to ensure a homogeneous protein product following tag cleavage. We illustrate the method by describing its stepwise application to the production of calmodulin samples with different stable isotope labeling patterns for NMR analysis

  7. Chemical Ligation of Folded Recombinant Proteins: Segmental Isotopic Labeling of Domains for NMR Studies

    Science.gov (United States)

    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. The method for production of high purity carrier free ortophosphoric acid labeled with isotopes Phosphorus-32 and Phosphorus-33

    International Nuclear Information System (INIS)

    Abdukayumov, M.N.; Abdusalyamov, A.N.; Chistyakov, P.G.; Yuldashev, B.S.

    2001-01-01

    Extensive application for various radioactive isotopes was found in an extremity of the 20-Th century in a science and production. Labeled compounds are used with growing effectiveness in a molecular biology, gene engineering, medicine and other areas. Phosphorus-32 and Phosphorus-33 isotopes as a different labeled compounds that are used mainly in molecular biology are produced at the Radiopreparat enterprise of the Institute of Nuclear Physics of Academy of Sciences of Uzbekistan Republic. The quality of labeled preparations is very high. The specifications for above mentioned preparations corresponds to demands most of customers in different countries. P-32 or P-33 labeled orthophosphoric acid has high radiochemical purity (more than 99 %) and specific radioactivity close to theoretical. Orthophosphoric acid prepared by the described above method has radiochemical purity about 95 % and output of the target product 99%

  9. Techniques for intrinsically labeling wheat with 65Zn

    International Nuclear Information System (INIS)

    Starks, T.L.; Johnson, P.E.

    1985-01-01

    Several techniques of intrinsically labeling wheat with 65 Zn were compared: stem injection of 65 Zn, stem injection of 65 Zn + ZnSO 4 , foliar application of 65 Zn, and the addition of 65 Zn to a hydroponic solution. Incorporation levels of 65 Zn into the grain were 62.6% stem injection, 45.2% stem-injected 65 Zn + ZnSO 4 , 57.5% foliar application, and 2.3% hydroponic solution. Four protein fractions were extracted from fat-free whole wheat flour. Distribution of 65 Zn into the protein fractions for all treatments, was 8.5-20.3% in albumins and globulins, 47.4-60.3% in glutenins, 1-2.6% in gliadins, and 9.8-28.3% in the remaining proteins. Separation of the fractions by gel chromatography showed that protein and Zn distributions were similar among the treatments and when compared to the controls. Zinc-65 distribution was similar to the natural Zn distribution. These data illustrate that stem-injected 65 Zn is incorporated in the same manner and ratios as Zn naturally utilized by wheat

  10. Sequencing of Isotope-Labeled Small RNA Using Femtosecond Laser Ablation Time-of-Flight Mass Spectrometry

    Science.gov (United States)

    Kurata-Nishimura, Mizuki; Ando, Yoshinari; Kobayashi, Tohru; Matsuo, Yukari; Suzuki, Harukazu; Hayashizaki, Yoshihide; Kawai, Jun

    2010-04-01

    A novel method for the analysis of sequences of small RNAs using nucleotide triphosphates labeled with stable isotopes has been developed using time-of-flight mass spectroscopy combined with femtosecond laser ablation (fsLA-TOF-MS). Small RNAs synthesized with nucleotides enriched in 13C and 15N were efficiently atomized and ionized by single-shot fsLA and the isotope ratios 13C/12C and 15N/14N were evaluated using the TOF-MS method. By comparing the isotope ratios among four different configurations, the number of nucleotide contents of the control RNA sample were successfully reproduced.

  11. Affordable uniform isotope labeling with {sup 2}H, {sup 13}C and {sup 15}N in insect cells

    Energy Technology Data Exchange (ETDEWEB)

    Sitarska, Agnieszka; Skora, Lukasz; Klopp, Julia; Roest, Susan; Fernández, César; Shrestha, Binesh; Gossert, Alvar D., E-mail: alvar.gossert@novartis.com [Novartis Institutes for BioMedical Research (Switzerland)

    2015-06-15

    For a wide range of proteins of high interest, the major obstacle for NMR studies is the lack of an affordable eukaryotic expression system for isotope labeling. Here, a simple and affordable protocol is presented to produce uniform labeled proteins in the most prevalent eukaryotic expression system for structural biology, namely Spodoptera frugiperda insect cells. Incorporation levels of 80 % can be achieved for {sup 15}N and {sup 13}C with yields comparable to expression in full media. For {sup 2}H,{sup 15}N and {sup 2}H,{sup 13}C,{sup 15}N labeling, incorporation is only slightly lower with 75 and 73 %, respectively, and yields are typically twofold reduced. The media were optimized for isotope incorporation, reproducibility, simplicity and cost. High isotope incorporation levels for all labeling patterns are achieved by using labeled algal amino acid extracts and exploiting well-known biochemical pathways. The final formulation consists of just five commercially available components, at costs 12-fold lower than labeling media from vendors. The approach was applied to several cytosolic and secreted target proteins.

  12. Study of groundwater recharge in Rechna Doab using isotope techniques

    International Nuclear Information System (INIS)

    Sajjad, M.I.; Tasneem, M.A.; Ahmed, M.; Hussain, S.D.; Khan, I.H.; Akram, W.

    1992-04-01

    Isotopic studies were performed in the Rechna Doab area to understand the recharge mechanism, investigate the relative contributions from various sources such as rainfall, rivers and canal system and to estimate the turn over times and replenishment rate of groundwater. The isotopic data suggest that the groundwater in the project area can be divided into different zones each having its own characteristic isotopic composition. The enriched isotopic values show rain recharge and depleted isotopic values are associated with river/canal system while the intermediate isotopic values show a mixing of two or more sources of water. The major contribution, however, comes from canal system. The isotopic data suggest that there is no quick movement of groundwater in the area. 18 figs. (author)

  13. Application of isotope techniques to investigate groundwater pollution in India

    International Nuclear Information System (INIS)

    Shivanna, K.; Navada, S.V.; Kulkarni, K.M.; Sinha, U.K.; Sharma, S.

    1998-01-01

    Environmental isotopes ( 2 H, 18 O, 34 S, 3 H, and 14 C) techniques have been used along with hydrogeology and hydrochemistry to investigate: (a). the source of salinity and origin of sulphate in groundwaters of coastal Orissa, Orissa State, India and (b) to study the source of salinity in deep saline groundwaters of charnockite terrain at Kokkilimedu, South of Chennai, India. In the first case, as a part of large drinking water supply project, thousands of hand pumps were installed from 1985. Many of them became quickly unacceptable for potable supply due to salinity, increased iron and sulphate contents of the groundwater. In this alluvial, multiaquifer system, fresh, brackish and saline groundwaters occur in a rather complicated fashion. The conditions change from phreatic to confined flowing type with increasing depth. The results of the isotope geochemical investigation indicate that the shallow groundwater (depth/<50m) is fresh and modern. Groundwater salinity in intermediate aquifer (50 - 100m) is due to the Flandrian transgression during Holocene period. Fresh and modern deep groundwater forms a well developed aquifer which receives recharge through weathered basement rock. The saline groundwater found below the fresh deep aquifer have marine water entrapped during late Pleistocene. The source of high sulphate in the groundwater is of marine origin. In the second case, under the host rock characterization programme, the charnockite rock formation at Kokkilimedu, Kalpakkam was evaluated to assess its suitability as host medium for location of a geological repository for high level radioactive waste. Four deep boreholes were drilled in this area, the depth varying from 200 to 618 m. In these boreholes, large variations in groundwater salinity were observed over a distance of only a few hundred meters and no regional pattern could be identified. The results of the isotope investigation show that there are two different sources of salinity in this area. Among

  14. Isotope techniques in studies of selenium deficiency and toxicity syndromes in farm animals

    International Nuclear Information System (INIS)

    Giese, W.W.

    1984-01-01

    In a brief review of the Se deficiency syndrome in ruminants, studies using non-isotopic methods are applied to an introductory description of the disease. They include methods currently applied for determining Se deficiency status in feed and in ruminant animals. Detection of potential white muscle disease in lambs and calves is discussed. The application of 75 Se in studies on absorption, tissue distribution and excretion under feeding regimes with different Se levels and partly with addition of SO 4 ions or vitamin E is reviewed. In vitro studies with 75 Se include a description of the 75 Se uptake test with red blood cells, the metabolism of selenite, selenate and selenomethionine in rumen microorganisms, and the distribution of 75 Se in cow's and goat's milk. Methods of Se supplementation in Se-deficient areas are summarized and tests with 75 Se-labelled ruminal pellets in sheep and cattle are described. The Se toxicity syndrome is surveyed with respect to causative agents, symptomatology and gross pathology. Special reference is made to the blind staggers and the alkali disease types of selenosis. Isotope techniques are found to be less frequently applied in studies on Se toxicity than in Se deficiency studies. (author)

  15. The Application of Isotope Techniques in Nutrient Assessment and Management in Riverine Systems. Present and Future

    Energy Technology Data Exchange (ETDEWEB)

    Ito, M.; Newman, B. D. [International Atomic Energy Agency, Isotope Hydrology Section, Vienna (Austria); Hadwen, W. L. [Australian Rivers Institute, Griffith School of Environment, Griffith University - Nathan Campus, Brisbane, Queensland (Australia); Rogers, K. [National Isotope Center, GNS Science, Lower Hutt (New Zealand); Mayer, B. [Department of Geoscience, University of Calgary, Calgary, Alberta (Canada); Hein, T. [Wasser Cluster Lunz, Interuniversitary Center for Aquatic Research, Lunz-See, and University of Natural Resources and Applied Life Sciences, Institute of Hydrobiology and Aquatic Ecosystem Management, Vienna (Austria); Stellato, L. [Centre for Isotopic Research on Cultural and Environmental Heritage (CIRCE), Seconda Universita degli Studi di Napoli, Caserta (Italy); Ohte, N. [Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo (Japan); Mclaughlin, K. [Southern California Coastal Water Research Project, Costa Mesa, California (United States)

    2013-05-15

    A variety of sources contribute to nutrients in rivers and nutrients may subsequently take various pathways and undergo different transformation processes. We first review representative types of isotopes and the roles of isotope techniques that have been or could be used for nutrient assessment and management. We then present technical, financial and logistical matters to be considered in selecting appropriate isotope techniques for nutrient assessment and management. Lastly we propose several approaches on the application of isotope techniques to make more effective the studies and management of nutrients in rivers in the near future. (author)

  16. Multiple stable isotope tracer technique for studying the metabolic kinetics of amino acids in hepatic failure

    Energy Technology Data Exchange (ETDEWEB)

    Zongqin, Xia; Tengchang, Dai; Jianhua, Zhang; Yaer, Hu; Bingyao, Yu; Xingrong, Xu; Guanlu, Huang; Gengrong, Shen; Yaqiu, Zhou; Hong, Yu

    1987-08-01

    In order to study the mechanism of the imbalance of amino acid metabolism during hepatic failure, a stable isotope tracer method for observing simultaneously the metabolic kinetics of several amino acids has been established. /sup 15/N-L-Ala, (2,3-D/sub 3/)-Leu and (2,3-D/sub 3/)-Phe were chosen as nonessential, branched chain and aromatic amino acids. A single iv injection of 40 mg N-Ala, 20 mg deuterated Leu and 20 mg deuterated Phe was given to each human subject. Blood samples were taken just before and at different times (up to 60 min) after the injection. Total free amino acids were isolated from the plasma with a small dowex 50 x 8 column and converted to trifluoroacetyl derivatives. Their abundances were then analyzed with a GC-MS system and typical double exponential time course curves were found for all the three labelled amino acids. A two-pool model was designed and applied for compartmental analysis. Significant changes were found in the kinetic parameters of Phe and Leu in patients with fulminant hepatitis or heptic cirrhosis. The half-lives of both Phe pools were longer and the pool sizes were larger than normal subjects, while the half-lives and pool sizes of Leu changes in the opposite direction. No marked change was found in Ala. The significance of intracellular imbalance of Phe and Leu metabolism was discussed. It is evident that the combination of GCMS technique and multiple-tracers labelled with stable isotopes is of great potential for similar purposes.

  17. Negative ion ESI-MS analysis of natural yellow dye flavonoids--An isotopic labelling study

    Science.gov (United States)

    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.

  18. Isotope labeling-based quantitative proteomics of developing seeds of castor oil seed (Ricinus communis L.)

    DEFF Research Database (Denmark)

    Nogueira, Fábio C S; Palmisano, Giuseppe; Schwämmle, Veit

    2013-01-01

    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...... 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......, 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...

  19. Extrinsic labelling of staple food crops with isotopic iron does not consistently result in full equilibration: Revisiting the methodology

    Science.gov (United States)

    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...

  20. Isotope labelling study of CO oxidation-assisted epoxidation of propene. Implications for oxygen activation on Au catalysts.

    Science.gov (United States)

    Jiang, Jian; Oxford, Sean M; Fu, Baosong; Kung, Mayfair C; Kung, Harold H; Ma, Jiantai

    2010-06-07

    (18)O isotope labelling studies of the CO oxidation-assisted epoxidation of propene, catalyzed by a mixture of Au/TiO(2) and TS-1, using a methanol-H(2)O solvent showed the O in the epoxide was exclusively from O(2) and not H(2)O or methanol.

  1. Isotope exchange reactions of the hydrogen H-5 of selected pyrimidine derivatives and the preparation of tritium-labeled pyrimidines

    Czech Academy of Sciences Publication Activity Database

    Dračínský, Martin; Jansa, Petr; Elbert, Tomáš

    2011-01-01

    Roč. 76, č. 12 (2011), s. 1567-1577 ISSN 0010-0765 R&D Projects: GA AV ČR KJB400550903; GA MŠk 1M0508 Institutional research plan: CEZ:AV0Z40550506 Keywords : isotopic labeling * NMR spectroscopy * nucleobases * pyrimidines Subject RIV: CC - Organic Chemistry Impact factor: 1.283, year: 2011

  2. Cross-Course Collaboration in the Undergraduate Chemistry Curriculum: Isotopic Labeling with Sodium Borodeuteride in the Introductory Organic Chemistry Laboratory

    Science.gov (United States)

    Kjonaas, Richard A.; Fitch, Richard W.; Noll, Robert J.

    2017-01-01

    A microscale isotopic labeling experiment is described for the introductory organic chemistry laboratory course wherein half of the students use sodium borohydride (NaBH[subscript 4]) and the other half use sodium borodeuteride (NaBD[subscript 4]) to reduce acetophenone to 1-phenylethanol and then compare spectral data. The cost is reasonable, and…

  3. Use of environmental isotope techniques in groundwater hydrology

    International Nuclear Information System (INIS)

    Tirumalesh, K.; Shivanna, K.

    2009-01-01

    Environmental isotopes (stable and radioactive) have been used as tracers for investigating various hydrological problems. Wide variation in isotopic distribution ( 2 H, 13 C, 18 O, 15 N, 35 S, 3 H and 14 C) in the environment help in identifying the source, origin, pathways and processes affecting the system under consideration. In this article, a few Indian case studies covering some of the very important isotope applications in groundwater hydrology are briefly summarized. (author)

  4. Agronomic evaluation of guano sources by means of isotope techniques

    International Nuclear Information System (INIS)

    Zapata, F.; Arrillaga, J.L.

    2002-01-01

    Many soils of the tropics and subtropics under continuous cultivation are very infertile, thus poor yields are obtained and little crop residues remain to protect the soils from degrading erosion. External nutrient inputs in the form of chemical fertilizers, organic materials and other nutrients sources are essential for developing sustainable agricultural production systems. As chemical fertilizers are costly for developing countries with insufficient foreign currency for their purchase and their supplies are limited and irregular for small landholders, alternative nutrient sources must be sought and evaluated for use in dominant agricultural production systems. Locally available organic materials of different origin are potential sources of nutrients. One such source with high agronomic potential is guano. The present study was carried out to evaluate the agronomic effectiveness of two guano materials of different origin (Zaire and Peru) as sources of nitrogen and phosphorus as compared to chemical fertilizers (ammonium sulfate and triple superphosphate) using isotopic ( 15 N and 32 P) techniques. Using the classical method of comparing dry matter weight and P uptake, no significant differences among the tested guano sources were found. The use of the isotopic techniques allowed a quantitative assessment of the N and P supply to crops. Both guano materials were found to be good sources of N but in contrast were poor sources of phosphorus. In addition, from the agronomic evaluation, it was found that the guano of Zaire and the ammonium sulfate were N sources of equivalent efficiency and the guano of Peru even slightly better than the ammonium sulfate. As expected, P in the single superphosphate was as available to the P in the triple superphosphate. However, the substitution ratios for the guano sources were relatively high. Thus, 1 kg P as single superphosphate was equivalent to 9.5 kg P as guano from Zaire or 12.5 kg P as guano from Peru. Further field trials in

  5. Mutual complementation between water chemistry and isotope techniques

    International Nuclear Information System (INIS)

    Matthess, G.

    1976-01-01

    In the water chemistry and isotope methods which applied together enable more extensive statements to be made than each on its own, the following regions of cooperation are brought out: 1) Isotopes as conservative indicators a) microbial decomposition of organic substances in the anaerobic and aerobic region; b) precipitation and coprecipitation; c) mechanical filtration, adsorption and coprecipitation; d) gas exchange; e) dilution by infiltration; 2) geochemical observations as additional basis for isotope investigations; 3) the investigation of the water content substances as additional help to isotope hydrology. (HK/LH) [de

  6. Demonstration of de novo synthesis of enzymes by density labelling with stable isotopes

    International Nuclear Information System (INIS)

    Huebner, G.; Hirschberg, K.

    1977-01-01

    The technique of in vivo density labelling of proteins with H 2 18 O and 2 H 2 O has been used to investigate hormonal regulation and developmental expression of enzymes in plant cells. Buoyant density data obtained from isopycnic equilibrium centrifugation demonstrated that the cytokinine-induced nitrate reductase activity and the gibberellic acid-induced phosphatase activity in isolated embryos of Agrostemma githago are activities of enzymes synthesized de novo. The increase in alanine-specific aminopeptidase in germinating A. githago seeds is not due to de novo synthesis but to the release of preformed enzyme. On the basis of this result it is possible to apply the enzyme aminopeptidase as an internal density standard in equilibrium centrifugation. Density labelling experiments on proteins in pea cotyledons have been used to study the change in the activity of acid phosphatase, alanine-specific aminopeptidase, and peroxidase during germination. The activities of these enzymes increase in cotyledons of Pisum sativum. Density labelling by 18 O and 2 H demonstrates de novo synthesis of these three enzymes. The differential time course of enzyme induction shows the advantage of using H 2 18 O as labelling substance in cases when the enzyme was synthesized immediately at the beginning of germination. At this stage of development the amino-acid pool available for synthesis is formed principally by means of hydrolysis of storage proteins. The incorporation of 2 H into the new proteins takes place in a measurable amount at a stage of growth in which the amino acids are also synthesized de novo. The enzyme acid phosphatase of pea cotyledons was chosen to demonstrate the possibility of using the density labelling technique to detect protein turnover. (author)

  7. iMS2Flux – a high–throughput processing tool for stable isotope labeled mass spectrometric data used for metabolic flux analysis

    Directory of Open Access Journals (Sweden)

    Poskar C Hart

    2012-11-01

    Full Text Available Abstract Background Metabolic flux analysis has become an established method in systems biology and functional genomics. The most common approach for determining intracellular metabolic fluxes is to utilize mass spectrometry in combination with stable isotope labeling experiments. However, before the mass spectrometric data can be used it has to be corrected for biases caused by naturally occurring stable isotopes, by the analytical technique(s employed, or by the biological sample itself. Finally the MS data and the labeling information it contains have to be assembled into a data format usable by flux analysis software (of which several dedicated packages exist. Currently the processing of mass spectrometric data is time-consuming and error-prone requiring peak by peak cut-and-paste analysis and manual curation. In order to facilitate high-throughput metabolic flux analysis, the automation of multiple steps in the analytical workflow is necessary. Results Here we describe iMS2Flux, software developed to automate, standardize and connect the data flow between mass spectrometric measurements and flux analysis programs. This tool streamlines the transfer of data from extraction via correction tools to 13C-Flux software by processing MS data from stable isotope labeling experiments. It allows the correction of large and heterogeneous MS datasets for the presence of naturally occurring stable isotopes, initial biomass and several mass spectrometry effects. Before and after data correction, several checks can be performed to ensure accurate data. The corrected data may be returned in a variety of formats including those used by metabolic flux analysis software such as 13CFLUX, OpenFLUX and 13CFLUX2. Conclusion iMS2Flux is a versatile, easy to use tool for the automated processing of mass spectrometric data containing isotope labeling information. It represents the core framework for a standardized workflow and data processing. Due to its flexibility

  8. The labelling of Nanocoll[reg] with [111In] for dual-isotope scanning

    International Nuclear Information System (INIS)

    Mitterhauser, Markus; Wadsak, Wolfgang; Key Mien, Leonhard-; Eidherr, Harald; Roka, Sebastian; Zettinig, Georg; Angelberger, Peter; Viernstein, Helmut; Kletter, Kurt; Dudczak, Robert

    2003-01-01

    Visualization and biopsy of sentinel lymph nodes play an important role in planning and controlling the therapy of breast cancer. Hitherto two methods--scintigraphy or gamma probe detection after injection of [ 99m Tc]-nanocolloids and visual detection after injection of patent blue dye--are used routinely. There are no conclusive publications elucidating such important parameters as injection site, injection method and colloidal parameters. The present work aims to label Nanocoll[reg] with [ 111 In] to provide an alternative method, a simultaneous one-compound dual-isotope application. Methods: [ 111 In]-Indiumchloride was buffered with acetate and transferred to the nanocolloid. The colloid labelling reaction was complete after 30 min and filtrated through 100 nm Nuclepore[reg] filters. Results: Incorporation yield of [ 111 In]-Indium into the nanocolloid was nearly quantitative, the step associated with the major loss of activity was the particle sizing with a mean yield of 55%. Conclusion: The presented method allows for the routine supply of [ 111 In]-nanocolloids. Size-filtered [ 111 In]-Nanocoll[reg] shows the same particle size range as [ 99m Tc]-Nanocoll[reg

  9. Highly efficient residue-selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain

    International Nuclear Information System (INIS)

    Miyanoiri, Yohei; Ishida, Yojiro; Takeda, Mitsuhiro; Terauchi, Tsutomu; Inouye, Masayori; Kainosho, Masatsune

    2016-01-01

    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.

  10. Highly efficient residue-selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain

    Energy Technology Data Exchange (ETDEWEB)

    Miyanoiri, Yohei [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan); Ishida, Yojiro [Rutgers University-Robert Wood Johnson Medical School, Center for Advanced Biotechnology and Medicine (United States); Takeda, Mitsuhiro [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan); Terauchi, Tsutomu [Tokyo Metropolitan University, Graduate School of Science and Engineering (Japan); Inouye, Masayori [Rutgers University-Robert Wood Johnson Medical School, Center for Advanced Biotechnology and Medicine (United States); Kainosho, Masatsune, E-mail: kainosho@tmu.ac.jp [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan)

    2016-06-15

    We recently developed a practical protocol for preparing proteins bearing stereo-selectively {sup 13}C-methyl labeled leucines and valines, instead of the commonly used {sup 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.

  11. Radiation oxidation of polypropylene: A solid-state 13C NMR study using selective isotopic labeling

    International Nuclear Information System (INIS)

    Mowery, Daniel M.; Assink, Roger A.; Derzon, Dora K.; Klamo, Sara B.; Bernstein, Robert; Clough, Roger L.

    2007-01-01

    Polypropylene samples, in which the three different carbon atoms along the chain were selectively labeled with carbon-13, were subjected to radiation under inert and air atmospheres, and to post-irradiation exposure in air at various temperatures. By using solid-state 13 C NMR measurements at room temperature, we have been able to identify and quantify the oxidation products. The isotopic labeling provides insight into chemical reaction mechanisms, since oxidation products can be traced back to their positions of origin on the macromolecule. The major products include peroxides and alcohols, both formed at tertiary carbon sites along the chain. Other products include methyl ketones, acids, esters, peresters, and hemiketals formed from reaction at the tertiary carbon, together with in-chain ketones and esters from reaction at the secondary chain carbon. No evidence is found of products arising from reactions at the methyl side chain. Significant temperature-dependent differences are apparent; for example much higher yields of chain-end methyl ketones, which are the indicator product of chain scission, are generated for both elevated temperature irradiation and for post-irradiation treatment at elevated temperatures. Time-dependent plots of yields of the various oxidation products have been obtained under a wide range of conditions, including the post-irradiation oxidation of a sample at room temperature in air that has been monitored for 2 years. Radiation-oxidation products of polypropylene are contrasted to products measured for 13 C-labeled polyethylene in an earlier investigation: the peroxides formed in irradiated polypropylene are remarkably longer lived, the non-peroxidic products are significantly different, and the overall ratios of oxidation products in polypropylene change relatively little as a function of the extent of oxidation

  12. Identification of degradation routes of metamitron in soil microcosms using 13C-isotope labeling.

    Science.gov (United States)

    Wang, Shizong; Miltner, Anja; Nowak, Karolina M

    2017-01-01

    Metamitron is one of the most commonly used herbicide in sugar beet and flower bulb cultures. Numerous laboratory and field studies on sorption and degradation of metamitron were performed. Detailed biodegradation studies in soil using 13 C-isotope labeling are still missing. Therefore, we aimed at providing a detailed turnover mass balance of 13 C 6 -metamitron in soil microcosms over 80 days. In the biotic system, metamitron mineralized rapidly, and 13 CO 2 finally constituted 60% of the initial 13 C 6 -metamitron equivalents. In abiotic control experiments CO 2 rose to only 7.4% of the initial 13 C 6 -metamitron equivalents. The 13 C label from 13 C 6 -metamitron was incorporated into microbial amino acids that were ultimately stabilized in the soil organic matter forming presumably harmless biogenic residues. Finally, 13 C label from 13 C 6 -metamitron was distributed between the 13 CO 2 and the 13 C-biogenic residues indicating nearly complete biodegradation. The parallel increase of 13 C-alanine, 13 C-glutamate and 13 CO 2 indicates that metamitron was initially biodegraded via the desamino-metamitron route suggesting its relevance in the growth metabolism. In later phases of biodegradation, the "Rhodococcus route" was indicated by the low 13 CO 2 evolution and the high relevance of the pyruvate pathway, which aims at biomolecule synthesis and seems to be related to starvation. This is a first report on the detailed degradation route of metamitron in soil. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Traceability technique of isotopic application in food safety

    International Nuclear Information System (INIS)

    Guo Boli; Wei Yimin; Pan Jiarong

    2006-01-01

    Epidemics such as BSE, mouth and foot disease, avian influenza have brought new pressure to food safety management, constituted a extreme threat to people health, and caused serious economic loss and social scare to countries with outbreaks of above diseases. Isotopic tracing technology is an effect tool for tracing food origin and implementing the preservation of production premise in the world at present, and it is promising in the field of food safety traceability, so some developed countries have put a lot of effort on establishment of isotopic technology for food traceability. In this paper, the basic principles of isotopic tracing technology and the recent research advancement were be expounded, and the differentiate and connection was be compared between isotopic tracing technology and others. Furthermore, the suggestion about study of isotopic tracing technology in China was put forward. The aim of the paper is to promote the establishment and improvement of food traceability system, and ensure the consumer health. (authors)

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

    Science.gov (United States)

    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

  15. Labeling of monoclonal antibodies with a 67Ga-phenolic aminocarboxylic acid chelate. Part I. Chemistry and labeling technique.

    Science.gov (United States)

    Schuhmacher, J; Matys, R; Hauser, H; Maier-Borst, W; Matzku, S

    1986-01-01

    As a chelating agent for labeling antibodies (Abs) with metallic radionuclides, a propionic acid substituted ethylenediamine N,N'-di-[(o-hydroxyphenyl) acetic acid] (P-EDDHA), which tightly complexes 67Ga, was synthesized. The 67Ga-P-EDDHA chelate was coupled in aqueous solution to IgG at a molar ratio of 1:1 via carbodiimide. The average coupling yield was 15%. A specific activity of 4 mCi/mg IgG could be obtained with commercially supplied 67Ga. In vitro stability was evaluated in human serum at 37 degrees C and showed a half-life of about 120 h for the release of 67Ga from the labeled Ab during the initial phase of incubation. This in vitro halflife is similar to that measured for 111In-DTPA labeled Abs. Because of the high stability of the 67Ga-P-EDDHA chelate, the in vivo formation of radioactive labeled transferrin by transchelation, as described for 111In-DTPA labeled Abs, should, however, be reduced by this labeling technique.

  16. Determining synthesis rates of individual proteins in zebrafish (Danio rerio) with low levels of a stable isotope labelled amino acid.

    Science.gov (United States)

    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. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Validation of 13CO2 breath analysis as a measurement of demethylation of stable isotope labeled aminopyrine in man

    International Nuclear Information System (INIS)

    Schneider, J.F.; Schoeller, D.A.; Nemchausky, B.; Bayer, J.L.; Klein, P.

    1978-01-01

    Interval sampling of expired breath as a simple, non-invasive assessment of the effect of liver disease upon hepatic microsomal drug metabolism, has been demonstrated with [ 14 C] dimethylaminoantipyrine (aminopyrine). In order to eliminate radiation risk the authors have validated the use of aminopyrine labeled with the stable, non-radioactive isotope 13 C. Simultaneous oral administration of both [ 14 C]- and [ 13 C] aminopyrine to five adult subjects without liver disease as well as five patients with known liver disease, resulted in the excretion of label at nearly identical rates in both individual time collections (r=0.94) as well as cumulative excretion for three hours (r=0.97). An oral dose of 2-mg/kg of [ 13 C) aminopyrine resulted in rates of production of 13 CO 2 significantly greater than baseline variations in 13 CO 2 production in the fasting, resting subject. Measurements of a single peak value at one half hour correlated closely with the determination of cumulative appearance over three hours (r=0.96). A consistent reproducible increase in the peak production of 13 CO 2 was observed when five patients received phenobarbital. Stable isotope labeled aminopyrine may be used to detect the effects of disease and treatment upon hepatic N-demethylation activity in human subjects without incurring any risk from radiation. Furthermore, the availability of another isotopic carbon label should make possible the study of direct drug-drug interaction utilizing CO 2 analysis. (Auth.)

  18. Isotope coded protein labeling coupled immunoprecipitation (ICPL-IP): a novel approach for quantitative protein complex analysis from native tissue.

    Science.gov (United States)

    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.

  19. Isotope Coded Protein Labeling Coupled Immunoprecipitation (ICPL-IP): A Novel Approach for Quantitative Protein Complex Analysis From Native Tissue*

    Science.gov (United States)

    Vogt, Andreas; Fuerholzner, Bettina; Kinkl, Norbert; Boldt, Karsten; Ueffing, Marius

    2013-01-01

    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. PMID:23268931

  20. Use of isotope techniques in water resources inventory planning and development

    International Nuclear Information System (INIS)

    1979-01-01

    The seminar, sponsored jointly by the IAEA, UNESCO, and WHO, was organized by the Isotope Hydrology Section for hydrologists and hydrogeologists from English-speaking African countries who have primary administrative and technical responsibility in planning, implementing, and supervising programmes in the field of water resources in their countries. The aim of the seminar was to discuss and to inform the participants of both the theoretical and applied aspects of isotope techniques in hydrology and their potential use in studies dealing with water resources inventory, planning, and development. A similar regional seminar was organized in 1973 by the IAEA in Mexico City for Latin- American countries. In 1979, such a seminar will be held for French-speaking African countries. The Nairobi seminar was held at the East African Institute for Meteorological Training and Research. It was opened by the Minister for Water Development of the Kenyan Government, Dr. Gikonyo Kiano, who stressed the importance of water development problems in the African region and who appreciated the IAEA/UNESCO/WHO initiative in holding the seminar on isotope techniques in water resources in Nairobi. The programme of seminar lectures and discussions included the following topics. 1 Basic principles of radioisotope techniques and stable isotope ratios in hydrology 2. Tritium and radiocarbon as environmental tracers for dating water bodies 3 Isotope techniques in studying the origin of groundwater, recharge, and flow of groundwater. 4. Isotope techniques for identification of surface and groundwater relationships. 5 Surface water studies including lake dynamics, discharge measurements and sediment transport 6. Isotope methods in aquifer characteristics. 7 Isotope methods in geothermal resources prospecting. 8. Isotope techniques in hydraulics engineering. Each topic was illustrated with detailed descriptions of case studies During discussions, participants presented important problems (of water

  1. Techniques and problems in studying intestinal absorption with radioactive isotopes in children

    International Nuclear Information System (INIS)

    James, W.P.T.; Waterlow, J.C.

    1976-01-01

    Radioactive isotopes give substantial promise for assisting the study of gastrointestinal absorption in children in that they allow reduction or elimination of the collection of blood, urine and faeces specimens. These operations are particularly difficult and unreliable in infants, on whom greatest interest in paediatric gastroenterology is centred in the tropics. Here intestinal malabsorption is most commonly associated with malnutrition, lactose intolerance, gastroenteritis, parasitic infestation and iron-deficiency anaemia. Two general techniques that have been employed are whole-body counting and analyses of 14 CO 2 exhaled in the breath after the feeding of 14 C-labelled nutrients. The former is advantageous if radionuclides suitable for the test at hand exist; the latter may be hard to interpret because of problems in the distribution and metabolism of the nutrient and intermediary products. Proper selection and understanding of the tests is particularly important in paediatric work, where the use of radioactive tracer techniques is unacceptable merely for the convenience of the investigator. (author)

  2. Synthesis of stable isotopically labeled peptides with filter-assisted enzymatic labeling for the diagnosis of hepatitis B virus infection utilizing mass spectrometry-based proteomics strategy

    International Nuclear Information System (INIS)

    Tsai, Hsing-Fen; Hsiao, He-Hsuan

    2017-01-01

    A facile method for the preparation of stable isotopically labeled peptides was developed by means of filter-assisted tryptic "1"6O/"1"8O water labeling, which could be directly applied to the determination of hepatitis B virus infection from human serum with tandem mass spectrometry. Tryptic peptides of hepatitis B surface antigen or hepatitis B e antigen from different subtypes of hepatitis B virus were synthesized with traditional solid-phase peptide synthesis as potential biomarkers. Trypsin catalyzed oxygen-18 exchange at their amidated c-terminus of arginine or lysine residue. The protease catalyzed oxygen-18 to oxygen-16 back exchange reaction was eliminated due to the complete removal of trypsin by the centrifugal filter containing a thin membrane associated with molecular weight cut-off of 10 KDa. The synthetic isotopic peptides were spiked into trichloroacetic acid/acetone precipitated human serum as internal standards and were selectively detected with multiplexed parallel reaction monitoring on a hybrid quadrupole-orbitrap mass spectrometer. The limit of detection for all synthetic peptides were in the range of 0.09 fmol–1.13 fmol. The results indicated that the peptide YLWEWASVR derived from hepatitis B surface antigen was quantified approximately 200 fmol per μl serum and may serve as a diagnostic biomarker for the detection of hepatitis B virus infected disease. - Highlights: • Facile synthesis of an inexpensive and highly reproducible stable isotopically labeled peptides. • Complete incorporation of two "1"8O atoms into synthesized peptides with filter-assisted enzymatic labeling. • Targeted analysis with parallel reaction monitoring assay for the disease diagnosis.

  3. Synthesis of stable isotopically labeled peptides with filter-assisted enzymatic labeling for the diagnosis of hepatitis B virus infection utilizing mass spectrometry-based proteomics strategy

    Energy Technology Data Exchange (ETDEWEB)

    Tsai, Hsing-Fen; Hsiao, He-Hsuan, E-mail: hhhsiao@dragon.nchu.edu.tw

    2017-03-01

    A facile method for the preparation of stable isotopically labeled peptides was developed by means of filter-assisted tryptic {sup 16}O/{sup 18}O water labeling, which could be directly applied to the determination of hepatitis B virus infection from human serum with tandem mass spectrometry. Tryptic peptides of hepatitis B surface antigen or hepatitis B e antigen from different subtypes of hepatitis B virus were synthesized with traditional solid-phase peptide synthesis as potential biomarkers. Trypsin catalyzed oxygen-18 exchange at their amidated c-terminus of arginine or lysine residue. The protease catalyzed oxygen-18 to oxygen-16 back exchange reaction was eliminated due to the complete removal of trypsin by the centrifugal filter containing a thin membrane associated with molecular weight cut-off of 10 KDa. The synthetic isotopic peptides were spiked into trichloroacetic acid/acetone precipitated human serum as internal standards and were selectively detected with multiplexed parallel reaction monitoring on a hybrid quadrupole-orbitrap mass spectrometer. The limit of detection for all synthetic peptides were in the range of 0.09 fmol–1.13 fmol. The results indicated that the peptide YLWEWASVR derived from hepatitis B surface antigen was quantified approximately 200 fmol per μl serum and may serve as a diagnostic biomarker for the detection of hepatitis B virus infected disease. - Highlights: • Facile synthesis of an inexpensive and highly reproducible stable isotopically labeled peptides. • Complete incorporation of two {sup 18}O atoms into synthesized peptides with filter-assisted enzymatic labeling. • Targeted analysis with parallel reaction monitoring assay for the disease diagnosis.

  4. Accurate and sensitive determination of molar fractions of "1"3C-Labeled intracellular metabolites in cell cultures grown in the presence of isotopically-labeled glucose

    International Nuclear Information System (INIS)

    Fernández-Fernández, Mario; Rodríguez-González, Pablo; Hevia Sánchez, David; González-Menéndez, Pedro; Sainz Menéndez, Rosa M.; García Alonso, J. Ignacio

    2017-01-01

    This work describes a methodology based on multiple linear regression and GC-MS for the determination of molar fractions of isotopically-labeled intracellular metabolites in cell cultures. Novel aspects of this work are: i) the calculation of theoretical isotopic distributions of the different isotopologues from an experimentally measured value of % 13C enrichment of the labeled precursor ii) the calculation of the contribution of lack of mass resolution of the mass spectrometer and different fragmentation mechanism such as the loss or gain of hydrogen atoms in the EI source to measure the purity of the selected cluster for each metabolite and iii) the validation of the methodology not only by the analysis of gravimetrically prepared mixtures of isotopologues but also by the comparison of the obtained molar fractions with experimental values obtained by GC-Combustion-IRMS based on "1"3C/"1"2C isotope ratio measurements. The method is able to measure molar fractions for twenty-eight intracellular metabolites derived from glucose metabolism in cell cultures grown in the presence of "1"3C-labeled Glucose. The validation strategies demonstrate a satisfactory accuracy and precision of the proposed procedure. Also, our results show that the minimum value of "1"3C incorporation that can be accurately quantified is significantly influenced by the calculation of the spectral purity of the measured cluster and the number of "1"3C atoms of the labeled precursor. The proposed procedure was able to accurately quantify gravimetrically prepared mixtures of natural and labeled glucose molar fractions of 0.07% and mixtures of natural and labeled glycine at molar fractions down to 0.7%. The method was applied to initial studies of glucose metabolism of different prostate cancer cell lines. - Highlights: • Determination of molar fractions of "1"3C-labeled metabolites in cell cultures. • The method is based on multiple linear regression and GC-MS. • Validation of the method by

  5. Accurate and sensitive determination of molar fractions of {sup 13}C-Labeled intracellular metabolites in cell cultures grown in the presence of isotopically-labeled glucose

    Energy Technology Data Exchange (ETDEWEB)

    Fernández-Fernández, Mario [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Julián Clavería 8, 33006 Oviedo (Spain); Rodríguez-González, Pablo, E-mail: rodriguezpablo@uniovi.es [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Julián Clavería 8, 33006 Oviedo (Spain); Hevia Sánchez, David; González-Menéndez, Pedro; Sainz Menéndez, Rosa M. [University Institute of Oncology (IUOPA), University of Oviedo, Julián Clavería 6, 33006 Oviedo (Spain); García Alonso, J. Ignacio [Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, Julián Clavería 8, 33006 Oviedo (Spain)

    2017-05-29

    This work describes a methodology based on multiple linear regression and GC-MS for the determination of molar fractions of isotopically-labeled intracellular metabolites in cell cultures. Novel aspects of this work are: i) the calculation of theoretical isotopic distributions of the different isotopologues from an experimentally measured value of % 13C enrichment of the labeled precursor ii) the calculation of the contribution of lack of mass resolution of the mass spectrometer and different fragmentation mechanism such as the loss or gain of hydrogen atoms in the EI source to measure the purity of the selected cluster for each metabolite and iii) the validation of the methodology not only by the analysis of gravimetrically prepared mixtures of isotopologues but also by the comparison of the obtained molar fractions with experimental values obtained by GC-Combustion-IRMS based on {sup 13}C/{sup 12}C isotope ratio measurements. The method is able to measure molar fractions for twenty-eight intracellular metabolites derived from glucose metabolism in cell cultures grown in the presence of {sup 13}C-labeled Glucose. The validation strategies demonstrate a satisfactory accuracy and precision of the proposed procedure. Also, our results show that the minimum value of {sup 13}C incorporation that can be accurately quantified is significantly influenced by the calculation of the spectral purity of the measured cluster and the number of {sup 13}C atoms of the labeled precursor. The proposed procedure was able to accurately quantify gravimetrically prepared mixtures of natural and labeled glucose molar fractions of 0.07% and mixtures of natural and labeled glycine at molar fractions down to 0.7%. The method was applied to initial studies of glucose metabolism of different prostate cancer cell lines. - Highlights: • Determination of molar fractions of {sup 13}C-labeled metabolites in cell cultures. • The method is based on multiple linear regression and GC-MS.

  6. Isotopically modified compounds

    International Nuclear Information System (INIS)

    Kuruc, J.

    2009-01-01

    In this chapter the nomenclature of isotopically modified compounds in Slovak language is described. This chapter consists of following parts: (1) Isotopically substituted compounds; (2) Specifically isotopically labelled compounds; (3) Selectively isotopically labelled compounds; (4) Non-selectively isotopically labelled compounds; (5) Isotopically deficient compounds.

  7. Use of radioactive double labelling technique in the chemical analysis of the mediators of cellular immunity

    International Nuclear Information System (INIS)

    Sorg, C.

    1978-01-01

    Radioactive double labelling was adapted for the analysis of mediators of cellular immunity. Two identical lymphocyte cultures were simultaneously labelled with [ 3 H]- or [ 14 C] leucine. Each of the cultures was stimulated with antigen or mitogen. The combined supernatants were then subjected to various fractionation procedures. By determining the isotope ratio in each fraction it is possible to identify those products of activated lymphocytes that have been produced either de novo or in increased amounts. The method proved sensitive enough to detect lymphocyte activation products in supernatants of activated lymphocyte cultures from guinea pig, mouse, and man

  8. Application of SAIL phenylalanine and tyrosine with alternative isotope-labeling patterns for protein structure determination

    Energy Technology Data Exchange (ETDEWEB)

    Takeda, Mitsuhiro [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan); Ono, Akira M.; Terauchi, Tsutomu [SAIL Technologies Co., Inc. (Japan); Kainosho, Masatsune, E-mail: kainosho@nmr.chem.metro-u.ac.j [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan)

    2010-01-15

    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}-{sup 13}C/{sup 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}-{sup 13}C/{sup 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 {sup 13}C, {sup 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.

  9. Application of SAIL phenylalanine and tyrosine with alternative isotope-labeling patterns for protein structure determination.

    Science.gov (United States)

    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. Application of SAIL phenylalanine and tyrosine with alternative isotope-labeling patterns for protein structure determination

    International Nuclear Information System (INIS)

    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 (ε- and ζ-SAIL Phe) and tyrosine (ε-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 δ-SAIL Phe and δ-SAIL Tyr, which allow us to observe and assign δ- 13 C/ 1 H signals very efficiently. Each of the various types of SAIL Phe and SAIL Tyr yields well-resolved resonances for the δ-, ε- or ζ- 13 C/ 1 H signals, respectively, which can readily be assigned by simple and robust pulse sequences. Since the δ-, ε-, and ζ-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 ζ-SAIL Phe and ε-SAIL Tyr would be practically the best choice for protein structural determinations.

  11. IsoMS: automated processing of LC-MS data generated by a chemical isotope labeling metabolomics platform.

    Science.gov (United States)

    Zhou, Ruokun; Tseng, Chiao-Li; Huan, Tao; Li, Liang

    2014-05-20

    A chemical isotope labeling or isotope coded derivatization (ICD) metabolomics platform uses a chemical derivatization method to introduce a mass tag to all of the metabolites having a common functional group (e.g., amine), followed by LC-MS analysis of the labeled metabolites. To apply this platform to metabolomics studies involving quantitative analysis of different groups of samples, automated data processing is required. Herein, we report a data processing method based on the use of a mass spectral feature unique to the chemical labeling approach, i.e., any differential-isotope-labeled metabolites are detected as peak pairs with a fixed mass difference in a mass spectrum. A software tool, IsoMS, has been developed to process the raw data generated from one or multiple LC-MS runs by peak picking, peak pairing, peak-pair filtering, and peak-pair intensity ratio calculation. The same peak pairs detected from multiple samples are then aligned to produce a CSV file that contains the metabolite information and peak ratios relative to a control (e.g., a pooled sample). This file can be readily exported for further data and statistical analysis, which is illustrated in an example of comparing the metabolomes of human urine samples collected before and after drinking coffee. To demonstrate that this method is reliable for data processing, five (13)C2-/(12)C2-dansyl labeled metabolite standards were analyzed by LC-MS. IsoMS was able to detect these metabolites correctly. In addition, in the analysis of a (13)C2-/(12)C2-dansyl labeled human urine, IsoMS detected 2044 peak pairs, and manual inspection of these peak pairs found 90 false peak pairs, representing a false positive rate of 4.4%. IsoMS for Windows running R is freely available for noncommercial use from www.mycompoundid.org/IsoMS.

  12. Isotope label-aided mass spectrometry reveals the influence of environmental factors on metabolism in single eggs of fruit fly.

    Directory of Open Access Journals (Sweden)

    Te-Wei Tseng

    Full Text Available In order to investigate the influence of light/dark cycle on the biosynthesis of metabolites during oogenesis, here we demonstrate a simple experimental protocol which combines in-vivo isotopic labeling of primary metabolites with mass spectrometric analysis of single eggs of fruit fly (Drosophila melanogaster. First, fruit flies were adapted to light/dark cycle using artificial white light. Second, female flies were incubated with an isotopically labeled sugar ((13C(6-glucose for 12 h--either during the circadian day or the circadian night, at light or at dark. Third, eggs were obtained from the incubated female flies, and analyzed individually by matrix-assisted laser desorption/ionization (MALDI mass spectrometry (MS: this yielded information about the extent of labeling with carbon-13. Since the incorporation of carbon-13 to uridine diphosphate glucose (UDP-glucose in fruit fly eggs is very fast, the labeling of this metabolite was used as an indicator of the biosynthesis of metabolites flies/eggs during 12-h periods, which correspond to circadian day or circadian night. The results reveal that once the flies adapted to the 12-h-light/12-h-dark cycle, the incorporation of carbon-13 to UDP-glucose present in fruit fly eggs was not markedly altered by an acute perturbation to this cycle. This effect may be due to a relationship between biosynthesis of primary metabolites in developing eggs and an alteration to the intake of the labeled substrate - possibly related to the change of the feeding habit. Overall, the study shows the possibility of using MALDI-MS in conjunction with isotopic labeling of small metazoans to unravel the influence of environmental cues on primary metabolism.

  13. Industrial scale production of stable isotopes employing the technique of plasma separation

    International Nuclear Information System (INIS)

    Stevenson, N.R.; Bigelow, T.S.; Tarallo, F.J.

    2003-01-01

    Calutrons, centrifuges, diffusion and distillation processes are some of the devices and techniques that have been employed to produce substantial quantities of enriched stable isotopes. Nevertheless, the availability of enriched isotopes in sufficient quantities for industrial applications remains very restricted. Industries such as those involved with medicine, semiconductors, nuclear fuel, propulsion, and national defense have identified the potential need for various enriched isotopes in large quantities. Economically producing most enriched (non-gaseous) isotopes in sufficient quantities has so far eluded commercial producers. The plasma separation process is a commercial technique now available for producing large quantities of a wide range of enriched isotopes. Until recently, this technique has mainly been explored with small-scale ('proof-of-principle') devices that have been built and operated at research institutes. The new Theragenics TM facility at Oak Ridge, TN houses the only existing commercial scale PSP system. This device, which successfully operated in the 1980's, has recently been re-commissioned and is planned to be used to produce a variety of isotopes. Progress and the capabilities of this device and it's potential for impacting the world's supply of stable isotopes in the future is summarized. This technique now holds promise of being able to open the door to allowing new and exciting applications of these isotopes in the future. (author)

  14. Auto-inducing media for uniform isotope labeling of proteins with 15N, 13C and 2H

    International Nuclear Information System (INIS)

    Guthertz, Nicolas; Klopp, Julia; Winterhalter, Aurélie; Fernández, César; Gossert, Alvar D.

    2015-01-01

    Auto-inducing media for protein expression offer many advantages like robust reproducibility, high yields of soluble protein and much reduced workload. Here, an auto-inducing medium for uniform isotope labelling of proteins with 15 N, 13 C and/or 2 H in E. coli is presented. So far, auto-inducing media have not found widespread application in the NMR field, because of the prohibitively high cost of labeled lactose, which is an essential ingredient of such media. Here, we propose using lactose that is only selectively labeled on the glucose moiety. It can be synthesized from inexpensive and readily available substrates: labeled glucose and unlabeled activated galactose. With this approach, uniformly isotope labeled proteins were expressed in unattended auto-inducing cultures with incorporation of 13 C, 15 N of 96.6 % and 2 H, 15 N of 98.8 %. With the present protocol, the NMR community could profit from the many advantages that auto-inducing media offer

  15. Developing powerful tritide technique: Organic and biological molecule labeling

    International Nuclear Information System (INIS)

    Anon.

    1991-01-01

    Complex hydrides are very important reagents in organic synthesis due to the range of reducing powers and selectivities available from different agents. Unfortunately, the availability of these compounds for radiosynthesis has been extremely limited due to the difficulty of making them with adequate levels of tritium. Investigators at the Lawrence Berkeley Laboratory (LBL) National Tritium Labeling Facility have developed a new addition to the repertoire of the tritium-labeling chemist. The new method allows site-specific incorporation of tritium into organic and biological molecules by efficient reduction processes. Exceptionally reactive and selective reducing agents are prepared and used for labeling in a on-pot process. Three new tritide reagents - supertritide (lithium triethyl borotritide), LiAlT 4 (lithium aluminum tritide), and L-Selectride (sterically hindered lithium tri-sec-butyl borotritide) - have been synthesized at carrier-free levels, and have been demonstrated to be fully reactive. The availability of these versatile and reactive reagents gives the tritium radiochemist great control over chemoselectivity and stereoselectivity. The LBL tritide reagents can drive numerous conventional chemical reactions, and have been used to reduce p-toluene sulfonates, amides, lactones, esters, and aldehydes. These reactions produce good yields and result in products with maximum specific activities. The reagents clearly exhibit superior reactivity and may be used in many more synthetic processes than sodium borohydride, which is the currently used reagent. In addition, tritide reagents such as L-selectride have been shown to give greater control over stereochemistry and selectivity than sodium borohydride

  16. Stable isotope labeling, in vivo, of D- and L-tryptophan pools in lemna gibba and the low incorporation of label into indole-3-acetic acid

    International Nuclear Information System (INIS)

    Baldi, B.G.; Maher, B.R.; Slovin, J.P.; Cohen, J.D.

    1991-01-01

    The authors present evidence that the role of tryptophan and other potential intermediates in the pathways that could lead to indole derivatives needs to be reexamined. Two lines of Lemna gibba were tested for uptake of [ 15 N-indole]-labeled tryptophan isomers and incorporation of that label into free indole-3-acetic acid (IAA). Both lines required levels of L-[ 15 N]tryptophan 2 to 3 orders of magnitude over endogenous levels in order to obtain measurable incorporation of label into IAA. Labeled L-tryptophan was extractable from plant tissue after feeding and showed no measurable isomerization into D-tryptophan. D-[ 15 N]trytophan supplied to Lemna at rates of approximately 400 times excess of endogenous D-tryptophan levels (to yield an isotopic enrichment equal to that which allowed detection of the incorporation of L-tryptophan into IAA), did not result in measurable incorporation of label into free IAA. These results demonstrate that L-tryptophan is a more direct precursor to IAA than the D isomer and suggest (a) that the availability of tryptophan in vivo is not a limiting factor in the biosynthesis of IAA, thus implying that other regulatory mechanisms are in operation and (b) that L-tryptophan also may not be a primary precursor to IAA in plants

  17. Isotopic techniques for measuring the biological activity in plant rhizosphere

    International Nuclear Information System (INIS)

    Warembourg, F.R.

    1975-01-01

    The use of 14 C made it possible to separate root respired CO 2 and microbial CO 2 resulting from exudates utilisation by the rhizosphere microflora. Measurements were done after wheat plants grown under axenic and non axenic conditions were placed during short period of time in an atmosphere contaning 14 CO 2 . Under axenic conditions evolution of 14 CO 2 follows a bell shaped curve due to the brief appearance of labelled compounds translocated from the aerial part of the plants to the roots. In the presence of microorganisms, the maximum of activity due to root respiration is identical but immediately followed by a second peak of 14 CO 2 evolution that was attributed to the decomposition of labelled exudates by the microflora. The same observations resulted from the labelling of a grassland vegetation sampled with its soil and placed in the laboratory. Preliminary results obtained using this method of short term labelling of plants are presented here [fr

  18. Labeling of antibodies with a /sup 67/Ga-phenolic aminocarboxylic acid chelate. Pt. 1. Chemistry and labeling technique

    Energy Technology Data Exchange (ETDEWEB)

    Schuhmacher, J.; Matys, R.; Hauser, H.; Maier-Borst, W.; Matzku, S.

    1986-11-01

    As a chelating agent for labeling antibodies (Abs) with metallic radionuclides, a propionic acid substituted ethylenediamine N, N'-di-((o-hydroxyphenyl) acetic acid) (P-EDDHA), which tighly complexes /sup 67/Ga, was synthetized. The /sup 67/Ga-P-EDDHA chelate was coupled in aqueous solution to IgG at a molar ratio of 1:1 via carbodiimide. The average coupling yield was 15%. A specific activity of 4 mCi/mg IgG could be obtained with commercially supplied /sup 67/Ga. In vitro stability was evaluated in human serum at 37/sup 0/C and showed a half-life of about 120 h for the release of /sup 67/Ga from the labeled Ab during the initial phase of incubation. This in vitro halflife is similar to that measured for /sup 111/In-DTPA labeled Abs. Because of the high stability of the /sup 67/Ga-P-EDDHA chelate, the in vivo formation of radioactive lebeled transferrin by transchelation, as described for /sup 111/In-DTPA labeled Abs, should, however, be reduced by this labeling technique.

  19. LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

    Science.gov (United States)

    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.

  20. Metabolic Flux Analysis in Isotope Labeling Experiments Using the Adjoint Approach.

    Science.gov (United States)

    Mottelet, Stephane; Gaullier, Gil; Sadaka, Georges

    2017-01-01

    Comprehension of metabolic pathways is considerably enhanced by metabolic flux analysis (MFA-ILE) in isotope labeling experiments. The balance equations are given by hundreds of algebraic (stationary MFA) or ordinary differential equations (nonstationary MFA), and reducing the number of operations is therefore a crucial part of reducing the computation cost. The main bottleneck for deterministic algorithms is the computation of derivatives, particularly for nonstationary MFA. In this article, we explain how the overall identification process may be speeded up by using the adjoint approach to compute the gradient of the residual sum of squares. The proposed approach shows significant improvements in terms of complexity and computation time when it is compared with the usual (direct) approach. Numerical results are obtained for the central metabolic pathways of Escherichia coli and are validated against reference software in the stationary case. The methods and algorithms described in this paper are included in the sysmetab software package distributed under an Open Source license at http://forge.scilab.org/index.php/p/sysmetab/.

  1. Progress of new label-free techniques for biosensors: a review.

    Science.gov (United States)

    Sang, Shengbo; Wang, Yajun; Feng, Qiliang; Wei, Ye; Ji, Jianlong; Zhang, Wendong

    2016-01-01

    The detection techniques used in biosensors can be broadly classified into label-based and label-free. Label-based detection relies on the specific properties of labels for detecting a particular target. In contrast, label-free detection is suitable for the target molecules that are not labeled or the screening of analytes which are not easy to tag. Also, more types of label-free biosensors have emerged with developments in biotechnology. The latest developed techniques in label-free biosensors, such as field-effect transistors-based biosensors including carbon nanotube field-effect transistor biosensors, graphene field-effect transistor biosensors and silicon nanowire field-effect transistor biosensors, magnetoelastic biosensors, optical-based biosensors, surface stress-based biosensors and other type of biosensors based on the nanotechnology are discussed. The sensing principles, configurations, sensing performance, applications, advantages and restriction of different label-free based biosensors are considered and discussed in this review. Most concepts included in this survey could certainly be applied to the development of this kind of biosensor in the future.

  2. Manufacture of anti-bogus label by track-etching technique

    International Nuclear Information System (INIS)

    He Xiangming; Wan Chunrong

    2006-01-01

    Anti-bogus label is manufactured by the track-etching technique. The apparent pattern on the label consists of track-etched pores on the membrane. The manufacture of the label depends on the intricate technology and the state strictly controls the sensitive nuclear facilities, ensuring that the label is not copied. The pattern on the label is specially characterized by permeability of liquid in order to distinguish it from a forged one. A genuine label can be distinguished from a sham one by a transparent liquid (e.g. water) or a colorful pen. Nowadays, the products of more than 100 famous brands (trade mark) have been protected from forgery by this technology in the market of China. This is a new method for the utilization of a research reactor

  3. Isotope correlation techniques for verifying input accountability measurements at a reprocessing plant

    International Nuclear Information System (INIS)

    Umezawa, H.; Nakahara, Y.

    1983-01-01

    Isotope correlation techniques were studied to verify input accountability measurements at a reprocessing plant. On the basis of a historical data bank, correlation between plutonium-to-uranium ratio and isotopic variables was derived as a function of burnup. The burnup was determined from the isotopic ratios of uranium and plutonium, too. Data treatment was therefore made in an iterative manner. The isotopic variables were defined to cover a wide spectrum of isotopes of uranium and plutonium. The isotope correlation techniques evaluated important parameters such as the fuel burnup, the most probable ratio of plutonium to uranium, and the amounts of uranium and plutonium in reprocessing batches in connection with fresh fuel fabrication data. In addition, the most probable values of isotope abundance of plutonium and uranium could be estimated from the plutonium-to-uranium ratio determined, being compared with the reported data for verification. A pocket-computer-based system was developed to enable inspectors to collect and evaluate data in a timely fashion at the input accountability measurement point by the isotope correlation techniques. The device is supported by battery power and completely independent of the operator's system. The software of the system was written in BASIC. The data input can be stored in a cassette tape and transferred into a higher level computer. The correlations used for the analysis were given as a form of analytical function. Coefficients for the function were provided relevant to the type of reactor and the initial enrichment of fuel. (author)

  4. Milking technique of sup(99m)Tc generators and labeling efficiencies

    International Nuclear Information System (INIS)

    Salehi, N.; Guignard, P.A.

    1985-01-01

    Increased levels of 99 Tc in generator produced sup(99m)Tc have an adverse effect on the labelling efficiency of red blood cells and human serum albumin. A two-step milking technique in which the first 1-2 ml of eluate is discarded has been found to produce higher and more constant labelling efficiency of lymphocytes and platelets than a one-step procedure. Binding efficiency of platelet-rich plasma and lymphocytes with sup(99m)Tc is greater in the two-step technique. High activity concentration in the eluate for critical labelling is between 1.5-3 ml. (U.K.)

  5. Absolute Ca Isotopic Measurement Using an Improved Double Spike Technique

    Directory of Open Access Journals (Sweden)

    Jason Jiun-San Shen

    2009-01-01

    Full Text Available A new vector analytical method has been developed in order to obtain the true isotopic composition of the 42Ca-48Ca double spike. This is achieved by using two different sample-spike mixtures combined with the double spike and natural Ca data. Be cause the natural sample (two mixtures and the spike should all lie on a single mixing line, we are able to con strain the true isotopic composition of our double spike using this new approach. Once the isotopic composition of the Ca double spike is established, we are able to obtain the true Ca isotopic composition of the NIST Ca standard SRM915a, 40Ca/44Ca = 46.537 ± 2 (2sm, n = 55, 42Ca/44Ca = 0.31031 ± 1, 43Ca/44Ca = 0.06474 ± 1, and 48Ca/44Ca = 0.08956 ± 1. De spite an off set of 1.3% in 40Ca/44Ca between our result and the previously re ported value (Russell et al. 1978, our data indicate an off set of 1.89__in 40Ca/44Ca between SRM915a and seawater, entirely consistent with the published results.

  6. Stable isotope labeling – Liquid chromatography/mass spectrometry for quantitative analysis of androgenic and progestagenic steroids

    International Nuclear Information System (INIS)

    Guo, Ning; Liu, Ping; Ding, Jun; Zheng, Shu-Jian; Yuan, Bi-Feng; Feng, Yu-Qi

    2016-01-01

    Steroid hormones play important roles in mammal at very low concentrations and are associated with numerous endocrinology and oncology diseases. Therefore, quantitative analysis of steroid hormones can provide crucial information for uncovering underlying mechanisms of steroid hormones related diseases. In the current study, we developed a sensitive method for the detection of steroid hormones (progesterone, dehydroepiandrosterone, testosterone, pregnenolone, 17-hydroxyprogesterone, androstenedione and 17α-hydroxypregnenolone) in body fluids by stable isotope labeling coupled with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis. In this respect, a pair of isotopes labeling reagents, Girard reagent P (GP) and d_5-Girard reagent P (d_5-GP), were synthesized and utilized to label steroid hormones in follicular fluid samples and steroid hormone standards, respectively. The heavy labeled standards were used as internal standards for quantification to minimize quantitation deviation in MS analysis due to the matrix and ion suppression effects. The ionization efficiencies of steroid hormones were greatly improved by 4–504 folds through the introduction of a permanent charged moiety of quaternary ammonium from GP. Using the developed method, we successfully quantified steroid hormones in human follicular fluid. We found that the contents of testosterone and androstenedione exhibited significant increase while the content of pregnenolone had significant decrease in follicular fluid of polycystic ovarian syndrome (PCOS) patients compared with healthy controls, indicating that these steroid hormones with significant change may contribute to the pathogenesis of PCOS. Taken together, the developed stable isotope labeling coupled LC-ESI-MS/MS analysis demonstrated to be a promising method for the sensitive and accurate determination of steroid hormones, which may facilitate the in-depth investigation of steroid hormones related

  7. Stable isotope labeling – Liquid chromatography/mass spectrometry for quantitative analysis of androgenic and progestagenic steroids

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Ning; Liu, Ping; Ding, Jun; Zheng, Shu-Jian; Yuan, Bi-Feng; Feng, Yu-Qi, E-mail: yqfeng@whu.edu.cn

    2016-01-28

    Steroid hormones play important roles in mammal at very low concentrations and are associated with numerous endocrinology and oncology diseases. Therefore, quantitative analysis of steroid hormones can provide crucial information for uncovering underlying mechanisms of steroid hormones related diseases. In the current study, we developed a sensitive method for the detection of steroid hormones (progesterone, dehydroepiandrosterone, testosterone, pregnenolone, 17-hydroxyprogesterone, androstenedione and 17α-hydroxypregnenolone) in body fluids by stable isotope labeling coupled with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis. In this respect, a pair of isotopes labeling reagents, Girard reagent P (GP) and d{sub 5}-Girard reagent P (d{sub 5}-GP), were synthesized and utilized to label steroid hormones in follicular fluid samples and steroid hormone standards, respectively. The heavy labeled standards were used as internal standards for quantification to minimize quantitation deviation in MS analysis due to the matrix and ion suppression effects. The ionization efficiencies of steroid hormones were greatly improved by 4–504 folds through the introduction of a permanent charged moiety of quaternary ammonium from GP. Using the developed method, we successfully quantified steroid hormones in human follicular fluid. We found that the contents of testosterone and androstenedione exhibited significant increase while the content of pregnenolone had significant decrease in follicular fluid of polycystic ovarian syndrome (PCOS) patients compared with healthy controls, indicating that these steroid hormones with significant change may contribute to the pathogenesis of PCOS. Taken together, the developed stable isotope labeling coupled LC-ESI-MS/MS analysis demonstrated to be a promising method for the sensitive and accurate determination of steroid hormones, which may facilitate the in-depth investigation of steroid hormones

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

    Energy Technology Data Exchange (ETDEWEB)

    Opitz, Christian; Isogai, Shin; Grzesiek, Stephan, E-mail: Stephan.Grzesiek@unibas.ch [University of Basel, Focal Area Structural Biology and Biophysics, Biozentrum (Switzerland)

    2015-07-15

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

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

    International Nuclear Information System (INIS)

    Opitz, Christian; Isogai, Shin; Grzesiek, Stephan

    2015-01-01

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

  10. Sulfur Isotope Exchange between S-35 Labeled Inorganic Sulfur-Compounds in Anoxic Marine-Sediments

    DEFF Research Database (Denmark)

    FOSSING, H.; THODEANDERSEN, S.; JØRGENSEN, BB

    1992-01-01

    of isotope exchange, specific radioactivities of the reduced sulfur pools were poorly defined and could not be used to calculate their rates of formation. Such isotope exchange reactions between the reduced inorganic sulfur compounds will affect the stable isotope distribution and are expected to decrease...

  11. Measurement of the viability of stored red cells by the single-isotope technique using 51Cr. Analysis of validity

    International Nuclear Information System (INIS)

    Beutler, E.; West, C.

    1984-01-01

    A single-isotope 51 Cr method often is used to evaluate the viability of stored red cells. In this technique, the red cell mass is measured by back-extrapolation to time zero (t0) of the radioactivity of the blood between 5 and 20 minutes after infusion of the sample. If there is early destruction of stored cells, this method provides an overestimate of the red cell mass and, hence, of the viability of the stored cells. Freshly drawn red cells from normal donors were labeled with /sup 99m/Tc, and cells from the same donor which had been stored in citrate-phosphate-dextrose-adenine-one (CPDA-1) for periods ranging from 7 to 49 days were labeled with 51 Cr. A comparison of the ''true red cell mass'' as determined with /sup 99m/Tc with the back-extrapolated red cell mass from stored 51 Cr-labeled cells has made it possible to define the magnitude of error introduced by early loss of red cells. The overestimation of red cell mass and viability was diminished if only the 51 Cr radioactivity between 5 and 15 minutes after infusion was used in back-extrapolating to t0. The degree of overestimation of red cell mass was greatest when the red cell viability had declined to very low levels. However, in the entire range of 10 to 80 percent viability, the overestimate of viability was usually less than 4 percent. The overestimate of viability proved to be quite similar for all samples and may be taken into account when using the single-isotope technique for measurement of red cell viability

  12. Comparative study of label and label-free techniques using shotgun proteomics for relative protein quantification.

    Science.gov (United States)

    Sjödin, Marcus O D; Wetterhall, Magnus; Kultima, Kim; Artemenko, Konstantin

    2013-06-01

    The analytical performance of three different strategies, iTRAQ (isobaric tag for relative and absolute quantification), dimethyl labeling (DML) and label free (LF) for relative protein quantification using shotgun proteomics have been evaluated. The methods have been explored using samples containing (i) Bovine proteins in known ratios and (ii) Bovine proteins in known ratios spiked into Escherichia coli. The latter case mimics the actual conditions in a typical biological sample with a few differentially expressed proteins and a bulk of proteins with unchanged ratios. Additionally, the evaluation was performed on both QStar and LTQ-FTICR mass spectrometers. LF LTQ-FTICR was found to have the highest proteome coverage while the highest accuracy based on the artificially regulated proteins was found for DML LTQ-FTICR (54%). A varying linearity (k: 0.55-1.16, r(2): 0.61-0.96) was shown for all methods within selected dynamic ranges. All methods were found to consistently underestimate Bovine protein ratios when matrix proteins were added. However, LF LTQ-FTICR was more tolerant toward a compression effect. A single peptide was demonstrated to be sufficient for a reliable quantification using iTRAQ. A ranking system utilizing several parameters important for quantitative proteomics demonstrated that the overall performance of the five different methods was; DML LTQ-FTICR>iTRAQ QStar>LF LTQ-FTICR>DML QStar>LF QStar. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Isotopes in day to day life

    Science.gov (United States)

    1984-06-01

    Developments are reported in the use of isotopic labeling and isotope irradiation in agriculture, medical science, hydrology, geochemistry, geophysics, environment pollution detection, and industries. Radioisotope instruments are described as well as techniques for gamma radiography, neutron radiography, and autoradiography. Isotope dating in geology and archaeology is covered. Basic scientific research topics in various areas are listed.

  14. Quality Assurance of technitium-labelled radiopharmaceuticals in the Radiation and Isotopes Centre of Khartoum (RICK)

    International Nuclear Information System (INIS)

    Adlan, A.A.

    2005-09-01

    This descriptive, exploratory study was conducted in the nuclear medicine department at the Radiation and Isotopes Center of Khartoum (RICK) during 2003-2005 the aim of the study was explore and define the dimensions of a problem which was regarded as urgent by the people working in the field of nuclear medicine in Sudan. The problem concerned the quality of technitium-labelled radiopharmaceuticals which are used in more than 90% of the nuclear medicine imaging studies performed in nuclear medicine. Impure 9 ''9''m Tc-labelled radiopharmaceuticals may create problems, and could lead to false diagnosis. These agents must be tested for determination of the levels of radionuclides, radiochemical and chemicals, before administration to patients. They should also be sterile and pyrogen-free. A number of data collection methods, were used by the researcher for adequate exploration of the dimensions of the problem including interviews, questionnaires and close observations to all activities related to the preparation of radiopharmaceuticals in the hot laboratory. Information concerning all the aspects of quality assurance were collected. These aspects were management and organisation of the work, equipment and tools, knowledge and practical experience of the staff members and methods of preparation and administration of the radioactive agents. Data from different sources were then compared with observation results for more validation and finally lead to the following results: All the quality control tests were not normally performed in the department, therefore the levels of impurities in these agents were not exactly determined, moreover these preparations were subject to contamination with microorganisms, due to low level of cleanliness at the work area. The study detected a number of defaults which were likely to be the causes behind these problems. These were, bad management and organisation, in availability of equipment, tools and materials necessary for testing

  15. The application of high-resolution IR spectroscopy and isotope labeling for detailed investigation of TiO2/gas interface reactions

    Czech Academy of Sciences Publication Activity Database

    Civiš, Svatopluk; Ferus, Martin; Zukalová, Markéta; Kavan, Ladislav; Zelinger, Zdeněk

    2013-01-01

    Roč. 36, č. 1 (2013), s. 159-162 ISSN 0925-3467 R&D Projects: GA ČR GAP208/10/2302; GA ČR GA13-07724S Institutional support: RVO:61388955 Keywords : isotope exchange * titania * isotope labeling Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.075, year: 2013

  16. The role and future of in-vitro isotopic techniques in molecular biology

    International Nuclear Information System (INIS)

    Dar, L.; Khan, B.K.

    2004-01-01

    In this review we discuss isotopic in-vitro molecular biology techniques, and their advantages and applications. Isotopic methods have helped to shape molecular biology since its early days. Despite the availability of non-isotopic alternatives, isotopic methods continue to be used in molecular biology due to certain advantages, especially related to sensitivity and cost-effectiveness. Numerous techniques involving the use of isotopes help in the characterization of genes, including the detection of single nucleotide polymorphisms (SNPs) or mutations. Other isotopic molecular methods are utilized to study the phenotypic expression of gene sequences and their mutation. Emerging branches of molecular biology like functional genomics and proteomics are extremely important for exploiting the rapidly growing data derived from whole genomic sequencing of human and microbial genomes. Recent molecular biology applications like the high-throughput array techniques are relevant in the context of both structural and functional genomics. In proteomics, stable isotope based technology has found applications in the analysis of protein structure and interactions. (author)

  17. Sampling technique for collection of expired COsub(2) in studies using naturally labelled sup(13)C in calves

    International Nuclear Information System (INIS)

    Chevalier, R.; Pelletier, G.; Gagnon, M.

    1984-01-01

    Natural isotopic labelling of animal metabolic processes presents great advantages over the use of artificially enriched compounds. These advantages include the use of natural diets, low cost and easiness of handling, sampling and disposal. Changes in the sup(13)C/sup(12)C ratio of expired COsub(2) after ingestion of a test meal containing a variable proportion of Csub(3) and/or Csub(4) plant-derived products permits measurements of the immediate oxidation of dietary carbohydrates. A breathing device for collection of expired COsub(2) and purification techniques are described. The breath-sampling device is composed of a polypropylene enclosure and a rubber bag. The rubber bag is blown up by the animal and a sample of the expired gas is transferred to a sampling glass holder in which a vacuum had been previously achieved. Sample purification of expired COsub(2) is done by differential freezing

  18. Utilization of carbon 13-labelled stable isotopes for studying drug toxicity on cellular metabolism

    International Nuclear Information System (INIS)

    Herve, M.; Wietzerbin, J.; Tran-Dinh, S.

    1994-01-01

    A new approach for studying the effects of two drugs, amphotericine B (AMB), an anti-fungal antibiotic, and 2-deoxy-D-glucose (DG), on the glucose metabolism in brewer yeast cells (Saccharomyces cerevisiae), is presented; AMB interacts with the membrane sterols, inducing formation of pores through which ions and small molecules can pass. DG may enter in the cytosol, where it is phosphoryled by hexokinase into deoxy-D-glucose 6-phosphate (DG6P) which disappears very slowly. DG slows down the glycolysis process and induces the formation of new substances. This paper shows the advantages of utilizing carbon 13-labelled substrates combined to the NMR-13C and NMR-1H techniques. 6 figs., 5 refs

  19. Automatic detection of adverse events to predict drug label changes using text and data mining techniques.

    Science.gov (United States)

    Gurulingappa, Harsha; Toldo, Luca; Rajput, Abdul Mateen; Kors, Jan A; Taweel, Adel; Tayrouz, Yorki

    2013-11-01

    The aim of this study was to assess the impact of automatically detected adverse event signals from text and open-source data on the prediction of drug label changes. Open-source adverse effect data were collected from FAERS, Yellow Cards and SIDER databases. A shallow linguistic relation extraction system (JSRE) was applied for extraction of adverse effects from MEDLINE case reports. Statistical approach was applied on the extracted datasets for signal detection and subsequent prediction of label changes issued for 29 drugs by the UK Regulatory Authority in 2009. 76% of drug label changes were automatically predicted. Out of these, 6% of drug label changes were detected only by text mining. JSRE enabled precise identification of four adverse drug events from MEDLINE that were undetectable otherwise. Changes in drug labels can be predicted automatically using data and text mining techniques. Text mining technology is mature and well-placed to support the pharmacovigilance tasks. Copyright © 2013 John Wiley & Sons, Ltd.

  20. Separation and sampling technique of light element isotopes by chemical exchange process

    International Nuclear Information System (INIS)

    Kato, Shunsaku; Oi, Kenta; Takagi, Norio; Hirotsu, Takafumi; Kano, Hirofumi; Sonoda, Akinari; Makita, Yoji

    2000-01-01

    Lithium and boron isotope separation technique were studied. Granulation of lithium isotope separation agent was carried out by cure covering in solution. Separation of lithium isotope was stepped up by ammonium carbonate used as elusion agent. Styrene and ester resin derived three kinds of agents such as 2-amino-1, 3-propanediol (1, 3-PD), 2-amino-2-methyl-1, 3-propanediol (Me-1,3-PD) and tris(2-hydroxyethyl)amine (Tris) were used as absorbent.The ester resin with Tris showed larger amount of adsorption (1.4 mmol/g) than other resins. However, all resins with agent indicated more large adsorption volume of boron than the objective value (0.5 mmol/g). Large isotope shift was shown by the unsymmetrical vibration mode of lithium ion on the basis of quantum chemical calculation of isotope effect on dehydration of hydrated lithium ion. (S.Y.)

  1. Application of Isotope techniques in the arid and semi-arid zone hydrology

    International Nuclear Information System (INIS)

    Gonfiantini, R.; Louvat, D.; Aranyossy, J.F.

    1993-01-01

    The paper presents a brief introduction on the scientific background of isotope hydrology,and after, explains the applications of environment isotope techniques in groundwater hydrology of the arid and semi-arid zones. It includes the study of aquifer recharge and discharge, identification of palaeorecharge, groundwater movement and age in unconfined and confined aquifers, and interconnections between aquifers. The contribution of isotopes is highlighted with many examples of field case studies, with emphasis on studies carried out with IAEA support. Finally, a short description of IAEA program on isotope hydrology is given, with a list of regional projects supported through the IAEA Technical Cooperation program, and of Coordinated Research Programs. The latter give the modern research trends in isotope application to hydrology and hydrosphere environmental studies. (author). 2 tabs., 25 figs., 69 refs

  2. Preparation of H3-labelled methyl ethers of saturated fatty acids by heterogeneous catalytic isotope exchange in solution with gaseous tritium

    International Nuclear Information System (INIS)

    Shevchenko, V.P.; Myasoedov, N.F.

    1980-01-01

    A simple method of preparing 3 H-labelled methyl ethers of saturated fatty acids in the dioxane solution using the method of isotopic heterogenous catalytic exchange with gaseous tritium, is suggested. 3 H-labelled natural fatty acids (C 12 -C 18 ) are prepared by alkaline hydrolysis [ru

  3. Salars evaporation rates evaluation using isotope techniques, Bellavista Salar, Chile

    International Nuclear Information System (INIS)

    Grilli, A.; Ortiz, J.

    1989-01-01

    Long term evaporation rates are evaluated in different soil conditions at Bellavista Salar, using environmental isotope profiles (oxygen-18) of the unsaturated soil zone. The Barnes and Allison model was adapted to stratified soils under non-saturation conditions and for a non-isothermal permanent regime. To apply the proposed model, field data of the different variables were used and the evaporation rates were obtained adjunting the δ 18 O values generated by the model to those experimentally measured in the water extracted from the soil profile of the unsaturated soil zone. (author). 13 refs, 8 figs

  4. The stable isotope fingerprinting technique for agricultural pesticide

    Science.gov (United States)

    Suto, N.; Kawashima, H.

    2014-12-01

    The compound specific isotope analysis (CSIA) is nowadays an important and powerful tool in geochemical, environmental, and forensics field. In particular, the stable isotope ratio of pesticide is applied to biological process and reaction in the soil and distribution channel as forensics science. The aim of this study is to measure the stable isotope ratios of pesticide using various analytical methodologies, GC/IRMS, EA/IRMS, and LC/IRMS under high accuracy and precision. Therefore, these methods seemed to be important knowledge as geological field. In particular case, we present the method to measure carbon isotope ratio of nine malathion emulsion pesticides using GC/IRMS with cryo-focusing system to identify the source. In December 2013, food poisoning occurred after eating frozen dumplings (i.e., pizza and chicken nuggets) in Japan. There was a very high concentration, maximum value 15,000ppm, of malathion (diethyl (dimethoxythiophosphorylthio) succinate) in products. This incident was caused by an employee of process, and threatened the food safety. We analyzed the δ13C of malathion ranged from -30.63‰ to -29.54‰ (S.D. 0.10‰), the differences less than 1.0‰. All malathion emulsion sold in Japan are imported from Cheminova India Lat., Denmark to Sumitomo Chemical Co. Ltd., Japan. After that, Japanese each manufacture buy from Sumitomo Chemical Co. Ltd. And blended malathion and organic solvent (ethylbenzene and xylene). Therefore, ethylbenzene and xylene may be important tool as source identification. We measured the δ13C of ethylbenzene and m-,p-xylene, too. As the results, the δ13C of ethylbenzene and m-,p-xylene ranged from -28.20‰ to -20.84‰ (S.D. 0.16‰), -28.69‰ to -25.15‰ (S.D. 0.13‰), respectively. The δ13C of ethylbenzene and m-,p-xylene can be identified manufacture, although the δ13C of malathion indicated same value. In addition, we measured five pesticides (acephate, acetamiprid, glufosinate, glyphosate, and oxamyl) using

  5. Appraisal of groundwater resources of Ziarat valley using isotopic techniques

    International Nuclear Information System (INIS)

    Ahmad, M.; Akram, W.; Tasneem, M.A.; Rafique, M.

    2009-07-01

    Study of water resources of Ziarat Valley was carried out to investigate groundwater recharge mechanism and effectiveness of delay action dams. Samples of precipitation (rain, snow), dam reservoirs and groundwater (dug wells, tube wells, karezes, springs) were periodically collected from different locations and analyzed for environmental isotopes (/sup 2/H, /sup 3/H, /sup 18/O, /sup 34/S). The data indicate that rainfall and snow samples show wide ranges of delta /sup 2/H and delta /sup 18/O. However, the mean values for these isotopes are -6.4% and -37% respectively. Mean tritium value of rain is 9TU. Delta /sup 2/H and delta /sup 18/O values of dam reservoirs range from -6.7 to +4.9% and -42 to +30% respectively. Average isotopic indices of all the karezes are close to each other. Mean delta /sup 18/O and delta /sup 2/H values of Sandaman Tangi, Faran Tangi and Quaid springs vary from -6.3 to -6% and -40 to -31%. Tritium concentration of Sandaman Tangi and Faran Tangi springs (7 TU) is less than Quaid spring (11TU). Ranges of mean delta /sup 18/O and delta /sup 2/H values of all the groundwater samples (wells, karezes, springs) are -6.6 to -2.2% and -40 to -16% respectively. Delta /sup 34/S values of dissolved sulphates in groundwater vary from -8.5 to -0.8%. In /sup 18/O vs. /sup 2/H plot, most of the groundwater samples lie close to LMWL indicating the meteoric origin. Reservoir water in Pechi Dam shows highly enriched isotopic values in summer due to evaporation. Such enriched values are not depicted by the groundwater in the wells and karezes downstream of the dam. This implies that there is no significant recharge from this dam. Similar is the case of Mana Dam. Vouch Ghouski Dam has some contribution towards groundwater recharge while Warchoom Dam is much effective and makes significant contribution. Results of tritium dating suggest that residence time of groundwater is quite short (only few years). (author)

  6. Isotopic labeling as a tool to establish intramolecular vibrational coupling: The reaction of 2-propanol on Mo(110)

    International Nuclear Information System (INIS)

    Uvdal, P.; Wiegand, B.C.; Serafin, J.G.; Friend, C.M.

    1992-01-01

    The reactions of 2-propanol on Mo(110) were investigated using temperature programmed reaction, high resolution electron energy loss, and x-ray photoelectron spectroscopies. 2-Propanol forms 2-propoxide upon adsorption at 120 K on Mo(110). The 2-propoxide intermediate deoxygenates via selective γ C--H bond scission to eliminate propene as well as C--O bond hydrogenolysis to form trace amounts of propane. The C--O bond of 2-propoxide is estimated to be nearly perpendicular to the surface. Selective isotopic labeling was used to establish the coupling between the C--O stretch and modes associated with the hydrocarbon framework. The degree of coupling was strongly affected by bonding to the surface, primarily due to weakening of the C--O bond when 2-propoxide is bound to Mo(110). Selective isotopic labeling was, therefore, essential in making vibrational assignments and in identifying key reaction steps. Only a small kinetic isotope effect was observed during reaction of (CD 3 )(CH 3 )CHOH, consistent with a substantial component of C--O bond breaking in the transition state for propene elimination. Coupling of the C--O stretch to motion of the methyl group is also suggested to be important in the transition state for propene elimination

  7. Stable Isotope Technique to Assess Intake of Human Milk in Breastfed Infants

    International Nuclear Information System (INIS)

    2014-01-01

    This publication was developed by an international group of experts as an integral part of the IAEA’s efforts to contribute to the transfer of technology and knowledge in nutrition. Its aim is to assist Member States in their efforts to combat malnutrition by facilitating the use of relevant nuclear techniques. The stable (non-radioactive) isotope technique has been developed to assess intake of human milk in breastfed infants. The practical application of the stable isotope technique, based on analysis of deuterium by Fourier transform infrared spectrometry (FTIR), is presented in this book

  8. Sample preparation techniques of biological material for isotope analysis

    International Nuclear Information System (INIS)

    Axmann, H.; Sebastianelli, A.; Arrillaga, J.L.

    1990-01-01

    Sample preparation is an essential step in all isotope-aided experiments but often it is not given enough attention. The methods of sample preparation are very important to obtain reliable and precise analytical data and for further interpretation of results. The size of a sample required for chemical analysis is usually very small (10mg-1500mg). On the other hand the amount of harvested plant material from plots in a field experiment is often bulky (several kilograms) and the entire sample is too large for processing. In addition, while approaching maturity many crops show not only differences in physical consistency but also a non-uniformity in 15 N content among plant parts, requiring a plant fractionation or separation into parts (vegetative and reproductive) e.g. shoots and spikes, in case of small grain cereals, shoots and pods in case of grain legumes and tops and roots or beets (including crown) in case of sugar beet, etc. In any case the ultimate goal of these procedures is to obtain representative subsample harvested from greenhouse or field experiments for chemical analysis. Before harvesting an isotopic-aided experiment the method of sampling has to be selected. It should be based on the type of information required in relation to the objectives of the research and the availability of resources (staff, sample preparation equipment, analytical facilities, chemicals and supplies, etc.). 10 refs, 3 figs, 3 tabs

  9. Chemical reaction on solid surface observed through isotope tracer technique

    International Nuclear Information System (INIS)

    Tanaka, Ken-ichi

    1983-01-01

    In order to know the role of atoms and ions on solid surfaces as the partners participating in elementary processes, the literatures related to the isomerization and hydrogen exchanging reaction of olefines, the hydrogenation of olefines, the metathesis reaction and homologation of olefines based on solid catalysts were reviewed. Various olefines, of which the hydrogen atoms were substituted with deuterium at desired positions, were reacted using various solid catalysts such as ZnO, K 2 CO 3 on C, MoS 2 (single crystal and powder) and molybdenum oxide (with various carriers), and the infra-red spectra of adsorbed olefines on catalysts, the isotope composition of reaction products and the production rate of the reaction products were measured. From the results, the bonding mode of reactant with the atoms and ions on solid surfaces, and the mechanism of the elementary process were considered. The author emphasized that the mechanism of the chemical reaction on solid surfaces and the role of active points or catalysts can be made clear to the considerable extent by combining isotopes suitably. (Yoshitake, I.)

  10. Synthesis of isotopically labelled angiotensin II receptor antagonist GR138950X

    International Nuclear Information System (INIS)

    Carr, R.M.; Cable, K.M.; Newman, J.J.; Sutherland, D.R.

    1996-01-01

    Syntheses of [ 13 C] and [ 14 C]-labelled versions of angiotensin II receptor antagonist GR138950X, labelled in the imidazole carboxamide residue, are described. These involved preparation of an iodoimidazole substrate by a novel iododecarboxylation procedure, followed by cyanation with a mixture of carbon-labelled potassium cyanide and copper (l) iodide in DMF at high temperature. The preparation of a mass-labelled (M+5) version of GR138950X is also described. This involved the synthesis of an [ 13 C 3 , 15 N 2 ]-labelled imidazole from a 1,2,3-tricarbonyl compound, [ 13 C 3 ]propionaldehyde and [ 15 N]ammonia. The labelled imidazole was further elaborated into multiply-labelled GR138950X. (Author)

  11. Identifying the sources of produced water in the oil field by isotopic techniques

    International Nuclear Information System (INIS)

    Nguyen Minh Quy; Hoang Long; Le Thi Thu Huong; Luong Van Huan; Vo Thi Tuong Hanh

    2014-01-01

    The objective of this study is to identify the sources of the formation water in the Southwest Su-Tu-Den (STD SW) basement reservoir. To achieve the objective, isotopic techniques along with geochemical analysis for chloride, bromide, strontium dissolved in the water were applied. The isotopic techniques used in this study were the determination of water stable isotopes signatures (δ 2 H and (δ 18 O) and of the 87 Sr/ 86 Sr ratio of strontium in rock cutting sample and that dissolved in the formation water. The obtained results showed that the stable isotopes compositions of water in the Lower Miocene was -3‰ and -23‰ for (δ 18 O and (δ 2 H, respectively indicating the primeval nature of seawater in the reservoir. Meanwhile, the isotopic composition of water in the basement was clustered in a range of alternated freshwater with (δ 18 O and (δ 2 H being -(3-4)‰ and -(54-60)‰, respectively). The strontium isotopes ratio for water in the Lower Miocene reservoir was lower compared to that for water in the basement confirming the different natures of the water in the two reservoirs. The obtained results are assured for the techniques applicability, and it is recommended that studies on identification of the flow-path of the formation water in the STD SW basement reservoir should be continued. (author)

  12. Groundwater salinity study in the Mekong Delta using isotope techniques

    International Nuclear Information System (INIS)

    Le Van Khoi, Nguyen Kien Chinh; Do Tien Hung

    2002-01-01

    Environmental isotopes D, 18 O and chemical composition were used for study of recharge and salinization of groundwater in the are located between Bassac and Mekong Rivers. The results showed that: (a) Pleistocene aquifers are recharged through flood plains and outcrops located at the same altitude. The sanility of groundwater in these aquifers is mostly due to dissolution of the aquifer material, (b) Pliocene and Miocene aquifers receive recharge through outcrops located at the higher altitude on the northeast extension of the Delta and Cambodia. The salinity of groundwater in the coastal region of the aquifer is attributable to sea water intrusion. There appears to be significant retention of sea water in the coastal sediment during intrusion. (Author)

  13. Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Applied to Quantitative Proteomics of Bacillus subtilis

    DEFF Research Database (Denmark)

    Soufi, Boumediene; Kumar, C.; Gnad, F.

    2010-01-01

    We applied stable isotope labeling by amino acids in cell culture (SILAC) to large-scale quantitative proteomics analyses of the model bacterium Bacillus subtilis in two physiological conditions: growth on succinate and growth under phosphate starvation. Using a B. subtilis strain auxotrophic...... of the most comprehensive quantitative proteomics studies in bacteria, covering more than 75% of the B. subtilis genes expressed in the log phase of growth. Furthermore, we detect and quantify dynamics of 35 Ser/Thr/Tyr phosphorylation sites under growth on succinate, and 10 phosphorylation sites under...

  14. A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae

    Czech Academy of Sciences Publication Activity Database

    Foldynová-Trantírková, Silvie; Matulová, J.; Dötsch, V.; Löhr, F.; Cirstea, I.; Alexandov, K.; Breitling, R.; Lukeš, Julius; Trantírek, Lukáš

    2009-01-01

    Roč. 26, č. 6 (2009), s. 755-761 ISSN 0739-1102 R&D Projects: GA ČR GP204/08/P585; GA AV ČR 1QS600220554; GA AV ČR KAN200100801; GA MŠk 2B06129 Institutional research plan: CEZ:AV0Z60220518 Keywords : NMR * isotope labeling * protein expression * Leishmania * low-level enrichment Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.124, year: 2009

  15. Determination of symbiotic nitrogen fixation by labelling the soil atmosphere with sup(15)N sub(2) at low isotope enrichment

    International Nuclear Information System (INIS)

    Trivelin, P.C.O.

    1982-01-01

    A direct method to determine the total symbiotic nitrogen fixation during the leguminous plants cycles has been, developed, by labelling the soil atmosphere with sup(15)N sub(2) at low isotope enrichment, of about 1 atom % excess. The soil explored by the root system of leguminous plants was confined by means of a chamber in the field and by sealed pots in greenhouse experiments in order to maintain the soil air labelled with sup(15)N sub(2). The average sup(15)N concentration in the soil atmosphere, necessary to calculate dinitrogen fixation, was obtained by integration of the exponential functions of isotope dilution. Those functions were obtained by periodic sampling and analysis of the N sub(2) in the soil atmosphere. The field experiment with labelled atmosphere was carried out from the 22 sup(nd) to the 31 sup(st) day of the bean crop cycle and 5.5 mg N/plant (24% of total plant N) was derived from fixation. In pot experiments, under greenhouse conditions, integrated determination of fixation was made in Phaseolus beans (from the 19 sup(th) to the 67 sup(th) day from planting) and in soybeans (from the 24 sup(th) to the 70 sup(th) day from planting). The soil atmosphere was labelled with sup(15)N sub(2) in both cases. Average fixation obtained for Phaseolus beans was 80 mg N/plant (65% of total plant N) and for soybeans 265 mg N/plant (71% of total plant N). Evaluation of the basic concept of the isotope dilution method to determine nitrogen fixation in pots experiments, as proposed by Fried and Middelboe (1977) has also been made in the present paper. Simultaneous determinations of fixation in soybeans, using the isotope dilution method of Fried and Middelboe, natural variation of the sup(15)N/ sup(14)N ratios, and total-N differences, indicated the same results for pot experiments, harvested at the end of the plant cycle. (author)

  16. Evaluation of a new component used for isotopic lymphography: colloidal rhenium sulfide sup(99m)Tc labelled

    International Nuclear Information System (INIS)

    Pecking, A.; Le Mercier, N.; Gobin, R.; Bardy, A.; Najean, Y.

    1978-01-01

    We have studied for lymphatic scintigraphy a new radiopharmaceutical, sup(99m)Tc-labelled rhenium sulfocolloid. This preliminary study includes 20 adults patients with lymphomas and lymphoedemas. The principal advantage of this drug is its absence of toxicity and local pain, so that a rapid sub-cutaneous injection without local anesthesia is made possible. Good results have been obtained, as well in morphological studies of para-aortic and mammary lymph nodes as for kinetic studies of lymphatic flow in lymphoedemas. No liver and spleen uptake of radio-isotope was observed after foot injection [fr

  17. Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility

    International Nuclear Information System (INIS)

    Kobayashi, Hiroshi; Swapna, G. V. T.; Wu, Kuen-Phon; Afinogenova, Yuliya; Conover, Kenith; Mao, Binchen; Montelione, Gaetano T.; Inouye, Masayori

    2012-01-01

    A common obstacle to NMR studies of proteins is sample preparation. In many cases, proteins targeted for NMR studies are poorly expressed and/or expressed in insoluble forms. Here, we describe a novel approach to overcome these problems. In the protein S tag-intein (PSTI) technology, two tandem 92-residue N-terminal domains of protein S (PrS 2 ) from Myxococcus xanthus is fused at the N-terminal end of a protein to enhance its expression and solubility. Using intein technology, the isotope-labeled PrS 2 -tag is replaced with non-isotope labeled PrS 2 -tag, silencing the NMR signals from PrS 2 -tag in isotope-filtered 1 H-detected NMR experiments. This method was applied to the E. coli ribosome binding factor A (RbfA), which aggregates and precipitates in the absence of a solubilization tag unless the C-terminal 25-residue segment is deleted (RbfAΔ25). Using the PrS 2 -tag, full-length well-behaved RbfA samples could be successfully prepared for NMR studies. PrS 2 (non-labeled)-tagged RbfA (isotope-labeled) was produced with the use of the intein approach. The well-resolved TROSY-HSQC spectrum of full-length PrS 2 -tagged RbfA superimposes with the TROSY-HSQC spectrum of RbfAΔ25, indicating that PrS 2 -tag does not affect the structure of the protein to which it is fused. Using a smaller PrS-tag, consisting of a single N-terminal domain of protein S, triple resonance experiments were performed, and most of the backbone 1 H, 15 N and 13 C resonance assignments for full-length E. coli RbfA were determined. Analysis of these chemical shift data with the Chemical Shift Index and heteronuclear 1 H– 15 N NOE measurements reveal the dynamic nature of the C-terminal segment of the full-length RbfA protein, which could not be inferred using the truncated RbfAΔ25 construct. CS-Rosetta calculations also demonstrate that the core structure of full-length RbfA is similar to that of the RbfAΔ25 construct.

  18. Simultaneous determination of the intravenous and oral pharmacokinetic parameters of D,L-verapamil using stable isotope-labelled verapamil.

    Science.gov (United States)

    Eichelbaum, M; Somogyi, A; von Unruh, G E; Dengler, H J

    1981-01-01

    Following i.v. administration, the plasma concentration-time curve of verapamil could best be described by either a mono- or biexponential equation. Total plasma clearance (1.26 1/min) approached liver blood flow (1.51/min), so it can be concluded that its clearance is liver blood flow-dependent. Although absorption was almost complete after oral administration, absolute bioavailability (20%) was low, due to extensive hepatic first-pass metabolism. The approach using stable isotope-labelled and unlabelled drug permits simultaneous administration by the intravascular and extravascular routes, thus allowing determination of absolute bioavailability in a single experiment.

  19. A solvent extraction technique for the isotopic measurement of dissolved copper in seawater

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, Claire M., E-mail: claire.thompson@anu.edu.au; Ellwood, Michael J., E-mail: michael.ellwood@anu.edu.au; Wille, Martin, E-mail: martin.wille@uni-tuebingen.de

    2013-05-02

    Graphical abstract: -- Highlights: •A new sample preparation method for seawater copper isotopic analysis (δ{sup 65}Cu). •Solvent-extraction was used to pre-concentrate metals from seawater samples. •Anion-exchange was used to purify copper from the metal-rich extract. •δ{sup 65}Cu was measured in the north Tasman Sea. •Seawater δ{sup 65}Cu may be linked to marine biological activity. -- Abstract: Stable copper (Cu) isotope geochemistry provides a new perspective for investigating and understanding Cu speciation and biogeochemical Cu cycling in seawater. In this work, sample preparation for isotopic analysis employed solvent-extraction with amino pyrollidine dithiocarbamate/diethyl dithiocarbamate (APDC/DDC), coupled with a nitric acid back-extraction, to concentrate Cu from seawater. This was followed by Cu-purification using anion-exchange. This straightforward technique is high yielding and fractionation free for Cu and allows precise measurement of the seawater Cu isotopic composition using multi-collector inductively coupled plasma mass-spectrometry. A deep-sea profile measured in the oligotrophic north Tasman Sea shows fractionation in the Cu isotopic signature in the photic zone but is relatively homogenised at depth. A minima in the Cu isotopic profile correlates with the chlorophyll a maximum at the site. These results indicate that a range of processes are likely to fractionate stable Cu isotopes in seawater.

  20. Techniques for combining isotopic images obtained at different energies

    International Nuclear Information System (INIS)

    Soussaline, F.; Di Paola, R.; Bazin, J.P.

    1976-01-01

    The technique described should be considered as a first step towards the classification of scintigraphic data where the energy is included. As in all such studies the interpretation of the resulting images is not necessarily at first evident, and certain experience needs to be established. This applies in particular to the images obtained with the higher factors. It is possible that the use of this technique may resolve, without requiring a priori information, the problem previously encountered using the other 'subtraction' type techniques [fr

  1. Phosphorus use efficiency by cotton measured through 32P isotope technique

    Science.gov (United States)

    Marcante, N. C.; Muraoka, T.; Camacho, M. A.; César, F. R. C. F.; Bruno, I. P.

    2012-04-01

    Deficiency of phosphorus (P) is the major limitation to agricultural production in the Brazilian Savannah (Cerrado), which is naturally poor in this nutrient. Most of the P applied by fertilizer in Cerrado soils are converted into low solubility forms and can not be easily absorbed by plants. This occurs for characteristics of adsorption, conditioned by the predominance of low pH and aluminum and iron oxides in the clay fraction. The development of genotypes and cultivars with greater capacity to grow up in soils with low P availability ('phosphorus efficiency') is interesting to improve the agriculture in these areas in a sustainable way. Cotton (Gossypium spp.) is the main product for the fibers used nationally and globally in the textile chain. This study aim was to evaluate the efficiency of absorption and utilization of P by cotton cultivars/genotypes grown in Cerrado soil by the isotopic dilution technique. The soil classified as Ultisols, was labeled with the radioisotope 32P.The experiment was conducted in a greenhouse in a completely randomized design factorial 2 x 17. Factors were considered two levels of P (insufficient = 20 mg kg-1 and sufficient = 120 mg kg-1) and 17 genetic materials of cotton recommended for Cerrado region. Phosphorus levels influenced significantly the shoots dry matter production, the P content and accumulation, the 32P specific activity, the L value and L value less seed cotton P by cultivars and genotypes. The hierarchical clustering analysis used to verify the similarities between the cultivars and genotypes of cotton, classified them into internally homogeneous groups and heterogeneous between different groups. Cultivars FMT 523, FM 910 and CNPA GO 2043 were the most responsive to phosphate fertilizer in sufficient level of P, while the genotype Barbadense 01 and cultivars FM 966LL, IPR Jataí, BRS Aroeira and BRS Buriti were most efficient absorbing P in soils with insufficient level.

  2. Development of high-performance chemical isotope labeling LC-MS for profiling the human fecal metabolome.

    Science.gov (United States)

    Xu, Wei; Chen, Deying; Wang, Nan; Zhang, Ting; Zhou, Ruokun; Huan, Tao; Lu, Yingfeng; Su, Xiaoling; Xie, Qing; Li, Liang; Li, Lanjuan

    2015-01-20

    Human fecal samples contain endogenous human metabolites, gut microbiota metabolites, and other compounds. Profiling the fecal metabolome can produce metabolic information that may be used not only for disease biomarker discovery, but also for providing an insight about the relationship of the gut microbiome and human health. In this work, we report a chemical isotope labeling liquid chromatography-mass spectrometry (LC-MS) method for comprehensive and quantitative analysis of the amine- and phenol-containing metabolites in fecal samples. Differential (13)C2/(12)C2-dansyl labeling of the amines and phenols was used to improve LC separation efficiency and MS detection sensitivity. Water, methanol, and acetonitrile were examined as an extraction solvent, and a sequential water-acetonitrile extraction method was found to be optimal. A step-gradient LC-UV setup and a fast LC-MS method were evaluated for measuring the total concentration of dansyl labeled metabolites that could be used for normalizing the sample amounts of individual samples for quantitative metabolomics. Knowing the total concentration was also useful for optimizing the sample injection amount into LC-MS to maximize the number of metabolites detectable while avoiding sample overloading. For the first time, dansylation isotope labeling LC-MS was performed in a simple time-of-flight mass spectrometer, instead of high-end equipment, demonstrating the feasibility of using a low-cost instrument for chemical isotope labeling metabolomics. The developed method was applied for profiling the amine/phenol submetabolome of fecal samples collected from three families. An average of 1785 peak pairs or putative metabolites were found from a 30 min LC-MS run. From 243 LC-MS runs of all the fecal samples, a total of 6200 peak pairs were detected. Among them, 67 could be positively identified based on the mass and retention time match to a dansyl standard library, while 581 and 3197 peak pairs could be putatively

  3. Enantio-specific C(sp3)-H activation catalyzed by ruthenium nanoparticles: application to isotopic labeling of molecules of biological interest

    International Nuclear Information System (INIS)

    Taglang, Celine

    2015-01-01

    Isotopic labeling with deuterium and tritium is extensively used in chemistry, biology and pharmaceutical research. Numerous methods of labeling by isotopic exchange allow high isotopic enrichments but generally require harsh conditions (high temperatures, acidity). As a consequence, a general, regioselective and smooth labeling method that might be applicable to a wide diversity of substrates remains to develop. In the first part of this thesis, we demonstrated that the use of ruthenium nanoparticles, synthesized by Pr. Bruno Chaudret's team (INSA Toulouse), allowed the mild (2 bar of deuterium gas at 55 C), effective and selective H/D exchange reaction of a large variety of nitrogen-containing compounds, such as pyridines, indoles and primary, secondary and tertiary alkyl amines. The usefulness and the efficiency of this novel methodology was demonstrated by the deuteration of eight nitrogen-containing molecules of biological interest without altering their chemical and stereochemical properties. However, the conservation of the original stereochemistry of an activated chiral C-H center remains a major issue. We studied the reactivity of RuNP(at)PVP on different categories of nitrogen-containing substrates (amines, aminoacids and peptides) in water or in organic solvents. Our results showed that C-H activation of chiral carbons C(sp3) took place efficiently, selectively and, in all cases, with total retention of configuration. The wide range of applications of this procedure was demonstrated by the labeling of three chiral amines, fourteen aminoacids, three aromatic amino esters and four peptides. Moreover, our collaboration with Pr. Romuald Poteau's team (INSA Toulouse) led to the identification of two mechanisms by ab initio simulation in agreement with our experimental results: the σ-bond metathesis mechanism and the oxidative addition mechanism. These two mechanisms imply two vicinal ruthenium atoms leading to the formation an original

  4. Uranium isotopes in ground water as a prospecting technique

    International Nuclear Information System (INIS)

    Cowart, J.B.; Osmond, J.K.

    1980-02-01

    The isotopic concentrations of dissolved uranium were determined for 300 ground water samples near eight known uranium accumulations to see if new approaches to prospecting could be developed. It is concluded that a plot of 234 U/ 238 U activity ratio (A.R.) versus uranium concentration (C) can be used to identify redox fronts, to locate uranium accumulations, and to determine whether such accumulations are being augmented or depleted by contemporary aquifer/ground water conditions. In aquifers exhibiting flow-through hydrologic systems, up-dip ground water samples are characterized by high uranium concentration values (> 1 to 4 ppB) and down-dip samples by low uranium concentration values (less than 1 ppB). The boundary between these two regimes can usually be identified as a redox front on the basis of regional water chemistry and known uranium accumulations. Close proximity to uranium accumulations is usually indicated either by very high uranium concentrations in the ground water or by a combination of high concentration and high activity ratio values. Ground waters down-dip from such accumulations often exhibit low uranium concentration values but retain their high A.R. values. This serves as a regional indicator of possible uranium accumulations where conditions favor the continued augmentation of the deposit by precipitation from ground water. Where the accumulation is being dispersed and depleted by the ground water system, low A.R. values are observed. Results from the Gulf Coast District of Texas and the Wyoming districts are presented

  5. Qualitative Metabolome Analysis of Human Cerebrospinal Fluid by 13C-/12C-Isotope Dansylation Labeling Combined with Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

    Science.gov (United States)

    Guo, Kevin; Bamforth, Fiona; Li, Liang

    2011-02-01

    Metabolome analysis of human cerebrospinal fluid (CSF) is challenging because of low abundance of metabolites present in a small volume of sample. We describe and apply a sensitive isotope labeling LC-MS technique for qualitative analysis of the CSF metabolome. After a CSF sample is divided into two aliquots, they are labeled by 13C-dansyl and 12C-dansyl chloride, respectively. The differentially labeled aliquots are then mixed and subjected to LC-MS using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR MS). Dansylation offers significant improvement in the performance of chromatography separation and detection sensitivity. Moreover, peaks detected in the mass spectra can be readily analyzed for ion pair recognition and database search based on accurate mass and/or retention time information. It is shown that about 14,000 features can be detected in a 25-min LC-FTICR MS run of a dansyl-labeled CSF sample, from which about 500 metabolites can be profiled. Results from four CSF samples are compared to gauge the detectability of metabolites by this method. About 261 metabolites are commonly detected in replicate runs of four samples. In total, 1132 unique metabolite ion pairs are detected and 347 pairs (31%) matched with at least one metabolite in the Human Metabolome Database. We also report a dansylation library of 220 standard compounds and, using this library, about 85 metabolites can be positively identified. Among them, 21 metabolites have never been reported to be associated with CSF. These results illustrate that the dansylation LC-FTICR MS method can be used to analyze the CSF metabolome in a more comprehensive manner.

  6. Qualitative metabolome analysis of human cerebrospinal fluid by 13C-/12C-isotope dansylation labeling combined with liquid chromatography Fourier transform ion cyclotron resonance mass spectrometry.

    Science.gov (United States)

    Guo, Kevin; Bamforth, Fiona; Li, Liang

    2011-02-01

    Metabolome analysis of human cerebrospinal fluid (CSF) is challenging because of low abundance of metabolites present in a small volume of sample. We describe and apply a sensitive isotope labeling LC-MS technique for qualitative analysis of the CSF metabolome. After a CSF sample is divided into two aliquots, they are labeled by (13)C-dansyl and (12)C-dansyl chloride, respectively. The differentially labeled aliquots are then mixed and subjected to LC-MS using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR MS). Dansylation offers significant improvement in the performance of chromatography separation and detection sensitivity. Moreover, peaks detected in the mass spectra can be readily analyzed for ion pair recognition and database search based on accurate mass and/or retention time information. It is shown that about 14,000 features can be detected in a 25-min LC-FTICR MS run of a dansyl-labeled CSF sample, from which about 500 metabolites can be profiled. Results from four CSF samples are compared to gauge the detectability of metabolites by this method. About 261 metabolites are commonly detected in replicate runs of four samples. In total, 1132 unique metabolite ion pairs are detected and 347 pairs (31%) matched with at least one metabolite in the Human Metabolome Database. We also report a dansylation library of 220 standard compounds and, using this library, about 85 metabolites can be positively identified. Among them, 21 metabolites have never been reported to be associated with CSF. These results illustrate that the dansylation LC-FTICR MS method can be used to analyze the CSF metabolome in a more comprehensive manner. © American Society for Mass Spectrometry, 2011

  7. A novel dual-isotope labelling method for distinguishing between soil sources of N2O

    NARCIS (Netherlands)

    Wrage, N.; Groenigen, van J.W.; Oenema, O.; Baggs, E.M.

    2005-01-01

    We present a novel O-18-N-15-enrichment method for the distinction between nitrous oxide (N2O) from nitrification, nitrifier denitrification and denitrification based on a method with single- and double-N-15-labelled ammonium nitrate. We added a new treatment with O-18-labelled water to quantify N2O

  8. Pharmacokinetics of lidocaine and bupivacaine and stable isotope labelled analogues : a study in healthy volunteers

    NARCIS (Netherlands)

    Burm, A.G.D.; de Boer, A G; van Kleef, J.W.; Vermeulen, N P; de Leede, L G; Spierdijk, J; Breimer, D D

    1988-01-01

    The pharmacokinetics of lidocaine and bupivacaine and tri-deuteromethyl-labelled lidocaine and bupivacaine were investigated in healthy volunteers. The deuterium-labelled and the unlabelled form of the drug to be investigated were simultaneously infused in 10 min. Plasma concentrations were

  9. Determination of trace quantities of uranium in rocks mass spectrometric isotope dilution technique

    International Nuclear Information System (INIS)

    Kakazu, Mauricio Hiromitu

    1980-01-01

    A detailed experimental investigation on the thermionic emission of uranium deposited on a single flat type rhenium filament has been carried out. The study was aimed at determining the influence of various forms of deposition on the emission sensitivity and thermal stability of U + , UO + and UO 2 + ions. Based on these investigations, a technique, involving an addition of a small quantity of colloidal suspension of graphite on top of the uranyl nitrate sample deposited, was chosen because of its higher, emission sensitivity for uranium metal ions. The experimental parameters of the technique were optimised and the technique was employed in the determination of trace quantities of uranium in rock samples using mass spectrometric isotope dilution method. For the mass spectrometric isotope dilution analysis National Bureau of Standards uranium isotopic standard NBS-U 970 was employed as a tracer, where as the mass discrimination effect in the uranium isotope analysis was corrected using the uranium isotopic standard NBS-U500. Uranium was determined in each of the seven granite samples from Wyoming, USA and two USGS standard rocks. The precision of the analysis was found to be ±1% . The uranium values obtained on the rock samples were compared with the analyses of other investigators. Influence of the sample splitting on the uranium analysis was discussed in the light of the analytical results obtained.(author)

  10. A technique for studying digestion by grasshoppers using a 103Ruthenium-labelled marker

    International Nuclear Information System (INIS)

    Gandar, M.V.

    1981-01-01

    103 Ru-labelled tris (I,10-phenanthroline) ruthenium (II) chloride has proved to be a convenient marker in investigating digestion in grasshoppers. Assimilation efficiencies and the retension time of the food in the gut were measured. It is surmised that the technique described in this paper will be applicable to studies of a variety of chewing herbivorous insects. (author)

  11. International conference on isotopic and nuclear analytical techniques for health and environment. Book of abstracts

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2003-07-01

    Decision makers and stakeholders are becoming increasingly dependent on reliable chemical measurements that serve as a basis for decisions related to health, consumer safety, commerce, environment protection and compliance to regulations. Several millions of analytical results are produced annually, thus consuming appreciable amounts of resources. Reliability of these data is of major concern if cost-benefit figures are applied. Nuclear and isotopic analytical techniques (NATs) have been supported by the IAEA as part of their mandate to foster the peaceful use of nuclear energy for many years. Nuclear analytical laboratories have been installed and upgraded through Technical Co-operation assistance in many Member State laboratories. These techniques, including INAA, XRF, PIXE, stable and radioisotopes, spectrometroscopy, etc. have been applied to a wide range of subjects with varying success. Nuclear analytical techniques, featuring some intrinsic quality control aspects, such as multi-nuclide analysis, frequently serve as 'reference methods' to cross-check critical results. As nuclear properties of elements are targeted, matrix problems seem to be negligible to a great extent. The International Conference on Isotopic and Nuclear Analytical Techniques for Health and Environment was held 10-13 June 2003 in Vienna, Austria. The main purpose of this Conference was to bring together scientists, technologists, representatives of industry and regulatory authorities to exchange information and review the status of current developments and applications of isotopic and nuclear analytical techniques, and to discuss future trends and developments. A further objective is to identify potential opportunities for developing countries for applying isotopic and nuclear analytical techniques in health and environmental studies, and to consider the promotion and transfer of such technology. International developments and trends in health care, nutrition, and environmental monitoring

  12. Classification-based quantitative analysis of stable isotope labeling by amino acids in cell culture (SILAC) data.

    Science.gov (United States)

    Kim, Seongho; Carruthers, Nicholas; Lee, Joohyoung; Chinni, Sreenivasa; Stemmer, Paul

    2016-12-01

    Stable isotope labeling by amino acids in cell culture (SILAC) is a practical and powerful approach for quantitative proteomic analysis. A key advantage of SILAC is the ability to simultaneously detect the isotopically labeled peptides in a single instrument run and so guarantee relative quantitation for a large number of peptides without introducing any variation caused by separate experiment. However, there are a few approaches available to assessing protein ratios and none of the existing algorithms pays considerable attention to the proteins having only one peptide hit. We introduce new quantitative approaches to dealing with SILAC protein-level summary using classification-based methodologies, such as Gaussian mixture models with EM algorithms and its Bayesian approach as well as K-means clustering. In addition, a new approach is developed using Gaussian mixture model and a stochastic, metaheuristic global optimization algorithm, particle swarm optimization (PSO), to avoid either a premature convergence or being stuck in a local optimum. Our simulation studies show that the newly developed PSO-based method performs the best among others in terms of F1 score and the proposed methods further demonstrate the ability of detecting potential markers through real SILAC experimental data. No matter how many peptide hits the protein has, the developed approach can be applicable, rescuing many proteins doomed to removal. Furthermore, no additional correction for multiple comparisons is necessary for the developed methods, enabling direct interpretation of the analysis outcomes. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  13. Addressing national priorities through nuclear technology: Application of stable isotope techniques in evaluating nutritional intervention programs

    International Nuclear Information System (INIS)

    Mwangi, C.; Ndemwa, P.

    2008-01-01

    The concept is a new concept in Kenya that need driven technology. A paradigm shift from the conventional methods of measuring breast milk intake by means of weighing infants before and after feeding. A validation tool against anthropometrical measures of body fat through body density and skin-fold measurements. A reliable, accurate and non-invasive tool for monitoring lean body mass changes in clinical assessments.Isotopes Techniques in Body composition assessment.Technique-based Parameters of efficacy and/or effect are: Isotope (deuterium) dose given orally to subject (about 30 grams),Saliva (or urine) samples collected after 3-4 hrs, Concentration of isotope in saliva is measured using Fourier Transformed Infra-red Spectrophotometer (FTIR), Concentration gives the Total Body Water (TBW) component in the body, TBW = 73% Fat Free Mass (FFM), Calculate FFM (kg) from equation and subtract from Total Body Weight (kg) to get value of Fat Mass (kg)

  14. An In vitro evaluation of the reliability of QR code denture labeling technique.

    Science.gov (United States)

    Poovannan, Sindhu; Jain, Ashish R; Krishnan, Cakku Jalliah Venkata; Chandran, Chitraa R

    2016-01-01

    Positive identification of the dead after accidents and disasters through labeled dentures plays a key role in forensic scenario. A number of denture labeling methods are available, and studies evaluating their reliability under drastic conditions are vital. This study was conducted to evaluate the reliability of QR (Quick Response) Code labeled at various depths in heat-cured acrylic blocks after acid treatment, heat treatment (burns), and fracture in forensics. It was an in vitro study. This study included 160 specimens of heat-cured acrylic blocks (1.8 cm × 1.8 cm) and these were divided into 4 groups (40 samples per group). QR Codes were incorporated in the samples using clear acrylic sheet and they were assessed for reliability under various depths, acid, heat, and fracture. Data were analyzed using Chi-square test, test of proportion. The QR Code inclusion technique was reliable under various depths of acrylic sheet, acid (sulfuric acid 99%, hydrochloric acid 40%) and heat (up to 370°C). Results were variable with fracture of QR Code labeled acrylic blocks. Within the limitations of the study, by analyzing the results, it was clearly indicated that the QR Code technique was reliable under various depths of acrylic sheet, acid, and heat (370°C). Effectiveness varied in fracture and depended on the level of distortion. This study thus suggests that QR Code is an effective and simpler denture labeling method.

  15. Liquid–liquid extraction combined with differential isotope dimethylaminophenacyl labeling for improved metabolomic profiling of organic acids

    International Nuclear Information System (INIS)

    Peng, Jun; Li, Liang

    2013-01-01

    Graphical abstract: -- Highlights: •An improved method for profiling the carboxylic acid sub-metabolome is reported. •Liquid–liquid extraction was used for separating the organic acids from the amines. • 12 C/ 13 C-p-dimethylaminophenacyl (DmPA) labeling of the organic acids was carried out on the extract. •Detection interference by amines and labeling efficiency reduction by water were reduced. •About 2500 12 C/ 13 C-peak pairs or putative metabolites could be detected from 20 μL of human urine. -- Abstract: A large fraction of the known human metabolome belong to organic acids. However, comprehensive profiling of the organic acid sub-metabolome is a major analytical challenge. In this work, we report an improved method for detecting organic acid metabolites. This method is based on the use of liquid–liquid extraction (LLE) to selectively extract the organic acids, followed by using differential isotope p-dimethylaminophenacyl (DmPA) labeling of the acid metabolites. The 12 C-/ 13 C-labeled samples are analyzed by liquid chromatography Fourier-transform ion cyclotron resonance mass spectrometry (LC–FTICR–MS). It is shown that this LLE DmPA labeling method offers superior performance over the method of direct DmPA labeling of biofluids such as human urine. LLE of organic acids reduces the interference of amine-containing metabolites that may also react with DmPA. It can also remove water in a biofluid that can reduce the labeling efficiency. Using human urine as an example, it is demonstrated that about 2500 peak pairs or putative metabolites could be detected in a 30-min gradient LC–MS run, which is about 3 times more than that detected in a sample prepared using direct DmPA labeling. About 95% of the 1000 or so matched metabolites to the Human Metabolome Database (HMDB) are organic acids. It is further shown that this method can be used to handle as small as 10 μL of urine. We believe that this method opens the possibility of generating a

  16. Liquid–liquid extraction combined with differential isotope dimethylaminophenacyl labeling for improved metabolomic profiling of organic acids

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Jun; Li, Liang, E-mail: Liang.Li@ualberta.ca

    2013-11-25

    Graphical abstract: -- Highlights: •An improved method for profiling the carboxylic acid sub-metabolome is reported. •Liquid–liquid extraction was used for separating the organic acids from the amines. •{sup 12}C/{sup 13}C-p-dimethylaminophenacyl (DmPA) labeling of the organic acids was carried out on the extract. •Detection interference by amines and labeling efficiency reduction by water were reduced. •About 2500 {sup 12}C/{sup 13}C-peak pairs or putative metabolites could be detected from 20 μL of human urine. -- Abstract: A large fraction of the known human metabolome belong to organic acids. However, comprehensive profiling of the organic acid sub-metabolome is a major analytical challenge. In this work, we report an improved method for detecting organic acid metabolites. This method is based on the use of liquid–liquid extraction (LLE) to selectively extract the organic acids, followed by using differential isotope p-dimethylaminophenacyl (DmPA) labeling of the acid metabolites. The {sup 12}C-/{sup 13}C-labeled samples are analyzed by liquid chromatography Fourier-transform ion cyclotron resonance mass spectrometry (LC–FTICR–MS). It is shown that this LLE DmPA labeling method offers superior performance over the method of direct DmPA labeling of biofluids such as human urine. LLE of organic acids reduces the interference of amine-containing metabolites that may also react with DmPA. It can also remove water in a biofluid that can reduce the labeling efficiency. Using human urine as an example, it is demonstrated that about 2500 peak pairs or putative metabolites could be detected in a 30-min gradient LC–MS run, which is about 3 times more than that detected in a sample prepared using direct DmPA labeling. About 95% of the 1000 or so matched metabolites to the Human Metabolome Database (HMDB) are organic acids. It is further shown that this method can be used to handle as small as 10 μL of urine. We believe that this method opens the

  17. A microfluidics-based technique for automated and rapid labeling of cells for flow cytometry

    International Nuclear Information System (INIS)

    Patibandla, Phani K; Estrada, Rosendo; Kannan, Manasaa; Sethu, Palaniappan

    2014-01-01

    Flow cytometry is a powerful technique capable of simultaneous multi-parametric analysis of heterogeneous cell populations for research and clinical applications. In recent years, the flow cytometer has been miniaturized and made portable for application in clinical- and resource-limited settings. The sample preparation procedure, i.e. labeling of cells with antibodies conjugated to fluorescent labels, is a time consuming (∼45 min) and labor-intensive procedure. Microfluidics provides enabling technologies to accomplish rapid and automated sample preparation. Using an integrated microfluidic device consisting of a labeling and washing module, we demonstrate a new protocol that can eliminate sample handling and accomplish sample and reagent metering, high-efficiency mixing, labeling and washing in rapid automated fashion. The labeling module consists of a long microfluidic channel with an integrated chaotic mixer. Samples and reagents are precisely metered into this device to accomplish rapid and high-efficiency mixing. The mixed sample and reagents are collected in a holding syringe and held for up to 8 min following which the mixture is introduced into an inertial washing module to obtain ‘analysis-ready’ samples. The washing module consists of a high aspect ratio channel capable of focusing cells to equilibrium positions close to the channel walls. By introducing the cells and labeling reagents in a narrow stream at the center of the channel flanked on both sides by a wash buffer, the elution of cells into the wash buffer away from the free unbound antibodies is accomplished. After initial calibration experiments to determine appropriate ‘holding time’ to allow antibody binding, both modules were used in conjunction to label MOLT-3 cells (T lymphoblast cell line) with three different antibodies simultaneously. Results confirm no significant difference in mean fluorescence intensity values for all three antibodies labels (p < 0.01) between the

  18. Limited proteolysis combined with isotope labeling and quantitative LC-MALDI MS for monitoring protein conformational changes: a study on calcium-binding sites of cardiac Troponin C

    International Nuclear Information System (INIS)

    McDonald, Chris; Li Liang

    2005-01-01

    Studies of protein-protein and protein-ligand interactions are important for understanding biological functions of proteins. A new technique based on the partial proteolysis of proteins combined with quantitative mass spectrometry is developed as a means of tracking structural changes after the formation of a protein-ligand complex. In this technique, a protein of interest with and without the binding of a ligand is digested with an enzyme to generate a set of peptides, followed by separation of the peptides by liquid chromatography. Matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) is used to identify chromatographically separated peptides, and locate their sequence alignments in the parent protein. Using an isotopically labeled protein as a sample against an unlabeled protein standard, quantitative information can be gathered. This overcomes the inherent lack of quantitative capability of MALDI MS. The utility of the technique to investigate protein-ligand interactions is demonstrated in a model system involving calcium binding to cardiac Troponin C (cTnC). Using this technique, the general location of the three calcium-binding sites of cTnC can be determined by using several different enzymes to generate overlapping peptide maps of cTnC

  19. From position-specific isotope labeling towards soil fluxomics: a novel toolbox to assess the microbial impact on biogeochemical cycles

    Science.gov (United States)

    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

  20. Studies on osteoporosis in Chile using isotope-related techniques

    International Nuclear Information System (INIS)

    Lobo, G.; Palma, T.; Cortes-Toro, E.

    1996-01-01

    Several studies on bone densitometry measurements in healthy individuals have been performed in Chile. However due to the fact that different techniques and no uniform protocols have been used to select patients, the results obtained are not suitable as reference values for a normal chilean population. Therefore, foreign reference values are used. This study will select healthy normal individuals, typical urban chilean residents, and measure bone density using the DEXA technique. The selection will be made according to a well defined protocol. Serum osteocalcin, a marker of bone remodeling, will be measured in all subjects as a means-a assessing bone metabolism. Bone trace element composition will be measured in selected subjects. Samples will be obtained by biopsy or through normal surgical procedures and will be analyzed by neutron activation analysis. (author)

  1. Direct infusion electrospray ionization–ion mobility–mass spectrometry for comparative profiling of fatty acids based on stable isotope labeling

    Energy Technology Data Exchange (ETDEWEB)

    Leng, Jiapeng, E-mail: jpleng@126.com [Department of Chemical Engineering, North University of China, Taiyuan 030051 (China); Guan, Qing [Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center (FUSCC), Shanghai 200032 (China); Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032 (China); Sun, Tuanqi, E-mail: tuanqisun@163.com [Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center (FUSCC), Shanghai 200032 (China); Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032 (China); Wang, Haoyang [National Center for Organic Mass Spectrometry in Shanghai, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032 (China); Cui, Jianlan; Liu, Qinghao [Department of Chemical Engineering, North University of China, Taiyuan 030051 (China); Zhang, Zhixu; Zhang, Manyu [Agilent Technologies China Co., Ltd, Shanghai 200080 (China); Guo, Yinlong, E-mail: ylguo@sioc.ac.cn [National Center for Organic Mass Spectrometry in Shanghai, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032 (China)

    2015-08-05

    A rapid method for fatty acids (FAs) comparative profiling based on carboxyl-specific stable isotope labeling (SIL) and direct infusion electrospray ionization–ion mobility–mass spectrometry (ESI–IM–MS) is established. The design of the method takes advantage of the three-dimensional characteristics of IM–MS including drift time, m/z and ion intensity, for comparison of d0-/d6-2,4-dimethoxy-6-piperazin-1-yl pyrimidine (DMPP)-labeled FAs. In particular, without chromatographic separation, the method allowed direct FAs profiling in complex samples due to the advantageous priority of DMPP in signal enhancement as well as the extra resolution that IM–MS offered. Additionally, the d0-/d6-DMPP-labeled FAs showed expected features, including very similar drift times, 6 Da mass deviations, specific reporter ions, similar MS responses, and adherence to the drift time rule regarding the influence of carbon chain length and unsaturation on relative drift times. Therefore, the introduction of isotope analogs minimized the matrix effect and variations in quantification and ensured accurate identification of non-targeted FAs by those typical features. Peak intensity ratios between d0-/d6-DMPP-labeled ions were subsequently used in relative quantification for the detected FAs. The established strategy has been applied successfully in the rapid profiling of trace free FAs between normal and cancerous human thyroid tissues. Sixteen free FAs were found with the increased level with a statistically significant difference (p < 0.05) compared to the normal tissue samples. The integrated SIL technique and ESI–IM–MS are expected to serve as an alternative tool for high-throughput analysis of FAs in complex samples. - Highlights: • A novel method based on IM–MS and SIL was developed for FAs comparative profiling. • Without LC separation, the method allowed direct infusion profiling of FAs in complex samples. • Both of the efficiency and accuracy for FAs analyses

  2. Managing water resources in Malaysia: the use of isotope technique and its potential

    International Nuclear Information System (INIS)

    Keizrul Abdullah

    2006-01-01

    This keynote address discusses the following subjects; state of Malaysia water resources, water related problem i.e floods, water shortage (droughts), water quality, river sedimentation, water resources management and the ongoing and potential application of isotope techniques in river management

  3. A two emulsion autoradiographic technique and the discriminating of the three different types of labelling after double labelling with 3H- and 14C-thymidine

    International Nuclear Information System (INIS)

    Schultze, B.; Maurer, W.; Hagenbusch, H.

    1976-01-01

    The first part of the paper deals with a two emulsion autoradiographic technique for double labelling experiments with 3 H- and 14 C-thymidine which permits a clear discrimination of the different types of labelling. In the second part the application of this technique to cell kinetics studies is discussed. Accurate discrimination between the different types of labelling, namely purely 3 H-, purely 14 C- and double ( 3 H + 14 C) labelling, is only possible if the activity ratio of 3 H- to 14 C-thymidine is sufficiently high. This condition is necessary for a reliable distinction between those grains in the first emulsion which are due to true 3 H-labelling and spurious grains which are simultaneously produced in the same emulsion by 14 C-β- particles. Experiments are described to determine the required activity ratio of 3 H- to 14 C-thymidine. (author)

  4. Comparative international studies of osteoporosis using isotope techniques

    International Nuclear Information System (INIS)

    Krishnan, S.; Sturtridge, W.C.

    1996-01-01

    In-Vivo Neutron Activation Analysis (IVNAA) and Dual Energy X-ray Absorptiometry (DEXA) are routinely used in our laboratory for clinical and research investigation of Bone Mineral Content (BMC) and Bone Mineral Density (BMD in g/sq.cm). These two techniques differ in many respects. IVNAA is an established technique where 49 Ca radioisotope induced by the 48 Ca(n,gamma) 49 Ca reaction is measured to determine BMC. In DEXA, two X-rays of different energies are used and based on the known attenuation characteristics of bone and soft tissue and the measured attenuated signals, the BMC is calculated. Very little radiation dose (<0.1 mSv) is given in DEXA compared to IVNAA (3 to 5 mSv). Site specific information can be obtained in the former. Although DEXA is easier to use both the techniques are in use in the investigation of Osteopenia. The BMD, a clinical useful parameter, is a function of age and not height. The body calcium content determined by IVNAA is a function of height and not age. These differences are not yet clearly understood. Work is underway to determine the influence of several factors e.g. age, weight on BMD. We are also conducting experiments to determine the peak BMC and BMD of Canadians. The role of various life styles e.g. exercise, diet, ethnicity are being investigated as a determinant of BMD and BMC. Also the effects and safety of many drugs in the treatment of osteopenia and osteoporosis are also being studied. (author)

  5. Isotope techniques in non-destructive testing of dynamic systems

    International Nuclear Information System (INIS)

    Singh, Gursharan; Pant, H.J.

    1996-01-01

    A few applications of gamma scanning and radiotracer techniques for Non-destructive Testing (NDT) of dynamic systems in chemical and petrochemical industries are briefly discussed in this paper. Examples of gamma scanning inspections carried out for troubleshooting of various types of columns such as vacuum, extraction, separator and rectifier, with trays and packed beds and having diameters from 1 meter to 8.4 meters are given. Radiotracer applications for Residence Time Distribution (RTD) studies on different systems like an aniline production reactor in a chemical industry and a laboratory scale solid-liquid fluidized bed column are mentioned. (author)

  6. Studies of osteoporosis in Shanghai, China, using isotope related techniques

    International Nuclear Information System (INIS)

    Zhang Yongpeng

    2000-01-01

    This paper deals with the osteoporosis model of New Zealand rabbits in order to study the functions of inorganic elements for women patients with osteoporosis. The thirty cases of iliac crest are collected and determined using Neutron Activation Analysis (NAA) technique. The results show that the Fluorine concentrations in experimental group are higher than those in control group and the Magnesium concentrations are lower significantly (P<0.001), but the concentrations of K, Na, Ca, Ba, Mn, Sr are not remarkable difference. Combined with the serum biochemical markers of bone formation and resorption, the physiological functions of some metal elements in bone are also discussed. (author)

  7. Quantitative twoplex glycan analysis using 12C6 and 13C6 stable isotope 2-aminobenzoic acid labelling and capillary electrophoresis mass spectrometry.

    Science.gov (United States)

    Váradi, Csaba; Mittermayr, Stefan; Millán-Martín, Silvia; Bones, Jonathan

    2016-12-01

    Capillary electrophoresis (CE) offers excellent efficiency and orthogonality to liquid chromatographic (LC) separations for oligosaccharide structural analysis. Combination of CE with high resolution mass spectrometry (MS) for glycan analysis remains a challenging task due to the MS incompatibility of background electrolyte buffers and additives commonly used in offline CE separations. Here, a novel method is presented for the analysis of 2-aminobenzoic acid (2-AA) labelled glycans by capillary electrophoresis coupled to mass spectrometry (CE-MS). To ensure maximum resolution and excellent precision without the requirement for excessive analysis times, CE separation conditions including the concentration and pH of the background electrolyte, the effect of applied pressure on the capillary inlet and the capillary length were evaluated. Using readily available 12/13 C 6 stable isotopologues of 2-AA, the developed method can be applied for quantitative glycan profiling in a twoplex manner based on the generation of extracted ion electropherograms (EIE) for 12 C 6 'light' and 13 C 6 'heavy' 2-AA labelled glycan isotope clusters. The twoplex quantitative CE-MS glycan analysis platform is ideally suited for comparability assessment of biopharmaceuticals, such as monoclonal antibodies, for differential glycomic analysis of clinical material for potential biomarker discovery or for quantitative microheterogeneity analysis of different glycosylation sites within a glycoprotein. Additionally, due to the low injection volume requirements of CE, subsequent LC-MS analysis of the same sample can be performed facilitating the use of orthogonal separation techniques for structural elucidation or verification of quantitative performance.

  8. Combining position-specific 13C labeling with compound-specific isotope analysis: first steps towards soil fluxomics

    Science.gov (United States)

    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 knowledge on the biochemical pathways and its controlling factors. Tracing C fate in soil by isotopes became on of the most applied and promising biogeochemistry tools. Up to now, studies on LMWOS were nearly exclusively based on uniformly labeled organic substances i.e. all C atoms in the molecules were labeled with 13C or 14C. However, this classical approach did not allow the differentiation between use of intact initial substances in any process, or whether they were transformed 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 LMWOS and quantification of 13CO2 and 13C in bulk soil enabled following the basic metabolic pathways of soil microorganisms. However, only the combination of position-specific 13C labeling with compound-specific isotope analysis of microbial biomarkers and metabolites allowed 1) tracing specific anabolic pathways in diverse microbial communities in soils and 2) identification of specific pathways of individual functional microbial groups. So, these are the prerequisites for soil fluxomics. Our studies combining position-specific labeled glucose with amino

  9. Lifetimes of organic photovoltaics: Using TOF-SIMS and 18O2 isotopic labelling to characterise chemical degradation mechanisms

    DEFF Research Database (Denmark)

    Norrman, K.; Krebs, Frederik C

    2006-01-01

    The lifetimes of organic photovoltaic cells based on conjugated polymer materials were studied. The device geometry was glass:ITO:PEDOT:PSS:C-12-PSV:C-60:aluminium. To characterise and elucidate the parts of the degradation mechanisms induced by molecular oxygen, 1802 isotopic labelling was emplo......The lifetimes of organic photovoltaic cells based on conjugated polymer materials were studied. The device geometry was glass:ITO:PEDOT:PSS:C-12-PSV:C-60:aluminium. To characterise and elucidate the parts of the degradation mechanisms induced by molecular oxygen, 1802 isotopic labelling...

  10. Direct methods and residue type specific isotope labeling in NMR structure determination and model-driven sequential assignment

    International Nuclear Information System (INIS)

    Schedlbauer, Andreas; Auer, Renate; Ledolter, Karin; Tollinger, Martin; Kloiber, Karin; Lichtenecker, Roman; Ruedisser, Simon; Hommel, Ulrich; Schmid, Walther; Konrat, Robert; Kontaxis, Georg

    2008-01-01

    Direct methods in NMR based structure determination start from an unassigned ensemble of unconnected gaseous hydrogen atoms. Under favorable conditions they can produce low resolution structures of proteins. Usually a prohibitively large number of NOEs is required, to solve a protein structure ab-initio, but even with a much smaller set of distance restraints low resolution models can be obtained which resemble a protein fold. One problem is that at such low resolution and in the absence of a force field it is impossible to distinguish the correct protein fold from its mirror image. In a hybrid approach these ambiguous models have the potential to aid in the process of sequential backbone chemical shift assignment when 13 C β and 13 C' shifts are not available for sensitivity reasons. Regardless of the overall fold they enhance the information content of the NOE spectra. These, combined with residue specific labeling and minimal triple-resonance data using 13 C α connectivity can provide almost complete sequential assignment. Strategies for residue type specific labeling with customized isotope labeling patterns are of great advantage in this context. Furthermore, this approach is to some extent error-tolerant with respect to data incompleteness, limited precision of the peak picking, and structural errors caused by misassignment of NOEs

  11. Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose.

    Directory of Open Access Journals (Sweden)

    Darren J Creek

    2015-03-01

    Full Text Available Metabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of Trypanosoma brucei, a parasite responsible for human African trypanosomiasis. Glucose enters many branches of metabolism beyond glycolysis, which has been widely held to be the sole route of glucose metabolism. Whilst pyruvate is the major end-product of glucose catabolism, its transamination product, alanine, is also produced in significant quantities. The oxidative branch of the pentose phosphate pathway is operative, although the non-oxidative branch is not. Ribose 5-phosphate generated through this pathway distributes widely into nucleotide synthesis and other branches of metabolism. Acetate, derived from glucose, is found associated with a range of acetylated amino acids and, to a lesser extent, fatty acids; while labelled glycerol is found in many glycerophospholipids. Glucose also enters inositol and several sugar nucleotides that serve as precursors to macromolecule biosynthesis. Although a Krebs cycle is not operative, malate, fumarate and succinate, primarily labelled in three carbons, were present, indicating an origin from phosphoenolpyruvate via oxaloacetate. Interestingly, the enzyme responsible for conversion of phosphoenolpyruvate to oxaloacetate, phosphoenolpyruvate carboxykinase, was shown to be essential to the bloodstream form trypanosomes, as demonstrated by the lethal phenotype induced by RNAi-mediated downregulation of its expression. In addition, glucose derivatives enter pyrimidine biosynthesis via oxaloacetate as a precursor to aspartate and orotate.

  12. Studies of osteoporosis in Croatia using isotope related techniques

    International Nuclear Information System (INIS)

    Dekanic, D.; Blanusa, M.; Cvijetic, S.; Prester, L.; Uzelac, V.

    1996-01-01

    A short overview is presented of research activities concerning osteoporosis and trace element exposure monitoring carried out by the Institute for Medical Research and Occupational Health in Zagreb during the last forty years. The overview includes: cross-sectional and Prospective epidemiological studies in districts with habitually low and high calcium intake; clinical studies concerning peak bone mass and bone mineral density in Postmenopausal women; studies on trace element exposure monitoring. The collaborating institutions, agencies and relevant recent publications are quoted. A survey is given of relevant methods which were used in earlier studies as well as the techniques which will be used in the current IAEA project. A brief description of the work which will be undertaken (particularly during the first year) within the framework of the IAEA CRP is given. (author)

  13. A Breast Cell Atlas: Organelle analysis of the MDA-MB-231 cell line by density-gradient fractionation using isotopic marking and label-free analysis

    Directory of Open Access Journals (Sweden)

    Marianne Sandin

    2015-09-01

    Full Text Available Protein translocation between organelles in the cell is an important process that regulates many cellular functions. However, organelles can rarely be isolated to purity so several methods have been developed to analyse the fractions obtained by density gradient centrifugation. We present an analysis of the distribution of proteins amongst organelles in the human breast cell line, MDA-MB-231 using two approaches: an isotopic labelling and a label-free approach.

  14. Isotope techniques in groundwater hydrology 1974, Vol. I. Proceedings of a symposium

    International Nuclear Information System (INIS)

    1974-01-01

    This symposium, held in Vienna on 11-15 March, 1974, was the fourth on the subject of isotope hydrology organized by the International Atomic Energy Agency. However, this one was limited to groundwater hydrology in view of the general increase of interest and activity in isotope hydrology since the previous meeting in 1970. The proceedings of this symposium are a good indicator of the present world status of these techniques. Thus one notes that many of the studies are in the developing areas of the world. Furthermore, there has been a shift to using these techniques as an additional applied tool in specific problems of development of water resources. Examples of such applications also give evidence of the closer collaboration between isotope specialists, who originally developed the methods, and hydrogeologists and geochemists. It is hoped that these proceedings will contribute to a wider appreciation of the potential use of isotope techniques to hydrological problems associated with the development of groundwater for agriculture, community water supply and industry

  15. Isotope techniques in human studies of zinc nutrition. Report of an IAEA consultants' meeting

    International Nuclear Information System (INIS)

    1997-01-01

    A Consultants' Meeting was convened by the IAEA in November 1996 to review current knowledge of the role of zinc in human nutrition - with particular reference to (1) relevant public health issues in developing countries, and (2) the role that isotope techniques can play in increasing our understanding of some of these matters. More specifically, the Consultants Group was invited to advise the Agency on its future programmes in this area. It was concluded that there is now abundant scientific evidence that zinc deficiency of nutritional origin is a widespread phenomenon with important health consequences, particularly in developing countries. Even mild zinc deficiency is now thought to contribute to pregnancy complications, low birth weight, impaired immune competence, maternal and infant mortality and morbidity, and growth failure in infancy and childhood. Hence zinc deficiency may have far-reaching consequences for maternal, infant and child health in many developing countries. Isotope techniques - mainly using stable isotopes of zinc, but in some cases using radioisotopes - provide many possibilities 'for improving our understanding of the basic biochemical and metabolic functions of zinc, and for increasing our knowledge of the roles of this essential micronutrient in human nutrition. Through the use of isotope techniques, we may also expect to be better able to assess, and to increase, the efficacy of dietary and other intervention programmes designed to improve the nutritional status of developing country populations whose intakes are below the levels needed to achieve optimal health. This report briefly reviews some of these isotope techniques and makes recommendations on their use in IAEA research and technical co-operation programmes

  16. Isotope techniques in human studies of zinc nutrition. Report of an IAEA consultants` meeting

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-12-31

    A Consultants` Meeting was convened by the IAEA in November 1996 to review current knowledge of the role of zinc in human nutrition - with particular reference to (1) relevant public health issues in developing countries, and (2) the role that isotope techniques can play in increasing our understanding of some of these matters. More specifically, the Consultants Group was invited to advise the Agency on its future programmes in this area. It was concluded that there is now abundant scientific evidence that zinc deficiency of nutritional origin is a widespread phenomenon with important health consequences, particularly in developing countries. Even mild zinc deficiency is now thought to contribute to pregnancy complications, low birth weight, impaired immune competence, maternal and infant mortality and morbidity, and growth failure in infancy and childhood. Hence zinc deficiency may have far-reaching consequences for maternal, infant and child health in many developing countries. Isotope techniques - mainly using stable isotopes of zinc, but in some cases using radioisotopes - provide many possibilities `for improving our understanding of the basic biochemical and metabolic functions of zinc, and for increasing our knowledge of the roles of this essential micronutrient in human nutrition. Through the use of isotope techniques, we may also expect to be better able to assess, and to increase, the efficacy of dietary and other intervention programmes designed to improve the nutritional status of developing country populations whose intakes are below the levels needed to achieve optimal health. This report briefly reviews some of these isotope techniques and makes recommendations on their use in IAEA research and technical co-operation programmes

  17. Use of the HRICP-MS technique for the evaluation of boron isotopes in Eucalitpus plants

    International Nuclear Information System (INIS)

    Mattiello, Edson Marcio; Ruiz, Hugo Alberto; Silva, Ivo Ribeiro da

    2011-01-01

    The 10 B isotope tracer technique is essential to study the B mobility in plants. Factors that can influence the quality of measured B isotope ratios were optimized experimentally using High Resolution Inductively Coupled Plasma Mass Spectrometry (HRICP-MS). An isotopically certified standard (NIST SRM-951) was used. The best combination was obtained using a resolution of 400, a RF power of 1250 W, followed by 15 measurements over a 10-s integration period each (15*10). Utilizing this approach it was possible to obtain a precision of 0.3 % in standard material and 2z % in the experimental samples. The results show the importance of establishing optimized work conditions before carrying out the analytical series. (author)

  18. Assessment of aquifer system using isotope techniques in urban centres Raipur, Calcutta and Jodhpur, India

    International Nuclear Information System (INIS)

    Sinha, U.K.; Kulkarni, K.M.; Sharma, S.; Ray, A.; Bodhankar, N.

    2002-01-01

    Three urban centres Raipur, Calcutta and Jodhpur were studied using isotope techniques ( 18 O, 2 H, 3 H, 13 C, and 14 C) and chemistry with different objectives. Groundwater in Raipur city is susceptible to contamination near waste disposal sites, landfills and dairy farms. Shallow groundwater is more affected by contamination than deeper zone groundwater. A few shallow zone groundwater samples in Jadavpur area of Calcutta city show arsenic concentration above permissible level. Stable isotope values of these groundwater samples indicate that they are depleted and tritium results show that they have less residence time. Deep groundwater is arsenic free and old. Seepage in the basement and rise of static water level of some parts of Jodhpur city has been observed from March 1998 onwards. Isotopic, hydrogeological and chemical analyses data has indicated that lake water is contributing to seepage water in the basement. (author)

  19. Groundwater problems studies in the Thar desert, India, using isotope techniques

    International Nuclear Information System (INIS)

    Navada, S.V.

    1999-01-01

    In groundwater management, particularly in arid regions like western Rajasthan, it is important to know the presence of modern recharge and to estimate the recharge rate to avoid over-exploitation of the groundwater resource. Isotopes can help to identify modern recharge and to estimate recharge rate to the aquifer. If modern recharge is absent, groundwater dating using radiocarbon could help to identify old groundwater or paleowaters. A number of isotope studies carried out in arid zones (particularly in the Sahara) have shown that the deep groundwater is generally very old. From these studies it was concluded that episodic large scale recharge corresponding to humid phases or pluvials occurred in these arid areas. The paper reviews our experiences on the application of isotope techniques in understanding groundwater recharge process in and western Rajasthan

  20. Using 15N isotope technique on evaluation of starter N effect on soybean-bacteria symbiosis intensity

    International Nuclear Information System (INIS)

    Sagheb, N.; Majd, F.; Porrdavai, H.; Yousefy, F.

    1994-02-01

    Nuclear techniques have solved many agricultural research problems during the last two decades, while resolving these problems with classical methods have been very difficult if not impossible. With this technique evaluation of unknown effects of biological nitrogen fixation became possible. The advantage of using 1 5N technique is its ability to e specify and identify exact amount of nitrogen from soil, fertilizer and that existing in the atmosphere as molecular nitrogen. Soybean crop (Elycine max(L.) Merill) is able to receive large portion of its nitrogen through biological nitrogen fixation from symbiosis of Rhizobium-Japonicum bacterium. In this study three soybean cultivars received 20 and 100 Kg of N ha -1 as starter nitrogen fertilizer and in form of 1 5N -labelled ammonium phosphate. In collected plant materials 1 5N /1 4N ratios were determined by the Kjeldahl procedure. By using 1 5N isotope method the amount of nitrogen derived from different sources (Soil, Fertilizer and atmosphere) at 20 and 100 Kg N ha -1 were evaluated. Significant differences were observed in percent and amounts of nitrogen fixed (Kg ha 1 ) from atmosphere. There were no significant differences in total N yield and total dry matter at two different levels of nitrogen fertilizer applied. (author)

  1. Differential isotope dansylation labeling combined with liquid chromatography mass spectrometry for quantification of intact and N-terminal truncated proteins

    International Nuclear Information System (INIS)

    Tang, Yanan; Li, Liang

    2013-01-01

    Graphical abstract: -- Highlights: •LC–MS was developed for quantifying protein mixtures containing both intact and N-terminal truncated proteins. • 12 C 2 -Dansylation of the N-terminal amino acid of proteins was done first, followed by microwave-assisted acid hydrolysis. •The released 12 C 2 -dansyl labeled N-terminal amino acid was quantified using 13 C 2 -dansyl labeled amino acid standards. •The method provided accurate and precise results for quantifying intact and N-terminal truncated proteins within 8 h. -- Abstract: The N-terminal amino acids of proteins are important structure units for maintaining the biological function, localization, and interaction networks of proteins. Under different biological conditions, one or several N-terminal amino acids could be cleaved from an intact protein due to processes, such as proteolysis, resulting in the change of protein properties. Thus, the ability to quantify the N-terminal truncated forms of proteins is of great importance, particularly in the area of development and production of protein-based drugs where the relative quantity of the intact protein and its truncated form needs to be monitored. In this work, we describe a rapid method for absolute quantification of protein mixtures containing intact and N-terminal truncated proteins. This method is based on dansylation labeling of the N-terminal amino acids of proteins, followed by microwave-assisted acid hydrolysis of the proteins into amino acids. It is shown that dansyl labeled amino acids are stable in acidic conditions and can be quantified by liquid chromatography mass spectrometry (LC–MS) with the use of isotope analog standards

  2. Isotopically labeled sulfur compounds and synthetic selenium and tellurium analogues to study sulfur metabolism in marine bacteria

    Directory of Open Access Journals (Sweden)

    Nelson L. Brock

    2013-05-01

    Full Text Available Members of the marine Roseobacter clade can degrade dimethylsulfoniopropionate (DMSP via competing pathways releasing either methanethiol (MeSH or dimethyl sulfide (DMS. Deuterium-labeled [2H6]DMSP and the synthetic DMSP analogue dimethyltelluriopropionate (DMTeP were used in feeding experiments with the Roseobacter clade members Phaeobacter gallaeciensis DSM 17395 and Ruegeria pomeroyi DSS-3, and their volatile metabolites were analyzed by closed-loop stripping and solid-phase microextraction coupled to GC–MS. Feeding experiments with [2H6]DMSP resulted in the incorporation of a deuterium label into MeSH and DMS. Knockout of relevant genes from the known DMSP demethylation pathway to MeSH showed in both species a residual production of [2H3]MeSH, suggesting that a second demethylation pathway is active. The role of DMSP degradation pathways for MeSH and DMS formation was further investigated by using the synthetic analogue DMTeP as a probe in feeding experiments with the wild-type strain and knockout mutants. Feeding of DMTeP to the R. pomeroyi knockout mutant resulted in a diminished, but not abolished production of demethylation pathway products. These results further corroborated the proposed second demethylation activity in R. pomeroyi. Isotopically labeled [2H3]methionine and 34SO42−, synthesized from elemental 34S8, were tested to identify alternative sulfur sources besides DMSP for the MeSH production in P. gallaeciensis. Methionine proved to be a viable sulfur source for the MeSH volatiles, whereas incorporation of labeling from sulfate was not observed. Moreover, the utilization of selenite and selenate salts by marine alphaproteobacteria for the production of methylated selenium volatiles was explored and resulted in the production of numerous methaneselenol-derived volatiles via reduction and methylation. The pathway of selenate/selenite reduction, however, proved to be strictly separated from sulfate reduction.

  3. Differential isotope dansylation labeling combined with liquid chromatography mass spectrometry for quantification of intact and N-terminal truncated proteins

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yanan; Li, Liang, E-mail: Liang.Li@ualberta.ca

    2013-08-20

    Graphical abstract: -- Highlights: •LC–MS was developed for quantifying protein mixtures containing both intact and N-terminal truncated proteins. •{sup 12}C{sub 2}-Dansylation of the N-terminal amino acid of proteins was done first, followed by microwave-assisted acid hydrolysis. •The released {sup 12}C{sub 2}-dansyl labeled N-terminal amino acid was quantified using {sup 13}C{sub 2}-dansyl labeled amino acid standards. •The method provided accurate and precise results for quantifying intact and N-terminal truncated proteins within 8 h. -- Abstract: The N-terminal amino acids of proteins are important structure units for maintaining the biological function, localization, and interaction networks of proteins. Under different biological conditions, one or several N-terminal amino acids could be cleaved from an intact protein due to processes, such as proteolysis, resulting in the change of protein properties. Thus, the ability to quantify the N-terminal truncated forms of proteins is of great importance, particularly in the area of development and production of protein-based drugs where the relative quantity of the intact protein and its truncated form needs to be monitored. In this work, we describe a rapid method for absolute quantification of protein mixtures containing intact and N-terminal truncated proteins. This method is based on dansylation labeling of the N-terminal amino acids of proteins, followed by microwave-assisted acid hydrolysis of the proteins into amino acids. It is shown that dansyl labeled amino acids are stable in acidic conditions and can be quantified by liquid chromatography mass spectrometry (LC–MS) with the use of isotope analog standards.

  4. Analysis of a benthic community food web using isotopically labelled potential food

    International Nuclear Information System (INIS)

    Beviss-Challinor, M.H.; Field, J.G.

    1982-01-01

    A series of experiments was designed to reveal the trophic structure of a benthic community using kelp holdfasts as microcosms within the kelp-bed community. The experimental food comprised zooplankton represented by 3 H 2 O-labelled Artemia sp. eggs and nauplii (200 to 300 μm), detritus derived from 14 C-labelled kelp (60 to 90 μm), and phytoplankton represented by 14 C-labelled Dunaliella primolecta (5 to 10 μm) cultures. Separate experiments of short duration (1 to 2 h) were used to indicate the primary consumers on each type of food, while longer experiments (4, 8 and 16 h) suggested the secondary consumers (coprophagous and carnivorous species). Several species were found to be omnivorous. (orig.)

  5. Report of the research co-ordination meeting on labelling techniques of biomolecules for targeted radiotherapy

    International Nuclear Information System (INIS)

    2000-01-01

    The CRP's focus is on the preparation of site-specific radiopharmaceuticals labelled with with β - emitters for the treatment of cancer. The radiopharmaceuticals should be designed in such a way to deliver the therapeutic doses with high specificity to tumour sites and with minimum dose to other organs. The 2nd Research Co-ordination Meeting (RCM) took place in Mumbai, India from 31 January to 4 February 2000. The present report includes the results of all the participants including the report of the participant from Pakistan who could not attend the meeting. During the second RCM, three main aspects were dealt with viz. the different therapeutic isotopes studied, different carrier molecules like peptides and antibody and the biological aspects of these preparations

  6. Report of the research co-ordination meeting on labelling techniques of biomolecules for targeted radiotherapy

    International Nuclear Information System (INIS)

    2000-01-01

    The CRP's focus is on the preparation of site-specific radiopharmaceuticals labelled with β-emitters for the treatment of cancer. The radiopharmaceuticals should be designed in such a way to deliver the therapeutic doses with high specificity to tumour sites and with minimum dose to other organs. The 2. Research Co-ordination Meeting (RCM) took place in Mumbai, India from 31 January to 4 February 2000. The present report includes the results of all the participants including the report of the participant from Pakistan who could not attend the meeting. During the second RCM, three main aspects were dealt with viz. the different therapeutic isotopes studied, different carrier molecules like peptides and antibody and the biological aspects of these preparations

  7. Report of the research co-ordination meeting on labelling techniques of biomolecules for targeted radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-07-01

    The CRP's focus is on the preparation of site-specific radiopharmaceuticals labelled with {beta}-emitters for the treatment of cancer. The radiopharmaceuticals should be designed in such a way to deliver the therapeutic doses with high specificity to tumour sites and with minimum dose to other organs. The 2. Research Co-ordination Meeting (RCM) took place in Mumbai, India from 31 January to 4 February 2000. The present report includes the results of all the participants including the report of the participant from Pakistan who could not attend the meeting. During the second RCM, three main aspects were dealt with viz. the different therapeutic isotopes studied, different carrier molecules like peptides and antibody and the biological aspects of these preparations.

  8. Dual-isotope technique for determination of in vivo ketone body kinetics

    International Nuclear Information System (INIS)

    Miles, J.M.; Schwenk, W.F.; McClean, K.L.; Haymond, M.W.

    1986-01-01

    Total ketone body specific activity has been widely used in studies of ketone body metabolism to circumvent so-called isotope disequilibrium between the two major ketone body pools, acetoacetate and beta-hydroxybutyrate. Recently, this approach has been criticized on theoretical grounds. In the present studies, [13C]acetoacetate and beta-[14C]hydroxybutyrate were simultaneously infused in nine mongrel dogs before and during an infusion of either unlabeled sodium acetoacetate or unlabeled sodium beta-hydroxybutyrate. Ketone body turnover was determined using total ketone body specific activity, total ketone body moles % enrichment, and an open two-pool model, both before and during the exogenous infusion of unlabeled ketone bodies. Basal ketone body turnover rates were significantly higher using [13C]acetoacetate than with either beta-[14C]hydroxybutyrate alone or the dual-isotope model (3.6 +/- 0.5 vs. 2.2 +/- 0.2 and 2.7 +/- 0.2 mumol X kg-1 X min-1, respectively, P less than 0.05). During exogenous infusion of unlabeled sodium acetoacetate, the dual-isotope model provided the best estimate of ketone body inflow, whereas 14C specific activity underestimated the known rate of acetoacetate infusion by 55% (P less than 0.02). During sodium beta-hydroxybutyrate infusion, [13C]-acetoacetate overestimated ketone body inflow by 55% (P = NS), while better results were obtained with 14C beta-hydroxybutyrate alone and the two-pool model. Ketone body interconversion as estimated by the dual-isotope technique increased markedly during exogenous ketone body infusion. In conclusion, significant errors in estimation of ketone body inflow were made using single-isotope techniques, whereas a dual-isotope model provided reasonably accurate estimates of ketone body inflow during infusion of exogenous acetoacetate and beta-hydroxybutyrate

  9. Analytical techniques for determination of framework oxygen isotope ratio of wairakite

    International Nuclear Information System (INIS)

    Noto, Masami; Kusakabe, Minoru; Uchida, Tetsuo.

    1990-01-01

    Dehydration techniques were developed for the analysis of isotopic ratios of framework oxygen of wairakite, one of calcium zeolites often encountered in geothermal systems. Channel water in wairakite were separated from aluminosilicate framework by dehydration in vacuum at 300 deg, 400 deg, 450 deg, 500 deg, 550 deg, 650 deg, 750 deg, 850 deg, and 950 degC, and by stepwise heating at temperatures from 300 deg to 700 degC. The oxygen isotopic analyses of the separated channel water and the residual aluminosilicate framework of wairakite indicated that dehydration at temperatures higher than 400 degC is accompanied by isotopic exchang between the framework oxygen and dehydrating water vapor. The isotopic exchange during the high temperature dehydration makes the δ 18 O of framework oxygen lower and that of channel water higher than those obtained by dehydration at 300 degC. These results are consistent with dehydration behavior of wairakite under vacuum that the maximum rate of dehydration of channel water is attained at about 400 degC. Consequently it is recommended to dehydrate wairakite at a temperature as low as possible in order to avoid the effect of the isotopic exchange. Time required to attain complete dehydration becomes longer with lowering the temperature of dehydration. To compromise these conflicting effects, the optimum conditions of dehydration have been found that most of the channel water is dehydrated at 300 degC for 24 hours, followed by stepwise heating for additional 17 hours up to 700 degC. We obtained a better than ± 0.1 reproducibility for the framework oxygen isotopic determinations with this technique. (author)

  10. Soil organic carbon assessments in cropping systems using isotopic techniques

    Science.gov (United States)

    Martín De Dios Herrero, Juan; Cruz Colazo, Juan; Guzman, María Laura; Saenz, Claudio; Sager, Ricardo; Sakadevan, Karuppan

    2016-04-01

    Introduction of improved farming practices are important to address the challenges of agricultural production, food security, climate change and resource use efficiency. The integration of livestock with crops provides many benefits including: (1) resource conservation, (2) ecosystem services, (3) soil quality improvements, and (4) risk reduction through diversification of enterprises. Integrated crop livestock systems (ICLS) with the combination of no-tillage and pastures are useful practices to enhance soil organic carbon (SOC) compared with continuous cropping systems (CCS). In this study, the SOC and its fractions in two cropping systems namely (1) ICLS, and (2) CCS were evaluated in Southern Santa Fe Province in Argentina, and the use of delta carbon-13 technique and soil physical fractionation were evaluated to identify sources of SOC in these systems. Two farms inside the same soil cartographic unit and landscape position in the region were compared. The ICLS farm produces lucerne (Medicago sativa Merrill) and oat (Avena sativa L.) grazed by cattle alternatively with grain summer crops sequence of soybean (Glicine max L.) and corn (Zea mays L.), and the farm under continuous cropping system (CCS) produces soybean and corn in a continuous sequence. The soil in the area is predominantly a Typic Hapludoll. Soil samples from 0-5 and 0-20 cm depths (n=4) after the harvest of grain crops were collected in each system and analyzed for total organic carbon (SOC, 0-2000 μm), particulate organic carbon (POC, 50-100 μm) and mineral organic carbon (MOC, is probably due to the presence of deep roots under pastures in ICLS. Delta carbon-13 values for 0-5 cm were -22.9, -21.2 and -19.9 per mil for REF, ICLS and CCS, respectively (Pis explained by the presence of tree species with high lignin content in natural vegetation. Lignin has lower delta carbon-13 compared to cellulose (dominating in crops and pastures), which is present in greater proportion in plant residues of

  11. Nutrition labelling, marketing techniques, nutrition claims and health claims on chip and biscuit packages from sixteen countries.

    Science.gov (United States)

    Mayhew, Alexandra J; Lock, Karen; Kelishadi, Roya; Swaminathan, Sumathi; Marcilio, Claudia S; Iqbal, Romaina; Dehghan, Mahshid; Yusuf, Salim; Chow, Clara K

    2016-04-01

    Food packages were objectively assessed to explore differences in nutrition labelling, selected promotional marketing techniques and health and nutrition claims between countries, in comparison to national regulations. Cross-sectional. Chip and sweet biscuit packages were collected from sixteen countries at different levels of economic development in the EPOCH (Environmental Profile of a Community's Health) study between 2008 and 2010. Seven hundred and thirty-seven food packages were systematically evaluated for nutrition labelling, selected promotional marketing techniques relevant to nutrition and health, and health and nutrition claims. We compared pack labelling in countries with labelling regulations, with voluntary regulations and no regulations. Overall 86 % of the packages had nutrition labels, 30 % had health or nutrition claims and 87 % displayed selected marketing techniques. On average, each package displayed two marketing techniques and one health or nutrition claim. In countries with mandatory nutrition labelling a greater proportion of packages displayed nutrition labels, had more of the seven required nutrients present, more total nutrients listed and higher readability compared with those with voluntary or no regulations. Countries with no health or nutrition claim regulations had fewer claims per package compared with countries with regulations. Nutrition label regulations were associated with increased prevalence and quality of nutrition labels. Health and nutrition claim regulations were unexpectedly associated with increased use of claims, suggesting that current regulations may not have the desired effect of protecting consumers. Of concern, lack of regulation was associated with increased promotional marketing techniques directed at children and misleadingly promoting broad concepts of health.

  12. Bioavailability of iron to rats from processed soybean fractions determined by intrinsic and extrinsic labeling techniques

    International Nuclear Information System (INIS)

    Weaver, C.M.; Nelson, N.; Elliott, J.G.

    1984-01-01

    Intrinsic and extrinsic labeling techniques were used to measure iron bioavailability from soybean fractions (isolated soy protein, defatted flour, soy hulls, insoluble material and whey) by iron-depleted and non-iron-depleted rats. As expected, absorption of iron was higher in the iron-depleted than in the non-iron-depleted rats. In the iron-depleted group, significantly more iron was absorbed from soy whey than from other fractions. No other significant difference in iron absorption associated with iron source was observed. The higher absorption rate of iron from whey by the iron-depleted rats probably was related to a lower quantity of food consumed during the test meal by this group. Intrinsic and extrinsic labeling techniques produced similar assessments of bioavailability of iron

  13. Synthesis of stereoarray isotope labeled (SAIL) lysine via the "head-to-tail" conversion of SAIL glutamic acid.

    Science.gov (United States)

    Terauchi, Tsutomu; Kamikawai, Tomoe; Vinogradov, Maxim G; Starodubtseva, Eugenia V; Takeda, Mitsuhiro; Kainosho, Masatsune

    2011-01-07

    A stereoarray isotope labeled (SAIL) lysine, (2S,3R,4R,5S,6R)-[3,4,5,6-(2)H(4);1,2,3,4,5,6-(13)C(6);2,6-(15)N(2)]lysine, was synthesized by the "head-to-tail" conversion of SAIL-Glu, (2S,3S,4R)-[3,4-(2)H(2);1,2,3,4,5-(13)C(5);2-(15)N]glutamic acid, with high stereospecificities for all five chiral centers. With the SAIL-Lys in hand, the unambiguous simultaneous stereospecific assignments were able to be established for each of the prochiral protons within the four methylene groups of the Lys side chains in proteins.

  14. Isotopic Determination of Nuclear Materials Using Nuclear Fission Track Registration Technique and Thermal Ionization Mass Spectrometric Technique

    International Nuclear Information System (INIS)

    Jeon, Young Sin; Pyo, Hyeong Yeol; Park, Yong Joon; Song, Kyu Seok; Kim, Won Ho; Jee, Kwang Yong

    2007-05-01

    It is very important to develope the technology for the determination of isotopic ratios of hot particles( 234 U, 235 U, 236 U etc.) detected from swipe samples of various nuclear facilities. This technology is highly competitive internationally and has to be established independently as long as our government maintains atomic energy and treats nuclear materials. In this text, sample pretreatment procedure, gamma-ray counting, alpha or fission track techniques, isotopic analysis of U and Pu, background problems and detection limits for mass determination, and their application to the real swipe sample were described with detailed procedure. This technology would contribute to the Korean economy's high growth rate as well as to superiority of government's leading research and development programs if successfully established

  15. A facile method for steroid labeling by heavy isotopes of hydrogen

    Czech Academy of Sciences Publication Activity Database

    Marek, Aleš; Klepetářová, Blanka; Elbert, Tomáš

    2015-01-01

    Roč. 71, č. 29 (2015), s. 4874-4882 ISSN 0040-4020 R&D Projects: GA AV ČR IAA400550801 Institutional support: RVO:61388963 Keywords : brassinosteroids * alpha-hydroxy ketones * reductive dehalogenation * tritium * labeled compounds Subject RIV: CC - Organic Chemistry Impact factor: 2.645, year: 2015

  16. Determination of the isotope distribution in 14C-labelled hydrocarbons by thermal fragmentation

    International Nuclear Information System (INIS)

    Kopinke, F.D.; Dermietzel, J.; Jockisch, W.; Raeuber, G.

    1986-01-01

    The gas chromatographic analysis of pyrolysis products of properly labelled hydrocarbons allows a definite and quantitative determination of the 14 C-distribution in those compounds. For this purpose a simple, fast, and versatilely applicable method has been developed and described

  17. A Practical Cryogen-Free CO2 Purification and Freezing Technique for Stable Isotope Analysis.

    Science.gov (United States)

    Sakai, Saburo; Matsuda, Shinichi

    2017-04-18

    Since isotopic analysis by mass spectrometry began in the early 1900s, sample gas for light-element isotopic measurements has been purified by the use of cryogens and vacuum-line systems. However, this conventional purification technique can achieve only certain temperatures that depend on the cryogens and can be sustained only as long as there is a continuous cryogen supply. Here, we demonstrate a practical cryogen-free CO 2 purification technique using an electrical operated cryocooler for stable isotope analysis. This approach is based on portable free-piston Stirling cooling technology and controls the temperature to an accuracy of 0.1 °C in a range from room temperature to -196 °C (liquid-nitrogen temperature). The lowest temperature can be achieved in as little as 10 min. We successfully purified CO 2 gas generated by carbonates and phosphoric acid reaction and found its sublimation point to be -155.6 °C at 0.1 Torr in the vacuum line. This means that the temperature required for CO 2 trapping is much higher than the liquid-nitrogen temperature. Our portable cooling system offers the ability to be free from the inconvenience of cryogen use for stable isotope analysis. It also offers a new cooling method applicable to a number of fields that use gas measurements.

  18. Automatic isotope gas analysis of tritium labelled organic materials Pt. 3

    International Nuclear Information System (INIS)

    Gacs, I.; Mlinko, S.; Payer, K.; Otvos, L.; Banfi, D.; Palagyi, T.

    1978-01-01

    An isotope analytical procedure and an automatic instrument developed for the determination of tritium in organic compounds and biological materials by internal gas counting are described. The sample is burnt in a stream of oxygen and the combustion products including water vapour carrying the tritium are led onto a column of molecular sieve-5A heated to 550 deg C. Tritium is retained temporarily on the column, then transferred into a stream of hydrogen by isotope exchange. After addition of butane, the tritiated hydrogen is led into an internal detector and enclosed there for radioactivity measurement. The procedure, providing quantitative recovery, is completed in five minutes. It is free of memory effect and suitable for the determination of tritium in a wide range of organic compounds and samples of biological origin. (author)

  19. Deciphering systemic wound responses of the pumpkin extrafascicular phloem by metabolomics and stable isotope-coded protein labeling.

    Science.gov (United States)

    Gaupels, Frank; Sarioglu, Hakan; Beckmann, Manfred; Hause, Bettina; Spannagl, Manuel; Draper, John; Lindermayr, Christian; Durner, Jörg

    2012-12-01

    In cucurbits, phloem latex exudes from cut sieve tubes of the extrafascicular phloem (EFP), serving in defense against herbivores. We analyzed inducible defense mechanisms in the EFP of pumpkin (Cucurbita maxima) after leaf damage. As an early systemic response, wounding elicited transient accumulation of jasmonates and a decrease in exudation probably due to partial sieve tube occlusion by callose. The energy status of the EFP was enhanced as indicated by increased levels of ATP, phosphate, and intermediates of the citric acid cycle. Gas chromatography coupled to mass spectrometry also revealed that sucrose transport, gluconeogenesis/glycolysis, and amino acid metabolism were up-regulated after wounding. Combining ProteoMiner technology for the enrichment of low-abundance proteins with stable isotope-coded protein labeling, we identified 51 wound-regulated phloem proteins. Two Sucrose-Nonfermenting1-related protein kinases and a 32-kD 14-3-3 protein are candidate central regulators of stress metabolism in the EFP. Other proteins, such as the Silverleaf Whitefly-Induced Protein1, Mitogen Activated Protein Kinase6, and Heat Shock Protein81, have known defensive functions. Isotope-coded protein labeling and western-blot analyses indicated that Cyclophilin18 is a reliable marker for stress responses of the EFP. As a hint toward the induction of redox signaling, we have observed delayed oxidation-triggered polymerization of the major Phloem Protein1 (PP1) and PP2, which correlated with a decline in carbonylation of PP2. In sum, wounding triggered transient sieve tube occlusion, enhanced energy metabolism, and accumulation of defense-related proteins in the pumpkin EFP. The systemic wound response was mediated by jasmonate and redox signaling.

  20. Evaluation of stable isotope labelling strategies for the quantitation of CP4 EPSPS in genetically modified soya

    Energy Technology Data Exchange (ETDEWEB)

    Ocana, Mireia Fernandez [Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of London, Egham TW20 0EX (United Kingdom)], E-mail: Mireia.FernandezOcana@pfizer.com; Fraser, Paul D. [Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of London, Egham TW20 0EX (United Kingdom); Patel, Raj K.P.; Halket, John M. [Specialist Bioanalytical Services Ltd., Royal Holloway, University of London, Egham TW20 0EX (United Kingdom); Bramley, Peter M. [Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of London, Egham TW20 0EX (United Kingdom)

    2009-02-16

    The introduction of genetically modified (GM) crops into the market has raised a general alertness relating to the control and safety of foods. The applicability of protein separation hyphenated to mass spectrometry to identify the bacterial enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein expressed in GM crops has been previously reported [M.F. Ocana, P.D. Fraser, R.K.P. Patel, J.M. Halket, P.M. Bramley, Rapid Commun. Mass Spectrom. 21 (2007) 319.]. Herein, we investigate the suitability of two strategies that employ heavy stable isotopes, i.e. AQUA and iTRAQ, to quantify different levels of CP4 EPSPS in up to four GM preparations. Both quantification strategies showed potential to determine whether the presence of GM material is above the limits established by the European Union. The AQUA quantification procedure involved protein solubilisation/fractionation and subsequent separation using SDS-PAGE. A segment of the gel in which the protein of interest was located was excised, the stable isotope labeled peptide added at a known concentration and proteolytic digestion initiated. Following recovery of the peptides, on-line separation and detection using LC-MS was carried out. A similar approach was used for the iTRAQ workflow with the exception that proteins were digested in solution and generated tryptic peptides were chemically tagged. Both procedures demonstrated the potential for quantitative detection at 0.5% (w/w) GM soya which is a level below the current European Union's threshold for food-labelling. In this context, a comparison between the two procedures is provided within the present study.

  1. Bacterial expression and one-step purification of an isotope-labeled heterotrimeric G-protein {alpha}-subunit

    Energy Technology Data Exchange (ETDEWEB)

    Abdulaev, Najmoutin G. [University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology (United States); Zhang Cheng; Dinh, Andy [University of Texas Health Science Center, Center for Membrane Biology, Department of Biochemistry and Molecular Biology (United States); Ngo, Tony; Bryan, Philip N. [University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology (United States); Brabazon, Danielle M. [Loyola College in Maryland, Department of Chemistry (United States); Marino, John P. [University of Maryland Biotechnology Institute, Center for Advanced Research in Biotechnology (United States)], E-mail: marino@carb.nist.gov; Ridge, Kevin D. [University of Texas Health Science Center, Center for Membrane Biology, Department of Biochemistry and Molecular Biology (United States)

    2005-05-15

    Heterologous expression systems are often employed to generate sufficient quantities of isotope-labeled proteins for high-resolution NMR studies. Recently, the interaction between the prodomain region of subtilisin and an active, mutant form of the mature enzyme has been exploited to develop a cleavable affinity tag fusion system for one-step generation and purification of full-length soluble proteins obtained by inducible prokaryotic expression. As a first step towards applying high-resolution NMR methods to study heterotrimeric G-protein {alpha}-subunit (G{sub {alpha}}) conformation and dynamics, the utility of this subtilisin prodomain fusion system for expressing and purifying an isotope-labeled G{sub {alpha}} chimera ({approx}40 kDa polypeptide) has been tested. The results show that a prodomain fused G{sub {alpha}} chimera can be expressed to levels approaching 6-8 mg/l in minimal media and that the processed, mature protein exhibits properties similar to those of G{sub {alpha}} isolated from natural sources. To assay for the functional integrity of the purified G{sub {alpha}} chimera at NMR concentrations and probe for changes in the structure and dynamics of G{sub {alpha}} that result from activation, {sup 15}N-HSQC spectra of the GDP/Mg{sup 2+} bound form of G{sub {alpha}} obtained in the absence and presence of aluminum fluoride, a well known activator of the GDP bound state, have been acquired. Comparisons of the {sup 15}N-HSQC spectra reveals a number of changes in chemical shifts of the {sup 1}HN, {sup 15}N crosspeaks that are discussed with respect to expected changes in the protein conformation associated with G{sub {alpha}} activation.

  2. Evaluation of stable isotope labelling strategies for the quantitation of CP4 EPSPS in genetically modified soya

    International Nuclear Information System (INIS)

    Ocana, Mireia Fernandez; Fraser, Paul D.; Patel, Raj K.P.; Halket, John M.; Bramley, Peter M.

    2009-01-01

    The introduction of genetically modified (GM) crops into the market has raised a general alertness relating to the control and safety of foods. The applicability of protein separation hyphenated to mass spectrometry to identify the bacterial enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) protein expressed in GM crops has been previously reported [M.F. Ocana, P.D. Fraser, R.K.P. Patel, J.M. Halket, P.M. Bramley, Rapid Commun. Mass Spectrom. 21 (2007) 319.]. Herein, we investigate the suitability of two strategies that employ heavy stable isotopes, i.e. AQUA and iTRAQ, to quantify different levels of CP4 EPSPS in up to four GM preparations. Both quantification strategies showed potential to determine whether the presence of GM material is above the limits established by the European Union. The AQUA quantification procedure involved protein solubilisation/fractionation and subsequent separation using SDS-PAGE. A segment of the gel in which the protein of interest was located was excised, the stable isotope labeled peptide added at a known concentration and proteolytic digestion initiated. Following recovery of the peptides, on-line separation and detection using LC-MS was carried out. A similar approach was used for the iTRAQ workflow with the exception that proteins were digested in solution and generated tryptic peptides were chemically tagged. Both procedures demonstrated the potential for quantitative detection at 0.5% (w/w) GM soya which is a level below the current European Union's threshold for food-labelling. In this context, a comparison between the two procedures is provided within the present study

  3. Bacterial expression and one-step purification of an isotope-labeled heterotrimeric G-protein α-subunit

    International Nuclear Information System (INIS)

    Abdulaev, Najmoutin G.; Zhang Cheng; Dinh, Andy; Ngo, Tony; Bryan, Philip N.; Brabazon, Danielle M.; Marino, John P.; Ridge, Kevin D.

    2005-01-01

    Heterologous expression systems are often employed to generate sufficient quantities of isotope-labeled proteins for high-resolution NMR studies. Recently, the interaction between the prodomain region of subtilisin and an active, mutant form of the mature enzyme has been exploited to develop a cleavable affinity tag fusion system for one-step generation and purification of full-length soluble proteins obtained by inducible prokaryotic expression. As a first step towards applying high-resolution NMR methods to study heterotrimeric G-protein α-subunit (G α ) conformation and dynamics, the utility of this subtilisin prodomain fusion system for expressing and purifying an isotope-labeled G α chimera (∼40 kDa polypeptide) has been tested. The results show that a prodomain fused G α chimera can be expressed to levels approaching 6-8 mg/l in minimal media and that the processed, mature protein exhibits properties similar to those of G α isolated from natural sources. To assay for the functional integrity of the purified G α chimera at NMR concentrations and probe for changes in the structure and dynamics of G α that result from activation, 15 N-HSQC spectra of the GDP/Mg 2+ bound form of G α obtained in the absence and presence of aluminum fluoride, a well known activator of the GDP bound state, have been acquired. Comparisons of the 15 N-HSQC spectra reveals a number of changes in chemical shifts of the 1 HN, 15 N crosspeaks that are discussed with respect to expected changes in the protein conformation associated with G α activation

  4. Isotope techniques in the study of the hydrology of fractured and fissured rocks

    International Nuclear Information System (INIS)

    1989-01-01

    It is generally agreed that the hydrology of fractured rocks refers to the occurrence and movement of groundwater in rocks whose porosity is due to cracks, fissures and fractures in compact rocks. Until recently, crystalline rocks were considered to be impervious (the role of karst reservoirs for groundwater storage is already known). Thus, although fractured rocks cover approximately one-third of the Earth's surface, knowledge of groundwater flow dynamics still needs to be substantively improved. The Proceedings include the papers presented at the Advisory Group Meeting on the Application of Isotope Techniques in the Study of the Hydrology of Fractured and Fissured Rocks, which took place in Vienna from 17 to 21 November 1986. The meeting was attended by 21 scientists from 10 Member States. The Group agreed that the following topics should be given the highest priority: (1) Protection of groundwater resources should be incorporated into all future activities. This implies that recharge areas have to be delineated, not only using nuclear techniques but also other hydrogeological and geochemical methods. Research efforts in this direction should be intensified. (2) The Group was aware that new isotope techniques are being developed in hydrogeology and agreed that their application in fractured and fissured rocks showed promise. It was therefore suggested that information on new isotopes such as 36 Cl and 129 I should be gathered, either in the form of the various techniques currently being used by different laboratories or through the various activities that are being undertaken. Ways of co-ordinating the work being done and exchanging information at the international level should be encouraged. The Proceedings should be of interest to scientists responsible for the evaluation of water resources in crystalline rocks. It is hoped that hydrologists and hydrogeologists working in such terrains who are not familiar with the use of isotope techniques will find ideas and

  5. Evaluation of the impact of matrix effect on quantification of pesticides in foods by gas chromatography-mass spectrometry using isotope-labeled internal standards.

    Science.gov (United States)

    Yarita, Takashi; Aoyagi, Yoshie; Otake, Takamitsu

    2015-05-29

    The impact of the matrix effect in GC-MS quantification of pesticides in food using the corresponding isotope-labeled internal standards was evaluated. A spike-and-recovery study of nine target pesticides was first conducted using paste samples of corn, green soybean, carrot, and pumpkin. The observed analytical values using isotope-labeled internal standards were more accurate for most target pesticides than that obtained using the external calibration method, but were still biased from the spiked concentrations when a matrix-free calibration solution was used for calibration. The respective calibration curves for each target pesticide were also prepared using matrix-free calibration solutions and matrix-matched calibration solutions with blank soybean extract. The intensity ratio of the peaks of most target pesticides to that of the corresponding isotope-labeled internal standards was influenced by the presence of the matrix in the calibration solution; therefore, the observed slope varied. The ratio was also influenced by the type of injection method (splitless or on-column). These results indicated that matrix-matching of the calibration solution is required for very accurate quantification, even if isotope-labeled internal standards were used for calibration. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Natural isotope technique for the exploration and exploitation of ground water

    International Nuclear Information System (INIS)

    Zainal Abidin; Hudi Hastowo; Aang Hanafiah

    2007-01-01

    In line with the condition of climate and hydrology, Indonesia has a fast amount of aquifers which are sources of ground water. In several areas large number of springs occurred with small to large debits which is a sign of ground water potential. Ground water is a potential reservoir to be use at maximum for several purposes such as drinking water, industry and tourism. Large cities such as Jakarta, Bandung and others depend on ground water for their industries and hotels. The exploitation of ground water use has to be controlled and monitoring of a management system have to be done. Research carried out only on the exploitation of geophysics and hydrology showed that the amount of ground water reservoirs is not enough to be used when it comes to justification to explore it. Other parameters are still be needed which are the origins and dating of the ground water, these last two factors mentioned have to be taken into consideration in the system of conversion and balance of water. An alternative technology to determine the two factors mentioned in a short time is the natural isotope technique of 18 O, 2 H and 14 C. This technique is used to determine the origin of water, and isotope 14 C is carried out to determine the age of ground water. Isotopes 18 H and 2 H are stable isotopes in the form of water and is integrated in the hydrological cycle. Their specific concentrations in rain water at several elevations are used as fingerprints to locate the area of ground water supplement and its origin. Isotope 14 C is a natural radioactive isotope with a half-life of 5.730 years and is found in the hydrology cycle and enters the ground water system through CO 2 gas which is dissolved in water. 14 C isotope could determine the age of ground water and is also able to indicate the potential/amount of ground water. Studies of exploration and exploration monitoring of ground water should be an integrated study by geohydrology, geophysics and isotope and could be a solution of

  7. Metabolic flux analysis of the phenylpropanoid pathway in wound-healing potato tuber tissue using stable isotope-labeled tracer and LC-MS spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Matsuda, Fumio; Morino, Keiko; Miyashita, Masahiro; Miyagawa, Hisashi [Kyoto Univ. (Japan). Department of Agriculture

    2003-05-01

    The metabolic flux of two phenylpropanoid metabolites, N-p-coumaroyloctopamine (p-CO) and chlorogenic acid (CGA), in the wound-healing potato tuber tissue was quantitatively analyzed by a newly developed method based upon the tracer experiment using stable isotope-labeled compounds and LC-MS. Tuber disks were treated with aqueous solution of L-phenyl-d{sub 5}-alanine, and the change in the ratio of stable isotope-labeled compound to non-labeled (isotope abundance) was monitored for p-CO and CGA in the tissue extract by LC-MS. The time-dependent change in the isotope abundance of each metabolite was fitted to an equation that was derived from the formation and conversion kinetics of each compound. Good correlations were obtained between the observed and calculated isotope abundances for both p-CO and CGA. The rates of p-CO formation and conversion (i.e. fluxes) were 1.15 and 0.96 nmol (g FW){sup -1}h{sup -1}, respectively, and for CGA, the rates 4.63 and 0.42 nmol (g FW){sup -1}h{sup -1}, respectively. This analysis enabled a direct comparison of the biosynthetic activity between these two compounds. (author)

  8. Validation of the doubly labeled water method using off-axis integrated cavity output spectroscopy and isotope ratio mass spectrometry.

    Science.gov (United States)

    Melanson, Edward L; Swibas, Tracy; Kohrt, Wendy M; Catenacci, Vicki A; Creasy, Seth A; Plasqui, Guy; Wouters, Loek; Speakman, John R; Berman, Elena S F

    2018-02-01

    When the doubly labeled water (DLW) method is used to measure total daily energy expenditure (TDEE), isotope measurements are typically performed using isotope ratio mass spectrometry (IRMS). New technologies, such as off-axis integrated cavity output spectroscopy (OA-ICOS) provide comparable isotopic measurements of standard waters and human urine samples, but the accuracy of carbon dioxide production (V̇co 2 ) determined with OA-ICOS has not been demonstrated. We compared simultaneous measurement V̇co 2 obtained using whole-room indirect calorimetry (IC) with DLW-based measurements from IRMS and OA-ICOS. Seventeen subjects (10 female; 22 to 63 yr) were studied for 7 consecutive days in the IC. Subjects consumed a dose of 0.25 g H 2 18 O (98% APE) and 0.14 g 2 H 2 O (99.8% APE) per kilogram of total body water, and urine samples were obtained on days 1 and 8 to measure average daily V̇co 2 using OA-ICOS and IRMS. V̇co 2 was calculated using both the plateau and intercept methods. There were no differences in V̇co 2 measured by OA-ICOS or IRMS compared with IC when the plateau method was used. When the intercept method was used, V̇co 2 using OA-ICOS did not differ from IC, but V̇co 2 measured using IRMS was significantly lower than IC. Accuracy (~1-5%), precision (~8%), intraclass correlation coefficients ( R = 0.87-90), and root mean squared error (30-40 liters/day) of V̇co 2 measured by OA-ICOS and IRMS were similar. Both OA-ICOS and IRMS produced measurements of V̇co 2 with comparable accuracy and precision compared with IC.

  9. Co-ordinated research programme on development and application of isotopic techniques in studies of vitamin A nutrition. Report of the first research co-ordination meeting

    International Nuclear Information System (INIS)

    1997-01-01

    In Vitamin A nutrition, evaluations to ascertain the efficacy of intervention strategies are becoming increasingly important. However, state-of-the-art methods for evaluating vitamin A status often do not provide enough quantitative information on vitamin A status and the bioconversion of carotenoids, particularly in people with subclinical vitamin A deficiency. These limitations have had programmatic consequences. The principal reason the new Coordinated Research programme (CRP) was formulated was to improve techniques for measuring vitamin A status and the bioconversion of carotenoids to vitamin A with the expectation that the new methods could contribute meaningfully to field-based evaluations of the efficacy of intervention strategies. The International Atomic Energy Agency (IAEA) is sponsoring programmes to develop and transfer isotopic techniques to improve nutrition monitoring in developing countries. The New CRP ''Development and Application of Isotopic Techniques in Studies of Vitamin A Nutrition'' has seven teams, six of which are working to develop methods based on orally administered isotopically labelled retinol which will be a valid measure of whole body retinol (mostly hepatic reserves) and useful under typical field conditions, particularly in women and children with marginal vitamin A deficiency. The seventh team is biosynthesizing uniformly deuterated β-carotene by growing foods in deuterated water. This report summarizes the research to be undertaken, as presented at the first Research Co-ordination Meeting

  10. Co-ordinated research programme on development and application of isotopic techniques in studies of vitamin A nutrition. Report of the first research co-ordination meeting

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-12-31

    In Vitamin A nutrition, evaluations to ascertain the efficacy of intervention strategies are becoming increasingly important. However, state-of-the-art methods for evaluating vitamin A status often do not provide enough quantitative information on vitamin A status and the bioconversion of carotenoids, particularly in people with subclinical vitamin A deficiency. These limitations have had programmatic consequences. The principal reason the new Coordinated Research programme (CRP) was formulated was to improve techniques for measuring vitamin A status and the bioconversion of carotenoids to vitamin A with the expectation that the new methods could contribute meaningfully to field-based evaluations of the efficacy of intervention strategies. The International Atomic Energy Agency (IAEA) is sponsoring programmes to develop and transfer isotopic techniques to improve nutrition monitoring in developing countries. The New CRP ``Development and Application of Isotopic Techniques in Studies of Vitamin A Nutrition`` has seven teams, six of which are working to develop methods based on orally administered isotopically labelled retinol which will be a valid measure of whole body retinol (mostly hepatic reserves) and useful under typical field conditions, particularly in women and children with marginal vitamin A deficiency. The seventh team is biosynthesizing uniformly deuterated {beta}-carotene by growing foods in deuterated water. This report summarizes the research to be undertaken, as presented at the first Research Co-ordination Meeting. Refs, figs, tabs.

  11. Determination of total uranium by mass spectrometry utilizing the isotopic dilution technique

    International Nuclear Information System (INIS)

    Cretella, R.F.; Noutary, C.J.; Servant, R.E.

    1981-01-01

    The isotopic dilution associated to mass spectrometry is a high-sensitivity technique that allows to work with microquantities of the sample, making it possible to analize the content in highly radioactive solutions with excellent accuracy and minimum risk. The proposed technique is described and its results are discussed through the analysis of: 1) A synthetic sample that simulates dissolved spent fuel elements; 2) Uranium dioxide of nuclear purity and 3) Uranium concentrate. 233 U(ORNL) was employed as a tracer and a Nuclide 12-90-SU mass spectrometer of simple magnetic focus as measurement instrument. The accuracy reached in the analyses is better than 0.5% with a reliability of 95%. The analysis of the errors shows that their main contributing source are the errors in the measurement of the isotopic ratios. (M.E.L.) [es

  12. Isotope techniques in water resource investigations in arid and semi-arid regions

    International Nuclear Information System (INIS)

    2001-03-01

    The Co-ordinated Research Project (CRP) on the Use of Isotope Techniques in Water Resources Investigations in Arid and Semi-arid Regions was initiated with the aim od contributing to the assessment of groundwater resources in arid areas through the use of environmental isotope techniques, and thereby to help in better management of these valuable fresh groundwater resources. The main emphases identified were in three key areas: (i) the evaluation of water balance components such as recharge rate estimation and recharge and discharge cycles at different spatial scales, (ii) paleohydrology and hydroclimatic change and, (iii) anthropogenic impacts and the assessment of the vulnerability of arid zone ground waters to salinisation and pollution impacts. This publication presents individual projects carried out within the frameworks of the CRP. Each paper has been indexed separately

  13. Anglo-Saxon animal husbandry techniques revealed though isotope and chemical variations in cattle teeth

    International Nuclear Information System (INIS)

    Evans, J.A.; Tatham, S.; Chenery, S.R.; Chenery, C.A.

    2007-01-01

    The Sr concentration and isotope composition of tooth enamel from domesticated animals from two neighbouring Anglo-Saxon settlements, at Empingham (6-7th century) and Ketton (10-12th century) in Rutland, central England, are compared both with each other, and with associated human populations. Data from the Empingham II site form discrete fields in Sr concentration and isotope composition space for cattle, pig and sheep with a partial overlap of the human and pig fields. By contrast there is significant overlap in all the animal and human data fields from the Ketton site. The differences in data distribution between the two sites are attributed to animal husbandry techniques, as the surface geology of the two areas is very similar, implying geological factors are an unlikely cause of the difference. It is suggested that the grazing and feeding patterns of animals at the Empingham II site were controlled and restricted, whereas at the Ketton site the animals grazed and foraged freely over a common area. Strontium isotope variation within cattle molars from the two settlements show marked differences that reflect the nature of their feeding and rearing. The enamel from a cattle molar from the Empingham II site has a well-defined, systematic variation of Sr isotope composition with Sr concentration, whereas no such patterns exist in a comparable cattle molar from Ketton. Chemical and O isotope variations in the cattle tooth from Empingham II show sympathetic variation of Sr and Ba concentrations with Sr isotope composition from cusp to cervix. The cusp has higher Sr and Ba concentrations log (Sr/Ca) ratio of -3.1 and 87 Sr/ 86 Sr ratio of 0.71151 where as the cervical region of the enamel has log (Sr/Ca) = -3.3 and 87 Sr/ 86 Sr = 0.71061

  14. Anglo-Saxon animal husbandry techniques revealed though isotope and chemical variations in cattle teeth

    Energy Technology Data Exchange (ETDEWEB)

    Evans, J.A. [NERC Isotope Geosciences Laboratory, BGS, Keyworth, Nottingham NG12 5GG (United Kingdom)], E-mail: je@nigl.nerc.ac.uk; Tatham, S. [School of Archaeology and Ancient History, University of Leicester, Leicester LE1 7RH (United Kingdom); Chenery, S.R. [British Geological Survey, Keyworth, Nottingham NG12 5GG (United Kingdom); Chenery, C.A. [NERC Isotope Geosciences Laboratory, BGS, Keyworth, Nottingham NG12 5GG (United Kingdom)

    2007-09-15

    The Sr concentration and isotope composition of tooth enamel from domesticated animals from two neighbouring Anglo-Saxon settlements, at Empingham (6-7th century) and Ketton (10-12th century) in Rutland, central England, are compared both with each other, and with associated human populations. Data from the Empingham II site form discrete fields in Sr concentration and isotope composition space for cattle, pig and sheep with a partial overlap of the human and pig fields. By contrast there is significant overlap in all the animal and human data fields from the Ketton site. The differences in data distribution between the two sites are attributed to animal husbandry techniques, as the surface geology of the two areas is very similar, implying geological factors are an unlikely cause of the difference. It is suggested that the grazing and feeding patterns of animals at the Empingham II site were controlled and restricted, whereas at the Ketton site the animals grazed and foraged freely over a common area. Strontium isotope variation within cattle molars from the two settlements show marked differences that reflect the nature of their feeding and rearing. The enamel from a cattle molar from the Empingham II site has a well-defined, systematic variation of Sr isotope composition with Sr concentration, whereas no such patterns exist in a comparable cattle molar from Ketton. Chemical and O isotope variations in the cattle tooth from Empingham II show sympathetic variation of Sr and Ba concentrations with Sr isotope composition from cusp to cervix. The cusp has higher Sr and Ba concentrations log (Sr/Ca) ratio of -3.1 and {sup 87}Sr/{sup 86}Sr ratio of 0.71151 where as the cervical region of the enamel has log (Sr/Ca) = -3.3 and {sup 87}Sr/{sup 86}Sr = 0.71061.

  15. Future Applications in Quantitative Isotopic Tracing using Homogeneously Carbon-13 Labelled Plant Material

    International Nuclear Information System (INIS)

    Slaets, Johanna I.F.; Chen, Janet; Resch, Christian; Mayr, Leopold; Weltin, Georg; Heiling, Maria; Gruber, Roman; Dercon, Gerd

    2017-01-01

    Carbon-13 ("1"3C) and nitrogen-15 ("1"5N) labelled plant material is increasingly being used to trace the fate of plant-derived C and N into the atmosphere, soil, water and organisms in many studies, including those investigating the potential of soils to store greenhouse gases belowground. Storage of C in soils can offset and even reduce atmospheric levels of the greenhouse gas, CO_2, and interest in such studies is growing due to problems associated with anthropogenic greenhouse gas emissions impacting climate change. Reduction of N loss in soils is also of great interest, as it reduces release of the greenhouse gas, N_2O, into the atmosphere. However, accurate quantitative tracing of plant-derived C and N in such research is only possible if plant material is labelled both homogeneously and in sufficient quantities.

  16. Labelling of long chain fatty acids by non isotopic nucleophilic halogen exchange

    International Nuclear Information System (INIS)

    Hallaba, E.; Al-Suhybani, A.A.; Zaki, F.S.

    1985-01-01

    The parameters of two exchange methods for preparing pure 97% labelled 17-Br-HDA in acetone and in benzene with dry NaI in a closed system are described. In aprotic solvents the need for a phase transfer catalyst up to 50 μg is necessary to dissolve the dry NaI. The use of aqueous medium in the exchange is totally prohibited. Energies of activation are calculated for both reactions. (author)

  17. Application of Isotope Techniques for Assessing Nutrient Dynamics in River Basins

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2013-05-15

    Nutrients are necessary for the growth and survival of animals, plants and other organisms. However, industrial, agricultural and urban development has dramatically increased nutrient levels in river systems, including nitrogen and phosphorus containing substances, degrading water quality, causing acidification and eutrophication and affecting aquatic ecosystems. Nutrient assessment and management in river systems has been an important part of water resource management for the past few decades, but the provision of appropriate and effective nutrient assessment and management continues to be a challenge for water resource managers and policy makers. Difficulties in assessment and management are due in part to the fact that nutrients in rivers may originate from a variety of sources, take numerous pathways and transform into other substances. This publication presents the application of isotope techniques as a powerful tool for evaluating the sources, pathways, transformation, and fate of nutrients in river systems, focusing on nitrogen, phosphorus and carbon containing substances. Eleven researchers using various isotope techniques for different aspects of nutrient studies and two IAEA officers met in a technical meeting and discussed a publication that could assist water resource managers in dealing with nutrient assessment and management issues in river systems. These researchers also recognized the need for careful consideration in selecting appropriate isotope techniques in view of not only technical, but also financial, human resources and logistical capabilities, among others. These contributors are listed as major authors in the later pages of this document. This publication aims at serving water resource managers as a guidebook on the application of isotope techniques in nutrient assessment and management, but it is also expected to be of practical aid for other interested and concerned individuals and organization.

  18. Application of Isotope Techniques for Assessing Nutrient Dynamics in River Basins

    International Nuclear Information System (INIS)

    2013-05-01

    Nutrients are necessary for the growth and survival of animals, plants and other organisms. However, industrial, agricultural and urban development has dramatically increased nutrient levels in river systems, including nitrogen and phosphorus containing substances, degrading water quality, causing acidification and eutrophication and affecting aquatic ecosystems. Nutrient assessment and management in river systems has been an important part of water resource management for the past few decades, but the provision of appropriate and effective nutrient assessment and management continues to be a challenge for water resource managers and policy makers. Difficulties in assessment and management are due in part to the fact that nutrients in rivers may originate from a variety of sources, take numerous pathways and transform into other substances. This publication presents the application of isotope techniques as a powerful tool for evaluating the sources, pathways, transformation, and fate of nutrients in river systems, focusing on nitrogen, phosphorus and carbon containing substances. Eleven researchers using various isotope techniques for different aspects of nutrient studies and two IAEA officers met in a technical meeting and discussed a publication that could assist water resource managers in dealing with nutrient assessment and management issues in river systems. These researchers also recognized the need for careful consideration in selecting appropriate isotope techniques in view of not only technical, but also financial, human resources and logistical capabilities, among others. These contributors are listed as major authors in the later pages of this document. This publication aims at serving water resource managers as a guidebook on the application of isotope techniques in nutrient assessment and management, but it is also expected to be of practical aid for other interested and concerned individuals and organization.

  19. Quantitative amino acid profiling and stable isotopically labeled amino acid tracer enrichment used for in vivo human systemic and tissue kinetics measurements

    DEFF Research Database (Denmark)

    Bornø, Andreas; van Hall, Gerrit

    2014-01-01

    An important area within clinical functional metabolomics is in vivo amino acid metabolism and protein turnover measurements for which accurate amino acid concentrations and stable isotopically labeled amino acid enrichments are mandatory not the least when tissue metabolomics is determined....... The present study describes a new sensitive liquid chromatography tandem mass-spectrometry method quantifying 20 amino acids and their tracer(s) ([ring-(13)C6]/D5Phenylalanine) in human plasma and skeletal muscle specimens. Before analysis amino acids were extracted and purified via deprotonization....../ion exchange, derivatized using a phenylisothiocyanate reagent and each amino acid was quantitated with its own stable isotopically labeled internal standard (uniformly labeled-(13)C/(15)N). The method was validated according to general recommendations for chromatographic analytical methods. The calibration...

  20. Dual isotope study of iodine-125 and indium-111-labeled antibody in athymic mice

    Energy Technology Data Exchange (ETDEWEB)

    Carney, P.L.; Rogers, P.E.; Johnson, D.K. (Abbott Laboratories, Abbott Park, IL (USA))

    1989-03-01

    Monoclonal antibody B72.3 was coupled to a benzylisothiocyanate derivative of diethylenetriaminepentaacetic acid (DTPA). The maximum substitution achievable without loss of immunoreactivity was three DTPA groups per immunoglobulin molecule. The resulting conjugate was labeled with {sup 111}In by brief incubation with {sup 111}InCl{sub 3}, giving a mean radiochemical yield of {sup 111}In-labeled antibody of 96%. The ({sup 111}In)B72.3 preparation was mixed with an ({sup 125}I) B72.3 preparation, obtained by the chloramine-T method, and the mixture administered to athymic mice bearing subcutaneous LS174T colon carcinoma xenografts. There were no significant differences (p greater than 0.1) in the biodistributions of the two labels at 1, 2, 5, and 7 days postinjection. These results are contrasted with prior studies showing elevated levels of {sup 111}In in liver, spleen, and kidneys using B72.3-DTPA conjugates prepared via the bicyclic anhydride. It is concluded that protein cross-linking and/or the formation of unstable chelate sites in anhydride coupled conjugates underlie these disparities.

  1. Dual isotope study of iodine-125 and indium-111-labeled antibody in athymic mice

    International Nuclear Information System (INIS)

    Carney, P.L.; Rogers, P.E.; Johnson, D.K.

    1989-01-01

    Monoclonal antibody B72.3 was coupled to a benzylisothiocyanate derivative of diethylenetriaminepentaacetic acid (DTPA). The maximum substitution achievable without loss of immunoreactivity was three DTPA groups per immunoglobulin molecule. The resulting conjugate was labeled with 111 In by brief incubation with 111 InCl 3 , giving a mean radiochemical yield of 111 In-labeled antibody of 96%. The [ 111 In]B72.3 preparation was mixed with an [ 125 I] B72.3 preparation, obtained by the chloramine-T method, and the mixture administered to athymic mice bearing subcutaneous LS174T colon carcinoma xenografts. There were no significant differences (p greater than 0.1) in the biodistributions of the two labels at 1, 2, 5, and 7 days postinjection. These results are contrasted with prior studies showing elevated levels of 111 In in liver, spleen, and kidneys using B72.3-DTPA conjugates prepared via the bicyclic anhydride. It is concluded that protein cross-linking and/or the formation of unstable chelate sites in anhydride coupled conjugates underlie these disparities

  2. Isotope-labeling studies on the formation pathway of acrolein during heat processing of oils.

    Science.gov (United States)

    Ewert, Alice; Granvogl, Michael; Schieberle, Peter

    2014-08-20

    Acrolein (2-propenal) is classified as a foodborne toxicant and was shown to be present in significant amounts in heated edible oils. Up to now, its formation was mainly suggested to be from the glycerol part of triacylglycerides, although a clear influence of the unsaturation of the fatty acid moiety was also obvious in previous studies. To unequivocally clarify the role of the glycerol and the fatty acid parts in acrolein formation, two series of labeled triacylglycerides were synthesized: [(13)C(3)]-triacylglycerides of stearic, oleic, linoleic, and linolenic acid and [(13)C(54)]-triacylglycerides with labeled stearic, oleic, and linoleic acid, but with unlabeled glycerol. Heating of each of the seven intermediates singly in silicon oil and measurement of the formed amounts of labeled and unlabeled acrolein clearly proved the fatty acid backbone as the key precursor structure. Enzymatically synthesized pure linoleic acid and linolenic acid hydroperoxides were shown to be the key intermediates in acrolein formation, thus allowing the discussion of a radical-induced reaction pathway leading to the formation of the aldehyde. Surprisingly, although several oils contained high amounts of acrolein after heating, deep-fried foods themselves, such as donuts or French fries, were low in the aldehyde.

  3. Secretory overexpression and isotopic labeling of the chimeric relaxin family peptide R3/I5 in Pichia pastoris.

    Science.gov (United States)

    Guo, Yu-Qi; Wu, Qing-Ping; Shao, Xiao-Xia; Shen, Ting; Liu, Ya-Li; Xu, Zeng-Guang; Guo, Zhan-Yun

    2015-06-01

    Relaxin family peptides are a group of peptide hormones with divergent biological functions. Mature relaxin family peptides are typically composed of two polypeptide chains with three disulfide linkages, rendering their preparation a challenging task. In the present study, we established an efficient approach for preparation of the chimeric relaxin family peptide R3/I5 through secretory overexpression in Pichia pastoris and in vitro enzymatic maturation. A designed single-chain R3/I5 precursor containing the B-chain of human relaxin-3 and the A-chain of human INSL5 was overexpressed in PichiaPink strain 1 by high-density fermentation in a two-liter fermenter, and approximately 200 mg of purified precursor was obtained from one liter of the fermentation supernatant. We also developed an economical approach for preparation of the uniformly (15)N-labeled R3/I5 precursor by culturing in shaking flasks, and approximately 15 mg of purified (15)N-labeled precursor was obtained from one liter of the culture supernatant. After purification by cation ion-exchange chromatography and reverse-phase high performance liquid chromatography, the R3/I5 precursor was converted to the mature two-chain form by sequential treatment with endoproteinase Lys-C and carboxypeptidase B. The mature R3/I5 peptide had an α-helix-dominated conformation and retained full receptor-binding and receptor activation activities. Thus, Pichia overexpression was an efficient approach for sample preparation and isotopic labeling of the chimeric R3/I5 peptide. This approach could also be extended to the preparation of other relaxin family peptides in future studies.

  4. Expression and isotopic labelling of the potassium channel blocker ShK toxin as a thioredoxin fusion protein in bacteria.

    Science.gov (United States)

    Chang, Shih Chieh; Galea, Charles A; Leung, Eleanor W W; Tajhya, Rajeev B; Beeton, Christine; Pennington, Michael W; Norton, Raymond S

    2012-10-01

    The polypeptide toxin ShK is a potent blocker of Kv1.3 potassium channels, which play a crucial role in the activation of human effector memory T-cells (T(EM)). Selective blockers constitute valuable therapeutic leads for the treatment of autoimmune diseases mediated by T(EM) cells, such as multiple sclerosis, rheumatoid arthritis, and type-1 diabetes. We have established a recombinant peptide expression system in order to generate isotopically-labelled ShK and various ShK analogues for in-depth biophysical and pharmacological studies. ShK was expressed as a thioredoxin fusion protein in Escherichia coli BL21 (DE3) cells and purified initially by Ni²⁺ iminodiacetic acid affinity chromatography. The fusion protein was cleaved with enterokinase and purified to homogeneity by reverse-phase HPLC. NMR spectra of ¹⁵N-labelled ShK were similar to those reported previously for the unlabelled synthetic peptide, confirming that recombinant ShK was correctly folded. Recombinant ShK blocked Kv1.3 channels with a K(d) of 25 pM and inhibited the proliferation of human and rat T lymphocytes with a preference for T(EM) cells, with similar potency to synthetic ShK in all assays. This expression system also enables the efficient production of ¹⁵N-labelled ShK for NMR studies of peptide dynamics and of the interaction of ShK with Kv1.3 channels. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Plasma Protein Turnover Rates in Rats Using Stable Isotope Labeling, Global Proteomics, and Activity-Based Protein Profiling

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Jordan N.; Tyrrell, Kimberly J.; Hansen, Joshua R.; Thomas, Dennis G.; Murphree, Taylor A.; Shukla, Anil K.; Luders, Teresa; Madden, James M.; Li, Yunying; Wright, Aaron T.; Piehowski, Paul D.

    2017-12-06

    Protein turnover is important for general health on cellular and organism scales providing a strategy to replace old, damaged, or dysfunctional proteins. Protein turnover also informs of biomarker kinetics, as a better understanding of synthesis and degradation of proteins increases the clinical utility of biomarkers. Here, turnover rates of plasma proteins in rats were measured in vivo using a pulse-chase stable isotope labeling experiment. During the pulse, rats (n=5) were fed 13C6-labeled lysine (“heavy”) feed for 23 days to label proteins. During the chase, feed was changed to an unlabeled equivalent feed (“light”), and blood was repeatedly sampled from rats over 10 time points for 28 days. Plasma samples were digested with trypsin, and analyzed with liquid chromatography-tandem mass spectrometry (LC-MS/MS). MaxQuant was used to identify peptides and proteins, and quantify heavy:light lysine ratios. A system of ordinary differential equations was used to calculate protein turnover rates. Using this approach, 273 proteins were identified, and turnover rates were quantified for 157 plasma proteins with half-lives ranging 0.3-103 days. For the ~70 most abundant proteins, variability in turnover rates among rats was low (median coefficient of variation: 0.09). Activity-based protein profiling was applied to pooled plasma samples to enrich serine hydrolases using a fluorophosphonate (FP2) activity-based probe. This enrichment resulted in turnover rates for an additional 17 proteins. This study is the first to measure global plasma protein turnover rates in rats in vivo, measure variability of protein turnover rates in any animal model, and utilize activity-based protein profiling for enhancing measurements of targeted, low-abundant proteins, such as those commonly used as biomarkers. Measured protein turnover rates will be important for understanding of the role of protein turnover in cellular and organism health as well as increasing the utility of protein

  6. Applications of isotopic labeling to the study of Friedel-Crafts reactions

    International Nuclear Information System (INIS)

    Roberts, R.M.; Gibson, T.L.

    1980-01-01

    A number of Friedel-Crafts processes that have been examined by the use of isotopic methods are considered: (1) the interaction of a Lewis acid with an acyl or alkyl halide; (2) the rearrangements of an alkylating species resulting from such an interaction, as observed in the arene products or in recovered starting materials; and (3) isomerizations of alkylation products which can occur in the presence of Lewis acids including (a) the alkylbenzene rearrangement (internal, of side chains), (b) disproportionation (intermolecular alkyl transfers), and (c) reorientations, which may be the results of either intra- or intermolecular processes. (Auth.)

  7. International conference on isotopic and nuclear analytical techniques for health and environment. Unedited papers

    International Nuclear Information System (INIS)

    2004-01-01

    The IAEA has been supporting nuclear and isotopic analytical techniques as part of its mandate to foster the peaceful uses of nuclear energy for many years. Nuclear analytical laboratories have been installed and upgraded through technical co-operation assistance in many laboratories of Member States. These techniques, including INAA, CRF, PIXE, stable isotopes and radioisotopes, α, β, γ spectrometry, Moessbauer spectrometry, etc., have been applied to a wide range of subjects with varying success. Nuclear analytical techniques, featuring some intrinsic quality control aspects, such as multi-nuclide analysis, frequently serve as 'reference methods' to crosscheck critical results. As nuclear properties of elements are targeted, matrix problems seem to be negligible. In light of its continued commitment and support, the IAEA organized the International Conference on Isotopic and Nuclear Analytical Techniques for Health and Environment. Out of 220 from 61 countries who applied for participation, 155 official participants and five observers from 47 countries were in attendance, with 67 from 32 developing countries and 21 from international organizations, including the World Health Organization. Eleven plenary sessions were held. Also conducted was a panel discussion on Human Capacity Development Needs in the Areas of Analytical Quality Control Services (AQCS), Radiochemistry and Nutrition. The scientific sessions were divided into several topics, which reflect some of the important activities of the IAEA's Department of Nuclear Sciences and Applications (NA): - Isotopic and nuclear techniques (general); - Metrology and quality assurance in nuclear measurements; - Nuclear analytical techniques for environmental monitoring; - Radioecology; - Environmental monitoring; - Radiological safety of food and water; - Methodological aspects of stable isotope techniques in health and environment; - Applications of isotopic techniques in health and environment; - New

  8. A new combined method of stable isotope-labeling derivatization-ultrasound-assisted dispersive liquid-liquid microextraction for the determination of neurotransmitters in rat brain microdialysates by ultra high performance liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Zheng, Longfang; Zhao, Xian-En; Zhu, Shuyun; Tao, Yanduo; Ji, Wenhua; Geng, Yanling; Wang, Xiao; Chen, Guang; You, Jinmao

    2017-06-01

    In this work, for the first time, a new hyphenated technique of stable isotope-labeling derivatization-ultrasound-assisted dispersive liquid-liquid microextraction has been developed for the simultaneous determination of monoamine neurotransmitters (MANTs) and their biosynthesis precursors and metabolites. The developed method was based on ultra high performance liquid chromatography tandem mass spectrometry detection using multiple-reaction monitoring mode. A pair of mass spectrometry sensitizing reagents, d 0 -10-methyl-acridone-2-sulfonyl chloride and d 3 -10-methyl-acridone-2-sulfonyl chloride, as stable isotope probes was utilized to facilely label neurotransmitters, respectively. The heavy labeled MANTs standards were prepared and used as internal standards for quantification to minimize the matrix effects in mass spectrometry analysis. Low toxic bromobenzene (extractant) and acetonitrile (dispersant) were utilized in microextraction procedure. Under the optimized conditions, good linearity was observed with the limits of detection (S/N>3) and limits of quantification (S/N>10) in the range of 0.002-0.010 and 0.015-0.040nmol/L, respectively. Meanwhile, it also brought acceptable precision (4.2-8.8%, peak area RSDs %) and accuracy (recovery, 96.9-104.1%) results. This method was successfully applied to the simultaneous determination of monoamine neurotransmitters and their biosynthesis precursors and metabolites in rat brain microdialysates of Parkinson's disease and normal rats. This provided a new method for the neurotransmitters related studies in the future. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Improvement of ground water management and protection through the use of isotope and Nuclear Techniques

    International Nuclear Information System (INIS)

    El Samad, O.

    2009-01-01

    To establish nuclear techniques for the study and management of water resources including technology transfer; to develop a national strategy for the use of isotope techniques in water management and development studies; to develop a water mangement framework; to solve problems related to water shortage, overexploitation, management and rapid quality deterioration; to evaluate the sources, recharge rates and renewal of ground water reservoires; to resolve the problems of mixed aquifers, the quantity of mixing and the exchange reactions between groundwater reservoirs and their matrix; to strengthen the role of the CNRS within national instituions and water authorities. (author)

  10. Application of Isotope Techniques in the Assessment of Groundwater Resource in Water Resources Region 10, Philippines

    International Nuclear Information System (INIS)

    Racadio, Charles Darwin T.; Mendoza, Norman DS.; Castañeda, Soledad S.; Abaño, Susan P.; Rongavilla, Luis S.; Castro, Joey

    2015-01-01

    Groundwater has been the primary source of drinking water of about 50% of the people in the Philippines and the numbers continue to rise. However, data and information on groundwater resources are generally spasmodic or sparse in the country. A specific remedy to address this gap is the use of isotope hydrological techniques. A pilot project utilizing this technique was done in Water Resources Region X with the aim of demonstrating the effectiveness and efficiency of these approach in groundwater resources assessment. When optimized, the technique will be replicated in other areas of the country. Groundwater samples from springs deep wells hand pumps and dug wells and river water were collected within the study area from September 2012 to June 2014. Monthly integrated precipitation samples were also collected at different points within the study area from October 2012 to March 2015. Samples were analyzed for stable isotope (δ”2H and δ”1”8O) using Laser Water Isotope Analyzer and tritium for groundwater dating. Results showed that aquifers in the study area are recharged by infiltrated rain during the heavy rainfall moths (May to November for Cagayan-Tagaloan Basin, and December to April for Agusan Basin). Water in Agusan Basin is isotopically enriched compared with the water in Cagayan-Tagaloan Basin. There appears to be interaction between shallow unconfined aquifer and deep semi-confined aquifer in Cagayan de Oro City. Shallow aquifers appear to be recharged by local precipitation. Groundwater in the study area is of Ca-Mg-HCO 3 type, which is characteristic of dynamic water with short residence time. Tritium-helium aging puts the water at ages between 18 to 72 years. Recharged rates of 422 to 625 mm/year were calculated for Cagayan de Oro City.(author)

  11. Application of isotope techniques in hydrology. A comprehensive report from a panel

    International Nuclear Information System (INIS)

    1962-01-01

    Tracers for water are not a new innovation in hydrology, but have been used on a rather limited scale in the past for various purposes. For instance various dyes or salts have been used in tracing underground water in karstic limestone areas and for computing discharge rates of small turbulent rivers like mountain streams. There are, however, several limitations to their use. Dyes employed in tracing underground water are often sorbed by the ground and hence results are qualitative. For discharge measurements in streams the amount of salt needed limits the method to small streams. With the advance of nuclear science, suitable radioactive, isotopes became available which, because of the sensitivity of their detection, could be added in relatively small amounts. Also, when tritium was discovered, it was realized that it is an ideal tracer for water as it can be incorporated into the water molecule. During recent years fractionation effects have been discovered in several stable isotopes present in water which seem promising as tools for obtaining useful hydrological information, especially on the origin of water. Finally, with the discovery of cosmic-ray-produced radioactive isotopes, especially tritium, a method has been found for determining the age of groundwater which is of great value for estimating the age and hence the recharge-rates of groundwater bodies. There is thus, at present, a great variety of isotope techniques available, some of them in the development stage, others more or less ready for application. These various techniques will be discussed below. The presentation of the information follows the pattern envisaged by the Panel. Thus, for each chapter the problem is first presented and then followed by a section called 'discussion' which summarizes the information given by the Panel members and also expresses their opinions. Finally, a section called 'conclusions and recommendations' sums up the present situation and the need for further development

  12. Sediment Origin Determination in the Sub-Catchment of Mistelbach (Austria) using Fatty Acids Biomarkers and Compound-Specific Stable Isotope Techniques

    International Nuclear Information System (INIS)

    Mabit, L.; Chen, X.; Resch, C.; Toloza, A.; Meusburger, K.; Alewell, C.; Gibbs, M.; Klik, A.; Eder, A.; Strauss, P.

    2016-01-01

    Compound-specific stable isotope (CSSI) signatures of inherent soil organic biomarkers allow discriminating and apportioning the source of soil contribution from different land uses. Plant communities label the soil where they grow by exuding organic biomarkers. Although all plants produce the same biomarkers, the stable isotopic signature of those biomarkers is different for each plant species. For agri-environmental investigations, the CSSI technique is based on the measurement of carbon-13 ( 13 C) natural abundance signatures of specific organic compounds such as natural fatty acids (FAs) in the soil. By linking fingerprints of land use to the sediment in deposition zones, this approach has been shown to be a useful technique for determining the source of eroded soil and thereby identifying areas prone to soil degradation. The authors have used this innovative technique to investigate a 3 hectares sub-catchment of Mistelbach situated 60 km north of Vienna. Using the 137 Cs technique, Mabit et al. (2009) reported a local maximum sedimentation rate reaching 20 to 50 t ha -1 yr -1 in the lowest part of this Austrian catchment. To test the ability of the CSSI technique to discriminate different sediment sources of these deposited sediments, representative soil samples from four main agricultural fields of the site were analyzed

  13. Development of new and improved labelling procedures for introducing isotopic hydrogen and carbon-11 into organic compounds

    International Nuclear Information System (INIS)

    Al-Qahtani, M.H.S.

    1999-10-01

    New and improved methods for introducing radioisotopic hydrogen (tritium) and carbon (positron-emitting short-lived carbon-11, t 1/ 2 = 20.4 min) into organic molecules for application in biological research have been explored. In Chapter 1 the applications of radioactive isotopes in biological and clinical research is surveyed, with particular emphasis on the value of β-emitting tritium and positron-emitting carbon-11. In Chapter 2 we report the use of the non-radioactive hydrogen isotope, deuterium, as a surrogate for tritium in the development of microwave-enhanced labelling procedures, based on catalytic hydrogen transfer to olefins (e.g. styrene, styrene derivatives, cinnamic acid and its derivatives). Hydrogen or deuterium donors (e.g. formate salts) were used alone or in combination with other sources (e.g. D 2 O). The method was found to give fully hydrogenated products using very short microwave irradiation times (∼ 2 min) and was highly reproducible. Importantly, the method is environmentally clean, as when extended to tritiated formates little or no radioactive waste is produced. In Chapter 3 we explored the labelling of CGP 62349 {3-[1-(R)-[3-(4-methoxybenzyl)phosphinyl-2-(S)-hydroxy-propyl- amino]ethyl]benzoic acid}, a γ-aminobutyric acid type B (GABA B ) receptor antagonist, with carbon-11 in order to provide a prospective radioligand for medical imaging with positron emission tomography (PET). Labelling agents, [ 11 C]iodomethane and [ 11 C]methyl triflate, prepared by improved methods, were used in the rapid methylation of desmethyl-CGP 62349. Substantially higher radiochemical yields (78%) of [ 11 C]CGP 62349 were achieved by the new methods compared to that produced in a previously published procedure (9%). In addition, the use of [ 11 C]methyl triflate rather than [ 11 C]iodomethane has the advantage of giving a high radiochemical yield and a lower amount of carrier. In Chapter 4 we report on the use of [ 11 C]carbon monoxide as a labelling

  14. Competition for phosphorus: differential uptake from dual-isotope-labeled soil interspaces between shrub and grass

    International Nuclear Information System (INIS)

    Caldwell, M.M.; Eissenstat, D.M.; Richards, J.H.; Allen, M.F.

    1985-01-01

    Two species of Agropyron grass differed strikingly in their capacity to compete for phosphate in soil interspaces shared with a common competitor, the sagebrush Artemisia tridentata. Of the total phosphorus-32 and -33 absorbed by Artemisia, 86% was from the interspace shared with Agropyron spicatum and only 14% from that shared with Agropyron desertorum. Actively absorbing mycorrhizal roots of Agropyron and Artemisia were present in both interspaces, where competition for the labeled phosphate occurred. The results have important implications about the way in which plants compete for resources below ground in both natural plant communities and agricultural intercropping systems

  15. Comparison of a Label-Free Quantitative Proteomic Method Based on Peptide Ion Current Area to the Isotope Coded Affinity Tag Method

    Directory of Open Access Journals (Sweden)

    Young Ah Goo

    2008-01-01

    Full Text Available Recently, several research groups have published methods for the determination of proteomic expression profiling by mass spectrometry without the use of exogenously added stable isotopes or stable isotope dilution theory. These so-called label-free, methods have the advantage of allowing data on each sample to be acquired independently from all other samples to which they can later be compared in silico for the purpose of measuring changes in protein expression between various biological states. We developed label free software based on direct measurement of peptide ion current area (PICA and compared it to two other methods, a simpler label free method known as spectral counting and the isotope coded affinity tag (ICAT method. Data analysis by these methods of a standard mixture containing proteins of known, but varying, concentrations showed that they performed similarly with a mean squared error of 0.09. Additionally, complex bacterial protein mixtures spiked with known concentrations of standard proteins were analyzed using the PICA label-free method. These results indicated that the PICA method detected all levels of standard spiked proteins at the 90% confidence level in this complex biological sample. This finding confirms that label-free methods, based on direct measurement of the area under a single ion current trace, performed as well as the standard ICAT method. Given the fact that the label-free methods provide ease in experimental design well beyond pair-wise comparison, label-free methods such as our PICA method are well suited for proteomic expression profiling of large numbers of samples as is needed in clinical analysis.

  16. Water stable isotope measurements of Antarctic samples by means of IRMS and WS-CRDS techniques

    Science.gov (United States)

    Michelini, Marzia; Bonazza, Mattia; Braida, Martina; Flora, Onelio; Dreossi, Giuliano; Stenni, Barbara

    2010-05-01

    In the last years in the scientific community there has been an increasing interest for the application of stable isotope techniques to several environmental problems such as drinking water safeguarding, groundwater management, climate change, soils and paleoclimate studies etc. For example, the water stable isotopes, being natural tracers of the hydrological cycle, have been extensively used as tools to characterize regional aquifers and to reconstruct past temperature changes from polar ice cores. Here the need for improvements in analytical techniques: the high request for information calls for technologies that can offer a great quantity of analyses in short times and with low costs. Furthermore, sometimes it is difficult to obtain big amount of samples (as is the case for Antarctic ice cores or interstitial water) preventing the possibility to replicate the analyses. Here, we present oxygen and hydrogen measurements performed on water samples covering a big range of isotopic values (from very negative antarctic precipitation to mid-latitude precipitation values) carried out with both the conventional Isotope Ratio Mass Spectrometry (IRMS) technique and with a new method based on laser absorption techniques, the Wavelenght Scanned Cavity Ringdown Spectroscopy (WS-CRDS). This study is focusing on improving the precision of the measurements carried out with WS-CRDS in order to extensively apply this method to Antarctic ice core paleoclimate studies. The WS-CRDS is a variation of the CRDS developed in 1988 by O'Keef and Deacon. In CRDS a pulse of light goes through a box with high reflective inner surfaces; when there is no sample in the box the light beam doesn't find any obstacle in its path, but the reflectivity of the walls is not perfect so eventually there will be an absorption of the light beam; when the sample is injected in the box there is absorption and the difference between the time of absorption without and with sample is proportional to the quantity

  17. Transfer Comparison Study Nitrogen on the Intact and Decapitated Legumes by Using the 15N Labeling Technique

    International Nuclear Information System (INIS)

    Widjayanto, Didik W.

    1998-01-01

    The experiment was done in order to evaluate the N transfer from the intact and decapitated legumes by using the 15 N labeling technique. Seven days after final labeling the above ground biomass from labeled legume species was removed and the remaining stalks capped to prevent regrowth. Twenty days after final labeling (fourteen days after capping) the all treatments were sample and analyzed. The decapitated legumes appeared to transfer more percentage N than the intact legumes. Although both decapitated and intact legumes transferred, the transfer of N did not incur a dry matter and N yield benefit

  18. In Vivo Isotopic Labeling of Symbiotic Bacteria Involved in Cellulose Degradation and Nitrogen Recycling within the Gut of the Forest Cockchafer (Melolontha hippocastani).

    Science.gov (United States)

    Alonso-Pernas, Pol; Bartram, Stefan; Arias-Cordero, Erika M; Novoselov, Alexey L; Halty-deLeon, Lorena; Shao, Yongqi; Boland, Wilhelm

    2017-01-01

    The guts of insects harbor symbiotic bacterial communities. However, due to their complexity, it is challenging to relate a specific symbiotic phylotype to its corresponding function. In the present study, we focused on the forest cockchafer ( Melolontha hippocastani ), a phytophagous insect with a dual life cycle, consisting of a root-feeding larval stage and a leaf-feeding adult stage. By combining in vivo stable isotope probing (SIP) with 13 C cellulose and 15 N urea as trophic links, with Illumina MiSeq (Illumina-SIP), we unraveled bacterial networks processing recalcitrant dietary components and recycling nitrogenous waste. The bacterial communities behind these processes change between larval and adult stages. In 13 C cellulose-fed insects, the bacterial families Lachnospiraceae and Enterobacteriaceae were isotopically labeled in larvae and adults, respectively. In 15 N urea-fed insects, the genera Burkholderia and Parabacteroides were isotopically labeled in larvae and adults, respectively. Additionally, the PICRUSt-predicted metagenome suggested a possible ability to degrade hemicellulose and to produce amino acids of, respectively, 13 C cellulose- and 15 N urea labeled bacteria. The incorporation of 15 N from ingested urea back into the insect body was confirmed, in larvae and adults, by isotope ratio mass spectrometry (IRMS). Besides highlighting key bacterial symbionts of the gut of M. hippocastani , this study provides example on how Illumina-SIP with multiple trophic links can be used to target microorganisms embracing different roles within an environment.

  19. In Vivo Isotopic Labeling of Symbiotic Bacteria Involved in Cellulose Degradation and Nitrogen Recycling within the Gut of the Forest Cockchafer (Melolontha hippocastani

    Directory of Open Access Journals (Sweden)

    Pol Alonso-Pernas

    2017-10-01

    Full Text Available The guts of insects harbor symbiotic bacterial communities. However, due to their complexity, it is challenging to relate a specific symbiotic phylotype to its corresponding function. In the present study, we focused on the forest cockchafer (Melolontha hippocastani, a phytophagous insect with a dual life cycle, consisting of a root-feeding larval stage and a leaf-feeding adult stage. By combining in vivo stable isotope probing (SIP with 13C cellulose and 15N urea as trophic links, with Illumina MiSeq (Illumina-SIP, we unraveled bacterial networks processing recalcitrant dietary components and recycling nitrogenous waste. The bacterial communities behind these processes change between larval and adult stages. In 13C cellulose-fed insects, the bacterial families Lachnospiraceae and Enterobacteriaceae were isotopically labeled in larvae and adults, respectively. In 15N urea-fed insects, the genera Burkholderia and Parabacteroides were isotopically labeled in larvae and adults, respectively. Additionally, the PICRUSt-predicted metagenome suggested a possible ability to degrade hemicellulose and to produce amino acids of, respectively, 13C cellulose- and 15N urea labeled bacteria. The incorporation of 15N from ingested urea back into the insect body was confirmed, in larvae and adults, by isotope ratio mass spectrometry (IRMS. Besides highlighting key bacterial symbionts of the gut of M. hippocastani, this study provides example on how Illumina-SIP with multiple trophic links can be used to target microorganisms embracing different roles within an environment.

  20. Determination of isoniazid concentration in rabbit vertebrae by isotope tracing technique in conjunction with HPLC.

    Science.gov (United States)

    Liu, Peng; Fu, Zhaozong; Jiang, Jianming; Yuan, Liang; Lin, Zhen

    2013-09-01

    Medications compounded with isoniazid (INH) are usually applied to surgical sites at the completion of surgery to locally kill postoperative residual tubercle bacilli. However, the distribution and elimination of INH in the vertebrae in vivo are not known. In this study, isotope tracing was used in conjunction with high-pressure liquid chromatography (HPLC) to address this. INH and technetium-99 m-labeled INH were applied to the vertebrae of rabbits. After 2 and 6 h, osseous tissues containing INH, as determined by radionuclide imaging, were collected for detection with HPLC. The results showed that INH mainly stayed around the vertebrae 6 h after its application and did not permeate widely into the blood or other organs, except for the kidneys. The standard deviations of INH concentrations in the technetium-99 m-INH group were approximately four-fold smaller than those in the INH group. This method of coupling isotope tracing and HPLC can effectively limit experimental error during sample collection, allowing accurate and reliable identification of the concentration levels of INH in osseous tissues in vivo. Copyright © 2013 John Wiley & Sons, Ltd.

  1. Proceedings of the international colloque on the use of isotopic techniques in water and soil resources field

    International Nuclear Information System (INIS)

    Laboratoire de geochimie isotopique et de paleoclimatologie de l'ecole nationale d'ingenieurs, Sfax

    1996-01-01

    This colloque deals with the use of isotopic techniques in water and soil field. 24 papers were read. Experiences of Tunisia, Algeria, Morocco, Mexico and African equator region were exposed. Articles selected for INIS

  2. Introduction to Body Composition Assessment Using the Deuterium Dilution Technique with Analysis of Urine Samples by Isotope Ratio Mass Spectrometry

    International Nuclear Information System (INIS)

    2010-01-01

    The IAEA has fostered the more widespread use of a stable isotope technique to assess body composition in different population groups to address priority areas in public health nutrition in Member States. It has done this by supporting national and regional nutrition projects through its technical cooperation programme and coordinated research projects over many years. This publication was developed by an international group of experts to provide practical hands-on guidance in the use of this technique in settings where analysis of stable isotope ratios in biological samples is to be made by isotope ratio mass spectrometry. The publication is targeted at new users of this technique, for example nutritionists, analytical chemists and other professionals. More detailed information on the theoretical background and the practical applications of state of the art methodologies to monitor changes in body composition can be found in IAEA Human Health Series No. 3, Assessment of Body Composition and Total Energy Expenditure in Humans by Stable Isotope Techniques

  3. Seepage investigation by using Isotope and Geophysical Techniques in Gumti Flood Embankment/Dyke, Comilla

    International Nuclear Information System (INIS)

    Ahmed, N.; Wallin, B. G.; Majumder, R. K.; Mikail, M.; Rahman, M. S.

    2004-06-01

    Gumti Flood Control Embankment/Dyke is vital for irrigation water supply and flood control. Water seepage/leakage and slope failures are the major issues in Gumti earthen dyke. The distinct seepage and slope failure zone were observed at three places (Farizpur, Kathalia and Ebdarpur) along the countryside of left dyke. The isotopic technique has been integrated in the conventional hydrologic investigations. The isotope methodology works essentially by developing a characteristics pattern of the isotopic composition to identify the sources and flow dynamics of seeping/leaking in the dykes. Two sampling campaigns were conducted; one was on October, 2002 and the other was on July, 2003; near the seepage/leakage site for chemical analysis and stable isotopic analysis (''2H and ''1 8 O). Both samplings were done after recession of peak water level in the Gumti river. Interpretation of the hydrochemical data implies that the groundwater near the investigated seepage zones is Na-Ca-HCO 3 type and the river water is Ca-Mg-HCO 3 type. The chlorides content of both groundwater and river water are found mostly similar, indicating mixing between the two water system. The stable isotopes (''2H and ''1 8 O) of groundwater fall on the Meteoric Water Line, ranging the oxygen-18 values from -4.98 to -5.46 per mil and deuterium values from -30.0 to -33.6 per mil. It indicates the recharge from the river water during peak water level in the river Gumti. On the other hand, the stable isotopes of the Gumti river show some evaporation effect, which might have occurred due to stagnation of flowing water in the river. The oxygen-18 and deuterium values for river water range from -3.61 to -4.43 per mil and from -22.30 to -28.48 per mil respectively. These isotope results reflect the hydraulic connectivity between the river water and groundwater through the base of dyke. The earth imaging resistivity survey was carried out in the dry period along the four above mentioned areas of the Gumti

  4. Validation of a simple isotopic technique for the measurement of global and separated renal function

    International Nuclear Information System (INIS)

    Chachati, A.; Meyers, A.; Rigo, P.; Godon, J.P.

    1986-01-01

    Schlegel and Gates described an isotopic method for the measurement of global and separated glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) based on the determination by scintillation camera of the fraction of the injected dose (99mTc-DTPA-[ 131 I]hippuran) present in the kidneys 1-3 min after its administration. This method requires counting of the injected dose and attenuation correction, but no blood or urine sampling. We validated this technique by the simultaneous infusion of inulin and para-amino hippuric acid (PAH) in patients with various levels of renal function (anuric to normal). To better define individual renal function we studied 9 kidneys in patients either nephrectomized or with a nephrostomy enabling separated function measurement. A good correlation between inulin, PAH clearance, and isotopic GFR-ERPF measurement for both global and separate renal function was observed

  5. Perspectives of application of isotope techniques in hydrology in the Syrian Arab Republic

    International Nuclear Information System (INIS)

    Safadi, Chafic

    1986-01-01

    The Syrian Arab territories are located in the warm moderate region, under arid and semi-arid conditions. In spite of the occurrence of main springs and permanent rivers, groundwater resources available are a limiting factor for socio-economic development in several Syrian regions. Investigation and development of these resources are facing serious problems, for which the application of isotope and nuclear techniques could be a significant process and a deterministic tool. So far, and on the light of current applications, the Syrian Arab Republic is already involved in the isotopic hydrology sector, and so it is needful to set up, in the near future, a relevant unit, on a centralized basis covering the concerned authorities demands, under co-ordinated and integrated conditions. The international co-operation would be of a concrete importance in this respect, to provide the mentioned unit with equipment, training of scientific and technical staff, and consultancy services during the period of establishment. 14 refs. (Author)

  6. Iron bioavailability studies as assessed by intrinsic and extrinsic labeling techniques

    International Nuclear Information System (INIS)

    Johnson, C.D.

    1985-01-01

    Although soybeans are a rich source of iron and incorporation of soy protein into diets is increasing, the presence of phytate or fiber endogenous to the seeds may inhibit total iron absorption from diets including soy protein. Four studies on iron bioavailability as assessed by intrinsic and extrinsic labeling techniques in rats were completed. The effect of previous dietary protein on the absorption of intrinsically 59 Fe labeled defatted soy flour was determined in rats. The results indicated that the type of dietary protein (animal vs. plant) in pre-test diets would have little influence on iron absorption from a single soy protein test meal. Therefore, adaptation of soy protein does not improve bioavailability of iron. Soybean hulls were investigated as a source of iron fortification in bread. The results indicated that retention of 59 Fe from white bread baked with soy hulls did not differ from white bread fortified with bakery grade ferrous sulfate. The effect of endogenous soybean phytate on iron absorption in rats was measured using seeds of varying phytate content and intrinsically labeled with 59 Fe. Increasing concentration of phytate in whole soybean flour had no significant effect on iron absorption

  7. Application of Environmental Isotope and Hydrogeochemical Techniques in Investigating the Geothermal Resources

    International Nuclear Information System (INIS)

    Kamarudin Samuding; Noor Akhmal Kamarudin; Mohd Shahrizal Mohamed Sharifodin; Azrul Arifin; Kamaruzaman Mohamad

    2016-01-01

    An investigation of geothermal resources at Ulu Slim has been carried out using integrated environmental isotope and hydro-geochemical techniques. Environmental isotope Oxygen-18 ("1"8O) and Deuterium ("2H) and Tritium ("3H) were used to identify the recharge zones and origin of the water, whereas the hydro-geochemical technique is used to determine the water type and the level of solutes in the geothermal waters out flowing at the surface as well as in shallow and deep groundwater system. The sampling programme includes precipitations, surface waters, hot springs, groundwater for isotopes and hydro-geochemical analyses. The plot graph of (δ"1"8O vs δ"2H) show that the stable isotope composition of hot spring is relatively depleted as compared to surface water and groundwater. This indicates that the recharge of the hot spring is likely to occur from farther and higher elevation areas of the geothermal system. Tritium content in hot spring, groundwater and surface water is ranged between 0.85 - 0.92 TU, 0.81- 1.05 TU, 1.60-2.07 TU respectively. The values of TU in hot spring and groundwater is seen similar suggests that these samples are older than the surface water. Based on the plot of Ternary Major Anion diagram (Cl-SO_4- HCO_3) and Tri-linear Piper diagram, all the water samples are identified from the type of bicarbonate (HCO_3). Nevertheless, the content of sodium (Na) in hot spring is detected relatively higher as compared to surface water. Tri-linear Piper diagram also shows that there is no mixing process between hot spring and surface water. (author)

  8. Use of isotopes techniques during the life cycle of dams and reservoirs: cases in Latin American

    International Nuclear Information System (INIS)

    Leon, S.H.

    2006-01-01

    In fact, the combined use of isotope and conventional techniques is considered a reliable tool for studying problems related to dam safety and has become a new culture for civil / dam engineers, hydro geologists and researchers who involve in water resource management fields. The use of natural (environmental) and artificial isotopes as tracers together with systematic analyses of the hydrochemistry, electrical conductivity and temperature profiles data during the investigation and monitoring of leakage and seepage in dams and reservoirs are now becoming popular among the dam owners in seeking the best solution for dam related problems. Many studies and experiences worldwide on effective dam management programmes have indicated that any investigation about leakages and seepages are not possible to be accomplished successfully without synergic application of the conventional technologies and isotopic techniques. The major advancement in this area is the measurements study for establishment of baseline hydrogeology at each hydraulic work project like dams and reservoirs. The parameters include hydro chemicals, isotopic and geologic in each basin, river, reservoir, dams, tunnels and groundwater which provide high value information for decision making during all the stages in the life cycle of the dams. Many hydroelectric and water supply projects in latin america apply these investigation strategies. The main target is to investigate and understand the water movement around the dam and its vicinity. Then the specialised work teams will decide for the effective and economic monitoring activities and the implementation of the recommended remedial measures to ensure high standards of safety and security of the large dams and reservoirs. A typical example of specific leakage investigation of la Honda dam is briefly discussed. (Author)

  9. Evaluation of salt tolerance in wheat genotypes using growth and carbon isotopes discrimination technique

    International Nuclear Information System (INIS)

    Shirazi, M.U.; Khan, M.A.; Mujtaba, S.M.; Shereen, A.

    2015-01-01

    Studies were conducted in green house to select suitable salt tolerant wheat genotypes on the basis of growth performance and carbon isotopes discrimination (CID) technique. Nine newly developed double haploids (DH) wheat genotypes were tested under gravel culture, along with salt tolerant (LU-26s) and high yielding (Sarsabz) checks. The crop was irrigated by non-saline (control) and saline (12dS/m) water and raised up to maturity, growth parameters (i.e. plant height, plant biomass, productive tillers, spike length, number of spiklets/spike, number of grains / spike, grain weight/ spike and grain yield/ 15 plants) were recorded after harvesting. Plant samples (straw) were collected and were analyzed for carbon isotopic ratio (C12/ C13) from IAEA laboratories Vienna Austria. The data showed that there was significant decrease in all the growth parameters due to salinity. On the basis of performance in different growth parameters it was found that wheat genotypes V3-DH, V9-DH, V10-DH, V13-DH, and LU-26s had good response at 12dSm-1, thus can be categorized as better performing genotypes. Studies on carbon isotopes discrimination (CID) showed a decreasing trend under salinity. Mean CID values were 20.86 and 17.49 under two environments (non saline and saline, respectively), showing an overall 19% decrease under salinity. Generally the wheat genotypes having higher grain yield also had high carbon isotopes discrimination (CID). The relationship between grain yield and CID was positive (R2 = 0.695). The genotypes V10-DH, V13-DH with lower decrease in CID (i.e. 1.2 and 11.0%, respectively), also had high grain yield under salinity. Therefore the studies suggest that we can include CID technique as one of the selection criteria for salt tolerance. (author)

  10. Understanding N timing in corn yield and fertilizer N recovery: An insight from an isotopic labeled-N determination

    Science.gov (United States)

    de Almeida, Rodrigo Estevam Munhoz; Pierozan Junior, Clovis; Lago, Bruno Cocco; Trivelin, Paulo Cesar Ocheuze

    2018-01-01

    Early fertilizer nitrogen (N) application on cover crops or their residues during the off-season is a practice adopted in Brazil subtropical conditions under no-tillage corn (Zea mays L.) systems. However, the effect of early N application on yield, plant N content, and N recovery efficiency (NRE) for corn is not yet well documented. Five fertilizer N timings in an oat-corn system were evaluated in two studies utilizing an isotopic-labeled N determination, 15N isotope. The N fertilization timings were: (i) oat tillering, (ii) 15 days before corn planting time, over the oat residues, (iii) at corn planting time, (iv) in-season at the three-leaf growth stage (V3), and (v) in-season split application at V3 and six-leaf (V6) growth stages. Based on the statistical analysis, the N fertilization timings were separated into three groups: 1) N-OATS, designated to N applied at oat; 2) N-PLANT, referred to pre-plant and planting N applications; and 3) N-CORN, designated to in-season corn N applications. Corn yield was not affected by the N fertilization timing. However, the N-CORN N fertilization timings enhanced NRE by 17% and 35% and final N recovery system (plant plus soil) by 16% and 24% all relative to N-OATS and N-PLANT groups, respectively. Overall, N-OATS resulted in the largest N derived from fertilizer (NDFF) amount in the deeper soil layer, in overall a delta of 10 kg N ha-1 relative to the rest of the groups. Notwithstanding corn yield was not affected, early N fertilization under subtropical conditions is not a viable option since NRE was diminished and the non-recovery N increased relative to the in-season N applications. PMID:29462178

  11. Understanding N timing in corn yield and fertilizer N recovery: An insight from an isotopic labeled-N determination.

    Science.gov (United States)

    Maciel de Oliveira, Silas; Almeida, Rodrigo Estevam Munhoz de; Ciampitti, Ignacio A; Pierozan Junior, Clovis; Lago, Bruno Cocco; Trivelin, Paulo Cesar Ocheuze; Favarin, José Laércio

    2018-01-01

    Early fertilizer nitrogen (N) application on cover crops or their residues during the off-season is a practice adopted in Brazil subtropical conditions under no-tillage corn (Zea mays L.) systems. However, the effect of early N application on yield, plant N content, and N recovery efficiency (NRE) for corn is not yet well documented. Five fertilizer N timings in an oat-corn system were evaluated in two studies utilizing an isotopic-labeled N determination, 15N isotope. The N fertilization timings were: (i) oat tillering, (ii) 15 days before corn planting time, over the oat residues, (iii) at corn planting time, (iv) in-season at the three-leaf growth stage (V3), and (v) in-season split application at V3 and six-leaf (V6) growth stages. Based on the statistical analysis, the N fertilization timings were separated into three groups: 1) N-OATS, designated to N applied at oat; 2) N-PLANT, referred to pre-plant and planting N applications; and 3) N-CORN, designated to in-season corn N applications. Corn yield was not affected by the N fertilization timing. However, the N-CORN N fertilization timings enhanced NRE by 17% and 35% and final N recovery system (plant plus soil) by 16% and 24% all relative to N-OATS and N-PLANT groups, respectively. Overall, N-OATS resulted in the largest N derived from fertilizer (NDFF) amount in the deeper soil layer, in overall a delta of 10 kg N ha-1 relative to the rest of the groups. Notwithstanding corn yield was not affected, early N fertilization under subtropical conditions is not a viable option since NRE was diminished and the non-recovery N increased relative to the in-season N applications.

  12. Can groundwater sampling techniques used in monitoring wells influence methane concentrations and isotopes?

    Science.gov (United States)

    Rivard, Christine; Bordeleau, Geneviève; Lavoie, Denis; Lefebvre, René; Malet, Xavier

    2018-03-06

    Methane concentrations and isotopic composition in groundwater are the focus of a growing number of studies. However, concerns are often expressed regarding the integrity of samples, as methane is very volatile and may partially exsolve during sample lifting in the well and transfer to sampling containers. While issues concerning bottle-filling techniques have already been documented, this paper documents a comparison of methane concentration and isotopic composition obtained with three devices commonly used to retrieve water samples from dedicated observation wells. This work lies within the framework of a larger project carried out in the Saint-Édouard area (southern Québec, Canada), whose objective was to assess the risk to shallow groundwater quality related to potential shale gas exploitation. The selected sampling devices, which were tested on ten wells during three sampling campaigns, consist of an impeller pump, a bladder pump, and disposable sampling bags (HydraSleeve). The sampling bags were used both before and after pumping, to verify the appropriateness of a no-purge approach, compared to the low-flow approach involving pumping until stabilization of field physicochemical parameters. Results show that methane concentrations obtained with the selected sampling techniques are usually similar and that there is no systematic bias related to a specific technique. Nonetheless, concentrations can sometimes vary quite significantly (up to 3.5 times) for a given well and sampling event. Methane isotopic composition obtained with all sampling techniques is very similar, except in some cases where sampling bags were used before pumping (no-purge approach), in wells where multiple groundwater sources enter the borehole.

  13. Various analytical techniques used for the measurement of isotopic purity of heavy water at Madras Atomic Power Station

    International Nuclear Information System (INIS)

    Satyanarayanan, V.; Umapathy, P.; Bhaskaran, R.; Nagarajan, J.; Pradeep, Jeena; Ayyar, S.R.

    2008-01-01

    The paper deals with the various techniques used for the measurement of isotopic purity of heavy water samples received from different sources viz. reactor systems, heavy water upgrading plant and fresh consignment from heavy water production plants. Heavy water is used in PHWRs as moderator and primary coolant. Isotopic Purity is an important parameter to be monitored/analysed regularly for both the systems. There is a minimum isotopic purity level to be maintained in the moderator system due to neutron economy/fuel burnup and in the case of coolant system the measurement is of paramount importance due to its safety considerations. The selection of the method of analysis depends on the isotopic range. The techniques used to measure the isotopic purity of heavy water are a) Infrared Spectrophotometry b) Refractometry c) Densitometry. Infrared spectrometer uses the property of molecular absorption of IR radiation by HOD species and the absorbance is the measure of isotopic purity. This technique is generally used for measuring high isotopic (80-99.98%) and low isotopic samples. Refractometer uses the property of refractive index of heavy water. The difference in refractive indices of light water and heavy water is 0.0048. A 1 % change in D 2 O concentration would thus equal to 0.000048 refractive index units. This method is used for determining the approximate isotopic value of a sample. Density meter uses the property of difference in densities of light and heavy water. The difference in density of 99.999% D 2 O and light water is 0.107540 which covers the whole range of interest. The experience gained with these techniques in the measurements of isotopic purity of various samples are presented in this paper. (author)

  14. Quantifying the effect of plant growth on litter decomposition using a novel, triple-isotope label approach

    Science.gov (United States)

    Ernakovich, J. G.; Baldock, J.; Carter, T.; Davis, R. A.; Kalbitz, K.; Sanderman, J.; Farrell, M.

    2017-12-01

    Microbial degradation of plant detritus is now accepted as a major stabilizing process of organic matter in soils. Most of our understanding of the dynamics of decomposition come from laboratory litter decay studies in the absence of plants, despite the fact that litter decays in the presence of plants in many native and managed systems. There is growing evidence that living plants significantly impact the degradation and stabilization of litter carbon (C) due to changes in the chemical and physical nature of soils in the rhizosphere. For example, mechanistic studies have observed stimulatory effects of root exudates on litter decomposition, and greenhouse studies have shown that living plants accelerate detrital decay. Despite this, we lack a quantitative understanding of the contribution of living plants to litter decomposition and how interactions of these two sources of C build soil organic matter (SOM). We used a novel triple-isotope approach to determine the effect of living plants on litter decomposition and C cycling. In the first stage of the experiment, we grew a temperate grass commonly used for forage, Poa labillardieri, in a continuously-labelled atmosphere of 14CO2 fertilized with K15NO3, such that the grass biomass was uniformly labelled with 14C and 15N. In the second stage, we constructed litter decomposition mescososms with and without a living plant to test for the effect of a growing plant on litter decomposition. The 14C/15N litter was decomposed in a sandy clay loam while a temperate forage grass, Lolium perenne, grew in an atmosphere of enriched 13CO2. The fate of the litter-14C/15N and plant-13C was traced into soil mineral fractions and dissolved organic matter (DOM) over the course of nine weeks using four destructive harvests of the mesocosms. Our preliminary results suggest that living plants play a major role in the degradation of plant litter, as litter decomposition was greater, both in rate and absolute amount, for soil mesocosms

  15. Nitrogen fixation by free-living microorganisms in tropical rice soils using labelled fertilizer. Part of a coordinated programme on isotope techniques in studies of biological nitrogen fixation for the dual purpose of increasing crop production and decreasing nitrogen fertilizer use to conserve the environment

    International Nuclear Information System (INIS)

    Rao, V.R.

    1981-11-01

    Both acetylene-reduction and 15 N techniques were used to study heterotrophic N fixation in the rhizosphere of rice plants. Soils subjected to flooding in 4 soil types in both greenhouse and the field were found to stimulate greater heterotrophic nitrogen fixation than moist soils. The addition of organic materials, in particular, cellulose and rice straw, in general, enhanced nitrogen fixed by heterotrophic organisms living in the rhizosphere of rice plants. The highest amount of N fixed was 38 kg N/ha, and was obtained in a flooded lateritic soil to which had been added cellulose. Heterotrophic nitrogen fixation was influenced by soil type. In this study, the lowest value for fixed N was recorded in an acid sulphate soil of low pH. The addition of increasing amounts of inorganic nitrogen fertilizer in the form of ammonium sulphate suppressed rhizospheric nitrogen fixation in all soils, but the extent of suppression differed in the different soils. Benomyl fungicide and methyl carbamate insecticide had a stimulatory effect on heterotrophic nitrogen fixation in soils under rice roots. Different rice cultivars stimulated strains of Azospirillum to varying extent, and thus did not fix nitrogen to the same extent. It is thus possible that varieties of rice could be selected on the basis of their ability to support non-symbiotic N fixation in their rhizosphere

  16. Auto-inducing media for uniform isotope labeling of proteins with {sup 15}N, {sup 13}C and {sup 2}H

    Energy Technology Data Exchange (ETDEWEB)

    Guthertz, Nicolas [Institute of Cancer Research, Division of Structural Biology (United Kingdom); Klopp, Julia; Winterhalter, Aurélie; Fernández, César; Gossert, Alvar D., E-mail: alvar.gossert@novartis.com [Novartis Institutes for BioMedical Research (Switzerland)

    2015-06-15

    Auto-inducing media for protein expression offer many advantages like robust reproducibility, high yields of soluble protein and much reduced workload. Here, an auto-inducing medium for uniform isotope labelling of proteins with {sup 15}N, {sup 13}C and/or {sup 2}H in E. coli is presented. So far, auto-inducing media have not found widespread application in the NMR field, because of the prohibitively high cost of labeled lactose, which is an essential ingredient of such media. Here, we propose using lactose that is only selectively labeled on the glucose moiety. It can be synthesized from inexpensive and readily available substrates: labeled glucose and unlabeled activated galactose. With this approach, uniformly isotope labeled proteins were expressed in unattended auto-inducing cultures with incorporation of {sup 13}C, {sup 15}N of 96.6 % and {sup 2}H, {sup 15}N of 98.8 %. With the present protocol, the NMR community could profit from the many advantages that auto-inducing media offer.

  17. Radiation Oxidation Mechanisms in Polyolefins Studied by C-13 Isotopic Labeling

    International Nuclear Information System (INIS)

    Clough, R.L.

    2006-01-01

    Control of oxidative degradation is a critical consideration in most applications involving polymers and radiation. In radiation crosslinking or sterilization, or in the use of polymers in radiation environments (such as nuclear plants), the objective is to minimize degradation as much as possible. In other applications, a controlled, partial degradation is desired to alter processing properties, or to enhance adhesion or solubility. To gain more understanding of the complex processes of radiation oxidation, samples of one important commercial polyolefin, polypropylene, were synthesized in which the three different carbon atoms along the chain were selectively labeled with carbon-13. These samples were subjected to radiation under inert and air atmospheres, and to post-irradiation thermal exposure in air at various temperatures. Analysis of macromolecular radiation-oxidation products was carried out using 13 C NMR and FTIR. Time-dependent plots of oxidation products have been obtained from the NMR measurements, including the post-irradiation oxidation of a sample held at room temperature in air that has been monitored for 2 years. Analysis of volatile oxidation products (CO, CO 2 , and small organic molecules) was accomplished with gas chromatography / mass spectroscopy. The position of the 13 C labels in the degradation products, have been traced back to their positions of origin on the macromolecule, providing insights into the chemical reaction mechanisms through which the products were formed. The major solid-phase products include peroxides and alcohols, both formed at tertiary carbon sites along the chain. Other products include methyl ketones, acids, esters, peresters, and hemiketals formed from reaction at the tertiary carbon, together with in-chain ketones and esters from reaction at the secondary chain carbon. No evidence is found of macromolecular products arising from reactions at the methyl side chain. Significant temperature-dependent differences are

  18. Effects of sample injection amount and time-of-flight mass spectrometric detection dynamic range on metabolome analysis by high-performance chemical isotope labeling LC-MS.

    Science.gov (United States)

    Zhou, Ruokun; Li, Liang

    2015-04-06

    The effect of sample injection amount on metabolome analysis in a chemical isotope labeling (CIL) liquid chromatography-mass spectrometry (LC-MS) platform was investigated. The performance of time-of-flight (TOF) mass spectrometers with and without a high-dynamic-range (HD) detection system was compared in the analysis of (12)C2/(13)C2-dansyl labeled human urine samples. An average of 1635 ± 21 (n = 3) peak pairs or putative metabolites was detected using the HD-TOF-MS, compared to 1429 ± 37 peak pairs from a conventional or non-HD TOF-MS. In both instruments, signal saturation was observed. However, in the HD-TOF-MS, signal saturation was mainly caused by the ionization process, while in the non-HD TOF-MS, it was caused by the detection process. To extend the MS detection range in the non-HD TOF-MS, an automated switching from using (12)C to (13)C-natural abundance peaks for peak ratio calculation when the (12)C peaks are saturated has been implemented in IsoMS, a software tool for processing CIL LC-MS data. This work illustrates that injecting an optimal sample amount is important to maximize the metabolome coverage while avoiding the sample carryover problem often associated with over-injection. A TOF mass spectrometer with an enhanced detection dynamic range can also significantly increase the number of peak pairs detected. In chemical isotope labeling (CIL) LC-MS, relative metabolite quantification is done by measuring the peak ratio of a (13)C2-/(12)C2-labeled peak pair for a given metabolite present in two comparative samples. The dynamic range of peak ratio measurement does not need to be very large, as only subtle changes of metabolite concentrations are encountered in most metabolomic studies where relative metabolome quantification of different groups of samples is performed. However, the absolute concentrations of different metabolites can be very different, requiring a technique to provide a wide detection dynamic range to allow the detection of as

  19. Uses of stable isotopes

    International Nuclear Information System (INIS)

    Axente, Damian

    1998-01-01

    The most important fields of stable isotope use with examples are presented. These are: 1. Isotope dilution analysis: trace analysis, measurements of volumes and masses; 2. Stable isotopes as tracers: transport phenomena, environmental studies, agricultural research, authentication of products and objects, archaeometry, studies of reaction mechanisms, structure and function determination of complex biological entities, studies of metabolism, breath test for diagnostic; 3. Isotope equilibrium effects: measurement of equilibrium effects, investigation of equilibrium conditions, mechanism of drug action, study of natural processes, water cycle, temperature measurements; 4. Stable isotope for advanced nuclear reactors: uranium nitride with 15 N as nuclear fuel, 157 Gd for reactor control. In spite of some difficulties of stable isotope use, particularly related to the analytical techniques, which are slow and expensive, the number of papers reporting on this subject is steadily growing as well as the number of scientific meetings organized by International Isotope Section and IAEA, Gordon Conferences, and regional meeting in Germany, France, etc. Stable isotope application development on large scale is determined by improving their production technologies as well as those of labeled compound and the analytical techniques. (author)

  20. Laser desorption ionization mass spectrometry: Recent progress in matrix-free and label-assisted techniques.

    Science.gov (United States)

    Mandal, Arundhoti; Singha, Monisha; Addy, Partha Sarathi; Basak, Amit

    2017-10-13

    The MALDI-based mass spectrometry, over the last three decades, has become an important analytical tool. It is a gentle ionization technique, usually applicable to detect and characterize analytes with high molecular weights like proteins and other macromolecules. The earlier difficulty of detection of analytes with low molecular weights like small organic molecules and metal ion complexes with this technique arose due to the cluster of peaks in the low molecular weight region generated from the matrix. To detect such molecules and metal ion complexes, a four-prong strategy has been developed. These include use of alternate matrix materials, employment of new surface materials that require no matrix, use of metabolites that directly absorb the laser light, and the laser-absorbing label-assisted LDI-MS (popularly known as LALDI-MS). This review will highlight the developments with all these strategies with a special emphasis on LALDI-MS. © 2017 Wiley Periodicals, Inc.

  1. Synthetic techniques of radiopharmaceuticals production labeled with C-11 for PET in cardiology

    Science.gov (United States)

    Dyubkov, V. S.; Ekaeva, I. V.; Katunina, T. A.; Rumyantsev, A. S.; Silchenkov, A. V.; Tuflina, T. V.

    2017-01-01

    Positron emission tomography (PET) and PET-Computerised Tomography (CT) are unique, non-invasive diagnostic techniques, in which the local, temporal and quantitative distributions of radioactive labelled substances are measured to investigate physiological processes. It is well known that PET centre of Bakulev Scientific Centre for Cardiovascular Surgery is the oldest one in Moscow. During more than fifteen years a large number of patients have received PET scans. Due to main stream of Scientific Centre, emphasis is placed on examining the heart functioning. For the diagnosis innervation of the heart muscle a number of radiopharmaceuticals are used, including PET radiopharmaceuticals such as 11C-CGP 12177, 11C-meta-hydroxyephedrine as well as its synthetic analogues labelled with other PET radionuclides (18F, 68Ga). 11C-meta-hydroxyephedrine is one of the most perspective radiopharmaceutical for an investigation of cardiac receptors function due to required materials availability for a radio synthesis in Russia. The main advantage of proposed 11C-meta-hydroxyephedrine synthesis technique is the use of a catalyst which allows one decrease reaction time from 5 minutes to 30 seconds. Obtained results allow one decrease reaction time of methylation and increase radiochemical and technological yields.

  2. Metabolic cartography: experimental quantification of metabolic fluxes from isotopic labelling studies.

    Science.gov (United States)

    O'Grady, John; Schwender, Jörg; Shachar-Hill, Yair; Morgan, John A

    2012-03-01

    For the past decade, flux maps have provided researchers with an in-depth perspective on plant metabolism. As a rapidly developing field, significant headway has been made recently in computation, experimentation, and overall understanding of metabolic flux analysis. These advances are particularly applicable to the study of plant metabolism. New dynamic computational methods such as non-stationary metabolic flux analysis are finding their place in the toolbox of metabolic engineering, allowing more organisms to be studied and decreasing the time necessary for experimentation, thereby opening new avenues by which to explore the vast diversity of plant metabolism. Also, improved methods of metabolite detection and measurement have been developed, enabling increasingly greater resolution of flux measurements and the analysis of a greater number of the multitude of plant metabolic pathways. Methods to deconvolute organelle-specific metabolism are employed with increasing effectiveness, elucidating the compartmental specificity inherent in plant metabolism. Advances in metabolite measurements have also enabled new types of experiments, such as the calculation of metabolic fluxes based on (13)CO(2) dynamic labelling data, and will continue to direct plant metabolic engineering. Newly calculated metabolic flux maps reveal surprising and useful information about plant metabolism, guiding future genetic engineering of crops to higher yields. Due to the significant level of complexity in plants, these methods in combination with other systems biology measurements are necessary to guide plant metabolic engineering in the future.

  3. Pu abundances, concentrations, and isotopics by x- and gamma-ray spectrometry assay techniques

    International Nuclear Information System (INIS)

    Camp, D.C.; Gunnink, R.; Ruhter, W.D.; Prindle, A.L.; Gomes, R.J.

    1986-01-01

    Two x- and gamma-ray systems were recently installed at-line in gloveboxes and will measure Pu solution concentrations from 5 to 105 g/L. These NDA technique, developed and refined over the past decade, are now used domestically and internationally for nuclear material process monitoring and accountability needs. In off- and at-line installations, they can measure solution concentrations to 0.2%. The K-XRFA systems use a transmission source to correct for solution density. The gamma-ray systems use peaks from 59- to 208-keV to determine solution concentrations and relative isotopics. A Pu check source monitors system stability. These two NDA techniques can be combined to form a new, NDA measurement methodology. With the instrument located outside of a glovebox, both relative Pu isotopics and absolute Pu abundances of a sample located inside a glovebox can be measured. The new technique works with either single or dual source excitation; the former for a detector 6 to 20 cm away with no geometric corrections needed; the latter requires geometric corrections or source movement if the sample cannot be measured at the calibration distance. 4 refs., 7 figs., 2 tabs

  4. Microspheres labelled with short-lived isotopes: Development and application for tumors treatment (Experimental study)

    International Nuclear Information System (INIS)

    Drozdovsky, B.Y.; Rosiev, R.A.; Goncharova, A.Y.; Skvortsov, V.G.; Petriev, V.M.; Grigoriev, A.N.; Schischkanov, N.G.

    1997-01-01

    Analysis of the conducted studies strongly suggests the possibility of usage of the domestic protein microspheres as a vehicle for radionuclide. The neutron-activating method of RPP production enables to utilize a broad spectrum of short-living isotopes that can be delivered into the target organ and anchored there for a long time. Good treatment results were obtained in case of the experimentally induced rheumatoid arthritis in rats after intraarticular loading of 165 Dy-hMSA. Mathematical calculations show that homogeneous distribution of RPP in human articulation cavity with the square of 100 cm 2 can be achieved when the quantity of administered particles exceeds 3000. On the example of 165 Dy-hMSA energy characteristic distribution we demonstrated that the absorbed dose for damaged cells at 2mm distance from the radioactive source is 7 times less than the one for a sphere of 2mm diameter. Analysis of dosimetric data in case of intratumoral loading of 165 Dy-hMSA also point out the necessity of the absorbed dose calculation methods taking into account the distance from the source and possible heterogeneity of RPP distribution inside the tumor to be employed. The prolonged RPP detention in the target causing no essential morphological and functional changes was achieved by embolization on the level of septal and interlobular arteries and of efferent arterioles in the animal's renal. The uniformity of microsphere distribution in the organ and their accumulation in tumors depends on the number of particles being administered. Investigations carried out suggest the efficacy of radionuclide therapy application for treatment of oncological and heavy somatic diseases. They also indicate the necessity of further investigations aimed to optimize the usage of microspheres as a radionuclide carrier usage and to work out the criteria of dosimetric planning

  5. Microspheres labelled with short-lived isotopes: Development and application for tumors treatment (Experimental study)

    Energy Technology Data Exchange (ETDEWEB)

    Drozdovsky, B.Y.; Rosiev, R.A.; Goncharova, A.Y.; Skvortsov, V.G.; Petriev, V.M.; Grigoriev, A.N.; Schischkanov, N.G. [Medical Radiological Research Centre RAMS, Kaluga Region, (Russian Federation)

    1997-10-01

    Analysis of the conducted studies strongly suggests the possibility of usage of the domestic protein microspheres as a vehicle for radionuclide. The neutron-activating method of RPP production enables to utilize a broad spectrum of short-living isotopes that can be delivered into the target organ and anchored there for a long time. Good treatment results were obtained in case of the experimentally induced rheumatoid arthritis in rats after intraarticular loading of {sup 165}Dy-hMSA. Mathematical calculations show that homogeneous distribution of RPP in human articulation cavity with the square of 100 cm{sup 2} can be achieved when the quantity of administered particles exceeds 3000. On the example of {sup 165}Dy-hMSA energy characteristic distribution we demonstrated that the absorbed dose for damaged cells at 2mm distance from the radioactive source is 7 times less than the one for a sphere of 2mm diameter. Analysis of dosimetric data in case of intratumoral loading of {sup 165}Dy-hMSA also point out the necessity of the absorbed dose calculation methods taking into account the distance from the source and possible heterogeneity of RPP distribution inside the tumor to be employed. The prolonged RPP detention in the target causing no essential morphological and functional changes was achieved by embolization on the level of septal and interlobular arteries and of efferent arterioles in the animal`s renal. The uniformity of microsphere distribution in the organ and their accumulation in tumors depends on the number of particles being administered. Investigations carried out suggest the efficacy of radionuclide therapy application for treatment of oncological and heavy somatic diseases. They also indicate the necessity of further investigations aimed to optimize the usage of microspheres as a radionuclide carrier usage and to work out the criteria of dosimetric planning 25 refs.

  6. Measurement of loss rates of organic compounds in snow using in situ experiments and isotopically labelled compounds

    Directory of Open Access Journals (Sweden)

    Erika von Schneidemesser

    2012-07-01

    Full Text Available Organic molecular marker compounds are widely used to identify emissions from anthropogenic and biogenic air pollution sources in atmospheric samples and in deposition. Specific organic compounds have been detected in polar regions, but their fate after deposition to snow is poorly characterized. Within this context, a series of exposure experiments were carried out to observe the post-depositional processing of organic compounds under real-world conditions in snow on the surface of the Greenland Ice Sheet, at the Summit research station. Snow was prepared from water spiked with isotopically labelled organic compounds, representative of typical molecular marker compounds emitted from anthropogenic activities. Reaction rate constants and reaction order were determined based on a decrease in concentration to a stable, non-zero, threshold concentration. Fluoranthene-d10, docosane-d46, hexadecanoic acid-d31, docosanoic acid-d43 and azelaic acid-d14 were estimated to have first order loss rates within surface snow with reaction rate constants of 0.068, 0.040, 0.070, 0.067 and 0.047 h−1, respectively. No loss of heptadecane-d36 was observed. Overall, these results suggest that organic contaminants are archived in polar snow, although significant post-depositional losses of specific organic compounds occur. This has implications for the environmental fate of organic contaminants, as well as for ice-core studies that seek to use organic molecular markers to infer past atmospheric loadings, and source emissions.

  7. Production, purification and detergent exchange of isotopically labeled Bacillussubtilis cytochrome b₅₅₈ (SdhC).

    Science.gov (United States)

    Baureder, Michael; Hederstedt, Lars

    2011-11-01

    Cytochrome b₅₅₈ of the gram-positive bacterium Bacillussubtilis is the membrane anchor subunit of the succinate:quinone oxidoreductase of the citric acid cycle. The cytochrome consists of the SdhC polypeptide (202 residues) and two protoheme IX groups that function in transmembrane electron transfer to menaquinone. The general structure of the cytochrome is known from extensive experimental studies and by comparison to Wolinellasuccinogenes fumarate reductase for which the X-ray crystal structure has been determined. Solution state NMR can potentially be used to identify the quinone binding site(s) and study, e.g. redox-linked, dynamics of cytochrome b₅₅₈. In this work we present an efficient procedure for the isolation of preparative amounts of isotopically labeled B. subtilis cytochrome b₅₅₈ produced in Escherichia coli. We have also evaluated several detergents suitable for NMR for their effectiveness in maintaining the cytochrome solubilized and intact for days at room temperature. Copyright © 2011 Elsevier Inc. All rights reserved.

  8. Preparation of radiopharmaceuticals labelled with bromine positron emitting isotopes for the study of dopaminergic receptors of the central nervous system using positron emission tomography

    International Nuclear Information System (INIS)

    Loc'h, C.

    1988-04-01

    The in vivo study of dopaminergic receptors of the central nervous system using positron emission tomography requires the preparation of radiopharmaceuticals labelled with β + emitting isotopes. The chemical and pharmacological properties of these ligands are evaluated. Cyclotron produced 75 and 76 bromine β + emitting isotopes are incorporated into dopaminergic ligands by electrophilic substitution using peracetic acid in a no-carrier added form. Purity, lipophilicity and specific activity are analyzed. Pharmacological criteria (specificity, saturability, displacement, localization) required for ligand-receptor binding studies are evaluated in vitro on striatal membranes and in vivo in the rat. Positron emission tomographic studies show that the study of dopaminergic D2 receptors is possible using 75 and 76 bromine labelled bromospiperone and bromolisuride. These ligands are used in physiological and pharmacological studies of the central nervous system [fr

  9. Development of uniformly stable isotope labeling system in higher plants for hetero-nuclear NMR experiments in vitro and in vivo

    International Nuclear Information System (INIS)

    Kikuchi, J.

    2005-01-01

    Full text: Novel methods for measurement of living systems are making new breakthroughs in life science. In the era of the metabolome (analysis of all measurable metabolites), a MS-based approach is considered to be the major technology, whereas a NMR-based method is recognized as minor technology due to its low sensitivity. Therefore, my laboratory is currently focusing to develop novel methodologies for an NMR-based metabolomics. This will be achieved by uniform stable isotope labeling of higher plants allowing application of multi-dimensional NMR experiments used in protein structure determination. Using these novel methods, I will analyze the dynamic molecular networks inside tissues. Especially, use of stable isotope labeling methods has enormous advantage for discrimination of incorporated or de novo synthesized compounds. Furthermore, potentiality of in vivo-NMR metabolomics will be discussed in the conference. (author)

  10. Acetylation and glycation of fibrinogen in vitro occur at specific lysine residues in a concentration dependent manner: A mass spectrometric and isotope labeling study

    International Nuclear Information System (INIS)

    Svensson, Jan; Bergman, Ann-Charlotte; Adamson, Ulf; Blombäck, Margareta; Wallén, Håkan; Jörneskog, Gun

    2012-01-01

    Highlights: ► Fibrinogen was incubated in vitro with glucose or aspirin. ► Acetylations and glycations were found at twelve lysine sites by mass spectrometry. ► The labeling by aspirin and glucose occurred dose-dependently. ► No competition between glucose and aspirin for binding to fibrinogen was found. -- Abstract: Aspirin may exert part of its antithrombotic effects through platelet-independent mechanisms. Diabetes is a condition in which the beneficial effects of aspirin are less prominent or absent – a phenomenon called “aspirin resistance”. We investigated whether acetylation and glycation occur at specific sites in fibrinogen and if competition between glucose and aspirin in binding to fibrinogen occurs. Our hypothesis was that such competition might be one explanation to “aspirin resistance” in diabetes. After incubation of fibrinogen in vitro with aspirin (0.8 mM, 24 h) or glucose (100 mM, 5–10 days), we found 12 modified sites with mass spectrometric techniques. Acetylations in the α-chain: αK191, αK208, αK224, αK429, αK457, αK539, αK562, in the β-chain: βK233, and in the γ-chain: γK170 and γK273. Glycations were found at βK133 and γK75, alternatively γK85. Notably, the lysine 539 is a site involved in FXIII-mediated cross-linking of fibrin. With isotope labeling in vitro, using [ 14 C-acetyl]salicylic acid and [ 14 C]glucose, a labeling of 0.013–0.084 and 0.12–0.5 mol of acetylated and glycated adduct/mol fibrinogen, respectively, was found for clinically (12.9–100 μM aspirin) and physiologically (2–8 mM glucose) relevant plasma concentrations. No competition between acetylation and glycation could be demonstrated. Thus, fibrinogen is acetylated at several lysine residues, some of which are involved in the cross-linking of fibrinogen. This may mechanistically explain why aspirin facilitates fibrin degradation. We find no support for the idea that glycation of fibrin(ogen) interferes with acetylation of

  11. Acetylation and glycation of fibrinogen in vitro occur at specific lysine residues in a concentration dependent manner: A mass spectrometric and isotope labeling study

    Energy Technology Data Exchange (ETDEWEB)

    Svensson, Jan, E-mail: jan.svensson@ki.se [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden); Bergman, Ann-Charlotte [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Adamson, Ulf [Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden); Blombaeck, Margareta [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Wallen, Hakan; Joerneskog, Gun [Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden)

    2012-05-04

    Highlights: Black-Right-Pointing-Pointer Fibrinogen was incubated in vitro with glucose or aspirin. Black-Right-Pointing-Pointer Acetylations and glycations were found at twelve lysine sites by mass spectrometry. Black-Right-Pointing-Pointer The labeling by aspirin and glucose occurred dose-dependently. Black-Right-Pointing-Pointer No competition between glucose and aspirin for binding to fibrinogen was found. -- Abstract: Aspirin may exert part of its antithrombotic effects through platelet-independent mechanisms. Diabetes is a condition in which the beneficial effects of aspirin are less prominent or absent - a phenomenon called 'aspirin resistance'. We investigated whether acetylation and glycation occur at specific sites in fibrinogen and if competition between glucose and aspirin in binding to fibrinogen occurs. Our hypothesis was that such competition might be one explanation to 'aspirin resistance' in diabetes. After incubation of fibrinogen in vitro with aspirin (0.8 mM, 24 h) or glucose (100 mM, 5-10 days), we found 12 modified sites with mass spectrometric techniques. Acetylations in the {alpha}-chain: {alpha}K191, {alpha}K208, {alpha}K224, {alpha}K429, {alpha}K457, {alpha}K539, {alpha}K562, in the {beta}-chain: {beta}K233, and in the {gamma}-chain: {gamma}K170 and {gamma}K273. Glycations were found at {beta}K133 and {gamma}K75, alternatively {gamma}K85. Notably, the lysine 539 is a site involved in FXIII-mediated cross-linking of fibrin. With isotope labeling in vitro, using [{sup 14}C-acetyl]salicylic acid and [{sup 14}C]glucose, a labeling of 0.013-0.084 and 0.12-0.5 mol of acetylated and glycated adduct/mol fibrinogen, respectively, was found for clinically (12.9-100 {mu}M aspirin) and physiologically (2-8 mM glucose) relevant plasma concentrations. No competition between acetylation and glycation could be demonstrated. Thus, fibrinogen is acetylated at several lysine residues, some of which are involved in the cross-linking of

  12. Quantitative Proteomics Analysis of VEGF-Responsive Endothelial Protein S-Nitrosylation Using Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and LC-MS/MS.

    Science.gov (United States)

    Zhang, Hong-Hai; Lechuga, Thomas J; Chen, Yuezhou; Yang, Yingying; Huang, Lan; Chen, Dong-Bao

    2016-05-01

    Adduction of a nitric oxide moiety (NO•) to cysteine(s), termed S-nitrosylation (SNO), is a novel mechanism for NO to regulate protein function directly. However, the endothelial SNO-protein network that is affected by endogenous and exogenous NO is obscure. This study was designed to develop a quantitative proteomics approach using stable isotope labeling by amino acids in cell culture for comparing vascular endothelial growth factor (VEGFA)- and NO donor-responsive endothelial nitroso-proteomes. Primary placental endothelial cells were labeled with "light" (L-(12)C6 (14)N4-Arg and L-(12)C6 (14)N2-Lys) or "heavy" (L-(13)C6 (15)N4-Arg and L-(13)C6 (15)N2-Lys) amino acids. The light cells were treated with an NO donor nitrosoglutathione (GSNO, 1 mM) or VEGFA (10 ng/ml) for 30 min, while the heavy cells received vehicle as control. Equal amounts of cellular proteins from the light (GSNO or VEGFA treated) and heavy cells were mixed for labeling SNO-proteins by the biotin switch technique and then trypsin digested. Biotinylated SNO-peptides were purified for identifying SNO-proteins by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Ratios of light to heavy SNO-peptides were calculated for determining the changes of the VEGFA- and GSNO-responsive endothelial nitroso-proteomes. A total of 387 light/heavy pairs of SNO-peptides were identified, corresponding to 213 SNO-proteins that include 125 common and 27 VEGFA- and 61 GSNO-responsive SNO-proteins. The specific SNO-cysteine(s) in each SNO-protein were simultaneously identified. Pathway analysis revealed that SNO-proteins are involved in various endothelial functions, including proliferation, motility, metabolism, and protein synthesis. We collectively conclude that endogenous NO on VEGFA stimulation and exogenous NO from GSNO affect common and different SNO-protein networks, implicating SNO as a critical mechanism for VEGFA stimulation of angiogenesis. © 2016 by the Society for the Study of Reproduction

  13. Stable isotope labeling confirms mixotrophic nature of streamer biofilm communities at alkaline hot springs

    Directory of Open Access Journals (Sweden)

    Florence eSchubotz

    2015-02-01

    Full Text Available Streamer biofilm communities (SBC are often observed within chemosynthetic zones of Yellowstone hot spring outflow channels, where temperatures exceed those conducive to photosynthesis. Nearest the hydrothermal source (75-88°C SBC comprise thermophilic Archaea and Bacteria, often mixed communities including Desulfurococcales and uncultured Crenarchaeota, as well as Aquificae, Thermus, each carrying diagnostic membrane lipid biomarkers. We tested the hypothesis that SBC can alternate their metabolism between autotrophy and heterotrophy depending on substrate availability. Feeding experiments were performed at two alkaline hot springs in Yellowstone National Park: Octopus Spring and ‘Bison Pool’, using various 13C-labeled substrates (bicarbonate, formate, acetate and glucose to determine the relative uptake of these different carbon sources. Highest 13C uptake, at both sites, was from acetate into almost all bacterial fatty acids, particularly into methyl-branched C15, C17 and C19 fatty acids that are diagnostic for Thermus/Meiothermus and some Firmicutes as well as into universally common C16:0 and C18:0 fatty acids. 13C-glucose showed a similar, but a 10 to 30 times lower uptake across most fatty acids. 13C bicarbonate uptake, signifying the presence of autotrophic communities was only significant at ‘Bison Pool’ and was observed predominantly in non-specific saturated C16, C18, C20 and C22 fatty acids. Incorporation of 13C-formate occurred only at very low rates at ‘Bison Pool’ and was almost undetectable at Octopus Spring, suggesting that formate is not an important carbon source for SBC. 13C uptake into archaeal lipids occurred predominantly with 13C acetate, suggesting also that archaeal communities at both springs have primarily heterotrophic carbon assimilation pathways. We hypothesize that these communities are energy-limited and predominantly nurtured by input of exogenous organic material, with only a small fraction being

  14. Simultaneous determination of glucose turnover, alanine turnover, and gluconeogenesis in human using a double stable-isotope-labeled tracer infusion and gas chromatography-mass spectrometry analysis

    International Nuclear Information System (INIS)

    Martineau, A.; Lecavalier, L.; Falardeau, P.; Chiasson, J.L.

    1985-01-01

    We have developed and validated a new method to measure simultaneously glucose turnover, alanine turnover, and gluconeogenesis in human, in steady and non-steady states, using a double stable-isotope-labeled tracer infusion and GC-MS analysis. The method is based on the concomitant infusion and dilution of D-[2,3,4,6,6-2H5]glucose and L-[1,2,3-13C3]alanine. The choice of the tracers was done on the basis of a minimal overlap between the ions of interest and those arising from natural isotopic abundances. Alanine was chosen as the gluconeogenic substrate because it is the major gluconeogenic amino acid extracted by the liver and, with lactate, constitutes the bulk of the gluconeogenic precursors. The method was validated by comparing the results obtained during simultaneous infusion of trace amounts of both stable isotope labeled compounds with the radioactive tracers (D-[3-3H]glucose and L-[1,2,3-14C3]alanine) in a normal and a diabetic subject; the radiolabeled tracers were used as the accepted reference procedure. A slight overestimation of glucose turnover (7.3 versus 6.8 in normal and 10.8 versus 9.2 mumol/kg min in diabetic subject) was noticed when the stable isotope-labeled tracers were used. For the basal turnover rate of alanine, similar values were obtained with both methods (6.2 mumol/kg min). For gluconeogenesis, higher values were observed in the basal state with the stable isotopes (0.42 versus 0.21 mumol/kg min); however, these differences disappeared in the postprandial period after the ingestion of a mixed meal. Despite those minor differences, the overall correlation with the reference method was excellent for glucose turnover (r = 0.87) and gluconeogenesis (r = 0.86)

  15. Analytical performance of reciprocal isotope labeling of proteome digests for quantitative proteomics and its application for comparative studies of aerobic and anaerobic Escherichia coli proteomes

    International Nuclear Information System (INIS)

    Lo, Andy; Weiner, Joel H.; Li, Liang

    2013-01-01

    Graphical abstract: -- Highlights: •Investigating a strategy of reciprocal isotope labeling of comparative samples. •Filtering out incorrect peptide identification or quantification values. •Analyzing the proteome changes of E. coli cells grown aerobically or anaerobically. •Presenting guidelines for reciprocal labeling experimental design. -- Abstract: Due to limited sample amounts, instrument time considerations, and reagent costs, only a small number of replicate experiments are typically performed for quantitative proteome analyses. Generation of reproducible data that can be readily assessed for consistency within a small number of datasets is critical for accurate quantification. We report our investigation of a strategy using reciprocal isotope labeling of two comparative samples as a tool for determining proteome changes. Reciprocal labeling was evaluated to determine the internal consistency of quantified proteome changes from Escherichia coli grown under aerobic and anaerobic conditions. Qualitatively, the peptide overlap between replicate analyses of the same sample and reverse labeled samples were found to be within 8%. Quantitatively, reciprocal analyses showed only a slight increase in average overall inconsistency when compared with replicate analyses (1.29 vs. 1.24-fold difference). Most importantly, reverse labeling was successfully used to identify spurious values resulting from incorrect peptide identifications and poor peak fitting. After removal of 5% of the peptide data with low reproducibility, a total of 275 differentially expressed proteins (>1.50-fold difference) were consistently identified and were then subjected to bioinformatics analysis. General considerations and guidelines for reciprocal labeling experimental design and biological significance of obtained results are discussed

  16. A study on Optical Labelling Techniques for All-Optical Networks

    DEFF Research Database (Denmark)

    Holm-Nielsen, Pablo Villanueva

    2005-01-01

    Optical switching has been proposed as an effective solution to overcoming the potential electronic bottleneck in all-optical network nodes carrying IP over WDM. The solution builds on the use of optical labelling as a mean to route packets or bursts of packets through the network. In addition...... of an intermediate wavelength between label erasure and label insertion. The above mentioned functionalities are assembled in whole network systems experiments that validates the different labelling schemes with respect to transmission, wavelength conversion, label swapping and retransmission. Optical labelling...... and specially the orthogonal schemes for optical labelling, are thus shown to be an effective solution to all-optical networks....

  17. Labelled compounds. (Pt. B)

    International Nuclear Information System (INIS)

    Buncel, E.; Jones, J.R.

    1991-01-01

    Since the end of World War II there has been a tremendous increase in the number of compounds that have been synthesized with radioactive or stable isotopes. They have found application in many diverse fields, so much so, that hardly a single area in pure and applied science has not benefited. Not surprisingly it has been reflected in appearance of related publications. The early proceedings of the Symposia on Advances in Trace Methodology were soon followed by various Euratom sponsored meetings in which methods of preparing and storing labelled compounds featured prominently. In due course a resurgence of interest in stable isotopes, brought about by their greater availability (also lower cost) and partly by development of new techniques such as gas chromatography - mass spectrometry (gc-ms), led to the publication of proceedings of several successful conferences. More recently conferences dealing with the synthesis and applications of isotopes and isotopically labelled compounds have been established on a regular basis. In addition to the proceedings of conferences and journal publications individuals left their mark by producing definitive texts, usually on specific nuclides. Only the classic two volume publication of Murray and Williams (Organic syntheses with isotopes, New York 1985), now over 30 years old and out of print, attempted to do justice to several nuclides. With the large amount of work that has been undertaken since then it seems unlikely that an updated edition could be produced. The alternative strategy was to ask scientists currently active to review specific areas and this is the approach adopted in the present series of monographs. In this way it is intended to cover the broad advances that have been made in the synthesis and applications of isotopes and isotopically labelled compounds in the physical and biomedical sciences. (author). refs.; figs.; tabs

  18. An isotope approach based on C-13 pulse-chase labelling vs. the root trenching method to separate heterotrophic and autotrophic respiration in cultivated peatlands

    Energy Technology Data Exchange (ETDEWEB)

    Biasi, C.; Pitkamaki, A. S.; Tavi, N. M.; Koponen, H. T.; Martikainen, P. J. [Univ.of Eastern Finland, Kuopio (Finland). Dept. of Environmental Science], e-mail: christina.biasi@uef.fi

    2012-11-01

    We tested an isotope method based on C-13 pulse-chase labelling for determining the fractional contribution of soil microbial respiration to overall soil respiration in an organic soil (cutaway peatland, eastern Finland), cultivated with the bioenergy crop, reed canary grass. The plants were exposed to CO{sub 2}-13 for five hours and the label was thereafter determined in CO{sub 2} derived from the soil-root system. A two-pool isotope mixing model was used to separate sources of respiration. The isotopic approach showed that a minimum of 50% of the total CO{sub 2} originated from soil-microbial respiration. Even though the method uses undisturbed soil-plant systems, it has limitations concerning the experimental determination of the true isotopic signal of all components contributing to autotrophic respiration. A trenching experiment which was comparatively conducted resulted in a 71% fractional contribution of soil-microbial respiration. This value was likely overestimated. Further studies are needed to evaluate critically the output from these two partitioning approaches. (orig.)

  19. Modification degrees at specific sites on heparan sulphate: an approach to measure chemical modifications on biological molecules with stable isotope labelling

    Science.gov (United States)

    Wu, Zhengliang L.; Lech, Miroslaw

    2005-01-01

    Chemical modification of biological molecules is a general mechanism for cellular regulation. A quantitative approach has been developed to measure the extent of modification on HS (heparan sulphates). Sulphation on HS by sulphotransferases leads to variable sulphation levels, which allows cells to tune their affinities to various extracellular proteins, including growth factors. With stable isotope labelling and HPLC-coupled MS, modification degrees at various O-sulphation sites could be determined. A bovine kidney HS sample was first saturated in vitro with 34S by an OST (O-sulphotransferase), then digested with nitrous acid and analysed with HPLC-coupled MS. The 34S-labelled oligosaccharides were identified based on their unique isotope clusters. The modification degrees at the sulphotransferase recognition sites were obtained by calculating the intensities of isotopic peaks in the isotope clusters. The modification degrees at 3-OST-1 and 6-OST-1 sites were examined in detail. This approach can also be used to study other types of chemical modifications on biological molecules. PMID:15743272

  20. The Potential for Zinc Stable Isotope Techniques and Modelling to Determine Optimal Zinc Supplementation

    Directory of Open Access Journals (Sweden)

    Cuong D. Tran

    2015-05-01

    Full Text Available It is well recognised that zinc deficiency is a major global public health issue, particularly in young children in low-income countries with diarrhoea and environmental enteropathy. Zinc supplementation is regarded as a powerful tool to correct zinc deficiency as well as to treat a variety of physiologic and pathologic conditions. However, the dose and frequency of its use as well as the choice of zinc salt are not clearly defined regardless of whether it is used to treat a disease or correct a nutritional deficiency. We discuss the application of zinc stable isotope tracer techniques to assess zinc physiology, metabolism and homeostasis and how these can address knowledge gaps in zinc supplementation pharmacokinetics. This may help to resolve optimal dose, frequency, length of administration, timing of delivery to food intake and choice of zinc compound. It appears that long-term preventive supplementation can be administered much less frequently than daily but more research needs to be undertaken to better understand how best to intervene with zinc in children at risk of zinc deficiency. Stable isotope techniques, linked with saturation response and compartmental modelling, also have the potential to assist in the continued search for simple markers of zinc status in health, malnutrition and disease.

  1. Application of isotope techniques to groundwater pollution research for Xiangshan uranium ore field, China

    International Nuclear Information System (INIS)

    Liu Fulin; Liu Peilun; Zhu Chuande; Wu Xiaowei; Zeng Yinsheng

    1998-01-01

    The investigation of groundwater pollution due to uranium deposits focused on the most important uranium metallogenic area-Zhoujiashan district of Xiangshan uranium ore field, China. Groundwater collected from five completed exploration boreholes in the area is regarded as the pollution source and is traced and analysed by using isotope as well as radio-hydrochemical techniques. In addition, the pollution situation of a small uranium ore pile for heap-leaching and a big uranium ore open pit are monitored by the same techniques. It has been experimentally proven that the uranium concentration and the uranium isotope ratio 234 U/ 238 U in natural waters are two sensitive indicators of radioactive pollution in natural waters. It was concluded that under present conditions, exploration of uranium deposits may not cause serious groundwater pollution of radioactive elements (U, Ra, Rn and Th), however, it is difficult to avoid the serious surface water pollution coming from the exploitation of uranium ore by a big open pit. (author)

  2. The Potential for Zinc Stable Isotope Techniques and Modelling to Determine Optimal Zinc Supplementation

    Science.gov (United States)

    Tran, Cuong D.; Gopalsamy, Geetha L.; Mortimer, Elissa K.; Young, Graeme P.

    2015-01-01

    It is well recognised that zinc deficiency is a major global public health issue, particularly in young children in low-income countries with diarrhoea and environmental enteropathy. Zinc supplementation is regarded as a powerful tool to correct zinc deficiency as well as to treat a variety of physiologic and pathologic conditions. However, the dose and frequency of its use as well as the choice of zinc salt are not clearly defined regardless of whether it is used to treat a disease or correct a nutritional deficiency. We discuss the application of zinc stable isotope tracer techniques to assess zinc physiology, metabolism and homeostasis and how these can address knowledge gaps in zinc supplementation pharmacokinetics. This may help to resolve optimal dose, frequency, length of administration, timing of delivery to food intake and choice of zinc compound. It appears that long-term preventive supplementation can be administered much less frequently than daily but more research needs to be undertaken to better understand how best to intervene with zinc in children at risk of zinc deficiency. Stable isotope techniques, linked with saturation response and compartmental modelling, also have the potential to assist in the continued search for simple markers of zinc status in health, malnutrition and disease. PMID:26035248

  3. The relation of plasma octopamine to hepatic encephalopathy studied with stable isotope dilution technique

    International Nuclear Information System (INIS)

    Zhang Jianhua

    1992-01-01

    In order to investigate the relationship between octopamine and hepatic encephalopathy, plasma octopamine content of normal adults and hepatic patients (compensated or de-compensated cirrhotic, patients with encephalopathy) were determined with isotope dilution and GCMS technique, 200 ng of [1, 1-D]-octopamine was added to each sample as internal standard. After the de-proteinized plasma was subjected to cation exchange chromatography, the amine fraction was derivatized with PFP and analyzed with GCMS using the selected ion monitoring technique. The result showed that the level of plasma octopamine was significantly elevated in patients with encephalopathy. Since this method is sensitive and specific, it is readily applicable to the analysis of a few nano grams of octopamine

  4. Development of isotope dilution-liquid chromatography/mass spectrometry combined with standard addition techniques for the accurate determination of tocopherols in infant formula

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Joonhee; Jang, Eun-Sil; Kim, Byungjoo, E-mail: byungjoo@kriss.re.kr

    2013-07-17

    Graphical abstract: -- Highlights: •ID-LC/MS method showed biased results for tocopherols analysis in infant formula. •H/D exchange of deuterated tocopherols in sample preparation was the source of bias. •Standard addition (SA)-ID-LC/MS was developed as an alternative to ID-LC/MS. •Details of calculation and uncertainty evaluation of the SA-IDMS were described. •SA-ID-LC/MS showed a higher-order metrological quality as a reference method. -- Abstract: During the development of isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) for tocopherol analysis in infant formula, biased measurement results were observed when deuterium-labeled tocopherols were used as internal standards. It turned out that the biases came from intermolecular H/D exchange and intramolecular H/D scrambling of internal standards in sample preparation processes. Degrees of H/D exchange and scrambling showed considerable dependence on sample matrix. Standard addition-isotope dilution mass spectrometry (SA-IDMS) based on LC/MS was developed in this study to overcome the shortcomings of using deuterium-labeled internal standards while the inherent advantage of isotope dilution techniques is utilized for the accurate recovery correction in sample preparation processes. Details of experimental scheme, calculation equation, and uncertainty evaluation scheme are described in this article. The proposed SA-IDMS method was applied to several infant formula samples to test its validity. The method was proven to have a higher-order metrological quality with providing very accurate and precise measurement results.

  5. Evaluation in vitro and in vivo of two labelling techniques of different 99mTc-dextrans for lymphoscintigraphy

    International Nuclear Information System (INIS)

    Wingardh, K.; Strand, S.E.

    1989-01-01

    Five dextrans with different molecular weights and charges were labelled with 99m Tc. The labelling methods presented by Henze et al. and Ercan et al. were compared. The labelling efficiency was tested with gel column chromatography scanning (GCS), gel chromatography (GC) combined with the Anthrone test, paper chromatography (PC) and thin layer chromatography (TLC). The GCS technique always indicated a lower labelling efficiency than the PC and TLC techniques, which was due to a more optimal separation of the radioactive components. Gel chromatography in combination with the Anthrone test made it easy to identify the different radiochemical components in contrast to the other methods. Dextran solutions were injected subcutaneously bilaterally at the xiphoid processes in rabbits. The injection sites were massaged for 30 s. Uptake in the parasternal lymph nodes was registered with a scintillation camera. The animals were killed and dissected at the end of the study. This investigation shows that the labelling method of Ercan et al. gives the highest labelling efficiency. Furthermore, the final pH (4.5) for the dextran solution makes it more useful for injection. For quality control of 99m Tc labelled dextran we recommend the Anthrone test as a complement to GC because it is a quick and simple method of determining the dextran content. (orig.)

  6. Application of the Microwave Discharge Modification of the Wilzbach Technique for the Tritium Labelling of some Organics of Biological Interest

    International Nuclear Information System (INIS)

    Gosztonyi, T.

    1969-01-01

    The modification of the Wilzbach technique using microwave discharge has been routinely used in our laboratory for rapid tritium labellings. The applicability of the method and the effects of some of the reaction parameters were studied and published previously. The main advantage of the method is its simplicity and rapidity and the low extent of decomposition of the compound to be labelled during the reaction. Specific activities obtained, however, are not high enough for some investigations. In this paper the labelling of dihydrostreptomycine, tetracycline and antipyrine will be described

  7. A technique developed for labeling the green manures (sunnhemp and velvet bean) with 15 N for nitrogen dynamic studies

    International Nuclear Information System (INIS)

    Ambrosano, Edmilson Jose

    1997-01-01

    A technique was developed for labeling the leguminous plant tissue with nitrogen ( 15 N) to obtain labelled material for nitrogen dynamic studies. Sunnhemp (crotalaria juncea L.) and velvet beans (Mucuna aterrima, sinonimia Stizolobium aterrimum Piper and Tracy) were grown in pots containing 10 kg of a Red Yellow Podzolic soil, under greenhouse conditions. The rate of 1.2 of nitrogen (ammonium sulphate with 11.37 atom % 15 N) per pot was applied three times. The labelled dried plant material showed 3.177 and 4.337 of atom % 15 N, respectively for velvet beans and sunnhemp. (author)

  8. Application of the Microwave Discharge Modification of the Wilzbach Technique for the Tritium Labelling of some Organics of Biological Interest

    Energy Technology Data Exchange (ETDEWEB)

    Gosztonyi, T

    1969-01-15

    The modification of the Wilzbach technique using microwave discharge has been routinely used in our laboratory for rapid tritium labellings. The applicability of the method and the effects of some of the reaction parameters were studied and published previously. The main advantage of the method is its simplicity and rapidity and the low extent of decomposition of the compound to be labelled during the reaction. Specific activities obtained, however, are not high enough for some investigations. In this paper the labelling of dihydrostreptomycine, tetracycline and antipyrine will be described.

  9. Bacterial diversity exploration in hydrocarbon polluted soil: metabolic potential and degrader community evolution revealed by isotope labeling

    International Nuclear Information System (INIS)

    Martin, F.

    2011-01-01

    Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous compounds produced by incomplete combustion of organic matter. They are a source of environmental pollution, especially associated to oil product exploitation, and represent a threat for living organisms including human beings because of their toxicity. Many bacteria capable of degrading PAHs have been isolated and studied. However, since less than 5% of soil bacteria can be cultivated in the laboratory, bacterial species able to degrade PAHs in situ have been poorly studied. The first goal of this study was to identify bacteria that degrade PAHs in soil using culture-independent molecular methods. To this end, a strategy known a stable isotope probing has been implemented based on the use of phenanthrene, a three rings PAH, in which the natural isotope of carbon was replaced by 13 C. This molecule has been introduced as a tracer in microcosms containing soil from a constructed wetlands collecting contaminated water from highway runoff. Bacteria having incorporated the 13 C were then identified by 16 S rRNA gene sequence analysis after PCR amplification from labeled genomic DNA extracted from soil. The results show that so far little studied Betaproteobacteria, belonging to the genera Acidovorax, Rhodoferax, Hydrogenophaga and Thiobacillus, as well as Rhodocyclaceae, were the key players in phenanthrene degradation. Predominance of Betaproteobacteries was established thanks to quantitative PCR measurements. A dynamic analysis of bacterial diversity also showed that the community structure of degraders depended on phenanthrene bioavailability. In addition, the phylogenetic diversity of ring-hydroxylating di-oxygenases, enzymes involved in the first step of PAH degradation, has been explored. We detected new sequences, mostly related to di-oxygenases from Sphingomonadales and Burkholderiales. For the first time, we were able to associate a catalytic activity for oxidation of PAHs to partial gene sequences

  10. Bioavailability of zinc from defatted soy flour, soy hulls and whole eggs as determined by intrinsic and extrinsic labeling techniques.

    Science.gov (United States)

    Meyer, N R; Stuart, M A; Weaver, C M

    1983-06-01

    Bioavailability of zinc from diets prepared from intrinsically and extrinsically labeled autoclaved, defatted soy flour and scrambled, freeze-dried egg was investigated in male rats marginally depleted in zinc. In one study, retention of zinc from intrinsically labeled soybean flour (73%) was significantly less than from 65ZnCl2 extrinsically added to a soy flour-based diet (80%). Zinc from intrinsically labeled soybean hulls and from soy flour diets containing 10% soybean hulls extrinsically labeled with 65ZnCl2 was as available as the zinc from the extrinsically labeled soy flour diet. In a second study, extrinsic and intrinsic labeling techniques gave a similar assessment of bioavailability of zinc from egg- and soy flour-based diets when extrinsic labeling was accomplished by thoroughly mixing 65ZnCl2 with the protein source prior to incorporation into the diet. Absorption of 65Zn was greater from egg diets than from soy flour diets and of intermediate value from mixed soy flour and egg (50:50, wt/wt) diets regardless of which protein source was labeled, indicating that the zinc entered a common pool.

  11. Vitamin A equivalency and apparent absorption of ß-carotene in ileostomy subjects using a dual-isotope dilution technique

    NARCIS (Netherlands)

    Bouwman, C.A.; Naber, T.H.J.; Breemen, van R.B.; Zhu, D.; Dicke, H.; Siebelink, E.; Hulshof, P.J.M.; Russel, F.G.M.; Schaafsma, G.; West, C.E.

    2010-01-01

    The objective was to quantify the vitamin A equivalency of ß-carotene in two diets using a dual-isotope dilution technique and the apparent ß-carotene absorption as measured by the oral–faecal balance technique. Seventeen healthy adults with an ileostomy completed the 4-week diet-controlled,

  12. Vitamin A equivalency and apparent absorption of beta-carotene in ileostomy subjects using a dual-isotope dilution technique.

    NARCIS (Netherlands)

    Loo-Bouwman, C.A. Van; Naber, T.H.; Breemen, R.B. van; Zhu, D.; Dicke, H.; Siebelink, E.; Hulshof, P.J.; Russel, F.G.M.; Schaafsma, G.; West, C.E.

    2010-01-01

    The objective was to quantify the vitamin A equivalency of beta-carotene in two diets using a dual-isotope dilution technique and the apparent beta-carotene absorption as measured by the oral-faecal balance technique. Seventeen healthy adults with an ileostomy completed the 4-week diet-controlled,

  13. Precise determination of Pb isotope ratios by simple double spike MC-ICP-MS technique without Tl addition

    Digital Repository Service at National Institute of Oceanography (India)

    Makishima, A.; Nath, B.N.; Nakamura, E.

    A double-spike multicollector ICP-MS (DS-MC-ICP-MS) technique for Pb isotope analysis without Tl addition is established and its analytical performance is examined in detail. This simple DS-MC-ICP-MS technique using 20 ng ml sup(-1) gave averages...

  14. ICT: isotope correction toolbox.

    Science.gov (United States)

    Jungreuthmayer, Christian; Neubauer, Stefan; Mairinger, Teresa; Zanghellini, Jürgen; Hann, Stephan

    2016-01-01

    Isotope tracer experiments are an invaluable technique to analyze and study the metabolism of biological systems. However, isotope labeling experiments are often affected by naturally abundant isotopes especially in cases where mass spectrometric methods make use of derivatization. The correction of these additive interferences--in particular for complex isotopic systems--is numerically challenging and still an emerging field of research. When positional information is generated via collision-induced dissociation, even more complex calculations for isotopic interference correction are necessary. So far, no freely available tools can handle tandem mass spectrometry data. We present isotope correction toolbox, a program that corrects tandem mass isotopomer data from tandem mass spectrometry experiments. Isotope correction toolbox is written in the multi-platform programming language Perl and, therefore, can be used on all commonly available computer platforms. Source code and documentation can be freely obtained under the Artistic License or the GNU General Public License from: https://github.com/jungreuc/isotope_correction_toolbox/ {christian.jungreuthmayer@boku.ac.at,juergen.zanghellini@boku.ac.at} Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  15. Magnetic resonance studies of isotopically labeled paramagnetic proteins: (2FE-2S) ferredoxins

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, H.; Xia, B.; Chae, Y.K.; Westler, W.M.; Markley, J.L. [Univ. of Wisconsin, Madison, WI (United States)

    1994-12-01

    Recent developments in NMR spectroscopy, especially multidimensional, multinuclear NMR techniques, have made NMR the most versatile tool available for studying protein structure and function in solution. Unlike diamagnetic proteins, paramagnetic proteins contain centers with unpaired electrons. These unpaired electrons interact with magnetic nuclei either through chemical bonds by a contact mechanism or through space by a pseudocontact mechanism. Such interactions make the acquisition and analysis of NMR spectra of paramagnetic proteins more challenging than those of diamagnetic proteins. Some NMR signals from paramagnetic proteins are shifted outside the chemical shift region characteristic of diamagnetic proteins; these {open_quotes}hyperfine-shifted{close_quotes} resonances originate from nuclei that interact with unpaired electrons from the paramagnetic center. The large chemical shift dispersion in spectra of paramagnetic proteins makes it difficult to excite the entire spectral window and leads to distortions in the baseline. Interactions with paramagnetic centers shorten T{sub 1} and T{sub 2} relaxation times of nuclei; the consequences are line broadening and lower spectral sensitivity. Scalar (through bond) and dipolar (through space) interactions between pairs of nuclei are what give rise to crosspeak signals in multi-dimensional NMR spectra of small diamagnetic proteins. When such interactions involve a nucleus that is strongly relaxed by interaction with a paramagnetic center, specialized methods may be needed for its detection or it may be completely undetectable by present nD NMR methods.

  16. Nitrogen Dynamic Study on Rice Mutant Lines Using 15N Isotope Techniques

    International Nuclear Information System (INIS)

    Ahmad Nazrul Abd Wahid; Shyful Azizi Abdul Rahman; Abdul Rahim Harun

    2015-01-01

    Malaysian Nuclear Agency in collaboration with UPM and MARDI has produced two types of rice mutant lines of MR219, viz. MR219-4 and MR219-9 developed under rice radiation mutagenenesis programme for adaptability to aerobic conditions. Aerobic cultivating is rice cultivation system on well drained soil and using minimal water input. At Malaysian Nuclear Agency, a nitrogen fertilization study in aerobic condition for the rice mutant lines was carried out in the shade house and field. The study is intended to examine and assess the dynamics of nitrogen by rice mutant lines through the different soil water management and nitrogen levels. Direct 15 N isotopic tracer method was used in this study, whereby the 15 N labeled urea fertilizer was utilized as a tracer for nitrogen nutrient uptake by the test crops. This paper is intended to highlight the progress that has been made in the study of the nitrogen dynamics on MR219-4 and MR219-9 rice mutant lines. (author)

  17. Raman spectroscopic studies of isotopic diatomic molecules and a technique for measuring stable isotope ratios using Raman scattering

    International Nuclear Information System (INIS)

    Harney, R.C.

    1976-01-01

    A method for measuring stable isotope ratios using Raman scattering has been developed. This method consists of simultaneously counting photons scattered out of a high-intensity laser beam by different isotopically-substituted molecules. A number of studies of isotopic diatomic molecules have been made. The Q-branches of the Raman spectra of the isotopic molecules 14 N 15 N and 16 O 18 O were observed at natural abundance in nitrogen and oxygen samples. Comparison of the ratios of the intensities of the Q-branches of the major nitrogen and oxygen isotopic molecules with mass spectrometric determinations of the isotopic compositions yielded scattering cross sections of 14 N 15 N relative to 14 N 14 N and 16 O 18 O relative to 16 O 16 O. These cross section ratios differ from unity, a difference which can be explained by considering nuclear mass effects on the Franck-Condon factors of the molecular transitions. The measured intensities of the 14 N 15 N and 16 O 18 O Q-branches provided the baseline data needed to make the previously-mentioned extrapolation. High-resolution (approximately 0.15 cm -1 ) spectra of the Q-branches of 14 N 14 N and 16 O 16 O yielded a direct determination of α/sub e/ (the difference between the rotational constant in the ground and first excited vibrational states) for these molecules. The measured values are in excellent agreement with those obtained by other means. Complete Raman spectra (pure rotation, rotation-vibration, and high-resolution Q-branch) were obtained on a sample of pure 18 O 18 O. Analysis of this data yielded the molecular parameters: the equilibrium internuclear separation r/sub e/, the moment of inertia I/sub e/, and the energy parameters α/sub e/, B/sub e/, and ΔG/sub 1 / 2 /. These are in good agreement with data obtained by microwave spectroscopy

  18. Methyl-coenzyme M reductase from methanogenic archaea: isotope effects on label exchange and ethane formation with the homologous substrate ethyl-coenzyme M.

    Science.gov (United States)

    Scheller, Silvan; Goenrich, Meike; Thauer, Rudolf K; Jaun, Bernhard

    2013-10-09

    Ethyl-coenzyme M (CH3CH2-S-CH2CH2-SO3(-), Et-S-CoM) serves as a homologous substrate for the enzyme methyl-coenzyme M reductase (MCR) resulting in the product ethane instead of methane. The catalytic reaction proceeds via an intermediate that already contains all six C-H bonds of the product. Because product release occurs after a second, rate-limiting step, many cycles of intermediate formation and reconversion to substrate occur before a substantial amount of ethane is released. In deuterated buffer, the intermediate becomes labeled, and C-H activation in the back reaction rapidly leads to labeled Et-S-CoM, which enables intermediate formation to be detected. Here, we present a comprehensive analysis of this pre-equilibrium. (2)H- and (13)C-labeled isotopologues of Et-S-CoM were used as the substrates, and the time course of each isotopologue was followed by NMR spectroscopy. A kinetic simulation including kinetic isotope effects allowed determination of the primary and α- and β-secondary isotope effects for intermediate formation and for the C-H/C-D bond activation in the ethane-containing intermediate. The values obtained are in accordance with those found for the native substrate Me-S-CoM (see preceding publication, Scheller, S.; Goenrich, M.; Thauer, R. K.; Jaun, B. J. Am. Chem. Soc. 2013, 135, DOI: 10.1021/ja406485z) and thus imply the same catalytic mechanism for both substrates. The experiment by Floss and co-workers, demonstrating a net inversion of configuration to chiral ethane with CH3CDT-S-CoM as the substrate, is compatible with the observed rapid isotope exchange if the isotope effects measured here are taken into account.

  19. Monitoring the biodegradation of polycyclic aromatic hydrocarbons in a co-contaminated soil using stable isotope labeling

    Science.gov (United States)

    Wawra, Anna; Friesl-Hanl, Wolfgang; Watzinger, Andrea; Soja, Gerhard; Puschenreiter, Markus

    2016-04-01

    Conventional remediation techniques like "dig and dump" are costly and limited in scale. Plant- and microbe-based alternatives, e.g. phytoremediation options, offer a cheap and environmentally friendly approach that can be applied on larger areas. However, the application of phytoremediation techniques to co-contaminated sites may be hindered due to a potential inhibition of biodegradation processes by the presence of heavy metals in soil. Therefore, the objective of this study is to test the hypothesis that the degradation of organic pollutants can be enhanced by immobilising potentially toxic heavy metals. This study aims to identify the influence of heavy metal immobilisation on the degradation of organic pollutants, and to determine chemical, physical and biological measures further accelerating these processes. The influence of heavy metals on organic pollutant degradation dynamics is assessed using 13C-phospholipid fatty acid analysis (13C-PLFA). Application of 13C-labeled phenanthrene allows the identification of microbial groups responsible for the degradation process. For metal immobilisation and enhanced biodegradation, distinct mineral and organic soil amendments (iron oxides, gravel sludge, biochar) are deployed, partly in combination with fast-growing and pollution-tolerant woody plants (willow, black locust and alder). Results of an incubation batch experiment show a fast degradation of the phenanthrene label within the first two weeks by various microbial groups (gram negative bacteria as indicated by the cy17:0 peak) resulting in a decrease by up to 80% of the total PAH concentration (Σ 16 EPA PAHs) measured in soil. A similar trend was observed in the greenhouse pot experiment, whereby heavy metal accumulation in the woody plants growing on the co-contaminated soil significantly varied with plant species (willow > black locust, alder).

  20. Molecular detection of drug resistance in microbes by isotopic techniques: The IAEA experience

    International Nuclear Information System (INIS)

    Dar, L.; Boussaha, A.; Padhy, A.K.; Khan, B.

    2003-01-01

    The International Atomic Energy Agency (IAEA) supports various programmes on the uses of radionuclide techniques in the management of human communicable diseases. An important issue, being addressed through several technology transfer projects, is the detection of drug resistance in microbes by radioisotope based molecular-biology diagnostic procedures. The techniques employed include dot blot hybridisation with P-32 labelled oligonucleotide probes to detect point mutations, associated with drug resistance, in microbial genes amplified by the polymerase chain reaction (PCR). Molecular methods have been used for the detection of drug resistance in the malarial parasite, Plasmodium falciparum, and in Mycobacterium tuberculosis. Radioisotope based molecular-biology methods have been demonstrated to have comparative advantages in being sensitive, specific, cost-effective, and suitable for application to large-scale molecular surveillance for drug resistance. (author)

  1. Undercovering the hidden links. Nuclear and isotope techniques target nutritional needs

    International Nuclear Information System (INIS)

    Iyengar, Venkatesh

    2001-01-01

    Global nutrition problems raise a host of questions and warrant action by the international community of scientists, nutritionists, physicians and other medical professionals. What steps should be taken to remedy this situation? How can this be accomplished economically? How can progress be monitored? What is the role of technology in the overall monitoring process? The last question, which is most relevant to this article, is of particular importance to the IAEA and its support of nutrition programmes. The IAEA's activities in human nutrition were initiated to apply nuclear and related isotopic techniques for solving problems prevalent in developing countries. Among the numerous applications available, isotopic techniques are uniquely well suited to targeting and tracking progress in food and nutrition development programmes. These are tools that help evaluate nutritional status of individuals and populations, measure nutrient requirements and the uptake and bio-availability of vitamins and minerals. The IAEA's efforts help to: verify the nature of the nutrition problem and the efficacy of specific interventions; implement nutrition intervention programmes by monitoring effectiveness and reducing programme costs; guide in the processing of local foods for optimal nutritional value; serve as early indicators of important long-term health improvements; and strengthen capacity building in developing countries. Among the numerous applications available, isotopic techniques are uniquely well suited to targeting and tracking progress in food and nutrition development programmes. These are tools that help evaluate nutritional status of individuals and populations, measure nutrient requirements and the uptake and bio-availability of vitamins and minerals. The IAEA's efforts help to: verify the nature of the nutrition problem and the efficacy of specific interventions; implement nutrition intervention programmes by monitoring effectiveness and reducing programme costs

  2. Stable isotopes - separation and application

    International Nuclear Information System (INIS)

    Lockhart, I.M.

    1980-01-01

    In this review, methods used for the separation of stable isotopes ( 12 C, 13 C, 14 N, 15 N, 16 O, 17 O, 18 O, 34 S) will be described. The synthesis of labelled compounds, techniques for detection and assay, and areas of application will also be discussed. Particular attention will be paid to the isotopes of carbon, nitrogen, and oxygen; to date, sulphur isotopes have only assumed a minor role. The field of deuterium chemistry is too extensive for adequate treatment; it will therefore be essentially excluded. (author)

  3. Inverse kinematics technique for the study of fission-fragment isotopic yields at GANIL energies

    International Nuclear Information System (INIS)

    Delaune, O.

    2012-01-01

    The characteristics of the fission-products distributions result of dynamical and quantum properties of the deformation process of the fissioning nucleus. These distributions have also an interest for the conception of new nuclear power plants or for the transmutation of the nuclear wastes. Up to now, our understanding of the nuclear fission remains restricted because of experimental limitations. In particular, yields of the heavy fission products are difficult to get with precision. In this work, an innovative experimental technique is presented. It is based on the use of inverse kinematics coupled to the use of a spectrometer, in which a 238 U beam at 6 or 24 A MeV impinges on light targets. Several actinides, from 238 U to 250 Cf, are produced by transfer or fusion reactions, with an excitation energy ranges from ten to few hundreds MeV depending on the reaction and the beam energy. The fission fragments of these actinides are detected by the VAMOS spectrometer or the LISE separator. The isotopic yields of fission products are completely measured for different fissioning systems. The neutron excess of the fragments is used to characterise the isotopic distributions. Its evolution with excitation energy gives important insights on the mechanisms of the compound-nucleus formation and its deexcitation. Neutron excess is also used to determine the multiplicity of neutrons evaporated by the fragments. The role of the proton and neutron shell effects into the formation of fission fragments is also discussed. (author) [fr

  4. Improved analytical techniques of sulfur isotopic composition in nanomole quantities by MC-ICP-MS.

    Science.gov (United States)

    Yu, Tsai-Luen; Wang, Bo-Shian; Shen, Chuan-Chou; Wang, Pei-Ling; Yang, Tsanyao Frank; Burr, George S; Chen, Yue-Gau

    2017-10-02

    We propose an improved method for precise sulfur isotopic measurements by multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS) in conjunction with a membrane desolvation nebulization system. The problems of sulfur loss through the membrane desolvation apparatus are carefully quantified and resolved. The method overcomes low intrinsic sulfur transmission through the instrument, which was initially 1% when operating at a desolvation temperature of 160 °C. Sulfur loss through the membrane desolvation apparatus was resolved by doping with sodium. A Na/S ratio of 2 mol mol -1 produced sulfur transmissions with 98% recovery. Samples of 3 nmol (100 ng) sulfur achieved an external precision of ±0.18‰ (2 SD) for δ 34 S and ±0.10‰ (2 SD) for Δ 33 S (uppercase delta expresses the extent of mass-independent isotopic fractionation). Measurements made on certified reference materials and in-house standards demonstrate analytical accuracy and reproducibility. We applied the method to examine microbial-induced sulfur transformation in marine sediment pore waters from the sulfate-methane transition zone. The technique is quite versatile, and can be applied to a range of materials, including natural waters and minerals. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Total energy expenditure in burned children using the doubly labeled water technique

    International Nuclear Information System (INIS)

    Goran, M.I.; Peters, E.J.; Herndon, D.N.; Wolfe, R.R.

    1990-01-01

    Total energy expenditure (TEE) was measured in 15 burned children with the doubly labeled water technique. Application of the technique in burned children required evaluation of potential errors resulting from nutritional intake altering background enrichments during studies and from the high rate of water turnover relative to CO2 production. Five studies were discarded because of these potential problems. TEE was 1.33 +/- 0.27 times predicted basal energy expenditure (BEE), and in studies where resting energy expenditure (REE) was simultaneously measured, TEE was 1.18 +/- 0.17 times REE, which in turn was 1.16 +/- 0.10 times predicted BEE. TEE was significantly correlated with measured REE (r2 = 0.92) but not with predicted BEE. These studies substantiate the advantage of measuring REE to predict TEE in severely burned patients as opposed to relying on standardized equations. Therefore we recommend that optimal nutritional support will be achieved in convalescent burned children by multiplying REE by an activity factor of 1.2

  6. New aspects of radioimmunochemical measurement of human parathyroid hormone using the labelled antibody technique

    International Nuclear Information System (INIS)

    Hesch, R.D.; McIntosh, C.H.S.; Woodhead, J.S.; Welsh National School of Medicine, Cardiff

    1975-01-01

    Two forms of heterogeneity of parathyroid hormone (PTH) have given rise to conflicting results: one due to the heterogeneity of the secreted species from the gland and their peripheral metabolism and the other representing the immunochemical heterogeneity of the available antibodies. We have developed sequence specific assays using the technique of labelled antibodies. Therefore, results of assays measuring the C-terminal part and the (1-34)-N-terminal part of the molecule could be compared to those of an assay for hormone bearing both N- and C-terminal antigenic determinants. This assay is supposed to detect predominantly (1-84)-intact hormone. The immunoradiometric assay of (1-34)-PTH has a sensitivity of 0.04 ng/ml. This technique avoids the critical iodination of the hormone fragment containing no tyrosine. There is the expected overlap between normal subjects and patients with primary and secondary hyperparathyroidism. The most important finding are results from patients undergoing neck catheterization. We demonstrated nonuniform secretion of different species of PTH by parathyroid adenomata and normal glands. This supports the hypothesis of cleavage of the (1-84)-molecule in the gland. (orig.) [de

  7. A committee report on the state-of-art of uranium isotope enrichment measurement techniques

    International Nuclear Information System (INIS)

    1982-06-01

    The activity of a sectional meeting (June, 1979 - February, 1982) for uranium enrichment measurement under the Committee on Analytical Chemistry of Nuclear Fuels and Reactor Materials is summarized. In the first part of this report, the object and present state of the measurement at the following organization are described; two development organizations (Power Reactor and Nuclear Fuel Development Corporation and Japan Nuclear Fuel Development), two nuclear fuel makers (Japan Nuclear Fuel and Mitsubishi Nuclear Fuel), one safeguards inspection organization (Nuclear Material Control Center) and one research organization (Japan Atomic Energy Research Institute). The second part contains the principle and technique of several measurement methods for uranium isotopic assay, such as mass spectrometry, passive and active assays and optical spectral method. Lastly, the concept of the reference materials and its practical information for mass spectrometry and non-destructive assay are described. (author)

  8. Estimation of endogenous faecal calcium in buffalo (BOS bubalis) by isotope dilution technique

    International Nuclear Information System (INIS)

    Singh, S.; Sareen, V.K.; Marwaha, S.R.; Sekhon, B.; Bhatia, I.S.

    1973-01-01

    Detailed investigations on the isotope-dilution technique for the estimation of endogenous faecal calcium were conducted with buffalo calves fed on growing ration. The ration consisted of wheat straw, green lucerne and concentrate mix. The endogenous faecal calcium was 3.71 g/day, which is 17.8 percent of the total faecal calcium. The apparent and true digestibilities of Ca were calculated as 51 and 60 percent respectively. The endogenous faecal calcium can be estimated in buffalo calves by giving single subcutaneous injection of Ca 45 and collecting blood samples on 12th and 21st days only, and representative sample from the faeces collected from 13th through 22nd day after the injection. (author)

  9. Study of ferrallitisation process in soil by application of isotopic dilution kinetic technique to iron

    International Nuclear Information System (INIS)

    Thomann, Christiane

    1978-01-01

    Isotopic dilution kinetic technique applied to iron may contribute to make clear the conditions of ''potential'' mobility of iron in soils, under the action of three factors: moisture, incubation period and organic matter imputs. Comparison between surface horizons of three tropical soils: leached ferruginous tropical soil, weakly ferrallitic red soil and ferrallitic soil shows that in the ferrallitisation process, weakly ferrallitic soil would take place between the two other types of soils with a maximum mobility of iron. This mobility decreases when organic matter rate decreases leading then to ''beige'' soil (ferruginous leached tropical soil), and when hydroxide rate increases, which leads to ferrallitic soil. In podzol (A 1 horizon), for the same rate of organic matter, potential mobility of iron is higher than in ferallitic soil, because it contains ten times more free iron than the podzol [fr

  10. Isotopic measurements (C,N,O) of detonation soot produced from labeled and unlabeled Composition B-3 indicate source of solid carbon residues

    Science.gov (United States)

    Podlesak, David; Manner, Virginia; Amato, Ronald; Dattelbaum, Dana; Gusavsen, Richard; Huber, Rachel

    2017-06-01

    Detonation of HE is an exothermic process whereby metastable complex molecules are converted to simple stable molecules such as H2 O, N2, CO, CO2, and solid carbon. The solid carbon contains various allotropes such as detonation nanodiamonds, graphite, and amorphous carbon. It is well known that certain HE formulations such as Composition B (60% RDX, 40% TNT) produce greater amounts of solid carbon than other more oxygen-balanced formulations. To develop a greater understanding of how formulation and environment influence solid carbon formation, we synthesized TNT and RDX with 13 C and 15 N at levels slightly above natural abundance levels. Synthesized RDX and TNT were mixed at a ratio of 60:40 to form Composition B and solid carbon residues were collected from detonations of isotopically-labeled as well as un-labelled Composition B. The raw HE and detonation residues were analyzed isotopically for C, N, O isotopic compositions. We will discuss differences between treatments groups as a function of formulation and environment. LA-UR - 17-21266.

  11. Application of the mass spectrometry-isotope dilution technique for the determination of uranium contents in rocks

    International Nuclear Information System (INIS)

    Kakazu, M.H.; Iyer, S.S.

    1980-01-01

    Application of the spectrometric isotope dilution technique for the accurate determination of parts per million range of uranium in rock samples is described. The various aspects of the method like sample dissolution, ion exchange separation, mass spectrometric procedures are discussed. A single filament ionization source was employed for the isotope analysis. A carbon reduction method was used to reduce uranium oxide ions to uranium metal ions. The tracer solution for isotope dilution was prepared from National Bureau of Standards uranium isotopic Standard NBS U-970. Uranium contents are meassured for nine rock samples and the values obtained are compared with the uranium values measured by others workers. Errors caused in the uranium determination due to sample splitting problems as well as the incomplete acid digestion of the samples are discussed. (Author) [pt

  12. Effect of novel inhaler technique reminder labels on the retention of inhaler technique skills in asthma: a single-blind randomized controlled trial.

    Science.gov (United States)

    Basheti, Iman A; Obeidat, Nathir M; Reddel, Helen K

    2017-02-09

    Inhaler technique can be corrected with training, but skills drop off quickly without repeated training. The aim of our study was to explore the effect of novel inhaler technique labels on the retention of correct inhaler technique. In this single-blind randomized parallel-group active-controlled study, clinical pharmacists enrolled asthma patients using controller medication by Accuhaler [Diskus] or Turbuhaler. Inhaler technique was assessed using published checklists (score 0-9). Symptom control was assessed by asthma control test. Patients were randomized into active (ACCa; THa) and control (ACCc; THc) groups. All patients received a "Show-and-Tell" inhaler technique counseling service. Active patients also received inhaler labels highlighting their initial errors. Baseline data were available for 95 patients, 68% females, mean age 44.9 (SD 15.2) years. Mean inhaler scores were ACCa:5.3 ± 1.0; THa:4.7 ± 0.9, ACCc:5.5 ± 1.1; THc:4.2 ± 1.0. Asthma was poorly controlled (mean ACT scores ACCa:13.9 ± 4.3; THa:12.1 ± 3.9; ACCc:12.7 ± 3.3; THc:14.3 ± 3.7). After training, all patients had correct technique (score 9/9). After 3 months, there was significantly less decline in inhaler technique scores for active than control groups (mean difference: Accuhaler -1.04 (95% confidence interval -1.92, -0.16, P = 0.022); Turbuhaler -1.61 (-2.63, -0.59, P = 0.003). Symptom control improved significantly, with no significant difference between active and control patients, but active patients used less reliever medication (active 2.19 (SD 1.78) vs. control 3.42 (1.83) puffs/day, P = 0.002). After inhaler training, novel inhaler technique labels improve retention of correct inhaler technique skills with dry powder inhalers. Inhaler technique labels represent a simple, scalable intervention that has the potential to extend the benefit of inhaler training on asthma outcomes. REMINDER LABELS IMPROVE INHALER TECHNIQUE: Personalized

  13. Evaluation of a novel educational strategy, including inhaler-based reminder labels, to improve asthma inhaler technique.

    Science.gov (United States)

    Basheti, Iman A; Armour, Carol L; Bosnic-Anticevich, Sinthia Z; Reddel, Helen K

    2008-07-01

    To evaluate the feasibility, acceptability and effectiveness of a brief intervention about inhaler technique, delivered by community pharmacists to asthma patients. Thirty-one pharmacists received brief workshop education (Active: n=16, CONTROL: n=15). Active Group pharmacists were trained to assess and teach dry powder inhaler technique, using patient-centered educational tools including novel Inhaler Technique Labels. Interventions were delivered to patients at four visits over 6 months. At baseline, patients (Active: 53, CONTROL: 44) demonstrated poor inhaler technique (mean+/-S.D. score out of 9, 5.7+/-1.6). At 6 months, improvement in inhaler technique score was significantly greater in Active cf. CONTROL patients (2.8+/-1.6 cf. 0.9+/-1.4, p<0.001), and asthma severity was significantly improved (p=0.015). Qualitative responses from patients and pharmacists indicated a high level of satisfaction with the intervention and educational tools, both for their effectiveness and for their impact on the patient-pharmacist relationship. A simple feasible intervention in community pharmacies, incorporating daily reminders via Inhaler Technique Labels on inhalers, can lead to improvement in inhaler technique and asthma outcomes. Brief training modules and simple educational tools, such as Inhaler Technique Labels, can provide a low-cost and sustainable way of changing patient behavior in asthma, using community pharmacists as educators.

  14. Reassessment of the NH4 NO3 thermal decomposition technique for calibration of the N2 O isotopic composition.

    Science.gov (United States)

    Mohn, Joachim; Gutjahr, Wilhelm; Toyoda, Sakae; Harris, Eliza; Ibraim, Erkan; Geilmann, Heike; Schleppi, Patrick; Kuhn, Thomas; Lehmann, Moritz F; Decock, Charlotte; Werner, Roland A; Yoshida, Naohiro; Brand, Willi A

    2016-09-08

    In the last few years, the study of N 2 O site-specific nitrogen isotope composition has been established as a powerful technique to disentangle N 2 O emission pathways. This trend has been accelerated by significant analytical progress in the field of isotope-ratio mass-spectrometry (IRMS) and more recently quantum cascade laser absorption spectroscopy (QCLAS). Methods The ammonium nitrate (NH 4 NO 3 ) decomposition technique provides a strategy to scale the 15 N site-specific (SP ≡ δ 15 N α - δ 15 N β ) and bulk (δ 15 N bulk  = (δ 15 N α  + δ 15 N β )/2) isotopic composition of N 2 O against the international standard for the 15 N/ 14 N isotope ratio (AIR-N 2 ). Within the current project 15 N fractionation effects during thermal decomposition of NH 4 NO 3 on the N 2 O site preference were studied using static and dynamic decomposition techniques. The validity of the NH 4 NO 3 decomposition technique to link NH 4 + and NO 3 - moiety-specific δ 15 N analysis by IRMS to the site-specific nitrogen isotopic composition of N 2 O was confirmed. However, the accuracy of this approach for the calibration of δ 15 N α and δ 15 N β values was found to be limited by non-quantitative NH 4 NO 3 decomposition in combination with substantially different isotope enrichment factors for the conversion of the NO 3 - or NH 4 + nitrogen atom into the α or β position of the N 2 O molecule. The study reveals that the completeness and reproducibility of the NH 4 NO 3 decomposition reaction currently confine the anchoring of N 2 O site-specific isotopic composition to the international isotope ratio scale AIR-N 2 . The authors suggest establishing a set of N 2 O isotope reference materials with appropriate site-specific isotopic composition, as community standards, to improve inter-laboratory compatibility. This article is protected by copyright. All rights reserved.

  15. Voltammetry coupled to mass spectrometry in the presence of isotope {sup 18}O labeled water for the prediction of oxidative transformation pathways of activated aromatic ethers: Acebutolol

    Energy Technology Data Exchange (ETDEWEB)

    Bussy, Ugo; Tea, Illa [LUNAM Université de Nantes, CNRS, Chimie et Interdisciplinarité: Synthèse, Analyse et Modélisation (CEISAM), UMR 6230, 2 rue de la Houssinière, BP 92208, F-44322 Nantes cedex 3 (France); Ferchaud-Roucher, Véronique; Krempf, Michel [Université de Nantes, Plateforme Spectrométrie de Masse, CRNH, SFR Santé F. Bonamy, Institut du Thorax, UMR S1087, IRT-UN, BP 70721, 8 Quai Moncousu, 44007 Nantes cedex 1 (France); Silvestre, Virginie; Galland, Nicolas [LUNAM Université de Nantes, CNRS, Chimie et Interdisciplinarité: Synthèse, Analyse et Modélisation (CEISAM), UMR 6230, 2 rue de la Houssinière, BP 92208, F-44322 Nantes cedex 3 (France); Jacquemin, Denis [LUNAM Université de Nantes, CNRS, Chimie et Interdisciplinarité: Synthèse, Analyse et Modélisation (CEISAM), UMR 6230, 2 rue de la Houssinière, BP 92208, F-44322 Nantes cedex 3 (France); Institut Universitaire de France, 103, Boulevard Saint-Michel, 75005 Cedex 5 France (France); Andresen-Bergström, Moa; Jurva, Ulrik [CVGI iMed DMPK, AstraZeneca R and D Mölndal, Mölndal (Sweden); and others

    2013-01-31

    Highlights: ► Voltammetry coupled to mass spectrometry method as a useful tool for on-line predictions of electrochemical transformations. ► Evidence of the O-dealkoxylation reaction pathway of acebutolol in the presence of labeled water. ► New approach for on line EC-MS applications. -- Abstract: The coupling between an electrochemical cell (EC) and a mass spectrometer (MS) is a useful screening tool (EC-MS) to study the oxidative transformation pathways of various electroactive species. For that purpose, we showed that the EC-MS method, carried out in the presence and absence of isotope {sup 18}O labeled water leads not only to a fast identification of oxidation products but also leads to a fast elucidation of the mechanism pathway reaction. We examined herein the case of the electrochemical hydrolysis of activated aromatic ether. Acebutolol (β-blockers) was selected herein as m