N-15 tracing helps explaining N leaching losses from contrasting forest ecosystems

Science.gov (United States)

Staelens, J.; Rütting, T.; Huygens, D.; Müller, C.; Verheyen, K.; Boeckx, P.

2009-04-01

Despite chronically enhanced nitrogen (N) deposition to forest ecosystems in Europe and NE America, considerable N retention by forests has been observed, reducing N leaching losses. Organic and mineral soil layers typically immobilize more N than the aboveground biomass, but it is unclear which factors determine N retention in forest ecoystems. However, this knowledge is crucial to assess the impact of changing anthropogenic N emissions on future N cycling and N loss of forests. For coniferous and deciduous forest stands at comparable sites, it is known that both N deposition onto the forest floor as well as N loss by leaching below the rooting zone are significantly higher in coniferous stands. In addition, the N loss in coniferous stands is often more enhanced than can be explained by the higher N input only. This suggests lower N retention by coniferous stands, and may be related to differences in litter and soil characteristics, microbial activity, and N uptake by plant roots. To test this hypothesis, we studied the effect of forest type on N retention using 15N tracing techniques: a field tracer experiment and a combination of in situ isotope pool dilution and a tracing model. The N dynamics were examined for two adjacent forest stands (pedunculate oak (Quercus robur L.) and Scots pine (Pinus sylvestris L.)) on a well-drained sandy soil and with a similar stand history, located in a region with high N deposition (Belgium). Input-output N budgets were established by quantifying atmospheric deposition and leaching below the rooting zone, and confirmed the above finding of higher N deposition and disproportionately higher N loss for the pine stand compared to the oak stand. First, the fate of inorganic N within the ecosystems was studied by spraying three pulses of dissolved 15N, either as ammonium or as nitrate, onto the forest floor in 12 plots of 25 m2. The organic and mineral soil layers, tree roots, soil water percolate, ferns, and tree foliage were sampled

Methodical investigation of the endogenous N excretion in feces by 15N-labelled rats

International Nuclear Information System (INIS)

Bergner, U.; Bergner, H.

1983-01-01

Wistar rats (approximately 100g live weight, n = 8) received a wheat diet and were labelled over a period of 7 days with 15 N-ammonium acetate. From day 1 - 5 of the experiment after the end of the labelling feces and urine were collected and analysed. After the animals were killed (day 5 of the experiment) the atom-% 15 N excess ( 15 N') in the contents of the digestive tract as well as in the tissues of stomach wall, intestinal wall, liver, pancreas and blood plasma was determined. The TCA-soluble fraction of the blood plasma showed 0.44 atom-% 15 N' on day 5 after the end of 15 N labelling. 3 hours before the killing fecal N also showed 0.44 and during the last collection period (24 hours before) an average of 0.51 atom-% 15 N'. Urine decreased in the same period from 0.71 to 0.59 atom-% 15 N'. The endogenous fecal N is calculated to 88%. As the tissues of the digestive tract are likely to supply the biggest part of the endogenous fecal protein, the values of atom-% 15 N' from the TCA-precipitable fraction of the intestinal wall and of the pancreas gland was calculed to an average of 0.526. According to this the calculation endogenous fecal N is 84%. It is probable that the quota of endogenous fecal N in the total amount of fecal N varies in dependence on the fermentable crude fiber in the diet as well as on the age of the test animals and thus the bacterial protein synthesis in the colon. As the N used by the bacteria is likely to come from the TCA-soluble fraction of the blood, the calculation formula suggested, which uses the TCA-soluble fraction of the blood plasma, achieves good approximate values also for higher bacterial protein synthesis in the colon. (author)

15N incorporation into organ proteins of newborn rats following single pulse-labelling with different tracers

International Nuclear Information System (INIS)

Wutzke, K.D.; Plath, C.; Richter, I.; Heine, W.; Zhukova, T.P.; Sorokina, E.G.; Friedrich, M.

1987-01-01

A short-chain 15 N-peptide mixture characterized by an average chain length of 2.3 was obtained when 15 N-labelled yeast protein was hydrolyzed enzymatically by thermitase from Thermoactinomyces vulgaris. Fifteen newborn Wistar rats were given a single pulse of [ 15 N]glycine. [ 15 N]H 4 Cl and [ 15 N]yeast protein thermitasehydrolysate (YPTH) in a dosage of 50 mg 15 N excess kg -1 by gastric tube. In comparison with [ 15 N]glycine the 15 N incorporation rates of brain, muscle and liver were approximately 150% higher after [ 15 N]YPTH application. Uniform labelling, high 15 N enrichment, almost complete absorption, avoidance of imbalances and the low price make this tracer substance superior to other tracers conventionally used for organ labelling. (author)

High-yield nitration of benzene in the synthesis of 15N-labelled nitrobenzene, acetanilide, and diphenylamine

International Nuclear Information System (INIS)

Konior, R.J.; Ling Yang; Walter, R.I.

1990-01-01

Labelled H 15 NO 3 was used as the least-cost source of nitrogen label to prepare nitrobenzene by reaction of acetyl nitrate with excess benzene. This labelled product was subsequently converted to acetanilide- 15 N and diphenylamine- 15 N. (author)

Effect of organic matter application on the fate of 15N-labeled ammonium fertilizer in an upland soil

International Nuclear Information System (INIS)

Nishio, T.; Oka, N.

2003-01-01

The effect of the application of organic matter on the fate of 15 N-labeled ammonium was investigated in a field. The organic materials incorporated into the experimental plots consisted of wheat straw, rape, pig compost, cow compost, plant manure. In May 2000, 10 g N m -2 of 15 N-labeled ammonium was applied to the field together with the organic materials, and maize and winter wheat were consecutively cultivated. The recovery of applied 15 N in soils and plants was determined after the harvest of each crop. Although only about 10% of the applied 15 N-labeled fertilizer remained in the 0-30 cm layer of the Control Plot and the Plant Manure Plot, more than 25% of the applied 15 N remained in the Pig Compost Plot. Amount and proportion of the immobilized 15 N to those of total N or microbial biomass N in soils were determined for the topsoil samples (0-10 cm layer). The amounts of both microbial biomass N and total immobilized 15 N in soil were highest in the Pig Compost Plot. Although the amount of microbial biomass N was comparable to the amount of immobilized 15 N-labeled fertilizer in soil, the amounts of 15 N-labeled fertilizer contained in the microbial biomass accounted for less than 10 % of the amount of total immobilized 15 N in soil. The ratio of the amount of 15 N-labeled fertilizer contained in biomass N to the total amount of biomass N was one order to magnitude higher than the ratio of the amount of immobilized 15 N-labeled fertilizer to the amount of total N in soil. No conspicuous changes in the amount of immobilized 15 N in soil were observed during the cultivation of winter wheat except for the Pig Compost Plot. No significant correlation was recognized between the amount of 15 N-labeled fertilizer contained in microbial biomass before wheat cultivation and that of 15 N-labeled fertilizer absorbed by wheat, indicating that microbial N immobilized during the growth period of the former crop (maize) was not a significant source of N for the latter

Nitrogen-15 labeling of Crotalaria juncea green manure Marcação do adubo verde Crotalaria juncea com 15N

Directory of Open Access Journals (Sweden)

Edmilson José Ambrosano

2003-02-01

Full Text Available Most studies dealing with the utilization of 15N labeled plant material do not present details about the labeling technique. This is especially relevant for legume species since biological nitrogen fixation difficults plant enrichment. A technique was developed for labeling leguminous plant tissue with 15N to obtain labeled material for nitrogen dynamics studies. Sun hemp (Crotalaria juncea L. was grown on a Paleudalf, under field conditions. An amount of 58.32 g of urea with 70.57 ± 0.04 atom % 15N was sprayed three times on plants grown on eight 6-m²-plots. The labelled material presented 2.412 atom % 15N in a total dry matter equivalent to 9 Mg ha-1 This degree of enrichment enables the use of the green manure in pot or field experiments requiring 15N-labeled material.A grande maioria dos estudos com a utilização de material vegetal marcado com o isótopo 15N não apresentam detalhes tão importantes sobre como foram obtidos esses materiais. Em se tratando de marcação de leguminosas as dificuldades em se obter material marcado com 15N são ainda maiores pelo fato de serem plantas fixadoras de nitrogênio. Isso posto foi estabelecida uma técnica de marcação de leguminosas com nitrogênio (15N, com o objetivo de obter material vegetal marcado isotopicamente para estudos de dinâmica do nitrogênio. Cultivou-se a leguminosa crotalária júncea (Crotalaria juncea L., em Argissolo Vermelho Amarelo distrófico, em campo. Ao se aplicarem via foliar 58,32 gramas de uréia em oito canteiros experimentais, (uréia com 70,57 ± 0,04% de átomos de 15N parceladas em três vezes, obteve-se material vegetal marcado seco que continha 2,412 % em átomos de 15N em uma massa seca equivalente a 9 Mg ha-1. Essa marcação permite o uso dessa massa vegetal em estudos de dinâmica de nitrogênio.

High-yield nitration of benzene in the synthesis of sup 15 N-labelled nitrobenzene, acetanilide, and diphenylamine

Energy Technology Data Exchange (ETDEWEB)

Konior, R.J.; Ling Yang; Walter, R.I. (Illinois Univ., Chicago, IL (USA). Dept. of Chemistry)

1990-11-01

Labelled H{sup 15}NO{sub 3} was used as the least-cost source of nitrogen label to prepare nitrobenzene by reaction of acetyl nitrate with excess benzene. This labelled product was subsequently converted to acetanilide-{sup 15}N and diphenylamine-{sup 15}N. (author).

Use of 15N enriched plant material for labelling of soil nitrogen in legume dinitrogen fixation experiments

International Nuclear Information System (INIS)

Jensen, E.S.

1989-06-01

The soil nitrogen in a field plot was labelled with nitrogen-15 (15N) by incorporating labelled plant material derived from previous experiments. The plot was used the following 3 years for determination of the amount of N2 fixed by different leguminous plants. The atom % 15N excess in grains of cereals grown as reference crops was 0.20, 0.05 and 0.03 in the 3 years, respectively. In the first year the level of enrichment was adequate for estimating symbiotic nitrogen fixation. In the second and third year lack of precision in determination of the 15N/14N ratios of legume N, may have caused an error in estimates of nitrogen fixation. About 23% of the labelled N was taken up by plants during the 3 years of cropping; after 4 years about 44% of the labelled N was found still to be present in the top soil. The labelling of the soil nitrogen with organic bound 15N, compared to adding mineral 15N at sowing, is advantageous because the labelled N is released by mineralization so that the enrichment of the plant available soil N pool become more uniform during the growth season; and high levels of mineral N, which may depress the fixation process, is avoided. (author) 7 tabs., 1 ill., 30 refs

Nitrogen assimilation and short term retention in a nutrient-rich tidal freshwater marsh – a whole ecosystem 15N enrichment study

Directory of Open Access Journals (Sweden)

B. Gribsholt

2007-01-01

Full Text Available An intact tidal freshwater marsh system (3477 m2 was labelled by adding 15N-ammonium as a tracer to the flood water inundating the ecosystem. The appearance and retention of 15N-label in different marsh components (leaves, roots, sediment, leaf litter and invertebrate fauna was followed over 15 days. This allowed us to elucidate the direct assimilation and dependence on creek-water nitrogen on a relatively short term and provided an unbiased assessment of the relative importance of the various compartments within the ecosystem. Two separate experiments were conducted, one in spring/early summer (May 2002 when plants were young and building up biomass; the other in late summer (September 2003 when macrophytes were in a flowering or early senescent state. Nitrogen assimilation rate (per hour inundated was >3 times faster in May compared to September. On both occasions, however, the results clearly revealed that the less conspicuous compartments such as leaf litter and ruderal vegetations are more important in nitrogen uptake and retention than the prominent reed (Phragmites australis meadows. Moreover, short-term nitrogen retention in these nutrient rich marshes occurs mainly via microbial pathways associated with the litter and sediment. Rather than direct uptake by macrophytes, it is the large reactive surface area provided by the tidal freshwater marsh vegetation that is most crucial for nitrogen transformation, assimilation and short term retention in nutrient rich tidal freshwater marshes. Our results clearly revealed the dominant role of microbes in initial nitrogen retention in marsh ecosystems.

Stabilization and plant uptake of N from 15N-labelled pea residue 16.5 years after incorporation in soil

DEFF Research Database (Denmark)

Laberge, G.; Ambus, P.; Hauggaard-Nielsen, H.

2006-01-01

The decline of N from N-15-labelled mature pea residues was followed in unplanted soil over 16.5 yr. Eight years after residue incorporation, 24% of the residue N-15 input was still present in the soil and, after 16.5 yr, 16% of the residue N-15 input remained. A double exponential model......-amended soils were obtaining 1.7% of their N from residue N. This is, to our knowledge, the longest study on decay of N in soils from N-15-labelled crop residues. The current study thus provides a unique data set for our empirical understanding of N-dynamics in agricultural systems, which is a prerequisite...

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

Science.gov (United States)

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

2014-01-01

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

Estimation of the endogenous N proportions in ileal digesta and faeces in 15N-labelled pigs

International Nuclear Information System (INIS)

Simon, O.; Bergner, H.

1987-01-01

4 castrated male pigs 40 kg fitted with simple 'T' cannulas in the terminal ileum were given 15 N-labelled ammonium salts, added to a low protein diet, for 6 days. Excretion of 15 N in urine and feces was monitored daily throughout the labelling and subsequent experimental periods. During the experimental period the pigs were given a diet based on wheat and fish meal, supplemented with varying levels of partially hydrolyzed straw meal to give crude fiber contents ranging from 40 to 132 g/kg. After adaptation to the particular levels of straw meal, feces and ileal digesta were collected during successive 24 h periods. N digestibility values were determined by the chromium oxide ratio method. The retention of 15 N-labelled non-specific N was 0.46 of the dose given. The validity of using urine values as a measure of 15 N abundance in endogenous N was demonstrated by the similarity of 15 N abundance in urine immediately before slaughter at the end of the experiment and in the digestive secretory organs thereafter. The average amount of endogenous N passing the terminal ileum was 3.4 g/day or 0.30-0.50 of total ileal N flow. This was not affected by dietary fiber level. The proportion of fecal N which was of endogenous origin was similar to that in ileal digesta, suggesting similar utilization of endogenous and residual dietary N by hindgut bacteria. Half the endogenous N entering the large intestine was reabsorbed there. Increasing dietary crude fiber from 40 to 132 g/kg increased fecal endogenous N excretion from 1.3 to 2.0 g/animal and day. (author)

The use of N-15 labelling to study the turnover and utilization of ruminant manure N

DEFF Research Database (Denmark)

Sørensen, P.; Jensen, E.S.

1998-01-01

An improved understanding of the cycling of animal manure N is a prerequisite for malting better use of this N source. A sheep was fed N-15-Iabelled grass in order to study the fate of N-15-Iabelled ruminant manure N in the plant-soil system. The uniformity of labelling was found to be satisfactory...

Nitrogen-15 labeling of Crotalaria juncea green manure

International Nuclear Information System (INIS)

Ambrosano, Edmilson Jose; Rossetto, Raffaella; Trivelin, Paulo Cesar Ocheuze; Muraoka, Takashi; Bendassolli, Jose Albertino; Cantarella, Heitor; Ambrosano, Glaucia Maria Bovi; Tamiso, Luciano Grassi; Vieira, Felipe de Campos; Prada Neto, Ithamar

2003-01-01

Most studies dealing with the utilization of 15 N labeled plant material do not present details about the labeling technique. This is especially relevant for legume species since biological nitrogen fixation difficult plant enrichment. A technique was developed for labeling leguminous plant tissue with 15 N to obtain labeled material for nitrogen dynamics studies. Sun hemp (Crotalaria juncea L.) was grown on a Paleudalf, under field conditions. An amount of 58.32 g of urea with 70.57± 0.04 atom % 15 N was sprayed three times on plants grown on eight 6-m2-plots. The labelled material presented 2.412 atom % 15 N in a total dry matter equivalent to 9 Mg ha -1 This degree of enrichment enables the use of the green manure in pot or field experiments requiring 15 N-labeled material. (author)

Utilization of 15N-labelled urea in laying hens. 8

International Nuclear Information System (INIS)

Gruhn, K.; Graf, H.

1987-01-01

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

Investigation of the metabolism of colostomized laying hens with 15N-labelled wheat. 6

International Nuclear Information System (INIS)

Gruhn, K.; Hennig, A.

1980-01-01

Three colostomized laving hens received 40 g 15 N-labelled wheat with 20.13 atom-% 15 N excess ( 15 N'), 19.18 atom-% 15 N'-lysine, 18.17 atom-% 15 N'-histidine and 20.43 atom-% 15 N'-arginine per day over a period of four days. After having received the same non-labelled feed ration on the following four days, the hens were slaughtered. The incorporation and distribution of 15 N' in the total nitrogen and the nitrogen of the basic amino acids was determined in liver, kidneys, muscles, bones and the remaining carcass (excluding blood, digestive tract and genital organs). The quota of nitrogen of natural isotope frequency ( 14 N) of the total 14 N of the hens' carcasses was 47% in the muscles, 14% in the bones and 20% in the feathers; the relative 15 N' values were 37%, 8% and 1%, resp. The atom-% 15 N' in the kidneys was twice as much as in the liver four days after the last 15 N' application. The average percentage of the nitrogen in the three basic amino acids of the total nitrogen in the tissues and organs (excluding feathers) is 25% concerning both 14 N and 15 N'. The 15N' balance revealed that in hen 1 100%, in hen 2 102% and in hen 3 101% of the consumed wheat 15 N' were found. (author)

Foliar absorption of 15N labeled urea by tea plant

International Nuclear Information System (INIS)

Hoshina, Tsuguo; Kozai, Shuji; Ishigaki, Kozo

1978-01-01

The effect of foliar application on the nitrogen nutrient status of tea shoots has been studied using 15 N labelled urea. Furthermore, the difference in nitrogen utilization by tea plant between foliar applied and top dressed nitrogen was investigated using 15 N labelled urea and ammonium sulfate. The foliar application of urea increased the amount of chlorophyll and total nitrogen in the new shoot, and the foliar application was more effective under shading condition. The urea sprayed upon old leaves prior to the opening of new leaf translocated to the new shoots. However, the foliar application after the opening of new leaf was more effective on nitrogen absorption by new shoots than one prior to that, and rather than top dressing for new shoots. It could be recognized that the foliar application of urea raises the nitrogen nutrient status of tea leaves in summer. (author)

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

Recovery of nitrogen by spring barley following incorporation of 15N-labelled straw and catch crop material

DEFF Research Database (Denmark)

Thomsen, I.K.; Jensen, E.S.

1994-01-01

The recovery by spring barley (Hordeum vulgare L.) of nitrogen mineralized from N-15-labelled straw and ryegrass material was followed for 3 years in the field. The effects of separate and combined applications of straw and ryegrass were studied using cross-labelling with N-15. Reference plots re...... mineral fertilizer was in the second and third barley crop similar to the recovery of N from incorporated plant residues.......The recovery by spring barley (Hordeum vulgare L.) of nitrogen mineralized from N-15-labelled straw and ryegrass material was followed for 3 years in the field. The effects of separate and combined applications of straw and ryegrass were studied using cross-labelling with N-15. Reference plots...... receiving (NH4NO3)-N-15-N-15 were included. Plant samples were taken every second week until maturity during the first growing season and at maturity in the two following years. Incorporation of plant material had no significant influence on the above-ground dry matter yield of the barley. The barley...

Tracing in situ amino acid uptake in plants and microbes with¹⁵N¹³C labelled compounds

DEFF Research Database (Denmark)

Andresen, Louise Christoffersen; Michelsen, Anders; Jonasson, Sven Evert

amino acids. Furthermore, tannin addition tended to reduce plant uptake of label. By combining data on 15N recovery after 1 day in shoots and roots (fine and coarse) of the dominant heathland plants: the evergreen dwarf shrub Calluna vulgaris and the graminoid Deschampsia flexuosa, in soil...... microorganisms (chloroform fumigation extraction) and in soil water, we discuss the relative importance of free amino acids and ammonium as plant nutrients and microbial substrates in natural N-limited ecosystems with a high proportion of soil N held in tannin-N complexes. ...

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)

Application of 15N labeling to topics in molecular hematology

International Nuclear Information System (INIS)

Lapidot, A.; Irving, C.S.

1975-01-01

The amount of information which can be obtained from many types of spectrometric analysis of compounds of hematological interest can be greatly enhanced when measurements are made on a series of isotopically labeled compounds. A murine Friend virus-induced erythroleukemic cell (FLC) culture was found to be a superior biosynthetic system for the preparation of highly and selectively 15 N and 13 C enriched hemoglobins. A mutant of Rhodopseudomonas spheroides was found suitable for the preparation of larger quantities of >90 percent enriched protoporphyrin-IX- 15 N and coproporphyrin-III-- 15 N. A comparison of the 15 N and 13 C NMR spectra of FLC carbomonoxy-[Gly- 15 N]-hemoglobin, carbomonoxy-[Gly- 13 C/sub alpha/]-hemoglobin, α and β globin-[Gly- 15 N] and globin-[Gly- 13 C/sub alpha/] demonstrated 1) 15 N peptide chemical shifts are sensitive to polypeptide sequence, whereas 13 C α-carbon chemical shifts are not, (2) variations in the solvation of the peptide N-H group can be detected in the 15 N spectra but not the 13 C spectra, (3) 15 N heme resonances could not be detected, whereas 13 C resonances could. These studies indicated that in hemoglobin the glycyl N-H resonances are either solvated by H 2 O or hydrogen bonded to peptide C=0 groups. In denatured globin, the majority of the glycyl residues are rapidly exchanging between these two states

Biosynthesis of agmatine in isolated mitochondria and perfused rat liver: studies with 15N-labelled arginine

Science.gov (United States)

2005-01-01

An important but unresolved question is whether mammalian mitochondria metabolize arginine to agmatine by the ADC (arginine decarboxylase) reaction. 15N-labelled arginine was used as a precursor to address this question and to determine the flux through the ADC reaction in isolated mitochondria obtained from rat liver. In addition, liver perfusion system was used to examine a possible action of insulin, glucagon or cAMP on a flux through the ADC reaction. In mitochondria and liver perfusion, 15N-labelled agmatine was generated from external 15N-labelled arginine. The production of 15N-labelled agmatine was time- and dose-dependent. The time-course of [U-15N4]agmatine formation from 2 mM [U-15N4]arginine was best fitted to a one-phase exponential curve with a production rate of approx. 29 pmol·min−1·(mg of protein)−1. Experiments with an increasing concentration (0– 40 mM) of [guanidino-15N2]arginine showed a Michaelis constant Km for arginine of 46 mM and a Vmax of 3.7 nmol·min−1·(mg of protein)−1 for flux through the ADC reaction. Experiments with broken mitochondria showed little changes in Vmax or Km values, suggesting that mitochondrial arginine uptake had little effect on the observed Vmax or Km values. Experiments with liver perfusion demonstrated that over 95% of the effluent agmatine was derived from perfusate [guanidino-15N2]arginine regardless of the experimental condition. However, the output of 15N-labelled agmatine (nmol·min−1·g−1) increased by approx. 2-fold (P<0.05) in perfusions with cAMP. The findings of the present study provide compelling evidence that mitochondrial ADC is present in the rat liver, and suggest that cAMP may stimulate flux through this pathway. PMID:15656789

Studies on the protein and amino acid metabolism of laying hens using 15N-labelled casein. 8

International Nuclear Information System (INIS)

Richter, G.

1977-01-01

Four colostomized Leghorn hens were fed, during 6 days, 15 N-labelled casein as sole protein source. Two animals were slaughtered 48 hours, the other two 144 hours after the last 15 N-application. The share of TCE-soluble N in total N averaged 16% for the body parts analysed, i.e. meat, bone, liver, kidneys, oviducts, residual viscera and other. The variation of the lysine, histidine and arginine levels in the body parts ranged from 3.6 to 7.9 g, 1.1 to 3.7 g and 6.4 to 7.4 g in 16.7 g hydrolysate N, respectively. Except for feathers, the analysed body parts contained an excess amount of heavy nitrogen. The degree of labelling was found to depend on the time of slaughtering after the tracer application. In the liver and in the oviduct being metabolically active organs, the 15 N-excess in the total N fraction decreased by 45% between the 2nd and the 6th days after 15 N-feeding, whilst in the meat it went down by 20%. The decline of the 15 N-concentration in the TCE-soluble N compounds was faster than in the total N-fraction. Out of the body samples analysed, the lysine of the liver having 0.26 atom% 15 N-excess was found to be more strongly labelled in hens 1 and 2. The amino acid arginine reached about the same level of labelling, the 15 N-frequency of histidine being the lowest. (author)

Dynamics of amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 1

International Nuclear Information System (INIS)

Hennig, A.; Gruhn, K.; Kirchner, E.

1987-01-01

In a 6-day preliminary period with a pelleted ration 12 colostomized laying hybrids received 15 N-labelled wheat protein over 4 days. The labelling of the wheat was 14.37 atom-% 15 N excess ( 15 N'). During the 4-day application of 15 N-labelled wheat protein each hen consumed 12.08 g N, 3.52 g lysine, 2.12 g histidine, 4.41 g arginine, of which were 540 mg 15 N', 18.1 mg lysine 15 N', 21.5 mg histidine 15 N' and 47.9 mg arginine 15 N'. Heavy nitrogen was determined in urine and its uric acid N in the daily urine samples of the individual animals. The average daily urine N excretion was 54% of the total nitrogen consumed with the ration. The labelling of the urine N reached a plateau on the fourth day of the experiment with 3.2 atom-% 15 N'. On an average of the total experiment the quota of heavy nitrogen of the uric acid in the total 15 N' of the urine was 83.4% and that of uric acid nitrogen in the total urine nitrogen 80.8%. (author)

15N-Labelling and structure determination of adamantylated azolo-azines in solution

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Sergey L. Deev

2017-11-01

Full Text Available Determining the accurate chemical structures of synthesized compounds is essential for biomedical studies and computer-assisted drug design. The unequivocal determination of N-adamantylation or N-arylation site(s in nitrogen-rich heterocycles, characterized by a low density of hydrogen atoms, using NMR methods at natural isotopic abundance is difficult. In these compounds, the heterocyclic moiety is covalently attached to the carbon atom of the substituent group that has no bound hydrogen atoms, and the connection between the two moieties of the compound cannot always be established via conventional 1H-1H and 1H-13C NMR correlation experiments (COSY and HMBC, respectively or nuclear Overhauser effect spectroscopy (NOESY or ROESY. The selective incorporation of 15N-labelled atoms in different positions of the heterocyclic core allowed for the use of 1H-15N (JHN and 13C-15N (JCN coupling constants for the structure determinations of N-alkylated nitrogen-containing heterocycles in solution. This method was tested on the N-adamantylated products in a series of azolo-1,2,4-triazines and 1,2,4-triazolo[1,5-a]pyrimidine. The syntheses of adamantylated azolo-azines were based on the interactions of azolo-azines and 1-adamatanol in TFA solution. For azolo-1,2,4-triazinones, the formation of mixtures of N-adamantyl derivatives was observed. The JHN and JCN values were measured using amplitude-modulated 1D 1H spin-echo experiments with the selective inversion of the 15N nuclei and line-shape analysis in the 1D 13С spectra acquired with selective 15N decoupling, respectively. Additional spin–spin interactions were detected in the 15N-HMBC spectra. NMR data and DFT (density functional theory calculations permitted to suggest a possible mechanism of isomerization for the adamantylated products of the azolo-1,2,4-triazines. The combined analysis of the JHN and JCN couplings in 15N-labelled compounds provides an efficient method for the structure

Synthesis of {sup 15}N labeled glyphosate; Sintese do glifosato enriquecido com {sup 15}N

Energy Technology Data Exchange (ETDEWEB)

Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Tavares, Glauco Arnold; Rossete, Alexssandra L.R.M.; Tagliassachi, Romulo Barbieri; Prestes, Cleuber Vieira [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Dept. de Isotopos Estaveis]. E-mail: crolivei@cena.usp.br

2005-07-01

Amongst the actually commercialized herbicides the Glyphosate is the most used in Brazil. Its efficiency as well as the others herbicides against undesirable weeds is harmed by its final composts left at the environment. Although studies has being carried out to improve the knowledge about the herbicides behavior at the environment its complexity has led them towards innumerous to new significant research work where the use of radiolabeled composts (radiative tracers) are recommended to evaluate their bio-availability in the soil. However is the use, the manipulation and the storage of radiolabeled composts is requires an extra care under chemical safety point of view. The use of non radiolabeled composts is a world tendency especially for field researches. Under this context the presented work describes a method for the synthesis of {sup 15}N labeled glyphosate. The {sup 15}N-herbicide was undertaken by phosphometilation with the phosphit dialquil and {sup 15}N-glycine. The tests where carried out through a micro scale production plant and of equimolars amounts. At these conditions it's was possible to reach approximately a 20% of yield. At the conclusion of a best operational condition its expected to offer another important toll that shall be used in glyphosate behavior at the environment and undesirably weeds. (author)

Sub-cellular localisation of a 15N-labelled peptide vector using NanoSIMS imaging

Science.gov (United States)

Römer, Winfried; Wu, Ting-Di; Duchambon, Patricia; Amessou, Mohamed; Carrez, Danièle; Johannes, Ludger; Guerquin-Kern, Jean-Luc

2006-07-01

Dynamic SIMS imaging is proposed to map sub-cellular distributions of isotopically labelled, exogenous compounds. NanoSIMS imaging allows the characterisation of the intracellular transport pathways of exogenous molecules, including peptide vectors employed in innovative therapies, using stable isotopes as molecular markers to detect the compound of interest. Shiga toxin B-subunit (STxB) was chosen as a representative peptide vector. The recombinant protein ( 15N-STxB) was synthesised in Escherichia coli using 15NH 4Cl as sole nitrogen source resulting in 15N enrichment in the molecule. Using the NanoSIMS 50 ion microprobe (Cameca), different ion species ( 12C 14N -, 12C 15N -, 31P -) originating from the same sputtered micro volume were simultaneously detected. High mass resolving power enabled the discrimination of 12C 15N - from its polyatomic isobars of mass 27. We imaged the membrane binding and internalisation of 15N-STxB in HeLa cells at spatial resolutions of less than 100 nm. Thus, the use of rare stable isotopes like 15N with dynamic SIMS imaging permits sub-cellular detection of isotopically labelled, exogenous molecules and imaging of their transport pathways at high mass and spatial resolution. Application of stable isotopes as markers can replace the large and chemically complex tags used for fluorescence microscopy, without altering the chemical and physical properties of the molecule.

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

Behavior of 15N-labelled amino acids in germinated corn

International Nuclear Information System (INIS)

Samukawa, Kisaburo; Yamaguchi, Masuro

1979-01-01

By investigating the rise and fall of 15 N-labelled amino acids in germinated corns, the behavior of amino radicals in free amino acids, the influence of the hydrolysis products of stored proteins on free amino acids and the change from heterotrophy to autotrophy of seeds were clarified. The amount of amino acid production depending on external nitrogen was very small in the early period of germination. 15 N incorporation into proline was not observed in the early period of germination, which suggested that the proline may be nitrogen-storing source. Most of the amino-state nitrogen of asparagine accumulated at the time of germination was internal nitrogen, and this fact suggested that aspartic acid serve as the acceptor of ammonia produced in the early stage of germination. 15 N content increased significantly on 9 th day after germination, and decreased on 12 th day. These facts prove that there are always active decomposition and production of protein in plant body. (Kobatake, H.)

Nitrogen turnover of three different agricultural soils determined by 15N triple labelling

Science.gov (United States)

Fiedler, Sebastian R.; Kleineidam, Kristina; Strasilla, Nicol; Schlüter, Steffen; Reent Köster, Jan; Well, Reinhard; Müller, Christoph; Wrage-Mönnig, Nicole

2017-04-01

To meet the demand for data to improve existing N turnover models and to evaluate the effect of different soil physical properties on gross nitrogen (N) transformation rates, we investigated two arable soils and a grassland soil after addition of ammonium nitrate (NH4NO3), where either ammonium (NH4+), or nitrate (NO3-), or both pools have been labelled with 15N at 60 atom% excess (triple 15N tracing method). Besides NH4+, NO3- and nitrite (NO2-) contents with their respective 15N enrichment, nitrous oxide (N2O) and dinitrogen (N2) fluxes have been determined. Each soil was adjusted to 60 % of maximum water holding capacity and pre-incubated at 20˚ C for two weeks. After application of the differently labelled N fertilizer, the soils were further incubated at 20˚ C under aerobic conditions in a He-N2-O2 atmosphere (21 % O2, 76 He, 2% N2) to increase the sensitivity of N2 rates via the 15N gas flux method. Over a 2 week period soil N pools were quantified by 2 M KCl extraction (adjusted to pH 7 to prevent nitrite losses) (Stevens and Laughlin, 1995) and N gas fluxes were measured by gas chromatography in combination with IRMS. Here, we present the pool sizes and fluxes as well as the 15N enrichments during the study. Results are discussed in light of the soil differences that were responsible for the difference in gross N dynamics quantified by the 15N tracing model Ntrace (Müller et al., 2007). References Müller, C., T. Rütting, J. Kattge, R.J. Laughlin, and R.J. Stevens, (2007) Estimation of parameters in complex 15N tracing models by Monte Carlo sampling. Soil Biology & Biochemistry. 39(3): p. 715-726. Stevens, R.J. and R.J. Laughlin, (1995) Nitrite transformations during soil extraction with potassium chloride. Soil Science Society of America Journal. 59(3): p. 933-938.

Combined solid state and solution NMR studies of α,ε-15N labeled bovine rhodopsin

International Nuclear Information System (INIS)

Werner, Karla; Lehner, Ines; Dhiman, Harpreet Kaur; Richter, Christian; Glaubitz, Clemens; Schwalbe, Harald; Klein-Seetharaman, Judith; Khorana, H. Gobind

2007-01-01

Rhodopsin is the visual pigment of the vertebrate rod photoreceptor cell and is the only member of the G protein coupled receptor family for which a crystal structure is available. Towards the study of dynamics in rhodopsin, we report NMR-spectroscopic investigations of α,ε- 15 N-tryptophan labeled rhodopsin in detergent micelles and reconstituted in phospholipids. Using a combination of solid state 13 C, 15 N-REDOR and HETCOR experiments of all possible 13 C' i-1 carbonyl/ 15 N i -tryptophan isotope labeled amide pairs, and H/D exchange 1 H, 15 N-HSQC experiments conducted in solution, we assigned chemical shifts to all five rhodopsin tryptophan backbone 15 N nuclei and partially to their bound protons. 1 H, 15 N chemical shift assignment was achieved for indole side chains of Trp35 1.30 and Trp175 4.65 . 15 N chemical shifts were found to be similar when comparing those obtained in the native like reconstituted lipid environment and those obtained in detergent micelles for all tryptophans except Trp175 4.65 at the membrane interface. The results suggest that the integrated solution and solid state NMR approach presented provides highly complementary information in the study of structure and dynamics of large membrane proteins like rhodopsin

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

O potencial da rotulação metabólica de 15N para a pesquisa de esquizofrenia The potential of 15N metabolic labeling for schizophrenia research

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Michaela D. Filiou

2013-01-01

Full Text Available Pesquisas em psiquiatria ainda necessitam de estudos não dirigidos por hipóteses para revelar fundamentos neurobiológicos e biomarcadores moleculares para distúrbios psiquiátricos. Metodologias proteômicas disponibilizam uma série de ferramentas para esses fins. Apresentamos o princípio de rotulação metabólica utilizando 15N para proteômica quantitativa e suas aplicações em modelos animais de fenótipos psiquiátricos com um foco particular em esquizofrenia. Exploramos o potencial de rotulação metabólica por 15N em diferentes tipos de experimentos, bem como suas considerações metodológicas.Psychiatric research is in need of non-hypothesis driven approaches to unravel the neurobiological underpinnings and identify molecular biomarkers for psychiatric disorders. Proteomics methodologies constitute a state-of-the-art toolbox for biomarker discovery in psychiatric research. Here we present the principle of in vivo 15N metabolic labeling for quantitative proteomics experiments and applications of this method in animal models of psychiatric phenotypes, with a particular focus on schizophrenia. Additionally we explore the potential of 15N metabolic labeling in different experimental set-ups as well as methodological considerations of 15N metabolic labeling-based quantification studies.

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.

Nitrogen budget of 15 N-Labelled urea applied to dry land wheat as affected by seasonal rainfall

International Nuclear Information System (INIS)

Abdel-Monem, M.A.; Ryan, J.; Christianson, B.

1999-01-01

Field trial was conducted at two locations of the semi-arid region of Morocco with an objective of estimating the N balance for the applied urea to wheat under two different agroecological zones. N-15 labelled urea was applied in microplots (1 m x 1 m) at tillering stage at rates of 0,40,80, and 120 kg/ha. Soil and plant samples were analyzed for N-15 excess. An average of 45% increase in grain yield of wheat was due to fertilization with 60 kg N/ha. Recovery of N-15 labelled N ranged between 28.8 and 45.1% for the rate of 40 kg N/ha. Results suggested that efficiency of the applied fertilizer is governed by seasonal rainfall and soil properties

The Contamination of Commercial 15N2 Gas Stocks with 15N–Labeled Nitrate and Ammonium and Consequences for Nitrogen Fixation Measurements

Science.gov (United States)

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

2014-01-01

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

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)

Ecosystem Nitrogen Retention Following Severe Bark Beetle and Salvage Logging Disturbance in Lodgepole Pine Forests: a 15N Enrichment Study

Science.gov (United States)

Avera, B.; Rhoades, C.; Paul, E. A.; Cotrufo, M. F.

2017-12-01

In recent decades, bark beetle outbreaks have caused high levels of tree mortality in lodgepole pine (Pinus contorta) dominated forests across western North America. Previous work has found increased soil mineral nitrogen (N) with tree mortality in beetle infested stands, but surprisingly little change in stream N export. These findings suggest an important role of residual live vegetation and altered soil microbial response for retaining surplus N and mitigating N losses from disturbed lodgepole forests. Post outbreak salvage of merchantable timber reduces fuel levels and promotes tree regeneration; however, the implications of the combined bark beetle and harvesting disturbances on ecosystem N retention and productivity are uncertain. To advance understanding of post-disturbance N retention we compare unlogged beetle-infested forests and salvage logged stands with post-harvest woody residue retention or removal. We applied 15N-labeled (2 atom%) and natural abundance ammonium sulfate to eight year old lodgepole pine seedlings in three replicate plots of the three forest management treatments. This approach allows us to quantify the relative contributions of N retention in soil, microbial biomass, and plant tissue. Our study targets gaps in understanding of the processes that regulate N utilization and transfer between soil and vegetation that result in effective N retention in lodgepole pine ecosystems. These findings will also help guide forest harvest and woody residue management practices in order to maintain soil productivity.

Experiments and strategies for the assignment of fully13 C/15N-labelled polypeptides by solid state NMR

International Nuclear Information System (INIS)

Straus, Suzana K.; Bremi, Tobias; Ernst, Richard R.

1998-01-01

High-resolution heteronuclear NMR correlation experiments and strategies are proposed for the assignment of fully 13 C/ 15 N-labelled polypeptides in the solid state. By the combination of intra-residue and inter-residue 13 C- 15 N correlation experiments with 13 C- 13 C spin-diffusion studies, it becomes feasible to partially assign backbone and side-chain resonances in solid proteins. The performance of sequences using 15 N instead of 13 C detection is evaluated regarding sensitivity and resolution for a labelled dipeptide (L-Val-L-Phe). The techniques are used for a partial assignment of the 15 N and 13 C resonances in human ubiquitin

A cost-effective approach to produce 15N-labelled amino acids employing Chlamydomonas reinhardtii CC503.

Science.gov (United States)

Nicolás Carcelén, Jesús; Marchante-Gayón, Juan Manuel; González, Pablo Rodríguez; Valledor, Luis; Cañal, María Jesús; Alonso, José Ignacio García

2017-08-18

The use of enriched stable isotopes is of outstanding importance in chemical metrology as it allows the application of isotope dilution mass spectrometry (IDMS). Primary methods based on IDMS ensure the quality of the analytical measurements and traceability of the results to the international system of units. However, the synthesis of isotopically labelled molecules from enriched stable isotopes is an expensive and a difficult task. Either chemical and biochemical methods to produce labelled molecules have been proposed, but so far, few cost-effective methods have been described. The aim of this study was to use the microalgae Chlamydomonas reinhardtii to produce, at laboratory scale, 15 N-labelled amino acids with a high isotopic enrichment. To do that, a culture media containing 15 NH 4 Cl was used. No kinetic isotope effect (KIE) was observed. The labelled proteins biosynthesized by the microorganism were extracted from the biomass and the 15 N-labelled amino acids were obtained after a protein hydrolysis with HCl. The use of the wall deficient strain CC503 cw92 mt+ is fit for purpose, as it only assimilates ammonia as nitrogen source, avoiding isotope contamination with nitrogen from the atmosphere or the reagents used in the culture medium, and enhancing the protein extraction efficiency compared to cell-walled wild type Chlamydomonas. The isotopic enrichment of the labelled amino acids was calculated from their isotopic composition measured by gas chromatography mass spectrometry (GC-MS). The average isotopic enrichment for the 16 amino acids characterized was 99.56 ± 0.05% and the concentration of the amino acids in the hydrolysate ranged from 18 to 90 µg/mL. Previously reported biochemical methods to produce isotopically labelled proteins have been applied in the fields of proteomics and fluxomics. For these approaches, low amounts of products are required and the isotopic enrichment of the molecules has never been properly determined. So far, only 13

Behavior of /sup 15/N-labelled amino acids in germinated corn

Energy Technology Data Exchange (ETDEWEB)

Samukawa, K; Yamaguchi, M [Osaka Prefectural Univ., Sakai (Japan). Coll. of Agriculture

1979-06-01

By investigating the rise and fall of /sup 15/N-labelled amino acids in germinated corns, the behavior of amino radicals in free amino acids, the influence of the hydrolysis products of stored proteins on free amino acids and the change from heterotrophy to autotrophy of seeds were clarified. The amount of amino acid production depending on external nitrogen was very small in the early period of germination. /sup 15/N incorporation into proline was not observed in the early period of germination, which suggested that the proline may be nitrogen-storing source. Most of the amino-state nitrogen of asparagine accumulated at the time of germination was internal nitrogen, and this fact suggested that aspartic acid serve as the acceptor of ammonia produced in the early stage of germination. /sup 15/N content increased significantly on 9 th day after germination, and decreased on 12 th day. These facts prove that there are always active decomposition and production of protein in plant body.

Quantifying nitrogen flux after application of 15N-labelled pig slurry on triticale in the late autumn

International Nuclear Information System (INIS)

Morvan, T.; Leterme, P.; Mary, B.

1996-01-01

Predicting nitrate leaching after spreading slurry in autumn is difficult because plant uptake, mineralization, immobilization, volatilization and denitrification occur and modify the nitrate pool available for leaching. To estimate these fluxes, pig slurry was labelled with (15NH4)2SO4 and spread in December (110 kg NH4-N.ha-1) on triticale. Soil microbial immobilization, crop uptake and soil inorganic nitrogen were measured at seven dates between day 2 and 63 after application. NH4-N in the slurry follows three ways of transformation: volatilization (38 kg N.ha-1 in 16 days), immobilization (29 kg N.ha-1 day 30) and nitrification (42 kg N.ha-1). This last one was achieved 48 days after spreading, despite the cold mean temperatures measured during the experiment. Gross mineralization of soil and slurry organic nitrogen was large (35 kg N.ha-1 for the 0-10 cm soil layer). The real utilization coefficient of 15N-labelled N was low, smaller than 4% at day 63. The leaching of nitrate was small because there was no rainfall after day 48. Thus, from the balance of 15N-labelled N, it is suggested that 22 kg NO3 N.ha-1 has been lost by denitrification [fr

Compound-specific δ15N amino acid measurements in littoral mussels in the California upwelling ecosystem: a new approach to generating baseline δ15N Isoscapes for coastal ecosystems.

Directory of Open Access Journals (Sweden)

Natasha L Vokhshoori

Full Text Available We explored δ(15N compound-specific amino acid isotope data (CSI-AA in filter-feeding intertidal mussels (Mytilus californianus as a new approach to construct integrated isoscapes of coastal primary production. We examined spatial δ(15N gradients in the California Upwelling Ecosystem (CUE, determining bulk δ(15N values of mussel tissue from 28 sites between Port Orford, Oregon and La Jolla, California, and applying CSI-AA at selected sites to decouple trophic effects from isotopic values at the base of the food web. Bulk δ(15N values showed a strong linear trend with latitude, increasing from North to South (from ∼ 7‰ to ∼ 12‰, R(2 = 0.759. In contrast, CSI-AA trophic position estimates showed no correlation with latitude. The δ(15N trend is therefore most consistent with a baseline δ(15N gradient, likely due to the mixing of two source waters: low δ(15N nitrate from the southward flowing surface California Current, and the northward transport of the California Undercurrent (CUC, with (15N-enriched nitrate. This interpretation is strongly supported by a similar linear gradient in δ(15N values of phenylalanine (δ(15NPhe, the best AA proxy for baseline δ(15N values. We hypothesize δ(15N(Phe values in intertidal mussels can approximate annual integrated δ(15N values of coastal phytoplankton primary production. We therefore used δ(15N(Phe values to generate the first compound-specific nitrogen isoscape for the coastal Northeast Pacific, which indicates a remarkably linear gradient in coastal primary production δ(15N values. We propose that δ(15N(Phe isoscapes derived from filter feeders can directly characterize baseline δ(15N values across major biochemical provinces, with potential applications for understanding migratory and feeding patterns of top predators, monitoring effects of climate change, and study of paleo- archives.

Transformation of {sup 15}N-Labelled Ammonium during Aerobic Decomposition of Plant Material

Energy Technology Data Exchange (ETDEWEB)

Danneberg, O. H.; Haunold, E.; Kaindl, K. [Institute for Biology and Agriculture. Reactor Centre, Seibersdorf (Austria)

1968-07-01

Plant material from maize leaves with the addition of {sup 15}N-labelled (NH{sub 4}){sub 2}SO{sub 4} was composted for periods of 10 to 180 d. The nitrogen of the decomposing samples was fractionated and the {sup 15}N enrichment in the fractions was determined by mass spectrometry. The added {sup 15}NH{sub 4}{sup +} was incorporated into organic compounds mainly during the first 10 d. The largest amount was found in the 'protein' fraction. The total nitrogen of this fraction increased up to 30 d, thus showing a marked synthesis of microbial protein. It decreased afterwards, when the microbial substances themselves were decomposed. Apart from this there was a marked synthesis of humic substances, especially in the first 10 d as indicated by an increase of the acid-insoluble ''humin'' fraction. A rather small amount of labelled ammonium was incorporated into this fraction within this time and this amount remained constant during the whole experiment. Because of the greater decay resistance the ''humin'' fraction was enriched towards the end of the experiment. (author)

Combined solid state and solution NMR studies of {alpha},{epsilon}-{sup 15}N labeled bovine rhodopsin

Energy Technology Data Exchange (ETDEWEB)

Werner, Karla; Lehner, Ines [Johann Wolfgang Goethe-Universitaet Frankfurt, Center for Biomolecular Magnetic Resonance (Germany); Dhiman, Harpreet Kaur [University of Pittsburgh School of Medicine, Department of Structural Biology (United States); Richter, Christian; Glaubitz, Clemens; Schwalbe, Harald, E-mail: schwalbe@nmr.uni-frankfurt.de; Klein-Seetharaman, Judith [Johann Wolfgang Goethe-Universitaet Frankfurt, Center for Biomolecular Magnetic Resonance (Germany); Khorana, H. Gobind [Massachusetts Institute of Technology, Departments of Biology and Chemistry (United States)], E-mail: khorana@mit.edu

2007-04-15

Rhodopsin is the visual pigment of the vertebrate rod photoreceptor cell and is the only member of the G protein coupled receptor family for which a crystal structure is available. Towards the study of dynamics in rhodopsin, we report NMR-spectroscopic investigations of {alpha},{epsilon}-{sup 15}N-tryptophan labeled rhodopsin in detergent micelles and reconstituted in phospholipids. Using a combination of solid state {sup 13}C,{sup 15}N-REDOR and HETCOR experiments of all possible {sup 13}C'{sub i-1} carbonyl/{sup 15}N{sub i}-tryptophan isotope labeled amide pairs, and H/D exchange {sup 1}H,{sup 15}N-HSQC experiments conducted in solution, we assigned chemical shifts to all five rhodopsin tryptophan backbone {sup 15}N nuclei and partially to their bound protons. {sup 1}H,{sup 15}N chemical shift assignment was achieved for indole side chains of Trp35{sup 1.30} and Trp175{sup 4.65}. {sup 15}N chemical shifts were found to be similar when comparing those obtained in the native like reconstituted lipid environment and those obtained in detergent micelles for all tryptophans except Trp175{sup 4.65} at the membrane interface. The results suggest that the integrated solution and solid state NMR approach presented provides highly complementary information in the study of structure and dynamics of large membrane proteins like rhodopsin.

Dynamics of the amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 7

International Nuclear Information System (INIS)

Gruhn, K.

1988-01-01

In a 15 N labelling experiment 12 colostomized laying hens received 15 N-labelled wheat with 14.37 atom-% 15 N excess ( 15 N') over 4 days. 3 hens each were butchered after 12 h, 36 h, 60 h and 108 h after the last 15 N' application. The gastrointestinal tract was divided into 3 parts (esophagus with crop and gizzard as well as glandular stomach, small intestine, large intestine). These parts and the pancreas were hydrolyzed with 6 N HCl and the individual basic as well as the sum of acid and neutral amino acids were determined in the hydrolyzed fractions. In addition, the amino acids and peptides were determined in the TCA soluble N fraction. The atom-% 15 N' was determined in the individual amino acid and peptide fractions. The labelling of the basic amino acids in the individual tract segments was lower than in the acid and neutral amino acids. In comparison to the peptides, a higher atom-% 15 N' could be determined in the free amino acids. (author)

Synthesis and biosynthesis of 13C, 15N labeled deoxynucleosides useful for biomolecular structural determinations

International Nuclear Information System (INIS)

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

1994-01-01

Currently, there is a great emphasis on elucidating the structure, function, and dynamics of DNA. Much of the research involved in this study utilizes nuclear magnetic resonance (NMR) spectroscopy. Effective use of NMR spectroscopy (more than 10,000 mw) in this arena requires stable isotope enrichment. Herein, the authors present strategies for the site-specific isotopic labeling of the purine bases adenosine and guanosine and the biosynthesis of [U- 13 C, 15 N] DNA from methylotrophic bacteria. With commercially available 6-chloropurine, an effective 2-step route leads to [6- 15 N]-2'-deoxadenosine (dA). The resulting [6- 15 N]-dA is used in a series of reactions to synthesize [2- 13 C, 1,2'- 15 N 2 ]-2'-deoxyguanosine or any combination thereof. An improved biosynthesis of labeled DNA has been accomplished using Methylobacterium extorquens AS1. Each liter of growth medium contains 4g of methanol to yield 1 gram of lyophilized cells. As much as 200 mg of RNA per liter of culture has been obtained. The authors are currently developing large scale isolation protocols. General synthetic pathways to oligomeric DNA are presented

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

International Nuclear Information System (INIS)

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

1994-01-01

Currently, there is a great emphasis on elucidating the structure, function, and dynamics of DNA. Much of the research involved in this study uses nuclear magnetic resonance (NMR) spectroscopy. Effective use of NMR spectroscopy for DNA molecules with mw > 10,000 requires stable isotope enrichment. We present strategies for site-specific isotopic labeling of the purine bases adenosine and guanosine and the biosynthesis of (U- 13 C, 15 N) DNA from methylotropic bacteria. With commercially available 6-chloropurine, an effective two-step route leads to 2'-deoxy-(amino- 15 N)adenosine (dA). The resulting d(amino- 15 N)A is used in a series of reactions to synthesize 2'-deoxy-(2- 13 C,1,amino- 15 N 2 )guanosine or any combination thereof. An improved biosynthesis of labeled DNA has been accomplished using Methylobacterium extorquens AS1. Each liter of growth medium contains 4 g of methanol to yield 1 g of lyophilized cells. As much as 200 mg of RNA per liter of culture has been obtained. We are currently developing large-scale isolation protocols. General synthetic pathways to oligomeric DNA will be presented

Distribution of 15N-labeled urea injected into field-grown corn plants

International Nuclear Information System (INIS)

Zhou, X.; Madrmootoo, C.A.; Mackenzie, A.F.; Smith, D.L.

1998-01-01

Nitrogen (N) assimilate supply to developing corn (Zea mays L.) ears plays a critical role in grain dry weight accumulation. The use of stem-perfused/injected 15N labeled compounds to determine the effects of an artificial N source on the subsequent distribution of injected N and grain weight of field-grown corn plants has not been reported previously. Our objective was to assess the distribution of N added via an artificial source. Three soil N fertilizer levels (0, 180, and 270 kg N ha-1) and three N solutions (distilled water control and 15N enriched urea at 15 and 30 mM N) were arranged in a split-plot design. Three N concentrations were injected using a pressurized stem injection technique. The injection started fifteen days after silking and continued until immediately prior to plant physiological maturity. The average uptake volume was 256 mL over the 30-day injection period. The N supplied via injection represented 1.5 to 3% of the total plant N. Neither soil applied N fertilizer nor injected N altered dry matter distribution among plant tissues. As the concentration of N in the injected solutions increased, N concentrations increased in the grain and upper stalks, and % 15N atom excess in ear+1 leaves and leaves increased. The relative degree of 15N enrichment for each of the tissues measured was injected internode grain upper stalks leaves lower stalks cob husk ear + 1 leaf ear leaf. This study indicated that the exogenous N supplied via stem-injection, was incorporated into all the measured plant parts, although not uniformly. The distribution of the injected 15N was affected both by the proximity of sinks to the point of injection and the strength of the various sinks

A novel medium for expression of proteins selectively labeled with 15N-amino acids in Spodoptera frugiperda (Sf9) insect cells

International Nuclear Information System (INIS)

Brueggert, Michael; Rehm, Till; Shanker, Sreejesh; Georgescu, Julia; Holak, Tad A.

2003-01-01

Whereas bacterial expression systems are widely used for production of uniformly or selectively 15 N-labeled proteins the usage of the baculovirus expression system for labeling is limited to very few examples in the literature. Here we present the complete formulations of the two insect media, IML406 and 455, for the high-yield production of selectively 15 N-labeled proteins in insect cells. The quantities of 15 N-amino acids utilized in the production of labeled GST were similar in the case of bacterial and viral expression. For the most studied amino acids essential for insect cells the 15 N-HSQC spectra, recorded with GST labeled in insect cells, showed no cross labeling and provided therefore spectra of better quality compared to NMR spectra of GST expressed in E. coli. Also in the case of amino acids not essential for Sf9 cells we were able to label a defined number of amino acid species. Therefore the selective labeling using the baculovirus expression vector system represents a complement or even an alternative to the bacterial expression system. Based on these findings we can provide a first simple overview of the network of the amino acid metabolism in E. coli and insect cells focused on nitrogen. For some amino acids the expression of labeled proteins in insect cells can replace the cell-free protein expression

Studies of the intermediary metabolism in cultured cells of the insect Spodoptera frugiperda using 13C- or 15N-labelled tracers

Directory of Open Access Journals (Sweden)

Bacher Adelbert

2005-11-01

Full Text Available Abstract Background Insect cells can serve as host systems for the recombinant expression of eukaryotic proteins. Using this platform, the controlled expression of 15N/13C labelled proteins requires the analysis of incorporation paths and rates of isotope-labelled precursors present in the medium into amino acids. For this purpose, Spodoptera frugiperda cells were grown in a complex medium containing [U-13C6]glucose. In a second experiment, cultures of S. frugiperda were grown in the presence of 15N-phenylalanine. Results Quantitative NMR analysis showed incorporation of the proffered [U-13C6]glucose into the ribose moiety of ribonucleosides (40 – 45% and into the amino acids, alanine (41%, glutamic acid/glutamine (C-4 and C-5, 30% and aspartate/asparagine (15%. Other amino acids and the purine ring of nucleosides were not formed from exogenous glucose in significant amounts (> 5%. Prior to the incorporation into protein the proffered 15N-phenylalanine lost about 70% of its label by transamination and the labelled compound was not converted into tyrosine to a significant extent. Conclusion Growth of S. frugiperda cells in the presence of [U-13C6]glucose is conducive to the fractional labelling of ribonucleosides, alanine, glutamic acid/glutamine and aspartic acid/asparagine. The isotopolog compositions of the ribonucleosides and of alanine indicate considerable recycling of carbohydrate intermediates in the reductive branch of the pentose phosphate pathway. The incorporation of 15N-labelled amino acids may be hampered by loss of the 15N-label by transamination.

Production of 15N-Labelled Liquid Organic Fertilisers Based on Manure and Crop Residue for Use in Fertigation Studies.

Directory of Open Access Journals (Sweden)

Belén Martínez-Alcántara

Full Text Available Large quantities of crop residue and animal manure from agricultural and livestock activities are annually produced worldwide. With proper management, these residues are potentially valuable sources of plant nutrients, mainly N. Recycling such subproducts in sustainably-based agricultural systems can minimise the use of mineral fertilisers, and hence reduce the potential risk of surface and groundwater pollution. Therefore, the purpose of this study was to obtain (small scale two liquid labelled-organic fertilisers, an animal- and a vegetal-based organic (AO and VO, respectively fertiliser, to be used as organic N sources in subsequent fertigation studies. Forage maize (Zea mays L. grown under 15N-labelled fertiliser supply was used as raw material for VO fertiliser production, and also as 15N-labelled sheep feed to obtain 15N-labelled manure. The labelled faeces fraction was used as raw material for the AO fertiliser. The VO fertiliser was obtained after an acidic and an enzyme-driven hydrolysis. The AO fertiliser was obtained after acidic hydrolysis. The VO liquid fertiliser presented an N concentration of 330 mg·L-1, 85% of total N was organic, while ammonium and nitrate N accounted for 55% and 45% of the mineral nitrogen fraction, respectively. This fertiliser also exhibited high K, Ca and S concentrations and notable values for the remaining macro- and micronutrients. The AO liquid fertiliser had a similar total N concentration (496 mg·L-1, 82% of total N in an organic form to that of VO, but its mineral N fraction significantly differed, which came in a predominantly (95% ammonia form. It also had a high content of N, P, K and other macronutrients, and sufficient Fe, Zn, Mn, Cu and B levels, which suggests its suitability as a potential fertiliser. The percentage of 15N enrichment in both VO and AO liquid fertilisers exceeded 2% 15N atom excess, which enabled their use in subsequent assays run to assess nitrogen uptake efficiency.

Decomposition and nitrogen dynamics of 15N-labeled leaf, root, and twig litter in temperate coniferous forests

Science.gov (United States)

van Huysen, Tiff L.; Harmon, Mark E.; Perakis, Steven S.; Chen, Hua

2013-01-01

Litter nutrient dynamics contribute significantly to biogeochemical cycling in forest ecosystems. We examined how site environment and initial substrate quality influence decomposition and nitrogen (N) dynamics of multiple litter types. A 2.5-year decomposition study was installed in the Oregon Coast Range and West Cascades using 15N-labeled litter from Acer macrophyllum, Picea sitchensis, and Pseudotsuga menziesii. Mass loss for leaf litter was similar between the two sites, while root and twig litter exhibited greater mass loss in the Coast Range. Mass loss was greatest from leaves and roots, and species differences in mass loss were more prominent in the Coast Range. All litter types and species mineralized N early in the decomposition process; only A. macrophyllum leaves exhibited a net N immobilization phase. There were no site differences with respect to litter N dynamics despite differences in site N availability, and litter N mineralization patterns were species-specific. For multiple litter × species combinations, the difference between gross and net N mineralization was significant, and gross mineralization was 7–20 % greater than net mineralization. The mineralization results suggest that initial litter chemistry may be an important driver of litter N dynamics. Our study demonstrates that greater amounts of N are cycling through these systems than may be quantified by only measuring net mineralization and challenges current leaf-based biogeochemical theory regarding patterns of N immobilization and mineralization.

Decomposition and nitrogen dynamics of (15)N-labeled leaf, root, and twig litter in temperate coniferous forests.

Science.gov (United States)

van Huysen, Tiff L; Harmon, Mark E; Perakis, Steven S; Chen, Hua

2013-12-01

Litter nutrient dynamics contribute significantly to biogeochemical cycling in forest ecosystems. We examined how site environment and initial substrate quality influence decomposition and nitrogen (N) dynamics of multiple litter types. A 2.5-year decomposition study was installed in the Oregon Coast Range and West Cascades using (15)N-labeled litter from Acer macrophyllum, Picea sitchensis, and Pseudotsuga menziesii. Mass loss for leaf litter was similar between the two sites, while root and twig litter exhibited greater mass loss in the Coast Range. Mass loss was greatest from leaves and roots, and species differences in mass loss were more prominent in the Coast Range. All litter types and species mineralized N early in the decomposition process; only A. macrophyllum leaves exhibited a net N immobilization phase. There were no site differences with respect to litter N dynamics despite differences in site N availability, and litter N mineralization patterns were species-specific. For multiple litter × species combinations, the difference between gross and net N mineralization was significant, and gross mineralization was 7-20 % greater than net mineralization. The mineralization results suggest that initial litter chemistry may be an important driver of litter N dynamics. Our study demonstrates that greater amounts of N are cycling through these systems than may be quantified by only measuring net mineralization and challenges current leaf-based biogeochemical theory regarding patterns of N immobilization and mineralization.

Investigation of the metabolism of colostomized laying hens with 15N-labelled wheat. 5

International Nuclear Information System (INIS)

Gruhn, K.

1980-01-01

In an experiment with 3 colostomized laying hybrids each animal received 80 g pelleted mixed feed and 40 g 15 N-labelled wheat with 20.13 atom-% 15 N excess ( 15 N') over a period of four days. On the following four days the hens received rations composed in the same way with unlabelled wheat, however in the tissues and organs of the slaughtered hens 15 N' was determined in the total N and the amino acids lysine, histidine and arginine in both the segments of the gastro intestinal tract and in its content. The amount of 15 N' stomach, small intestine and colon was 43.7%, 27.2% and 29.1%, respectively. The tissue of the small intestine contained, on an average, the highest 15 N' in lysine of all the basic amino acids. It was 0.82 atom-% 15 N' for lysine, 0.55% for histidine and 0.63% for arginine. The percentage of the 15 N' of the basic amino acids from the corresponding total 15 N' amount of the charges was 20.5% in the contents of the gastrointestinal tract, 28.0% in the stomach tissue and in the tissues of the small intestine 24.4% of the cecum 21.5% and of the rectum 25.7%. (author)

Uptake of pulse injected nitrogen by soil microbes and mycorrhizal and non-mycorrhizal plants in a species-diverse subarctic heath ecosystem

DEFF Research Database (Denmark)

Andresen, Louise Christoffersen; Jonasson, Sven; Strom, Lena

2008-01-01

15N labeled ammonium, glycine or glutamic acid was injected into subarctic heath soil in situ, with the purpose of investigating how the nitrogen added in these pulses was subsequently utilized and cycled in the ecosystem. We analyzed the acquisition of 15N label in mycorrhizal and non-mycorrhiza......15N labeled ammonium, glycine or glutamic acid was injected into subarctic heath soil in situ, with the purpose of investigating how the nitrogen added in these pulses was subsequently utilized and cycled in the ecosystem. We analyzed the acquisition of 15N label in mycorrhizal and non...

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.

Distribution, and uptake by rice plants of 15N-labeled ammonium applied in mudballs in paddy soils

International Nuclear Information System (INIS)

Ventura, Wilbur; Yoshida, Tomio

1978-01-01

A 1974 field experiment determined the distribution, and uptake by rice plants, of ammonium fertilizer at 60 kg N/ha applied in mudballs into the reduced layer of paddy soil. The fertilizer-carrying mudballs were placed at the center of four hills. At the center of the plot, one 15 N-labeled mudball was applied and the 15 N content of the plants surrounding the site of placement were determined. For comparison, labeled ammonium fertilizer was basally incorporated with the entire puddled layer and a topdress application was made 39 days before heading. There was little movement of the ammonium nitrogen horizontally from the site of placement so that the distribution of 15 N was restricted to the four adjacent plant hills. The distribution of incorporated ammonium fertilizer with the puddled layer was likewise restricted to the four adjacent rice plants but topdressing, with the unavoidable disturbance of the floodwater, resulted to a wide distribution of the 15 N-labeled fertilizer. In all the methods of application, there was an uneven uptake of 15 N among four plants adjacent to the site of placement. An increase of at least 10% in the efficiency of ammonium fertilizer was obtained by the deep placement of ammoniated mudballs as compared to the common practice of incorporating the fertilizer with the puddled soil layer. Topdressing at 39 days before heading, however, was as efficient as mudballs applied at the same stage of growth. There was no significant increase in grain yield by deep placement of fertilizer because of the high initial nitrogen content of the soil. (author)

Artifact suppression in electron paramagnetic resonance imaging of 14N- and 15N-labeled nitroxyl radicals with asymmetric absorption spectra

Science.gov (United States)

Takahashi, Wataru; Miyake, Yusuke; Hirata, Hiroshi

2014-10-01

This article describes an improved method for suppressing image artifacts in the visualization of 14N- and 15N-labeled nitroxyl radicals in a single image scan using electron paramagnetic resonance (EPR). The purpose of this work was to solve the problem of asymmetric EPR absorption spectra in spectral processing. A hybrid function of Gaussian and Lorentzian lineshapes was used to perform spectral line-fitting to successfully separate the two kinds of nitroxyl radicals. This approach can process the asymmetric EPR absorption spectra of the nitroxyl radicals being measured, and can suppress image artifacts due to spectral asymmetry. With this improved visualization method and a 750-MHz continuous-wave EPR imager, a temporal change in the distributions of a two-phase paraffin oil and water/glycerin solution system was visualized using lipophilic and hydrophilic nitroxyl radicals, i.e., 2-(14-carboxytetradecyl)-2-ethyl-4,4-dimethyl-3-oxazolidinyloxy (16-DOXYL stearic acid) and 4-hydroxyl-2,2,6,6-tetramethylpiperidine-d17-1-15N-1-oxyl (TEMPOL-d17-15N). The results of the two-phase separation experiment verified that reasonable artifact suppression could be achieved by the present method that deals with asymmetric absorption spectra in the EPR imaging of 14N- and 15N-labeled nitroxyl radicals.

A novel medium for expression of proteins selectively labeled with {sup 15}N-amino acids in Spodoptera frugiperda (Sf9) insect cells

Energy Technology Data Exchange (ETDEWEB)

Brueggert, Michael; Rehm, Till; Shanker, Sreejesh; Georgescu, Julia; Holak, Tad A. [Max Planck Institute for Biochemistry (Germany)], E-mail: holak.biochem@mpg.de

2003-04-15

Whereas bacterial expression systems are widely used for production of uniformly or selectively {sup 15}N-labeled proteins the usage of the baculovirus expression system for labeling is limited to very few examples in the literature. Here we present the complete formulations of the two insect media, IML406 and 455, for the high-yield production of selectively {sup 15}N-labeled proteins in insect cells. The quantities of {sup 15}N-amino acids utilized in the production of labeled GST were similar in the case of bacterial and viral expression. For the most studied amino acids essential for insect cells the {sup 15}N-HSQC spectra, recorded with GST labeled in insect cells, showed no cross labeling and provided therefore spectra of better quality compared to NMR spectra of GST expressed in E. coli. Also in the case of amino acids not essential for Sf9 cells we were able to label a defined number of amino acid species. Therefore the selective labeling using the baculovirus expression vector system represents a complement or even an alternative to the bacterial expression system. Based on these findings we can provide a first simple overview of the network of the amino acid metabolism in E. coli and insect cells focused on nitrogen. For some amino acids the expression of labeled proteins in insect cells can replace the cell-free protein expression.

Nitrate Leaching From a Mountain Forest Ecosystem with Gleysols Subjected to Experimentally Increased N Deposition

International Nuclear Information System (INIS)

Schleppi, Patrick; Hagedorn, Frank; Providoli, Isabelle

2004-01-01

Nitrate leaching was measured over seven years of nitrogen (N) addition in a paired-catchment experiment in Alptal, central Switzerland (altitude: 1200 m, bulk N deposition: 12 kg ha -1 a -1 ). Two forested catchments (1500 m 2 each) dominated by Picea abies) were delimited by trenches in the Gleysols. NH 4 NO 3 was added to one of the catchments using sprinklers. During the first year, the N addition was labelled with 15 N. Additionally, soil N transformations were studied in replicated plots. Pre-treatment NO 3 - -N leaching was 4 kg ha -1 a -1 from both catchments, and remained between 2.5 and 4.8 kg ha -1 a -1 in the control catchment. The first year of treatment induced an additional leaching of 3.1 kg ha -1 , almost 90% of which was labelled with 15 N, indicating that it did not cycle through the large N pools of the ecosystem (soil organic matter and plants). These losses partly correspond to NO 3 - from precipitation bypassing the soil due to preferential flow. During rain or snowmelt events, NO 3 - concentration peaks as the water table is rising, indicating flushing from the soil. Nitrification occurs temporarily along the water flow paths in the soil and can be the source of NO 3 - flushing. Its isotopic signature however, shows that this release mainly affects recently applied N, stored only between runoff events or up to a few weeks. At first, the ecosystem retained 90% of the added N (2/3 in the soil), but NO 3 - losses increased from 10 to 30% within 7 yr, indicating that the ecosystem became progressively N saturated

Dynamics of the amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 5

International Nuclear Information System (INIS)

Gruhn, K.; Zander, R.; Kirchner, E.

1987-01-01

12 colostomized laying hens which received 15 N-labelled wheat over 4 days were butchered 12 h, 36 h, and 108 h (3 animals each) after the last 15 N application. The intake of 15 N exess ( 15 N') from the wheat amounted to 540 mg 15 N' during the application period. The 15 N' in the blood plasma decreased after the last 15 N' application from 0.76 atom-% to 0.55 atom-% after 108 h, the labelling of the corpuscular components at the same measuring points increased from 0.28 to 0.50 atom-% 15 N'. 96.6% of the plasma 15 N' and 93,8% of that in the corpuscles is precipitable in trichloroacetic acid. The atom-% 15 N' of histidine in the total blood remained unchanged in dependence on the butchering time. The 15 N amount in lysine and arginine and that in the non-basic amino acids decreased inconsiderably in the period between 12 h and 108 h after the last 15 N' wheat feeding. (author)

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

DEFF Research Database (Denmark)

Truhlar, Stephanie M E; Cervantes, Carla F; Torpey, Justin W

2008-01-01

. MALDI TOF-TOF MS/MS data provide additional information that shows where the "extra" (15)N labels are incorporated, which can be useful in confirming ambiguous assignments. The described procedure provides a rapid technique to monitor the fidelity of selective labeling that does not require a lot...

The use of 15N-labelled dinitrogen in the study of nitrogen fixation by blue-green algae

International Nuclear Information System (INIS)

Jones, J.

1985-01-01

Prior to the development of the acetylene reduction technique 15 N was used as the main qualitative and quantitative measure of nitrogen fixation by free-living cyanobacteria in a variety of aquatic and terrestrial habitats. Despite its expense and the technical difficulty, 15 N is a major tool in the study of cyanobacteria, for example, incorporation of 15 N 2 is the definitive test for nitrogen fixation; it is used in the determination of the correct ratio of acetylene reduction to nitrogen fixation, in in situ nitrogen fixation assays, in tracing the formation and fate of extra-cellular nitrogen and in measuring the turnover and grazing rates of cyanobacterial intra-cellular nitrogen. These latter studies show that 15 N-labelled extra-cellular nitrogen can serve as nitrogen sources for a variety of bacteria, fungi, algae and higher plants, and that cyanobacteria are graced and digested by a variety of animals. The turnover rates of cyanobacterial 15 N-labelled cells are dependent on the type of cell, species, environmental conditions and the availability of degrading organisms. The breakdown products are rapidly mineralised and used as nitrogen sources by higher plants. (author)

Selective 15N labeling and direct observation by NMR of the active-site glutamine of Fe-containing superoxide dismutase

International Nuclear Information System (INIS)

Vance, Carrie K.; Kang, Young M.; Miller, Anne-Frances

1997-01-01

The glutamine in position 69 is one of only three conserved active-site amino acid differences between Fe- and Mn-containing superoxide dismutases (SODs). We have refined the conditions for extremely selective labeling of the side chains of glutamine with 15N, and thus obtained dramatically simplified spectra, despite the large size of Fe-SOD. The improved resolution afforded by such highly specific labeling permits the use of direct 15N detection to observe and assign Gln 69, even though its distance to the paramagnetic Fe2+ is only 5A. Selective glutamine side-chain labeling is inexpensive and has general utility for large (and paramagnet-containing) proteins

Dynamics of the amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 11

International Nuclear Information System (INIS)

Gruhn, K.; Zander, R.

1989-01-01

Over a period of 4 days 12 colostomized laying hens daily received 36 g 15 N-labelled wheat with 15 N excess ( 15 N') of 14.37 atom-% together with a conventional feed mixture for laying hens. The labelling of the lysine N in the wheat was 13.58 atom-%, that of histidine N 14.38 and that of arginine 15 N' 13.63 atom-% 15 N'. Three hens each were butchered 12, 36, 60 and 108 h after the last 15 N' feeding. The first three hens did not receive any feed before being butchered. The following three hens each received the unlabelled feed ration for another 1, 2 or 4 days, resp., after the main period until they were butchered. The total of skeleton muscles, heart and stomach muscle (without inner skin) of each hen were combined into one sample, cut thinly, drenched with fluid nitrogen and pulverized. N, 15 N' and the basic and non-basic amino acids as well as their 15 N' were determined in the individual samples. In contrast to the organs, the proteins in the muscle tissue have a long half-life so that a slight decrease of atom-% 15 N' in the muscles could only be detected after 108 h. The 14 N and 15 N' quota of the non-basic amino acids in the total nitrogen of the muscles is 50 %. The 14 N quota of the basic amino acids is 30% and the 15 N' quota only 22.5% in the total muscle N. The heavy nitrogen of the free lysine in the TCA soluble N fraction is hardly detectable 36 h and 60 h after the last 15 N' supply and not at all after 108 h. In contrast to this, the other two free basic amino acids remain significantly higher labelled in dependence on the last butchering time. (author)

Quantitative analysis of 15N labeled positional isomers of glutamine and citrulline via electrospray ionization tandem mass spectrometry of their dansyl derivatives

Science.gov (United States)

The enteral metabolism of glutamine and citrulline are intertwined because, while glutamine is one of the main fuel sources for the enterocyte, citrulline is one of its products. It has been shown that the administration of 15N labeled glutamine results in the incorporation of the 15N label into cit...

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

Energy Technology Data Exchange (ETDEWEB)

Ashburn, D.A.; Garcia, K.; Hanners, J.L.; Silks, L.A. III; Unkefer, C.J. [Los Alamos National Laboratory, NM (United States)

1994-12-01

Currently, there is a great emphasis on elucidating the structure, function, and dynamics of DNA. Much of the research involved in this study uses nuclear magnetic resonance (NMR) spectroscopy. Effective use of NMR spectroscopy for DNA molecules with mw > 10,000 requires stable isotope enrichment. We present strategies for site-specific isotopic labeling of the purine bases adenosine and guanosine and the biosynthesis of (U-{sup 13}C, {sup 15}N) DNA from methylotropic bacteria. With commercially available 6-chloropurine, an effective two-step route leads to 2{prime}-deoxy-(amino-{sup 15}N)adenosine (dA). The resulting d(amino-{sup 15}N)A is used in a series of reactions to synthesize 2{prime}-deoxy-(2-{sup 13}C,1,amino-{sup 15}N{sub 2})guanosine or any combination thereof. An improved biosynthesis of labeled DNA has been accomplished using Methylobacterium extorquens AS1. Each liter of growth medium contains 4 g of methanol to yield 1 g of lyophilized cells. As much as 200 mg of RNA per liter of culture has been obtained. We are currently developing large-scale isolation protocols. General synthetic pathways to oligomeric DNA will be presented.

δ15N of seagrass leaves for monitoring anthropogenic nutrient increases in coral reef ecosystems

International Nuclear Information System (INIS)

Yamamuro, M.; Kayanne, H.; Yamano, H.

2003-01-01

In a coral reef environment, a slight increase in dissolved inorganic nitrogen (DIN;≥1.0 μM) can alter the ecosystem via macroalgal blooms. We collected seagrass leaves from the tropical and subtropical Pacific Ocean in five countries and examined the interactions between nutrient concentrations (C, N, P), molar ratios of nutrients, and δ 15 N to find a possible indicator of the DIN conditions. Within most sites, the concentrations of nutrients and their molar ratios showed large variations owing to species-specific values. On the other hand, almost identical δ 15 N values were found in seagrass leaves of several species at each site. The correlations between δ 15 N and nutrient concentrations and between δ 15 N and molar ratios of nutrients suggested that nutrient availability did not affect the δ 15 N value of seagrass leaves by altering the physiological condition of the plants. Increases in δ 15 N of seagrass leaves mostly matched increases in DIN concentrations in the bottom water. We suggest that δ 15 N in seagrass leaves can be a good tool to monitor time-integrated decrease/increase of DIN concentrations at a site, both in the water column and the interstitial water

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)

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...

Synthesis of hydroxylamine-15 N.HCl

International Nuclear Information System (INIS)

Baldea, Aurel

2001-01-01

15 N labelled hydroxylamine is one of the starting substance for synthesis of labelled oximes. Industrial procedure was chosen to prepare hydroxylamine- 15 N. Sodium nitrite reduced by sodium bisulfite and sulfur dioxide, at temperature of 0-2 deg. C, produces sodium hydroxylamine disulfonate. The reaction mixture is treated with acetone and the resulting acetoxime is distilled. In order to obtain crystalline hydroxylamine hydrochloride, hydrochloric acid is added to the distillate and the solution is evaporated to dryness. The crude product was purified by recristallization, yielding 62-65% of theoretical amount. Labelled ammonium chloride formed as byproduct can be recovered improving 15 N balance. IR spectra is used for chemical analysis and mass spectrometry for isotopic analysis. For this purpose hydroxylamine- 15 N is converted into molecular nitrogen. (author)

Application of amino acid type-specific 1H- and 14N-labeling in a 2H-, 15N-labeled background to a 47 kDa homodimer: Potential for NMR structure determination of large proteins

International Nuclear Information System (INIS)

Kelly, Mark J.S.; Krieger, Cornelia; Ball, Linda J.; Yu Yihua; Richter, Gerald; Schmieder, Peter; Bacher, Adelbert; Oschkinat, Hartmut

1999-01-01

NMR investigations of larger macromolecules (>20 kDa) are severely hindered by rapid 1H and 13C transverse relaxation. Replacement of non-exchangeable protons with deuterium removes many efficient 1H-1H and 1H-13C relaxation pathways. The main disadvantage of deuteration is that many of the protons which would normally be the source of NOE-based distance restraints are removed. We report the development of a novel labeling strategy which is based on specific protonation and 14N-labeling of the residues phenylalanine, tyrosine, threonine, isoleucine and valine in a fully deuterated, 15N-labeled background. This allows the application of heteronuclear half-filters, 15N-editing and 1H-TOCSY experiments to select for particular magnetization transfer pathways. Results from investigations of a 47 kDa dimeric protein labeled in this way demonstrated that the method provides useful information for the structure determination of large proteins

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

Energy Technology Data Exchange (ETDEWEB)

Strzelecka, T.; Gronenborn, A.M.; Clore, G.M. [National Institutes of Health, Bethesda, MD (United States)

1994-12-01

The Ner protein is a small (74-amino acid) DNA-binding protein that regulates a switch between the lysogenic and lytic stages of phage Mu. It inhibits expression of the C repressor gene and down-regulates its own expression. Two-dimensional NMR experiments on uniformly {sup 15}N-labeled protein provided most of the backbone and some of the sidechain proton assignments. The secondary structure determination using two-dimensional NOESY experiments showed that Ner consists of five {alpha}-helices. However, because most of the sidechain protons could not be assigned, the full structure was not determined. Using uniformly {sup 13}C/{sup 15}N-labeled Ner and a set of three-dimensional experiments, we were able to assign all of the backbone and 98% of the sidechain protons. In particular, the CBCANH and CBCA(CO)NH experiments were used to sequentially assign the C{alpha} and C{beta} resonances; the HCCH-CTOCSY and HCCH-COSY were used to assign sidechain carbon and proton resonances.

15N-labeled nitrogen from green manure and ammonium sulfate utilization by the sugarcane ratoon

International Nuclear Information System (INIS)

Ambrosano, Edmilson Jose; Rossi, Fabricio; Trivelin, Paulo Cesar Ocheuze; Cantarella, Heitor; Ambrosano, Glaucia Maria Bovi; Schammass, Eliana Aparecida; Muraoka, Takashi

2011-01-01

Legumes as green manure are alternative sources of nitrogen (N) for crops and can supplement or even replace mineral nitrogen fertilization due to their potential for biological nitrogen fixation (BNF). The utilization of nitrogen by sugarcane (Saccharum spp.) fertilized with sunn hemp (Crotalaria juncea L.) and ammonium sulfate (AS) was evaluated using the 15 N tracer technique. N was added at the rate of 196 and 70 kg ha -1 as 15 N-labeled sunn hemp green manure (SH) and as ammonium sulfate (AS), respectively. Treatments were: (I) Control; (II) AS 15 N; (III) SH 15 N + AS; (IV) SH 15 N; and (V) AS 15 N + SH. Sugarcane was cultivated for five years and was harvested three times. 15 N recovery was evaluated in the two first harvests. In the sum of the three harvests, the highest stalk yields were obtained with a combination of green manure and inorganic N fertilizer; however, in the second cutting the yields were higher where SH was used than in plots with AS. The recovery of N by the first two consecutive harvests accounted for 19 to 21% of the N applied as leguminous green manure and 46 to 49% of the N applied as AS. The amounts of inorganic N, derived from both N sources, present in the 0-0.4 m layer of soil in the first season after N application and were below 1 kg ha -1 . (author)

Nitrogen mineralization from selected 15N-labelled crop residues and humus as affected by inorganic nitrogen

International Nuclear Information System (INIS)

Santos, J.A.

1987-01-01

The use of cover crops or crop residues as a source of N to succeeding crops has become a matter of increasing importance for economic and environmental reason. Greenhouse and field studies were conducted to determine the N contribution of four 15 N labelled crop residues, rye (Secale cereale L.), wheat (Triticum aestivum L.), crimson clover (Trifolium encarnatum L.), and hairy vetch (Vicia sativa L.), to successive crops and to evaluate the effect of different organic (ON) and inorganic N (IN) combinations on mineralization of the above residues. Total 15 N recovery from the residues ranged from 51% to 85% and 4% to 74% for the greenhouse and field studies, respectively

Distribution of total nitrogen and N-15 labelled nitrogen applied to apple trees

International Nuclear Information System (INIS)

Calvache, Marcelo.

1990-01-01

The efficiency of nitrogen fertilization from one year's application was studied in apple trees. Urea enriched with 1,5% N-15 a.e. was applied to 2 years old apple trees. Two irrigation treatments were studied, Al approx. 200mm/week and A2 approx. 100 mm/week. The distribution of N in the different parts of the trees was determined after 2 months of fertilization and after the experimental trees were excavated. The recovery of labelled fertilizer N was different in the trees in both treatments (Al = 1,2% and A2 = 3,1%). However, the distribution in the tree's parts was similar: 46% in leaves, 34% in branches and 20% in roots. We also determined that sampling only 20% of leaves at the beginning and the end of the experiment it is possible to know the quantity of nitrogen from fertilizer, without the excavation trees

A technique developed for labeling the green manures (sunnhemp and velvet bean) with {sup 15} N for nitrogen dynamic studies; Tecnica para marcacao dos adubos verdes crotalaria juncea e mucuna-preta com {sup 15} N para estudos de dinamica do nitrogenio

Energy Technology Data Exchange (ETDEWEB)

Ambrosano, Edmilson Jose [Instituto Agronomico de Campinas, SP (Brazil). Secao de Leguminosas; Trivelin, Paulo Cesar Ocheuze; Muruoka, Takashi [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil)

1997-07-01

A technique was developed for labeling the leguminous plant tissue with nitrogen ({sup 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 % {sup 15} N) per pot was applied three times. The labelled dried plant material showed 3.177 and 4.337 of atom % {sup 15} N, respectively for velvet beans and sunnhemp. (author)

Tracer experiments with 15N-labelled wheat to determine the endogenous and exogenous fecal N-proportion

International Nuclear Information System (INIS)

Krawielitzki, K.; Timm, E.

1978-01-01

In an experiment with growing Wistar rats of 100 g live weight the N-values and the 15 N-frequency of the nitrogen in feces, urine and the experimental carcasses were determined after feeding 15 N-labelled wheat. Proceeding from Czarnetzki's multicompartment model (1969) for N-metabolism in monogastric animals, the measured data were used to calculate the endogenous and exogenous fecal N-proportion of total nitrogen. In agreement with earlier studies the intestinal nitrogen loss was found to rise as the protein intake increased. In this experiment, the intestinal nitrogen loss went up from 8.2 mg N/animal and day (N-free diet) to 33.9 mg N/animal and day at a daily nitrogen intake of 240 mg/animal and day. The true digestibility of the wheat protein (determined by taking into account the rise of fecal N loss) was 97.2% this value being 8.4 units higher than the true digestibility calculated by the conventional regressive method of fecal analysis with a constant value being taken for fecal N loss. In connection with earlier findings, this experiment allows to draw the conclusion that the true digestibility determined conventionally by regression analysis does not reflect the actual digestibility of the protein. (author)

Foliar fertilization of sugarcane (Saccharum spp): absorption and translocation of 15-N-labeled urea

International Nuclear Information System (INIS)

Trivelin, P.C.O.; Carvalho, J.G. de; Silva, A.Q. da; Primavesi, A.C.P.A.; Camacho, E.; Eimori, I.E.; Guilherme, M.R.

1988-01-01

The absorption and translocation of foliar applied nitrogen as urea solution to sugar cane plants was evaluated. An experiment using the isotope dilution technique with 15 N labeled urea was carried out in green house condition. Seedlings of sugarcane variety IAC 53-150 were planted in pots with 5KG of top soil''latossolo vermelho amarelo, fase arenosa'' (Haplustox). (M.A.C.) [pt

Dynamics of the amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 10

International Nuclear Information System (INIS)

Gruhn, K.; Hennig, A.

1989-01-01

Over a period of 4 days 12 colostomized laying hens daily received 36 g coarse wheat meal containing 14.37 atom-% 15 N excess ( 15 N') together with a conventional ration. After the homogenisation of each oviduct N and 15 N' were determined. After the precipitation with TCA the 15 N' of the amino acids was analysed in both the precipitate and the supernatant. In addition, the free amino acids and the peptides were determined in the TCA soluble fraction. The atom-% 15 N' in the total N and in the non-basic amino acid N showed a parallel decrease; it diminshed from 1.75 atom-% 15 N' to 0.64. Of the three basic amino acids, lysine shows the lowest labelling at all four measuring points. The quotas of non-basic amino acid 14 N and 15 N' in the total 14 N and 15 N' of the oviduct are the same and amount to 53%. In contrast to this, the quota of the 14 N of the basic amino acids in the total 14 N of the oviduct only amounts to 21.6% and that of 15 N' only to 15.4%. The average atom-% 15 N' of the free amino acids 12 h after the last 15 N application is 1.54 and is considerably above that of the peptides with 1.15 atom-% 15 N'. 36 h after the last 15 N application the ascertained value of 1.25 is identical in both fractions. The labelling of the free amino acids decreases more quickly than that of the peptides the more time has passed after the last 15 N application. (author)

Stereospecific assignments of glycine in proteins by stereospecific deuteration and {sup 15}N labeling

Energy Technology Data Exchange (ETDEWEB)

Hansen, A.P.; Curley, R.W. Jr.; Panigot, M.J.; Fesik, S.W. [Ohio State Univ., Columbus, OH (United States)

1994-12-01

Stereospecific assignments are important for accurately determining the three-dimensional structures of proteins through the use of multidimensional NMR techniques. It is especially important to stereospecifically assign the glycine {alpha}-protons in proteins because of the potential for different backbone conformations of this residue. These stereospecific assignments are critical for interpreting the {sup 3}J{sub NH,{alpha}H} coupling constants and NOEs involving the glycine {alpha}-protons that determine the conformation of this part of the protein. However, it is often difficult to unambiguously obtain the stereospecific assignments for glycine residues by using only NOE data. In this poster, we present a method for unambiguous, stereospecific assignment of the {alpha}-protons of glycine residues. This method involves synthesis of stereo-specifically deuterated and {sup 15}N-labeled Gly using a slightly modified procedure originally described by Woodard and coworkers for the stereoselective deuteration of glycine. The stereospecifically deuterated and {sup 15}N-labeled Gy has been incorporated into recombinant proteins expressed in both bacterial systems (FKBP) and mammalian cells (u-PA). Two- and three-dimensional isotope-filtered and isotope-edited NMR experiments were used to obtain the stereospecific assignments of the glycine {alpha}-protons for these proteins.

Transformation of /sup 14/C labelled plant components in soil in relation to immobilization and remineralization of /sup 15/N fertilizer

Energy Technology Data Exchange (ETDEWEB)

Azam, F.; Haider, K.; Malik, K.A.

1985-01-01

Uniformly /sup 14/C labeled glucose, cellulose and wheat straw and specifically /sup 14/C labeled lignin component in corn stalks were aerobically incubated for 12 weeks in a chernozem soil along with /sup 15/N labeled ammonium sulfate. Glucose was most readily decomposed, followed in order by cellulose, wheat straw and corn stalk lignins labeled at methoxyl-, side chain 2- and ring-C. More than 50% of /sup 14/C applied as glucose, cellulose and wheat straw evolved as CO/sub 2/ during the first week. Lignin however, decomposed relatively slowly. A higher proportion of /sup 14/C was transformed into microbial biomass whereas lignins contributed a little to this fraction. After 12 weeks of incubation nearly 60% of the lignin /sup 14/C was found in humic compounds of which more than 70% was resistant to hydrolysis with 6N HCl. Maximum incorporation of /sup 15/N in humic compounds was observed in cellulose amended soil. However, in this case more than 80% of the /sup 15/N was in hydrolysable forms. Immobilization-remineralization of applied /sup 15/N was most rapid in glucose treated soil and a complete immobilization followed by remineralization was observed after 3 days. The process was much slow in soil treated with cellulose, wheat straw or corn stalks. More than 70% of the newly immobilized N was in hydrolysable forms mainly representing the microbial component. Serial hydrolysis of soil at different incubation intervals showed a greater proportion of 6N HCl hydrolysable /sup 14/C and /sup 15/N in fractions representing microbial material. /sup 14/C from lignin carbons was relatively more uniformly distributed in different fractions as compared to glucose, cellulose and wheat straw where a major portion of /sup 14/C was in easily hydrolysable fractions. 25 refs., 3 figs., 4 tabs.

{delta}{sup 15}N of seagrass leaves for monitoring anthropogenic nutrient increases in coral reef ecosystems

Energy Technology Data Exchange (ETDEWEB)

Yamamuro, M.; Kayanne, H.; Yamano, H

2003-04-01

In a coral reef environment, a slight increase in dissolved inorganic nitrogen (DIN;{>=}1.0 {mu}M) can alter the ecosystem via macroalgal blooms. We collected seagrass leaves from the tropical and subtropical Pacific Ocean in five countries and examined the interactions between nutrient concentrations (C, N, P), molar ratios of nutrients, and {delta}{sup 15}N to find a possible indicator of the DIN conditions. Within most sites, the concentrations of nutrients and their molar ratios showed large variations owing to species-specific values. On the other hand, almost identical {delta}{sup 15}N values were found in seagrass leaves of several species at each site. The correlations between {delta}{sup 15}N and nutrient concentrations and between {delta}{sup 15}N and molar ratios of nutrients suggested that nutrient availability did not affect the {delta}{sup 15}N value of seagrass leaves by altering the physiological condition of the plants. Increases in {delta}{sup 15}N of seagrass leaves mostly matched increases in DIN concentrations in the bottom water. We suggest that {delta}{sup 15}N in seagrass leaves can be a good tool to monitor time-integrated decrease/increase of DIN concentrations at a site, both in the water column and the interstitial water.

47 CFR 15.19 - Labelling requirements.

Science.gov (United States)

2010-10-01

... 47 Telecommunication 1 2010-10-01 2010-10-01 false Labelling requirements. 15.19 Section 15.19 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.19 Labelling... label shall be located in a conspicuous location on the device and shall contain the unique...

Nitrogen immobilization and mineralization during initial decomposition of 15N-labelled pea and barley residues

DEFF Research Database (Denmark)

Jensen, E.S.

1997-01-01

The immobilization and mineralization of N following plant residue incorporation were studied in a sandy loam soil using N-15-labelled field pea (Pisum sativum L.) and spring barley (Hordeum vulgare L.) straw. Both crop residues caused a net immobilization of soil-derived inorganic N during...... the complete incubation period of 84 days. The maximum rate of N immobilization was found to 12 and 18 mg soil-derived N g(-1) added C after incorporation of pea and barley residues, respectively. After 7 days of incubation, 21% of the pea and 17% of the barley residue N were assimilated by the soil microbial...... the decomposition of the barley residue. The net mineralization of residue-derived N was 2% in the barley and 22% in the pea residue treatment after 84 days of incubation. The results demonstrated that even if crop residues have a relative low C/N ratio (15), transient immobilization of soil N in the microbial...

Nitrogen accumulation and partitioning in a High Arctic tundra ecosystem from extreme atmospheric N deposition events

International Nuclear Information System (INIS)

Choudhary, Sonal; Blaud, Aimeric; Osborn, A. Mark; Press, Malcolm C.; Phoenix, Gareth K.

2016-01-01

Arctic ecosystems are threatened by pollution from recently detected extreme atmospheric nitrogen (N) deposition events in which up to 90% of the annual N deposition can occur in just a few days. We undertook the first assessment of the fate of N from extreme deposition in High Arctic tundra and are presenting the results from the whole ecosystem "1"5N labelling experiment. In 2010, we simulated N depositions at rates of 0, 0.04, 0.4 and 1.2 g N m"−"2 yr"−"1, applied as "1"5NH_4"1"5NO_3 in Svalbard (79"°N), during the summer. Separate applications of "1"5NO_3"− and "1"5NH_4"+ were also made to determine the importance of N form in their retention. More than 95% of the total "1"5N applied was recovered after one growing season (~ 90% after two), demonstrating a considerable capacity of Arctic tundra to retain N from these deposition events. Important sinks for the deposited N, regardless of its application rate or form, were non-vascular plants > vascular plants > organic soil > litter > mineral soil, suggesting that non-vascular plants could be the primary component of this ecosystem to undergo measurable changes due to N enrichment from extreme deposition events. Substantial retention of N by soil microbial biomass (70% and 39% of "1"5N in organic and mineral horizon, respectively) during the initial partitioning demonstrated their capacity to act as effective buffers for N leaching. Between the two N forms, vascular plants (Salix polaris) in particular showed difference in their N recovery, incorporating four times greater "1"5NO_3"− than "1"5NH_4"+, suggesting deposition rich in nitrate will impact them more. Overall, these findings show that despite the deposition rates being extreme in statistical terms, biologically they do not exceed the capacity of tundra to sequester pollutant N during the growing season. Therefore, current and future extreme events may represent a major source of eutrophication. - Highlights: • High Arctic tundra demonstrated a

{sup 15}N-labeled nitrogen from green manure and ammonium sulfate utilization by the sugarcane ratoon

Energy Technology Data Exchange (ETDEWEB)

Ambrosano, Edmilson Jose; Rossi, Fabricio, E-mail: ambrosano@apta.sp.gov.b [Agencia Paulista de Tecnologia dos Agronegocios (APTA), Piracicapa, SP (Brazil). Polo Rigional Centro Sul; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis; Cantarella, Heitor [Agencia Paulista de Tecnologia dos Agronegocios (APTA/IAC), Campinas, SP (Brazil). Instituto Agronomico de Campinas. Centro de Solos e Recursos Agroambientais; Ambrosano, Glaucia Maria Bovi [Universidade de Campinas (UNICAMP/FOP), Piracicaba, SP (Brazil). Fac. de Odontologia de Piracicaba. Dept. de Odontologia Social, Bioestatistica; Schammass, Eliana Aparecida [Agencia Paulista de Tecnologia dos Agronegocios (APTA/IZ), Nova Odessa, SP (Brazil). Instituto de Zootecnia; Muraoka, Takashi [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Fertilidade do solo

2011-05-15

Legumes as green manure are alternative sources of nitrogen (N) for crops and can supplement or even replace mineral nitrogen fertilization due to their potential for biological nitrogen fixation (BNF). The utilization of nitrogen by sugarcane (Saccharum spp.) fertilized with sunn hemp (Crotalaria juncea L.) and ammonium sulfate (AS) was evaluated using the {sup 15}N tracer technique. N was added at the rate of 196 and 70 kg ha{sup -1} as {sup 15}N-labeled sunn hemp green manure (SH) and as ammonium sulfate (AS), respectively. Treatments were: (I) Control; (II) AS{sup 15}N; (III) SH{sup 15}N + AS; (IV) SH{sup 15}N; and (V) AS{sup 15}N + SH. Sugarcane was cultivated for five years and was harvested three times. {sup 15}N recovery was evaluated in the two first harvests. In the sum of the three harvests, the highest stalk yields were obtained with a combination of green manure and inorganic N fertilizer; however, in the second cutting the yields were higher where SH was used than in plots with AS. The recovery of N by the first two consecutive harvests accounted for 19 to 21% of the N applied as leguminous green manure and 46 to 49% of the N applied as AS. The amounts of inorganic N, derived from both N sources, present in the 0-0.4 m layer of soil in the first season after N application and were below 1 kg ha{sup -1}. (author)

Effect of nitrificide in temporally varied /sup 15/N-labelled slurry fertilization

Energy Technology Data Exchange (ETDEWEB)

Peschke, H. (Humboldt-Universitaet, Berlin (German Democratic Republic). Sektion Pflanzenproduktion)

1983-01-01

The fertilizing effect of /sup 15/N-labelled semi-liquid cattle manure (application in August, November and March) with nitrification inhibitor 'N-Serve' added in amounts of 1 and 3 percent of the fertilizer N on perennial ryegrass with feed oats as follow-up seed was tested in an open-air pot experiment using sand of a sandy-rusty soil. A combination of semi-liquid manure and straw and winter rape-seed, respectively, as green manure (August) and ammonium sulfate with and without 1 per cent N-Serve (November and March) were used as comparative variants. Manuring in August had no effect on the yield, the 3 per cent N-Serve application in November and of 1 and 3 percent in March resulted in an increase of the perennial ryegrass yield. By analogy, the N utilization of the fertilizers was increased with later fertilizing dates and the amount of the nitrificide applied. In the case of semi-liquid manure application, N-Serve promoted the mineralization of the nitrogen in the soil.

Isobutylidene diurea as NPN-source for ruminants. 2. Digestion of /sup 15/N-labelled isobutylidene diurea in lactating dairy cows

Energy Technology Data Exchange (ETDEWEB)

Bergner, H; Goersch, R; Adam, K; Piatkowski, B; Voigt, J [Humboldt-Universitaet, Berlin (German Democratic Republic). Sektion Tierproduktion und Veterinaermedizin; Akademie der Landwirtschaftswissenschaften der DDR, Dummerstorf-Rostock. Forschungszentrum fuer Tierproduktion)

1981-04-01

Two cows with rumen cannulae and duodenal re-entrance cannulae received a conventional diet on the basis of a mixture of maize silage, hay and concentrated feed and after a three-week adaptation to isobutylidene diurea (IBDU) 138 g IBDU with 3.865 mg /sup 15/N excess as a single supplementation to their first meal. After a 6-week break without IBDU adaptation the same cows served for a repeated experiment. Irrespective of the adaptation, a reincrease of the /sup 15/N labelling in the TCA-soluble N in the rumen could be proved between the tsup(th) and the 8sup(th) hour after the intake of the isotopes, which resulted from the backflow of /sup 15/N to the rumen. In the duodenal digesta the maximum labelling of the TCA - soluble N fraction appeared 12 hours after the intake of the isotopes. At that moment a labelling plateau began in the protein fraction, which lasted to the 36sup(th) hour. On an average of all 4 cows approximately 30% of the /sup 15/N taken up in the TCA-precipitable fraction and 55 to 60% in the TCA-soluble fraction had passed the duodenum up to the 72sup(nd) hour after the beginning of the experiment approximately 15% were excreted in urine, 16% in feces and 7% in the milk. This shows that roughly one half of the 90% /sup 15/N amount measured at its passage in the re-entrance cannula (related to the intake) was metabolized in the rumen at least twice, and after the first passage through the duodenum it originated from the intermediary metabolism, resp. Negative correlations could be ascertained between the pH value of the rumen fluid and the /sup 15/N incorporation into the rumen proteins as well as the N excretion through the TCA-soluble and -precipitable quota of feces. An adaptation to IBDU is not necessary obviously.

13C- and 15N-labelled non-biogenic compounds used as stable isotope drugs for human liver function tests

International Nuclear Information System (INIS)

Krumbiegel, P.

1989-01-01

As a result of liver diseases, the elimination of certain drugs is retarded. After labelling a suitable drug with 13 C, the 13 CO 2 elimination rate serves as a liver function parameter. Current contributions to the 13 CO 2 breath test method are reviewed and related to the 14 CO 2 breath test proposals. In spite of several advantages of 13 C-labelled agents, some dissatisfaction has remained with the tests, especially at using them with infants. It is the necessity of face masks and the uncertainty to consider endogeneous CO 2 contributions diluting the exhaled 13 CO 2 . The problems are avoided if the other molecule site of the drug is labelled which is known to be eliminated via urine. With 15 N as a tracer, a suitable urine test using [ 15 N]-methacetin as agent has been proposed and put into practice. (author)

Nitrogen incorporation and retention by bacteria, algae, and fauna in a subtropical, intertidal sediment: An in situ 15N-labeling study

NARCIS (Netherlands)

Veuger, B.; Eyre, B.D.; Maher, D.; Middelburg, J.J.

2007-01-01

We performed a 15N-labeling study to investigate nitrogen incorporation and retention by the benthic microbial community (bacteria and benthic microalgae) and fauna in the intertidal sediment of the subtropical Australian Brunswick Estuary. The main experiment involved an in situ 15N pulse–chase

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

Science.gov (United States)

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

2015-01-01

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

Nitrogen accumulation and partitioning in a High Arctic tundra ecosystem from extreme atmospheric N deposition events

Energy Technology Data Exchange (ETDEWEB)

Choudhary, Sonal, E-mail: S.Choudhary@sheffield.ac.uk [Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Management School, University of Sheffield, Conduit Road, Sheffield S10 1FL (United Kingdom); Blaud, Aimeric [Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); Osborn, A. Mark [Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom); School of Applied Sciences, RMIT University, PO Box 71, Bundoora, VIC 3083 (Australia); Press, Malcolm C. [School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom); Manchester Metropolitan University, Manchester, M15 6BH (United Kingdom); Phoenix, Gareth K. [Department of Animal and Plant Sciences, University of Sheffield, Western Bank, Sheffield S10 2TN (United Kingdom)

2016-06-01

Arctic ecosystems are threatened by pollution from recently detected extreme atmospheric nitrogen (N) deposition events in which up to 90% of the annual N deposition can occur in just a few days. We undertook the first assessment of the fate of N from extreme deposition in High Arctic tundra and are presenting the results from the whole ecosystem {sup 15}N labelling experiment. In 2010, we simulated N depositions at rates of 0, 0.04, 0.4 and 1.2 g N m{sup −2} yr{sup −1}, applied as {sup 15}NH{sub 4}{sup 15}NO{sub 3} in Svalbard (79{sup °}N), during the summer. Separate applications of {sup 15}NO{sub 3}{sup −} and {sup 15}NH{sub 4}{sup +} were also made to determine the importance of N form in their retention. More than 95% of the total {sup 15}N applied was recovered after one growing season (~ 90% after two), demonstrating a considerable capacity of Arctic tundra to retain N from these deposition events. Important sinks for the deposited N, regardless of its application rate or form, were non-vascular plants > vascular plants > organic soil > litter > mineral soil, suggesting that non-vascular plants could be the primary component of this ecosystem to undergo measurable changes due to N enrichment from extreme deposition events. Substantial retention of N by soil microbial biomass (70% and 39% of {sup 15}N in organic and mineral horizon, respectively) during the initial partitioning demonstrated their capacity to act as effective buffers for N leaching. Between the two N forms, vascular plants (Salix polaris) in particular showed difference in their N recovery, incorporating four times greater {sup 15}NO{sub 3}{sup −} than {sup 15}NH{sub 4}{sup +}, suggesting deposition rich in nitrate will impact them more. Overall, these findings show that despite the deposition rates being extreme in statistical terms, biologically they do not exceed the capacity of tundra to sequester pollutant N during the growing season. Therefore, current and future extreme events

Plot-size for 15N-fertilizer recovery studies by tanzania-grass

International Nuclear Information System (INIS)

Martha Junior, Geraldo Bueno; Trivelin, Paulo Cesar Ocheuze; Corsi, Moacyr

2009-01-01

The understanding of the N dynamics in pasture ecosystems can be improved by studies using the 15 N tracer technique. However, in these experiments it must be ensured that the lateral movement of the labeled fertilizer does not interfere with the results. In this study the plot-size requirements for 15 N-fertilizer recovery experiments with irrigated Panicum maximum cv. Tanzania was determined. Three grazing intensities (light, moderate and intensive grazing) in the winter, spring and summer seasons were considered. A 1 m 2 plot-size, with a grass tussock in the center, was adequate, irrespective of the grazing intensity or season of the year. Increasing the distance from the area fertilized with 15 N negatively affected the N derived from fertilizer (Npfm) recovered in herbage.The lowest decline in Npfm values were observed for moderate and light grazing intensities. This fact might be explained by the vigorous growth characteristics of these plants. Increasing the grazing intensity decreased the tussock mass and, the smaller the tussock mass, the greater was the dependence on fertilizer nitrogen. (author)

Plot-size for {sup 15}N-fertilizer recovery studies by tanzania-grass; Tamanho da parcela para estudos de recuperacao de fertilizante-{sup 15}N por capim-tanzania

Energy Technology Data Exchange (ETDEWEB)

Martha Junior, Geraldo Bueno [EMBRAPA Cerrados, Planaltina, DF (Brazil)], e-mail: gbmartha@cpac.embrapa.br; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis], e-mail: pcotrive@cena.usp.br; Corsi, Moacyr [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Zootecnia], e-mail: moa@esalq.usp.br

2009-07-01

The understanding of the N dynamics in pasture ecosystems can be improved by studies using the {sup 15}N tracer technique. However, in these experiments it must be ensured that the lateral movement of the labeled fertilizer does not interfere with the results. In this study the plot-size requirements for {sup 15}N-fertilizer recovery experiments with irrigated Panicum maximum cv. Tanzania was determined. Three grazing intensities (light, moderate and intensive grazing) in the winter, spring and summer seasons were considered. A 1 m{sup 2} plot-size, with a grass tussock in the center, was adequate, irrespective of the grazing intensity or season of the year. Increasing the distance from the area fertilized with {sup 15}N negatively affected the N derived from fertilizer (Npfm) recovered in herbage.The lowest decline in Npfm values were observed for moderate and light grazing intensities. This fact might be explained by the vigorous growth characteristics of these plants. Increasing the grazing intensity decreased the tussock mass and, the smaller the tussock mass, the greater was the dependence on fertilizer nitrogen. (author)

Synthesis of [1,3 - 15 N2] uracil

International Nuclear Information System (INIS)

Chiriac, M.; Axente, D.

2001-01-01

The synthesis of 15 N labelled uracil, using CO( 15 NH 2 ) 2 as starting material, is presented. The experimental procedure is an adaptation of the synthesis methods for the corresponding unlabelled compounds. Urea- 15 N 2 used as starting material was obtained from H 15 NO 3 (99 at.% 15 N) produced at National Institute for Research and Development of Isotopic and Molecular Technologies, Cluj-Napoca.The uracil structure was determined using the mass spectrometry method and the isotopic labelling was determined by the same method on the molecular compound. The synthesis scheme of (1,3- 15 N 2 ) uracil is presented. (authors)

Cyanobacteria-derived nitrogen uptake by benthic invertebrates in Lake Taihu: a mesocosm study using 15N labeling

Directory of Open Access Journals (Sweden)

Yu J.

2014-01-01

Full Text Available Eutrophication of lakes can lead to dominance by cyanobacteria, which are hardly used by zooplankton due to their low nutrition value. However, sedimented cyanobacterial detritus may be a useful source for benthic invertebrates. We studied the Microcystis-derived nitrogen incorporation in benthic invertebrates in Lake Taihu using stable isotopic nitrogen (15N as a tracer. The δ15N of all organisms increased significantly with time after addition of the labeled Microcystis detritus. δ15N values of POM and periphyton peaked earlier than for benthic invertebrates, and the maximum levels were also higher than bivalves, snails and worms (Limnodrilus spp.. Among benthic invertebrates, Radix swinhoei peaked later than other invertebrates, but the maximum level and the excess 15N of the last sampling day were higher. At the end of the experiment, approximately 70% of the added 15N was retained in the benthic food web, while only a small fraction (less than 1% of the added detritus 15N occurred in the pelagic food web. Our results suggest that nitrogen from cyanobacteria can be incorporated more in benthic than pelagic food webs and cyanobacterial blooms may contribute to the development of benthic animals.

Labelling of animal manure nitrogen with ¹⁵N

DEFF Research Database (Denmark)

Sørensen, P.; Jensen, E.S.; Nielsen, N.E.

1994-01-01

A sheep was fed on N-15-labelled ryegrass hay during a period of 9 days in order to obtain N-15-labelled manure. After 9 days of feeding, the total N in faeces contained 3.70 atom % N-15 excess, which was equivalent to 82% of the N-15 enrichment of the hay N. The easily-decomposable fraction...

Bicarbonate as tracer for plant assimilated C and homogeneity of 14C and 15N distribution in ryegrass and white clover tissue by alternative labeling approaches

DEFF Research Database (Denmark)

Rasmussen, Jim; Kusliene, Gedrime; Jacobsen, Ole Stig

2013-01-01

that 15N also had a heterogeneous distribution (up to two orders of magnitude). Conclusion Bicarbonate can efficiently be used to introduce 14C or 13C into plant via the leaf-labeling method. Both 14C and 15N showed heterogeneous distribution in the plant, although the distribution of 15N was more even......Aims: Application of carbon (C) and nitrogen (N) isotopes is an essential tool to study C and N flows in plant-soil-microorganisms systems. When targeting single plants in a community the tracers need to be added via e.g., leaf-labeling or stem-feeding approaches. In this study we: (i) investigated...... if bicarbonate can be used to introduce 14C (or 13C) into white clover and ryegrass, and (ii) compared the patterns of 14C and 15N allocation in white clover and ryegrass to evaluate the homogeneity of tracer distribution after two alternative labeling approaches. Methods Perennial ryegrass and white clover were...

Medium-term response of microbial community to rhizodeposits of white clover and ryegrass and tracing of active processes induced by 13C and 15N labelled exudates

DEFF Research Database (Denmark)

Kusliene, Gedrime; Rasmussen, Jim; Kuzyakov, Yakov

2014-01-01

and actinomycetes was unaffected by plant species, but pool of Gram-negative and Gram-positive bacteria was greater under white clover at the 10 percent significance level. In the short term, microorganisms more actively utilised fresh exudates (13C-labelled) of ryegrass than of white clover. We expected ryegrass...... microbial groups in soil under white clover (Trifolium repens L.) and ryegrass (Lolium perenne L.) following leaf-labelling with 13C-bicarbonate and 15N-urea. In this way microbial N and 15N and the composition of PLFAs reflect the medium-term (two months) response of microorganisms to rhizodeposits......, whereas the 13C-label of the PLFAs reflects the short-term (one week) utilisation of root exudates following labelling of shoots. In the medium term, microbial biomass N and 15N were greater under the ryegrass, whereas total PLFA was higher under white clover. The relative abundance of fungi...

Unusually negative nitrogen isotopic compositions (δ15N) of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

Science.gov (United States)

Fogel, M. L.; Wooller, M. J.; Cheeseman, J.; Smallwood, B. J.; Roberts, Q.; Romero, I.; Meyers, M. J.

2008-12-01

Extremes in δ15N values in mangrove tissues and lichens (range =+4 to -22‰) were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, and atmospheric ammonia, and the δ15N of lichens, mangrove leaves, roots, stems, and wood were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Dwarfed Rhizophora mangle trees had the most negative δ15N, whereas fringing Rhizophora trees, the most positive δ15N values. Porewater ammonium concentrations had little relationship to N isotopic fractionation in mangrove tissues. In dwarfed mangroves, the δ15N of fine and coarse roots were 6-9‰ more positive than leaf tissue from the same tree, indicating different sources of N for root and leaf tissues. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year from -12‰ to -2‰, approaching the δ15N of porewater ammonium (δ15N=+4‰). Isotopically depleted ammonia in the atmosphere (δ15N=-19‰) and in rainwater (δ15N=-10‰) were found on Twin Cays. We propose that foliar uptake of these atmospheric sources by P-stressed, dwarfed mangrove trees and lichens can explain their very negative δ15N values. In environments where P is limiting for growth, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

Synthesis of puric bases labelled with carbon 14 and nitrogen 15

International Nuclear Information System (INIS)

Lamorre, Yves

1975-01-01

In this report for graduation in organic chemistry engineering, the author reports the synthesis of adenine 14 C-2 et 14 C-6 by two different chemical ways from two derivatives of imidazole. He has used adenine 14 C-6 to obtain hypoxanthine 14 C-6, and then, by enzymatic processing, uric acid 14 C-6. He reports the study of the production of guanine 14 C-2 by cyclization of silylated derivative of imidazole with the carbon 14 C sulphur. However, a method of complete synthesis of this same compound revealed to be more practical. This complete synthesis way allowed the labelling of guanine in positions 1, 2 and 3 by the 96 per cent isotopic nitrogen. Nitrogen in positions 7 and 9 could have been labelled by the same way from the ethyl cyanoacetate 15 N and from the sodium nitrite 15 N. The study of the mass spectrum of these compounds labelled with nitrogen 15 N allowed most of fragments obtained during this analysis to be identified [fr

Recovery of 15N-labelled fertilizers applied to bromegrass on a thin black chernozem soil

International Nuclear Information System (INIS)

Malhi, S.S.

1995-01-01

The availability of N fertilizers on established grass stands is a function of such processes as immobilization, gaseous loss, leaching and position of applied N. A field experiment was conducted on a Thin Black Chernozem soil at Crossfield, Alberta to determine the effect of source, time and method of application on the recovery of 15 N-labelled fertilizers applied to smooth bromegrass (Bromus inermis Leyss.). The treatments included two sources of N [urea and ammonium nitrate (AN)], four application times (early autumn, late autumn, early spring and late spring) and two methods of placement (surface-broadcast and subsurface banding). In most cases the 15 N recovery in soil did not differ much between urea and AN. However, when urea was surface-broadcast, there was, on average, 10.2% less 15 N recovery in plants than AN. The N recovery for late spring > early spring > late autumn = early autumn. When urea was banded 4 cm deep into the soil, N recovery in plants increased significantly compared with its surface-broadcast application. However, this was not observed when the source of N was AN. Banding generally increased the amount of immobilized N present in the soil and N recovery. We concluded that the N recovery in plants and in plants plus soil was less for urea than for AN and was less with autumn broadcast N application than with spring broadcast application. (author). 23 refs., 3 tabs

Preparation of 15N-13C-fulminic acid

International Nuclear Information System (INIS)

Wilmes, R.; Winnewisser, M.

1993-01-01

The precursor for the title compound was prepared in a three-step synthesis. The 13 C-label was incorporated in the first step employing 2- 13 C-ethyl acetate and the 15 N-label in the last step, using 15 N-sodium nitrite. Upon pyrolysis the precursor forms three fragments, one of them being the title compound. (Author)

North Atlantic ecosystem shifts revealed by cod otolith δ¹⁵N and δ¹³C chronologies

DEFF Research Database (Denmark)

Pedersen, Jens Brøgger; Nielsen, Jens Munk; Steingrund, Petur

. To study the link between environmental changes and ecosystem trophic structure we developed δ15N and δ13C chronologies by analyzing the organic matrix of cod otoliths from the Faroe Shelf cod population (1950-2010) and the Nuuk Fjord cod population (1927-2009). Significant correlations between δ15N & δ13C...... of organic matrix of otolith core material (Nuuk Fjord) and annual growth increments in Ocean Quahog (A. Islandica) shells will be included.......Changes in climate and exploitation have caused large fluctuations in the productivity of many North Atlantic cod populations and the collapse of many cod fisheries. These fluctuations are most likely due to a combined effect of physical processes and changes in ecosystem trophic structure...

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

Dynamics of nitrogen in an oxic paleudalf soil with the incorporation of 15N-tagged organic nitrogen (maize straw) and 15N-tagged mineral nitrogen (ammonium sulphate)

International Nuclear Information System (INIS)

Freitas, J.R. de.

1984-12-01

An experiment, carried out under field conditions in 12 lysimeters, each containing 3.0 ton of Oxic Paleudalf soil with four replicates, is described. This objective is labelling soil organic N. Nitrogen was incorporated into soil as maize straw, non-labelled and labelled with 15 N and ammonium sulphate - 15 N. The soil was sampled every 15 days in three different depths. N as NH + 4 , NO - 3 , total-N and (%)C and (%) moisture was analysed. (M.A.C.) [pt

Cyclotron production of molecules labelled with short-lived radioisotopes β+ emitters (15O, 13N, 11C) and their clinical uses

International Nuclear Information System (INIS)

Bougharouat, B.

1981-01-01

Clinical use of three short-lived radioisotopes: 15 O, 13 N and 11 C is studied on two complementary aspects. A production and purification system is realized; detection instruments in medical use are studied. The production of labelled molecules with the three radiotracers 15 O, 13 N, 11 C from the target bombardment with charged and accelerated particles was studied [fr

Unusually negative nitrogen isotopic compositions (δ15N of mangroves and lichens in an oligotrophic, microbially-influenced ecosystem

Directory of Open Access Journals (Sweden)

I. Romero

2008-12-01

Full Text Available Extremes in δ15N values in mangrove tissues and lichens (range =+4 to −22‰ were measured from a mangrove forest ecosystem located on Twin Cays, offshore islands in Belize, Central America. The N isotopic compositions and concentrations of NH4+/NH3 in porewater, rainwater, and atmospheric ammonia, and the δ15N of lichens, mangrove leaves, roots, stems, and wood were examined to study the biogeochemical processes important for establishing these unusual N isotopic ratios. Dwarfed Rhizophora mangle trees had the most negative δ15N, whereas fringing Rhizophora trees, the most positive δ15N values. Porewater ammonium concentrations had little relationship to N isotopic fractionation in mangrove tissues. In dwarfed mangroves, the δ15N of fine and coarse roots were 6–9‰ more positive than leaf tissue from the same tree, indicating different sources of N for root and leaf tissues. When P was added to dwarfed mangrove trees without added N, δ15N increased within one year from −12‰ to −2‰, approaching the δ15N of porewater ammonium (δ15N=+4‰. Isotopically depleted ammonia in the atmosphere (δ15N=−19‰ and in rainwater (δ15N=−10‰ were found on Twin Cays. We propose that foliar uptake of these atmospheric sources by P-stressed, dwarfed mangrove trees and lichens can explain their very negative δ15N values. In environments where P is limiting for growth, uptake of atmospheric N by Rhizophora mangle may be an important adaptive strategy.

Olive and citrus tree crops and their fertilization in Greece: Field studies with 15N labelled fertilizers

International Nuclear Information System (INIS)

Papanicolaou, E.P.

1982-01-01

Fertilizer use for tree crops in Greece is increasing rapidly, however, fertilizer experiments with olive and citrus tree crops have given results which were often inconclusive. The value of using isotopically labelled fertilizers to directly measure fertilizer uptake is thus obvious. A preliminary experiment determined that relatively low enriched 15 N fertilizers (1.0-1.5% atom excess) could be accurately detected in citrus trees. The concentration of added N was higher in the leaves than in the wood and fruit. In a second study foliarly applied 15 N urea was found to be taken up more efficiently than broadcast urea or ammonium sulphate. In an initial sampling distribution of N among the plant parts was similar to that of the initial study, although a final harvest and analysis must still be conducted. (author)

Nitrate removal in stream ecosystems measured by 15N addition experiments: Denitrification

Science.gov (United States)

Mulholland, P.J.; Hall, R.O.; Sobota, D.J.; Dodds, W.K.; Findlay, S.E.G.; Grimm, N. B.; Hamilton, S.K.; McDowell, W.H.; O'Brien, J. M.; Tank, J.L.; Ashkenas, L.R.; Cooper, L.W.; Dahm, Clifford N.; Gregory, S.V.; Johnson, S.L.; Meyer, J.L.; Peterson, B.J.; Poole, G.C.; Valett, H.M.; Webster, J.R.; Arango, C.P.; Beaulieu, J.J.; Bernot, M.J.; Burgin, A.J.; Crenshaw, C.L.; Helton, A.M.; Johnson, L.T.; Niederlehner, B.R.; Potter, J.D.; Sheibley, R.W.; Thomasn, S.M.

2009-01-01

We measured denitrification rates using a field 15N-NO- 3 tracer-addition approach in a large, cross-site study of nitrate uptake in reference, agricultural, and suburban-urban streams. We measured denitrification rates in 49 of 72 streams studied. Uptake length due to denitrification (SWden) ranged from 89 m to 184 km (median of 9050 m) and there were no significant differences among regions or land-use categories, likely because of the wide range of conditions within each region and land use. N2 production rates far exceeded N2O production rates in all streams. The fraction of total NO-3 removal from water due to denitrification ranged from 0.5% to 100% among streams (median of 16%), and was related to NHz 4 concentration and ecosystem respiration rate (ER). Multivariate approaches showed that the most important factors controlling SWden were specific discharge (discharge / width) and NO-3 concentration (positive effects), and ER and transient storage zones (negative effects). The relationship between areal denitrification rate (Uden) and NO- 3 concentration indicated a partial saturation effect. A power function with an exponent of 0.5 described this relationship better than a Michaelis-Menten equation. Although Uden increased with increasing NO- 3 concentration, the efficiency of NO-3 removal from water via denitrification declined, resulting in a smaller proportion of streamwater NO-3 load removed over a given length of stream. Regional differences in stream denitrification rates were small relative to the proximate factors of NO-3 concentration and ecosystem respiration rate, and land use was an important but indirect control on denitrification in streams, primarily via its effect on NO-3 concentration. ?? 2009.

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

Science.gov (United States)

Averill, Colin; Finzi, Adrien

2011-04-01

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

Utilization of 15N-labelled nitrogen fertilizer in dependence on organic manuring and carbon and nitrogen contents of loess chernozem profiles with different stratification

International Nuclear Information System (INIS)

Greilich, J.

1988-01-01

In an outdoor model experiment with different total C and N contents in five profile variants of loess chernozem, the utilization of 15 N-labelled mineral fertilizer N by maize was investigated over three years. The total nitrogen uptake in the variants correlated with the yields at nearly uniform nitrogen contents in dry matter. Total C and N contents of the profile variants and one organic manure application per year had no statistically significant effects on the 15 N-labelled fertilizer N proportion in total N content of biomass. As a result of the low yields obtained from the variants with low total C and N contents of soil, mineral fertilizer utilization was found to be lower, too, in most of these variants. Organic manuring had no essential effect on mineral fertilizer N utilization. (author)

Fate of N and relative efficiency of 15N-labeled organic materials applied to transplanted rice in northern Kyushu region of Japan

International Nuclear Information System (INIS)

Nishida, Mizuhiko; Tsuchiya, Kazunari; Yamamuro, Shigekazu

2004-01-01

Seven kinds of 15 N-labeled organic materials were applied to transplanted rice to investigate their N fate and relative efficiency in the northern Kyushu region of Japan. The 15 N-labeled organic materials examined in a micro-plot experiment were cattle manure compost, poultry manure compost, swine feces, rice straw compost, rice bran, rice straw, and wheat straw. Regarding swine feces, rice bran, and wheat straw, the direct evaluation of their N fate in paddy fields using 15 N organic materials has not been reported. A significant difference in the N fate in response to the type of organic materials was observed in the uptake rate by rice plants. The uptake rate at the maturity stage was significantly higher in poultry manure (29%), swine feces (25%), and rice bran (26%) than for the other organic materials (6-13%). Cattle manure compost showed the lowest value, namely 6-7%. Using the uptake rate of ( 15 NH 4 ) 2 SO 4 observed earlier, the relative efficiency of organ nic materials (relative uptake rate of organic material N to chemical fertilizer N) was calculated as the index of the organic material N efficiency. These relative efficiencies of organic materials derived from animal wastes were 16-19, 81, 72, and 71% for cattle manure compost, poultry manure compost, poultry manure compost without inherent NH 4 -H, and swine feces, respectively, and were similar to those estimated by indirect way. The relative efficiencies of organic materials derived from plant residues were 25-31, 73, 33 and 34% for rice straw compost, rice bran, rice straw, and wheat straw, respectively. The N uptake from the organic materials (OM-N uptake) in swine feces and cattle manure compost continued throughout the rice growth period, whereas the OM-N uptake of the other organic materials declined remarkably after 54 DAT. No significant difference was observed in the residual rate and the loss rate among the organic materials. However, some tendencies that might be related to the

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)

Determination of endogenous nitrogen in feces using 15N tracers

International Nuclear Information System (INIS)

Herrmann, U.; Krawielitzki, K.; Schadereit, R.; Smulikowska, S.

1986-01-01

A ration consisting of wheat gluten and N-free components was supplemented with L-lysine and L-leucine and fed to two groups of growing Wistar rats. Group 1 received 15 N Lys and unlabelled Leu, group 2 received unlabelled Lys and 15 N Leu in order to study the influence of the utilization of the 15 N marker on the labelling quota of feces and urine as well as various fractions of the body. The good utilization of Lys in group 1 results in a higher 15 N excess in feces and a reduced 15 N abundance in urine in comparison to group 2 with a lower utilization of 15 N Leu. The results show that the 15 N abundance in urine is unsuitable as an indicator of the 15 N labelling quota of endogenous metabolic fecal nitrogen. (author)

Resolution of the 15N balance enigma?

International Nuclear Information System (INIS)

Clough, T.J.; Sherlock, R.R.; Cameron, K.C.; Stevens, R.J.; Laughlin, R.J.; Mueller, C.

2001-01-01

The enigma of soil nitrogen balance sheets has been discussed for over 40 years. Many reasons have been considered for the incomplete recovery of 15 N applied to soils, including sampling uncertainty, gaseous N losses from plants, and entrapment of soil gases. The entrapment of soil gases has been well documented for rice paddy and marshy soils but little or no work appears to have been done to determine entrapment in drained pasture soils. In this study 15 N-labelled nitrate was applied to a soil core in a gas-tight glovebox. Water was applied, inducing drainage, which was immediately collected. Dinitrogen and N -2 were determined in the flux through the soil surface, and in the gases released into the glovebox as a result of irrigation or physical destruction of the core. Other components of the N balance were also measured, including soil inorganic-N and organic-N. Quantitative recovery of the applied 15 N was achieved when the experiment was terminated 484 h after the 15 N-labelled material was applied. Nearly 23% of the 15 N was recovered in the glovebox atmosphere as N 2 and N 2 O due to diffusion from the base of the soil core, convective flow after irrigation, and destructive soil sampling. This 15 N would normally be unaccounted for using the sampling methodology typically employed in 15 N recovery experiments. Copyright (2001) CSIRO Publishing

Oxygen-15 labelled water production for positron emission tomography

International Nuclear Information System (INIS)

Janus, A.; Sachinidis, J.I.; Chan, J.G.; Tochon-Danguy, H.J.

1998-01-01

Full text: Functional imaging using positron emission tomography (PET) and 15 O-labelled compounds is both scientifically and clinically challenging. The short half-life of oxygen-15 (t 1/2 = 2 min) allows for multiple administration to a patient without exceeding acceptable levels of absorbed radiation dose and without excessive delay between administrations. The clinical usefulness of [ 15 O]-labelled water for cerebral blood flow measurements has been well established. Here we report the development and construction of a [ 15 O]water generator based on an earlier design from Hammersmith Hospital, London. The cyclotron produces a continuous flow of [ 15 O]O 2 gas by the irradiation of a natural nitrogen target (1% O 2 in N 2 ) with a 5 MeV deuteron beam, via the nuclear reaction ( 14 N(d,n) 15 O). The radioactive gas is then mixed with 5% hydrogen in nitrogen and piped to the water generator located in the scanner room. The O 2 /N 2 gas mixture is reacted over a palladium catalyst at 1500 deg C to produce [ 15 O]H 2 O vapour. The vapour passes through an exchanger where it diffuses across a semi-permeable membrane (cellulose acetate) into saline solution. At the optimum gas flow- rate of 500 mL/min, more than 95% of the radioactive oxygen is converted to radioactive water. Waste radioactive gas is piped back to the cyclotron vault to decay before release into the atmosphere. The saline solution (0.9% NaCl) is pumped continuously through the system at 6 mL/min with an infusion pump (3M AVI470). The present system has been in operation for more than a year and has been used for clinical evaluation of stroke patients and for brain activation research studies

Studies on the process of attachment of diazotroph alcaligenes faecalis and its Tn5 mutants to rice roots using 15N-labelling technique

International Nuclear Information System (INIS)

Fang Xuanjun; Lin Min; You Chongbiao

1993-09-01

By using 15 N-labelling technique and Tn5-induced mutants the attachment of associative diazotroph Alcaligenes faecalis to intact rice plants was examined in vitro. Three distinguished modes of attachment of Alcaligenes faecalis: adsorption, anchoring and colonization were proposed by using 15 N-labelling bacterial cells and Tn5-induced mutants. Che - mutants affected on adsorption, but not on anchoring. Exo - Che - mutant is defective in both adsorption and anchoring. Exo - or exo ++ mutants are only defective in anchoring. Effective colonization is benefit for establishment on the associative system. The data also indicated that EPS (exopolysaccharide) play rather important roles in the association between the host plant and bacteria

Studies of 15N transamination following application of various tracer substances. 1

International Nuclear Information System (INIS)

Schadereit, R.; Krawielitzki, K.; Herrmann, U.

1986-01-01

4 groups of 3 growing Wistar rats each were orally given 15 N-labelled methionine, lysine, glycine and ammonia sulphate, resp., over 10 days. Measuring the 15 N accumulation in the amino acids (AA) of the body protein, the transamination of the individual 15 N substances and thus their suitability as tracer substances for studies of N metabolism was determined. None of the tested 15 N-AA achieved a proportionate labelling of all AA of the body protein. The AA used as tracer in each case showed the highest 15 N labelling. Of the amino- 15 N detected in the animal body, about 19% were found in Met after 15 N Met application, 88% in Lys after 15 N Lys application and 50% in Gly after 15 N Gly application. After the application of 15 N-ammonia sulphate about 42% of the body amino- 15 N are apportioned to the essential and 58% to the non-essential AA. Thus, this substance produces a more proportional labelling of the essential and non-essential AA of the body protein than 15 N-Gly. The following quotas of the 15 N amounts applied were found in the AA of the animal bodies: tracer substance lysine 52%, glycine 32%, ammonia sulphate 24%, methionine 21%. After summing up the amino acid 15 N amounts in the animal body, eliminating in each case the tracer AA and taking into account the molecular weight of the AA, there was a good agreement of the intensity of the accumulation of 15 N in the individual AA, irrespective of the applied tracer substance. (author)

Using N-15 Technique for Assessing Organic.N Turnover in Sandy Soil

International Nuclear Information System (INIS)

Soliman, S.; El-Akel, E.A.; Ismail, M.M.; El-Sherbiny, E.; Awad, E.E.

2008-01-01

Turnover of organic-N was traced under greenhouse condition. 15 N-labelled wheat and/or soybean residues were used as organic additives which applied individually or in combinations. These residues were applied at rates of 100, 75 and 25μg N g - 1 soil. Also, labelled ammonium sulfate with 2% 15 N atom excess, was applied either alone or in combination with the plant residues, at rates of 100, 75 and 25μg N g - 1 soil as single dose after 10 days from planting. Relative positive effect of the nitrogen plant residues on N-uptake and yield components can be arranged as follows: Soybean > wheat + > soybean > wheat residues. Tracer technique indicated that the mixture of labeled residues and ammonium sulfate at rates of (*50 + 50) and (*25 + 75), was effective on dry matter and N uptake. Effect of organic and inorganic nitrogen sources on portions N derived from residue (Ndfr) and N derived from fertilizer (Ndff) to wheat could be arranged as following: ammonium sulfate > soybean > mixture > wheat. Higher 15 N recovery percentage was noticed in grains as affected by addition of soybean residues combined with ordinary ammonium sulfate at rates of (*25 + 75) and (*50 + 50), respectively

Quantitative analysis of 15N-labeled positional isomers of glutamine and citrulline via electrospray ionization tandem mass spectrometry of their dansyl derivatives.

Science.gov (United States)

Marini, Juan C

2011-05-15

The enteral metabolisms of glutamine and citrulline are intertwined because, while glutamine is one of the main fuel sources for the enterocyte, citrulline is one of its products. It has been shown that the administration of (15)N-labeled glutamine results in the incorporation of the (15)N label into citrulline, but it is not clear which of the three nitrogen groups of citrulline is actually labeled. To determine the (15)N-enrichment of the positional isomers of glutamine and citrulline, a rapid liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed. The amino acids were analyzed as their dansyl derivatives. The product ion resulting from the loss of NH(3) from the omega carbon allows for the determination of the enrichment of the ureido (citrulline) or amido groups (glutamine). The protonated pyrrolidine (citrulline) or 5-oxopyrrolidine (glutamine) product ion contains the 2-N (amino group) and is used to determine its enrichment. The method described showed no ion suppression and a wide dynamic range ranging from 1.3 picomoles to 2 nanomoles for citrulline. Background samples and standards resulted in enrichments not different from those theoretically expected. The enrichment curves for the different glutamine and citrulline isotopomers were linear (R(2)  > 0.998) over the range of enrichments studied. The method developed provides an additional insight into the metabolism of glutamine and citrulline tracing the precursor-product relationship between these two amino acids. Copyright © 2011 John Wiley & Sons, Ltd.

δ15N constraints on long-term nitrogen balances in temperate forests

Science.gov (United States)

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

2011-01-01

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

A liver-function test using 15N-labelled ammonium chloride

International Nuclear Information System (INIS)

Jung, K.; Hirscherg, K.; Faust, H.; Matkowitz, R.

1985-01-01

Malfunction of the liver involves disturbances of urea synthesis and ammonia detoxification. These phenomena became apparent, especially during ammonia loading of patients. The functional state of the liver can be assessed by oral administration of 15 NH 4 Cl and subsequent analysis of 15 N-urea and 15 N-ammonia in urine by emission spectrometry. Clinical tests based on the ratio of the excess abundances (see Appendix) of 15 N-ammonia to 15 N-urea excreted in urine 3 h after oral administration gave values for patients with liver disease which differed significantly from those for healthy subjects. Absorption disturbances, which often accompany liver diseases, do not influence the effectiveness of the method. (orig.)

Liver-function test using /sup 15/N-labelled ammonium chloride

Energy Technology Data Exchange (ETDEWEB)

Jung, K; Hirscherg, K; Faust, H; Matkowitz, R

1985-08-01

Malfunction of the liver involves disturbances of urea synthesis and ammonia detoxification. These phenomena became apparent, especially during ammonia loading of patients. The functional state of the liver can be assessed by oral administration of /sup 15/NH/sub 4/Cl and subsequent analysis of /sup 15/N-urea and /sup 15/N-ammonia in urine by emission spectrometry. Clinical tests based on the ratio of the excess abundances (see Appendix) of /sup 15/N-ammonia to /sup 15/N-urea excreted in urine 3 h after oral administration gave values for patients with liver disease which differed significantly from those for healthy subjects. Absorption disturbances, which often accompany liver diseases, do not influence the effectiveness of the method.

The use of N-15 in the measurement of symbiotic nitrogen fixation by legumes under field condition

International Nuclear Information System (INIS)

Impithuksa, Viroj

1982-01-01

The amount of N fixation by legume crop in field condition by using 15 N can determine by the addition of labelled 15 N fertilizer into the soil and measuring the amount of labelled 15 N, soil N, and fixed N taken up by legume crop. This requires a standard crop (reference crop) as a control to determine labelled 15 N and soil N taken up by this crop. In case the same rate of labelled 15 N fertilizer is added to the legume crop and a standard crop

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

Energy Technology Data Exchange (ETDEWEB)

Wiedemann, Christoph; Goradia, Nishit; Häfner, Sabine [Leibniz Institute for Age Research, Fritz Lipmann Institute, Research Group Biomolecular NMR Spectroscopy (Germany); Herbst, Christian [Ubon Ratchathani University, Department of Physics, Faculty of Science (Thailand); Görlach, Matthias; Ohlenschläger, Oliver; Ramachandran, Ramadurai, E-mail: raman@fli-leibniz.de [Leibniz Institute for Age Research, Fritz Lipmann Institute, Research Group Biomolecular NMR Spectroscopy (Germany)

2015-10-15

A simple triple resonance NMR experiment that leads to the correlation of the backbone amide resonances of each amino acid residue ‘i’ with that of residues ‘i−1’ and ‘i+1’ in ({sup 13}C, {sup 15}N) labelled intrinsically disordered proteins (IDPs) is presented. The experimental scheme, {HN-NCA heteronuclear TOCSY-NH}, exploits the favourable relaxation properties of IDPs and the presence of {sup 1}J{sub CαN} and {sup 2}J{sub CαN} couplings to transfer the {sup 15}N{sub x} magnetisation from amino acid residue ‘i’ to adjacent residues via the application of a band-selective {sup 15}N–{sup 13}C{sup α} heteronuclear cross-polarisation sequence of ∼100 ms duration. Employing non-uniform sampling in the indirect dimensions, the efficacy of the approach has been demonstrated by the acquisition of 3D HNN chemical shift correlation spectra of α-synuclein. The experimental performance of the RF pulse sequence has been compared with that of the conventional INEPT-based HN(CA)NH pulse scheme. As the availability of data from both the HCCNH and HNN experiments will make it possible to use the information extracted from one experiment to simplify the analysis of the data of the other and lead to a robust approach for unambiguous backbone and side-chain resonance assignments, a time-saving strategy for the simultaneous collection of HCCNH and HNN data is also described.

Uptake of 15N-labelled urea and 32P-labelled phosphate from acid-based urea phosphate and granular fertilizers

International Nuclear Information System (INIS)

Bole, J.B.

1986-01-01

The availability of nitrogen and phosphorus in fertilizer products labelled with both 32 P and 15 N was measured in a growth chamber experiment. The uptake of N and P by soft white spring wheat (Triticum aestivum L.) from a solution of acid urea phosphate fertilizer did not differ significantly from that of a mixture of granular urea and monammonium phosphate fertilizer. The fertilizer-P uptake efficiency of both sources was higher in a neutral soil than in acid or calcareous soils. Banding either fertilizer increased the uptake of fertilizer P compared with sources mixed with the soil, but did not significantly affect fertilizer-N uptake. The increase in fertilizer-P efficiency due to banding was significantly greater for the urea-monammonium phosphate than for the acid urea phosphate solution. Banding fertilizer did not increase the uptake of fertilizer P in the calcareous soil, and decreased the uptake of fertilizer N in that soil compared with mixed treatments. It is suggested that soluble Ca formed from the reaction of acid with naturally occurring lime may have reduced the availability of fertilizer P in the band

Exploring the nitrogen ingestion of aphids--a new method using electrical penetration graph and (15N labelling.

Directory of Open Access Journals (Sweden)

Franziska Kuhlmann

Full Text Available Studying plant-aphid interactions is challenging as aphid feeding is a complex process hidden in the plant tissue. Here we propose a combination of two well established methods to study nutrient acquisition by aphids focusing on the uptake of isotopically labelled nitrogen ((15N. We combined the Electrical Penetration Graph (EPG technique that allows detailed recording of aphid feeding behaviour and stable isotope ratio mass spectrometry (IRMS to precisely measure the uptake of nitrogen. Bird cherry-oat aphids Rhopalosiphum padi L. (Hemiptera, Aphididae fed for 24 h on barley plants (Hordeum vulgare L., cultivar Lina, Poaceae that were cultivated with a (15N enriched nutrient solution. The time aphids fed in the phloem was strongly positive correlated with their (15N uptake. All other single behavioural phases were not correlated with (15N enrichment in the aphids, which corroborates their classification as non-feeding EPG phases. In addition, phloem-feeding and (15N enrichment of aphids was divided into two groups. One group spent only short time in the phloem phase and was unsuccessful in nitrogen acquisition, while the other group displayed longer phloem-feeding phases and was successful in nitrogen acquisition. This suggests that several factors such as the right feeding site, time span of feeding and individual conditions play a role for the aphids to acquire nutrients successfully. The power of this combination of methods for studying plant-aphid interactions is discussed.

Ammonium absorption mechanism of rice seedling roots and 15N-labelling pattern of their glutamine-amide group, 2

International Nuclear Information System (INIS)

Arima, Yasuhiro; Kumazawa, Kikuo

1975-01-01

The processes of producing glutamine and asparagine at the initial stage of the absorption and assimilation of ammonia in rice seedling roots were examined in relation to glutamic acid, aspartic acid and ammonia by 15 N-labelling method. When ( 15 NH 4 ) 2 SO 4 was absorbed into the roots, 15 N concentration appeared very high in glutamine-amide radical and ammonia. It was also higher in amide radical than in amino radical in both glutamine and asparagine, while 15 N concentration in the amino radical of glutamine and asparagine were far lower than that of corresponding glutamine acid and aspartic acid. From these facts, glutamine-amide radical seems to be produced directly from the ammonia in culture media at the contact point of root cells and the culture media, while there is some possibility that asparagine-amide radical is formed from other amino compounds than ammonia. Also the amino radical of aspartic acid seems to be produced not only by the transamination from glutamic acid but also by the reductive amination of oxalautic acid by ammonium. (Kobatake, H.)

Natural 15N abundance of soil N pools and N2O reflect the nitrogen dynamics of forest soils

DEFF Research Database (Denmark)

Pörtl, K.; Zechmeister-Boltenstern, S.; Wanek, W.

2007-01-01

Natural N-15 abundance measurements of ecosystem nitrogen (N) pools and N-15 pool dilution assays of gross N transformation rates were applied to investigate the potential of delta N-15 signatures of soil N pools to reflect the dynamics in the forest soil N cycle. Intact soil cores were collected...

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.

Dynamics of the amino acid and protein metabolism of laying hens after the application of 15N-labelled wheat protein. 6

International Nuclear Information System (INIS)

Gruhn, K.; Kirchner, E.

1988-01-01

12 colostomized laying hens received, together with a conventional feed ration, 15 N-labelled wheat with a 15 N excess ( 15 N') of 14.37 atom-% over 4 days. 3 animals each were butchered after 12 h, 36 h, 60 h and 108 h after the last 15 N' application and, apart from various organs, the contents and the tissue of the gastrointestinal tract of each hen was divided into 3 fractions. TCA precipitation was carried out with the contents and the tissue of the 3 fractions. Nitrogen and its atom-% 15 N' were determined in the supernatant and the precipitate. The 15 N' amount in the contents of the crop and the stomachs, the small and large intestines is still considerable 12 h after the last 15 N wheat feeding and still clearly detectable 108 h after it. The TCA precipitable amounts of 14 N and 15 N' of the contents of crop and stomach and that of the small intestine agree well; they are 75% and 50% resp. of the total N. The amount of atom-% 15 N' of the contents of the small and large intestines remains the same up to 36 h after the last 15 N' application and is higher at the following measuring points in the contents of the large intestine. A close correlation could be ascertained between the atom-% 15 N' in the contents and tissue of the small and large intestines. The TCA soluble N quotas of both 14 N and 15 N' in the pancreas are above 50%. (author)

General method of preparation of uniformly 13C, 15N-labeled DNA fragments for NMR analysis of DNA structures

International Nuclear Information System (INIS)

Rene, Brigitte; Masliah, Gregoire; Zargarian, Loussine; Mauffret, Olivier; Fermandjian, Serge

2006-01-01

Summary 13 C, 15 N labeling of biomolecules allows easier assignments of NMR resonances and provides a larger number of NMR parameters, which greatly improves the quality of DNA structures. However, there is no general DNA-labeling procedure, like those employed for proteins and RNAs. Here, we describe a general and widely applicable approach designed for preparation of isotopically labeled DNA fragments that can be used for NMR studies. The procedure is based on the PCR amplification of oligonucleotides in the presence of labeled deoxynucleotides triphosphates. It allows great flexibility thanks to insertion of a short DNA sequence (linker) between two repeats of DNA sequence to study. Size and sequence of the linker are designed as to create restriction sites at the junctions with DNA of interest. DNA duplex with desired sequence and size is released upon enzymatic digestion of the PCR product. The suitability of the procedure is validated through the preparation of two biological relevant DNA fragments

Functional Single-Cell Approach to Probing Nitrogen-Fixing Bacteria in Soil Communities by Resonance Raman Spectroscopy with 15N2 Labeling.

Science.gov (United States)

Cui, Li; Yang, Kai; Li, Hong-Zhe; Zhang, Han; Su, Jian-Qiang; Paraskevaidi, Maria; Martin, Francis L; Ren, Bin; Zhu, Yong-Guan

2018-04-17

Nitrogen (N) fixation is the conversion of inert nitrogen gas (N 2 ) to bioavailable N essential for all forms of life. N 2 -fixing microorganisms (diazotrophs), which play a key role in global N cycling, remain largely obscure because a large majority are uncultured. Direct probing of active diazotrophs in the environment is still a major challenge. Herein, a novel culture-independent single-cell approach combining resonance Raman (RR) spectroscopy with 15 N 2 stable isotope probing (SIP) was developed to discern N 2 -fixing bacteria in a complex soil community. Strong RR signals of cytochrome c (Cyt c, frequently present in diverse N 2 -fixing bacteria), along with a marked 15 N 2 -induced Cyt c band shift, generated a highly distinguishable biomarker for N 2 fixation. 15 N 2 -induced shift was consistent well with 15 N abundance in cell determined by isotope ratio mass spectroscopy. By applying this biomarker and Raman imaging, N 2 -fixing bacteria in both artificial and complex soil communities were discerned and imaged at the single-cell level. The linear band shift of Cyt c versus 15 N 2 percentage allowed quantification of N 2 fixation extent of diverse soil bacteria. This single-cell approach will advance the exploration of hitherto uncultured diazotrophs in diverse ecosystems.

Effect of organic fertilization of soil, differentiated over many years, on the utilization of 15N-labelled urea

International Nuclear Information System (INIS)

Markgraf, G.; Winterfeld, C.

1983-01-01

Topsoil from samples with different organic fertilization of a soil fertility experiment started in 1936 at the Thyrow experimental site of the Berlin Humboldt University Crop Production Department was used for a pot experiment to study the N utilization of equal amounts of mineral fertilizer N (1.380 mg N/pot in the form of 15 N-labelled urea with an N frequency of 50 +- 0.5 atom per cent). The results showed that combined application of mineral NPK fertilization and high amounts of farmyard manure over a period of about 40 years on deep-loam fallow soil-sandy-rusty soil (diluvial sand to loamy-sandy soil) will lead to better utilization of the applied urea N as well as to higher N availability from the N pool of the soil. (author)

Metabolic studies in colostomized laying hens using 15N-labelled wheat. 4

International Nuclear Information System (INIS)

Gruhn, K.; Glotz, D.

1979-01-01

3 colostomized laying hybrids received over 4 days a dosage of 672 mg 15 N excess ( 15 N'), 20.3 mg lysine 15 N', 23.0 mg histidine 15 N' and 66.7 mg arginine 15 N' with a ration customary in production. After feeding the same unlabelled ration for another 4 days the hens were killed and the N content of the blood as well as of its fractions (cells, plasma, free amino acids of the plasma) was determined. The 15 N' was determined in the total blood, the corpuscles, the plasma, the nonprotein-N (NPN) fraction as well as in the amino acids lysine, histidine and arginine. The average amount of the blood cell N in the total blood N was 58.5% and that of the plasma 40.3%; the corresponding 15 N' values amounted to 66.1% and 33.9%, respectively. The sum of the 15 N' of the basic amino acids of the blood cells, on an average, amounted to 39.7% of the total cell 15 N'; the corresponding average value for the total 15 N' in lysine, histidine and arginine of the blood plasma 15 N' was 23.6.% and the quota of the three free amino acids of the total NP 15 N' of the plasma was 6.2%. (author)

The fate of fresh and stored 15N-labelled sheep urine and urea applied to a sandy and a sandy loam soil using different application strategies

DEFF Research Database (Denmark)

Sørensen, P.; Jensen, E.S.

1996-01-01

The fate of nitrogen from N-15-labelled sheep urine and urea applied to two soils was studied under field conditions. Labelled and stored urine equivalent to 204 kg N ha(-1) was either incorporated in soil or applied to the soil surface prior to sowing of Italian ryegrass (Lolium multiflorum L...... and soil was not significantly different for incorporated urine and urea. Almost all the supplied labelled N was accounted for in soil and herbage in the sandy loam soil, whereas 33-34% of the labelled N was unaccounted for in the sandy soil. When the stored urine was applied to the soil surface, 20...... was applied to growing ryegrass at the sandy loam soil, the immobilization of urine-derived N was significantly reduced compared to application prior to sowing. The results indicated that the net mineralization of urine N was similar to that of urea in the sandy soil, but only about 75% of the urine N was net...

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

Bioavailability of nitrogen from sewage sludge using 15N-labelled ammonium sulphate

International Nuclear Information System (INIS)

El-Motaium, R.A.

2001-01-01

The high nutrient nitrogen and organic matter contents of sewage sludge (SS) make it a potential organic fertilizer for sandy soil. In this study, 15 N-labelled ammonium sulphate was used to investigate the availability of nitrogen from irradiated and non-irradiated sewage sludge to tomato plants. The application of sewage sludge to sandy soil increased dry matter production (DMP), nitrogen yield (NY) and nitrogen recovery (NR) over two successive years. A positive relationship was found between sludge application rate and DMP and NY. The increase was significantly higher (P=0.05) in irradiated than non-irradiated sewage sludge. Total nitrogen derived from non-irradiated sewage sludge are : 48.0, 63.7, 73.5, 105.2 Kg/ha, whereas, the total nitrogen derived from irradiated sewage sludge are: 55.1, 72.5, 88.9, 141.4 Kg/ha corresponding to application rates of 10 t/ha, 20 t/ha, 30 t/ha, respectively. This was attributed to higher dry matter production in the later than the former. A highly significant correlation (0.945**) was found between dry matter production and sludge nitrogen yield (i.e. nitrogen derived from sewage sludge). Fertilizer nitrogen yield (total nitrogen derived from fertilizer) was high in treatment receiving mineral fertilizer, however, the 15 N recovery by tomato was only 13.8%. Soil did not contribute well towards total nitrogen yield in tomato and most nitrogen was derived from sewage sludge. Percent nitrogen derived from sewage sludge was in the range 88-92%, depending on the application rate

15N liver function tests - concept, validity, clinical use

International Nuclear Information System (INIS)

Faust, H.; Jung, K.; Krumbiegel, P.; Hirschberg, K.; Reinhardt, R.; Junghans, P.

1987-01-01

Several liver function tests using the oral application of a nitrogen compound labelled with 15 N and the subsequent determination of 15 N in a certain fraction of urine by emission spectrometry are described. Because of the key position of the liver in the metabolism of nitrogen compounds the results of these tests allow conclusions concerning disturbances of special liver functions. Instructions for the clinical use of the '[ 15 N]Ammonium Test', '[ 15 N]Hippurate Test' the '[ 15 N]Methacetin Test', and the '[ 15 N]Glycine Test' are given. (author)

Understanding the Fate of Applied Nitrogen in Pine Plantations of the Southeastern United States Using 15N Enriched Fertilizers

Directory of Open Access Journals (Sweden)

Jay E. Raymond

2016-11-01

Full Text Available This study was conducted to determine the efficacy of using enhanced efficiency fertilizer (EEFs products compared to urea to improve fertilizer nitrogen use efficiency (FNUE in forest plantations. All fertilizer treatments were labeled with 15N (0.5 atom percent and applied to 100 m2 circular plots at 12 loblolly pine stands (Pinus taeda L. across the southeastern United States. Total fertilizer N recovery for fertilizer treatments was determined by sampling all primary ecosystem components and using a mass balance calculation. Significantly more fertilizer N was recovered for all EEFs compared to urea, but there were generally no differences among EEFs. The total fertilizer N ecosystem recovery ranged from 81.9% to 84.2% for EEFs compared to 65.2% for urea. The largest amount of fertilizer N recovered for all treatments was in the loblolly pine trees (EEFs: 38.5%–49.9%, urea: 34.8% and soil (EEFs: 30.6%–38.8%, urea: 28.4%. This research indicates that a greater ecosystem fertilizer N recovery for EEFs compared to urea in southeastern pine plantations can potentially lead to increased FNUE in these systems.

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

International Nuclear Information System (INIS)

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

2012-01-01

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

Measuring the BNF of Soybean Using 15N-Labelled Urea with Different Atom Excess (A.E. Content

Directory of Open Access Journals (Sweden)

A. Citraresmini

2012-12-01

Full Text Available The soybean is a legume which has an ability to supply its major nitrogen need by the biological nitrogen fixation (BNF process. This process is made possible by nodules formed in their roots, colonized by Rhizobium sp.bacteria. An accurate estimation of N gained by BNF is necessary to predict the increase or decrease of chemical fertilizer-N requirements to increase soybean production. Among several methods, the 15N method was used to estimate the ability of legumes to perform BNF. The study involved soybean var. Willis (W and a completely non-BNF soybean var. CV, which is termed as a standard crop. The standard crop is non-nodulated soybean, but it has the same main physiological traits with var. Willis. The aim of this study was to determine whether15N-labelled fertilizer with different %a.e. given to nodulated and non-nodulated soybean would not be of significant consequences for the calculation of N-BNF of W. The treatments applied were different rates of urea (20 kg N/ha and 100 kg N/ha combined with different atom excess percentages (%a.e.15N (2% and 10%. Thus, the combination of treatments were as follows:(1 W-ll (20 kg N; 2% a.e; (2 CV-hl (100 kg N; 2% a.e; (3 W-lh (20 kg N; 10% a.e; (4 CV-hh (100 kg N; 10% a.e; (5 CV-ll (20 kg N; 2% a.e; (6 W-hl (100 kg N; 2% a.e; (7 CV-lh (20 kg N; 10% a.e; (8 W-hh (100 kg N; 10% a.e. The result of the experiment showed that a high %a.e. with a low rate of 15N and a low %a.e. with a high rate of N should be used to study the %N-BNF of nodulated plants.

Marcação de fitomassa de cana-de-açúcar com aplicação de solução de uréia marcada com15N Sugarcane phytomass labeling with application of 15N-urea solution

Directory of Open Access Journals (Sweden)

Carlos Eduardo Faroni

2007-06-01

Full Text Available O objetivo deste trabalho foi comparar três métodos de aplicação de solução de uréia marcada com15N (15N-uréia : pulverização foliar, injeção na base do colmo e imersão radicular, a fim de se definir qual seria o mais eficiente na marcação de fitomassa de cana-de-açúcar. O experimento foi instalado na Estação Experimental Apta - Pólo Regional Centro Sul, em Piracicaba, SP. A cana-de-açúcar, variedade SP80 3280, foi plantada em vasos preenchidos com aproximadamente 120 dm³ de Neossolo Quartzarênico de textura arenosa. O delineamento experimental foi inteiramente casualizado, com quatro repetições. A fitomassa de cana-de-açúcar dos três tratamentos, no 11º mês de desenvolvimento, não diferiu estatisticamente, e suas abundâncias de 15N foram superiores à natural, tendo-se verificado a seguinte ordem decrescente de marcação com 15N: parte aérea > rizoma > rizomas+raízes na camada de 0,0-0,2 m > raízes na camada de 0,2-0,4 m > raízes em profundidade maior que 0,4 m. Entre os métodos de aplicação de 15N-uréia, a injeção na base de colmos é o de mais fácil execução, o mais efetivo na marcação da fitomassa e o que apresentou a maior recuperação do traçador (96%. A aplicação foliar é comparável à injeção, somente na marcação e na recuperação do traçador no sistema radicular.The objective of this research was to compare three methods of 15N-urea solutions application: spray on leaf, injection in the plant base stem and root immersion, in order to define the most efficient labeling sugarcane phytomass with 15N method. The experiment was carried out at APTA - Pólo Regional Centro Sul, in Piracicaba, SP, Brazil, and the sugarcane variety SP80 3280 was planted in pots filled out with approximately 120 dm³ of a Typic Quartzipsamment soil. The experiment was conducted in a completely randomized design with four replicates. There were no difference between the methods in the plant parts dry

Nitrogen Uptake During Fall, Winter and Spring Differs Among Plant Functional Groups in a Subarctic Heath Ecosystem

DEFF Research Database (Denmark)

Larsen, Klaus Steenberg; Michelsen, Anders; Jonasson, Sven

2012-01-01

Nitrogen (N) is a critical resource for plant growth in tundra ecosystems, and species differences in the timing of N uptake may be an important feature regulating community composition and ecosystem productivity. We added 15N-labelled glycine to a subarctic heath tundra dominated by dwarf shrubs...... to 37 ± 7% by April indicating significant microbial N turnover prior to spring thaw. Only the evergreen dwarf shrubs showed active 15N acquisition before early May indicating that they had the highest potential of all functional groups for acquiring nutrients that became available in early spring....... The faster-growing deciduous shrubs did not resume 15N acquisition until after early May indicating that they relied more on nitrogen made available later during the spring/early summer. The graminoids and mosses had no significant increases in 15N tracer recovery or tissue 15N tracer concentrations after...

Precision analysis of 15N-labelled samples with the emission spectrometer NOI-5 for nitrogen balance in field trials

International Nuclear Information System (INIS)

Lippold, H.

1984-01-01

A technique was adapted for the preparation of samples with 15 N to be analyzed with the emission spectrometer NOI-5. This technique is based on methods of analyzing 15 N labelled gas samples in denitrification experiments. Nitrogen released from ammonium compounds by using hypobromite is injected into a repeatedly usable gaseous discharge tube where it is freed from water traces by means of the molecular sieve 5A. The described procedure of activating the molecular sieve allows to record spectra of reproducible quality thus promising an accuracy of analysis of +- 0.003 at% in the range of natural isotope frequency and the possibility of soil nitrogen analysis in field trials with fertilizers of low nitrogen content (3 to 6.5 at%; corresponding with 0.055 to 0.14% N/sub t/ of soils) without being dependent on mass spectrometers. (author)

Transformations of sup(15)N labelled urea applied to a soil with maize strain incorporation under continuous flood

International Nuclear Information System (INIS)

Colaco, W.

1991-01-01

A greenhouse experiment with deep placed sup(15)N-labelled urea was conducted in an Aluvial soil in which maize straw was incorporated. The transformations of total-N, NH sub(4) sup(+) -N, and NO sub(3) sup(-) -N, - were investigated under continuous flooded conditions in soil planted or not to rice. Total -N and inorganic-N levels decreased in both conditions. The rapid inorganic decrease in planted soil would be mainly due to assimilation by the rice plants. However losses occurred in all cases. Such losses increased in the absence of rice plants and seemed to be mainly to nitrification - denitrification mechanisms although NH sub(3) volatilization could also be involved. Dry matter yield and N-uptake by rice increased with increasing rate of N-application. Maize straw was an important source of N for rice. N losses are modified by the incorporation of maize straw. (author)

Balance study of the fate of 15N fertilizer

International Nuclear Information System (INIS)

Korte, F.; Sotiriou, N.

1980-01-01

An interim report is presented on a series of experiments with wooden box-type lysimeters (60 cm x 60 cm x 70 cm) loaded with a sandy soil, a loess soil and straw-amended soil. The lysimeters support crops rotated over a five-year period to be studied - potato, barley, sugar-beet, barley (with winter rape) and finally (1979) potato. Each lysimeter received split applications of urea at total rates of 0, 50 or 100 kg.ha -1 . The effects of soil residues of the herbicide monolinuron were also studied. The report deals with data collected during the first three years of the planned experiments (1975 - 1977 inclusive). 15 N-labelled urea (47 atom 15 N% excess) was initially used but in some experiments this was followed by applications of unlabelled urea in order to study the fate of the residual 15 N in the subsequent years. The results to date indicated that in the first year highest recoveries in the plant of the applied 15 N obtained on the sandy soil. The low recoveries of 15 N in the subsequent years when unlabelled urea was supplied also indicated significant storage by soil or root organic matter of the applied 15 N. Compared with the control (zero application of urea nitrogen), potato took up more total nitrogen in the presence of fertilizer including more of the unlabelled soil pool nitrogen. Analyses of the soil profiles in terms of total soil nitrogen and fertilizer-derived nitrogen (on the basis of 15 N assays) indicated leaching of the labelled nitrogen down the soil profile in all cases during the three-year period. Analysis of NO 3 -N in leachates confirmed the presence of labelled urea-derived nitrogen. (author)

EFFECT OF IRRIGATION WITH SEWAGE WASTEWATER ON NITROGEN UPTAKE AND TRANSLOCATION BY NAVEL ORANGE TRANSPLANTS USING 15N-LABELLED AMMONIUM SULPHATE

International Nuclear Information System (INIS)

EL-MOTAIUM, R.A.; SHARAF, A.N.; HASHEM, M.E.; HEGAZI, E.S.; BADAWY, S.H.

2009-01-01

An isotope aided study using 15 N-labelled ammonium sulphate was applied to investigate nitrogen uptake and translocation from sewage water by one year old Navel orange transplants. The investigation was conducted in a pot experiment at the greenhouse for two successive seasons (2005 and 2006). Ammonium sulphate was applied to the soil at a rate of 6 g/pot. The treatments included irrigation with canal water and sewage water. Half strength Hoagland nutrient solution, nitrogen depleted, was applied to provide the required nutrients for healthy growth of the transplants. The pots were irrigated to maintain the soil moisture content at field capacity.The results indicated that there is an increase in the transplants biomass irrigated with sewage water than using canal water. A substantial increase in mineral fertilizer nitrogen ( 15 N) uptake by plant roots and translocation into shoots was observed using canal water than sewage water. The nitrogen percent derived from sewage water was higher (26.1-49.5%) than that derived from 15 N-labelled ammonium sulphate (4.9-12.7%). Fertilizer nitrogen recovery was higher under canal water than under sewage water irrigation regime, although the plant biomass was higher under sewage water. Using sewage water alone can provide the orange transplants with their nitrogen requirements with no need for application of mineral fertilizer

Investigations on the transformation of N-fertilizer and the mineralization of organic N using 15N Pt. 1

International Nuclear Information System (INIS)

Latkovics, Gy.-ne

1979-01-01

A composting experiment was set up on chernozem-type brown forest soil to investigate the transformation of nitrogen fertilizer and the mineralization of organic N. For the average soil sample from the ploughed layer the pH value was 7.1, the mineral N content 2.85 mg, the fixed ammonium content 15.98 mg and the total N 140.8 mg100/g soil. The humus content was 1.91%. In the experiment 15 N labelled ammonium nitrate was used, and, as 15 N labelled organic matter, ground, air-dried rye-grass and bean stalks and with approximately the same N content as the 0.4% of the soil quantity measured. The values obtained by chemical methods and isotope indication show that the N-loss during composting was negligible and that the methods tested are suitable for the investigation of the transformation processes of nitrogen. (author)

Uniform 15N- and 15N/13C-labeling of proteins in mammalian cells and solution structure of the amino terminal fragment of u-PA

International Nuclear Information System (INIS)

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

1994-01-01

Urokinase-type plasminogen activator (u-PA) is a 54-kDa glycoprotein that catalyzes the conversion of plasminogen to plasmin, a broad-specificity protease responsible for the degradation of fibrin clots and extracellular matrix components. The u-PA protein consists of three individual modules: a growth factor domain (GFD), a kringle, and a serine protease domain. The amino terminal fragment (ATF) includes the GFD-responsible for u-PA binding to its receptor-and the kringle domains. This protein was expressed and uniformly 15 N-and 15 N/ 13 C-labeled in mammalian cells by methods that will be described. In addition, we present the three-dimensional structure of ATF that was derived from 1299 NOE-derived distance restraints along with the φ angle and hydrogen bonding restraints. Although the individual domains in the structures were highly converged, the two domains are structurally independent. The overall structures of the individual domains are very similar to the structures of homologous proteins. However, important structural differences between the growth factor domain of u-PA and other homologous proteins were observed in the region that has been implicated in binding the urokinase receptor. These results may explain, in part, why other growth factors show no appreciable affinity for the urokinase receptor

Metabolism-oriented amino acid requirement determination by means of the catabolic rates of 14C- and 15N-labelled lysine under maintenance

International Nuclear Information System (INIS)

Simon, O.; Bergner, H.; Adam, K.

1977-01-01

Male Wistar rats (of 60 g live weight) allotted in 10 groups were fed diets with gradually increasing lysine levels ranging from 1.4 to 7.4 g lysine/16 g N. Feed intake was restricted so much that the experimental animals did not change their live weights during the last 3 days of the 8-day experimental period. On the 7the experimental day, 4 animals of each group were injected, i. p. 14 C-L-lysine, the 14 CO 2 -excretion being subsequently measured over a period of 2 hours. On the next day, 6 animals of each group were applied an i. p. injection of 15 N-L-lysine, the urine being collected over the following 24-hour period to measure the 15 N-frequency. Applying both labelling methods, an increased catabolisation of the amino acid was observed after the metabolically necessary lysine requirement had been covered. The methods are very sensitive and revealed, under the experimental conditions chosen, a lysine requirement coverage of about 3 g lysine/16 g N. The possibility of using also 15 N-labelled compounds in the metabolism-oriented amino acid requirement determination is likely to facilitate the transfer of the methodology to farm animals would thus allow to study the amino acid requirement of man. The metabolism-oriented amino acid requirement determination will likewise allow to estimate exact amino acid requirement data under conditions that cannot be rated on the basis of productive yields. (author)

Metabolism of [15N]alanine in the ectomycorrhizal fungus Paxillus involutus

International Nuclear Information System (INIS)

Chalot, M.; Finlay, R.D.; Ek, H.; Söderström, B.

1995-01-01

Chalot, M., Finlay, R. D., Ek, H., and Söderström, B. 1995. Metabolism of [ 15 N]alanine in the ectomycorrhizal fungus Paxillus involutus. Experimental Mycology 19, 297-304. Alanine metabolism in the ectomycorrhizal fungus Paxillus involutus was investigated using [ 15 N]alanine. Short-term exposure of mycelial discs to [ 15 N]alanine showed that the greatest flow of 15 N was to glutamate and to aspartate. Levels of enrichment were as high as 15-20% for glutamate and 13-18% for aspartate, whereas that of alanine reached 30%. Label was also detected in the amino-N of glutamine and in serine and glycine, although at lower levels. Preincubation of mycelia with aminooxyacetate, an inhibitor of transamination reactions. resulted in complete inhibition of the flow of the label to glutamate, aspartate, and amino-N of glutamine, whereas [ 15 N]alanine rapidly accumulated. This evidence indicates the direct involvement of alanine aminotransferase for translocation of 15 N from alanine to glutamate. Alanine may be a convenient reservoir of both nitrogen and carbon. (author)

Regional assessment of N saturation using foliar and root δ15N

Science.gov (United States)

L.H. Pardo; P.H. Templer; C.L. Goodale; S. Duke; P.M. Groffman; M.B. Adams; P. Boeckx; J. Boggs; J. Campbell; B. Colman; J. Compton; B. Emmett; P. Gundersen; J. Kjonaas; G. Lovett; M. Mack; A. Magill; M. Mbila; M.J. Mitchell; G. McGee; S. McNulty; K. Nadelhoffer; S. Ollinger; D. Ross; H. Rueth; L. Rustad; P. Schaberg; S. Schiff; P. Schleppi; J. Spoelstra; W. Wessel

2006-01-01

N saturation induced by atmospheric N deposition can have serious consequences for forest health in many regions. In order to evaluate whether foliar δ15N may be a robust, regional-scale measure of the onset of N saturation in forest ecosystems, we assembled a large dataset on atmospheric N deposition, foliar and root δ

15N tracer techniques in pediatric research

International Nuclear Information System (INIS)

Heine, W.; Richter, I.; Plath, C.; Wutzke, K.; Stolpe, H.J.; Tiess, M.; Toewe, J.

1983-01-01

The main topics of the review comprise mathematical fundamentals of the determination of N metabolism parameters using the 3-pool method, the value of different 15 N tracer substances for the determination of whole-body protein parameters, the utilization of parenterally applied D-amino acids, studies on the influence of different diets on the N metabolism of premature infants with the 15 N tracer technique, the application of the 15 N-glycine-STH-test for the evaluation of the therapeutic effect of STH in children suffering from hypothalamico-hypophyseal dwarfism, in vivo studies on urea utilization by the infant intestinal flora under various dietary regimens as well as in vitro investigations on the utilization of 15 N-labelled urea and NH 4 Cl, resp., by the intestinal flora

Turnover of manure _¹⁵N-labelled ammonium during composting and soil application as affected by lime and superphosphate addition

DEFF Research Database (Denmark)

Tran, Tien Minh; Luxhøi, Jesper; Jensen, Lars Stoumann

2012-01-01

). The NH4-N pool in the pig manure was initially labeled with 15N to determine the fate of manure NH4-N during composting. The composts were subsequently applied to soil to investigate the effects on soil mineral N and to trace the 15N during 60 d of incubation at 25°C. Of the initial manure 15NH4-N......, approximately 30, 90, and 20% was lost by NH3 volatilization during composting in the Straw, Lime, and SSP treatments, respectively. Concurrently, 62, 16, and 41% of initial 15NH4-N was immobilized in the respective treatments. When the composts were applied to soil, the mineral N in soil with SSP compost...... or effectively in balance. In soil with Lime compost, net N immobilization was strong in the fi rst 10 d, but then net N mineralization dominated the remaining period of soil incubation. Overall, adding lime before composting reduced the NH4-N content in the compost and the amount available in soil, while adding...

Does labelling frequency affect N rhizodeposition assessment using the cotton-wick method?

DEFF Research Database (Denmark)

Mahieu, S.; Fustec, J.; Jensen, Erik Steen

2009-01-01

The aim of the present study was to test and improve the reliability of the 15N cotton-wick method for measuring soil N derived from plant rhizodeposition, a critical value for assessing belowground nitrogen input in field-grown legumes. The effects of the concentration of the 15N labelling...... solution and the feeding frequency on assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.). Neither the method nor the feeding frequency altered plant biomass and N partitioning, and the method appeared well adapted for assessing...... the belowground contribution of field-grown legumes to the soil N pool. However, nitrogen rhizodeposition assessment was strongly influenced by the feeding frequency and the concentration of labelling solution. At pod-filling and maturity, despite similar root 15N enrichment, the fraction of plants' belowground...

Passage and absorption of dietary and endogenous nitrogen in different regions of the digestive tract of rats given a single meal of 15N-labelled barley

International Nuclear Information System (INIS)

Partridge, I.G.; Simon, O.; Bergner, H.

1985-01-01

Young male Wistar rats (86.9 +- 0.96 g) were fasted for 24 hours and then offered a single meal (intake of 1 to 2.5 g) of 15 N-labelled barley (5.34 atom% 15 N excess). The test meal also contained Cr 2 O 3 (20 mg/g). Groups of five animals were killed 0.5; 1; 1.5; 2; 2.5; 4; 6 and 8 hours after removal of food. The contents of different regions of the digestive tract (stomach, proximal, middle and distal third of small intestine, large intestine) and feces were analyzed for Cr 2 O 3 and for N and 15 N abundance in both a TCA soluble and a TCA precipitable fraction. The distribution patterns of Cr 2 O 3 and 15 N along the digestive tract were very similar. If the disappearance of 15 N from the contents of the small and of the large intestines was expressed as a proportion of the gastric outflow of 15 N, a disappearance rate of 90% was found. On the basis of isotopic dilution the proportion of dietary nitrogen in digesta was calculated. The results illustrated the intensive dilution of dietary nitrogen by endogenous secretions in all regions of the digestive tract. In the distal small intestine endogenous nitrogen accounted for 70% of total nitrogen. 17 mg endogenous N were produced by the stomach within 8 hours after the single meal. The results show the value of the method in determining the true digestibility of nitrogen in 15 N-labelled feedstuffs more accurately than with classical methods and in providing an insight into the dynamics of nitrogen absorption and secretion in the digestive tract. (author)

Localization of 15N uptake in a Tibetan alpine Kobresia pasture

Science.gov (United States)

Schleuß, Per-Marten; Kuzyakov, Yakov

2014-05-01

The Kobresia Pygmea ecotone covers approximately 450.000 km2 and is of large global and regional importance due several socio-ecological aspects. For instance Kobresia pastures store high amounts of carbon, nitrogen and other nutrients, represent large grazing areas for herbivores, provide a fast regrowth after grazing events and protect against mechanical degradation and soil erosion. However, Kobresia pastures are assumed to be a grazing induced and are accompanied with distinct root mats varying in thickness between 5-30 cm. Yet, less is known about the morphology and the functions of this root mats, especially in the background of a progressing degradation due to changes of climate and management. Thus we aimed to identify the importance of single soil layers for plant nutrition. Accordingly, nitrogen uptake from different soil depths and its remain in above-ground biomass (AGB), belowground biomass (BGB) and soil were determined by using a 15N pulse labeling approach during the vegetation period in summer 2012. 15N urea was injected into six different soil depths (0.5 cm, 2.5 cm, 7.5 cm, 12.5 cm, 17.5 cm, 22.5 cm / for each 4 replicates) and plots were sampled 45 days after the labeling. For soil and BGB samples were taken in strict sample intervals of 0-1 cm, 1-5 cm, 5-10 cm, 10-15 cm, 15-20 cm, 20-25 cm. Results indicate that total recovery (including AGB, BGB and soil) was highest, if tracer was injected into the top 5 cm and subsequently decreased with decreasing injection depth. This is especially the case for the 15N recovery of BGB, which is clearly attributed to the root density and strongly decreased with soil depth. In contrast, the root activity derived from the 15N content of roots increased with soil depth, which is primary associated to a proportionate increase of living roots related to dead roots. However, most 15N was captured in plant biomass (67.5-85.3 % of total recovery), indicating high 15N uptake efficiency possibly due to N limitation

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

International Nuclear Information System (INIS)

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

2011-01-01

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

Adsorption, translocation and redistribution of nitrogen (15N) in orange trees

International Nuclear Information System (INIS)

Fenilli, Tatiele Anete Bergamo; Boaretto, Antonio Enedi Boaretto; Bendassolli, Jose Albertino; Trivelin, Paulo Cesar Ocheuze; Muraoka, Takashi

2002-01-01

The objective was to evaluate the absorption of 15 N from nutrient solution by young orange trees and the translocation and the redistribution of the absorbed N. The treatments were constituted by four periods of 15 N labelling (spring, summer, autumn and winter). In the first treatment, the young orange trees received 15 N in the nutrient solution during the spring and five replicates of the plants were picked at the end of the period. The new part, which was developed during the 15 N labelling period, was separated from the other part (old part) in branch and leaf, and also in flower and fruit when they were. The old part was separated in leaf, stem and root. This same procedure was followed in the other treatments. The total N and the isotope ratios 15 N/ 14 N were performed by mass spectrometry. The major part of absorbed N during the spring and summer was translocated to the new part of the orange trees, but in autumn and winter the absorbed N was concentrated in the old plant part. The redistribution of N from of old plant parts was more intensive during the autumn and winter. (author)

Applications of stable isotopes of 2H, 13C and 15N to clinical problems

International Nuclear Information System (INIS)

Klein, P.D.; Szczepanik, P.A.; Hachey, D.L.

1974-01-01

The function of the Argonne Program is to provide synthetic, analytical instrumental capability in a core facility for the clinical investigator who needs to use 2 H, 13 C, or 15 N labelled compounds for metabolic or clinical research on pregnant women, newborn infants, young children, or for mass screening. To carry out such application development, there were six stages which were recurrent steps in every application. Five fundamental strategies should be adopted to establish the use of stable isotopes in clinical work. The instrument required for measurements was a combined gas chromatograph-mass spectrometer, and its use was schematically illustrated. Some of the successful experiences with compounds labelled by stable isotopes, such as deuterium labelled chenodeoxycholic acid, and respective 13 C and 15 N-labelled glycine were described. Deutrium labelled bile acid enabled easy and safe determination of the size of the bile acid pool and the replacement rate, providing clearer diagnoses for cholestatic liver disease and gallstones. 13 C and 15 N labelled compounds were used in clinical studies, of children with genetic disorders of amino acid metabolism, i.e., non ketotic hyperflycinemia, B 12 -responsive methyl malonic acidemia, and Lesch-Nyhan syndrome. 15 N-labelled glycine was also studied in a child with Lesch-Nyhan syndrome. (Mukohata, S.)

Distribution of complemented 15N - (NH4)2SO4 in an ethanolic fermentation process on insolube-N and solube-N fractions

International Nuclear Information System (INIS)

Lara Cabejas, W.A.R.; Trivelin, P.C.O.

1990-01-01

Looking for stillage labeling with 15 N for further utilization in studies of mineral fertilization of sugar-cane, 15 N-(NH 4 ) 2 SO 4 (43.5ppm, 45.401 atoms% 15 N) was supplemented in a single fermentative cycle, in a laboratory scale. A nitrogen fractionation was made between insoluble-N and soluble-N in several componentes of the fermentative process (yeast, sugar-cane juice, centrifugate wine, centrifugate yeast and stillage) with the objective of studying the added nitrogen distribution and its isotopic abundance composition. The nitrogen fractionation, and the isotopic analysis by mass spectrometry of 15 N, in the fractions of the several components of the fermentative process, showed 81.1% of N recovery, being 3.2% in stillage and mainly in a soluble-N fraction (71.4%), and the rest found in centrifugate yeast (77.9%), distributed mainly in a insoluble-N fraction (92.0%). Desuniform isotopic label was found in stillage, between soluble-N (1.333 atoms% 15 N) and insoluble-N fractions (0.744 atoms% 15 N). Means to improve the isotopic uniformity in these fractions is discussed. (autor) [pt

Changes in mineral 15 N from soils treated with 15 N-urea and 15 N-vinasse incorporated or not to sugar cane straw

International Nuclear Information System (INIS)

Silva, Vilma M.; Colaco, Waldeciro; Encarnacao, Fernando A.F.

1999-01-01

Changes in N derived from 15 N sources (urea and vinasse), applied to two soils differing in texture (PV sandy, LR clayey), incorporated or not to sugar cane straw (dry leaves and sheathes) and incubated in an open system for 35 days, were evaluated through an isotope technique. Soil samples were collected 7, 14, 21, 28 and 35 days after applications to determine nitrogen fractions (total-N, N H 4 + - N and NO 3 - - N) derived from the labelled sources. Mineral N was taken as the sum of N H 4 + - N and N H 3 - -N. 15 N-abundances were determined in the concentrated extracts of these fractions. The mineral N net transformation rates were found from the mineral N obtained by taking the difference between the values of two subsequent incubation times. The results showed that mineral N transformation rates were initially positives in the treatments of 15 N-urea, and significantly higher (10,30 mg kg -1 d -1 , PV and 8,08 mg kg -1 d -1 , LR) than those obtained in the treatments with 15 N vinasse (1,11 mg kg -1 dia -1 , PV and 0,55 mg kg -1 dia -1 , LR). In general terms, mineral-N net transformation rates were negative (0,06 and 0,26 mg.kg -1 d -1 , PV; -1,44 and 0,07 mg.kg -1 .d -1 , LR, respective;y for urea and vinasse) indicating prevalence of immobilization. The results also showed small fluctuations among treatments at some of the incubation periods, which reflects the influence of characteristics and properties of both soils. (author)

Dynamic effects of soil bulk density on denitrification and mineralisation by 15N labelled lettuce residue and paper wastes

International Nuclear Information System (INIS)

Hua Luo; Cheng Qing; Vinten, A.J.A.

1997-10-01

Two laboratory incubation experiments aimed to study the denitrification and mineralisation influenced by different additives ( 15 N labelled lettuce residue, paper wastes and mixture of both) and soil bulk densities were carried out by means of acetylene inhibition at the constant 15 degree C for 107 and 90 days, respectively. The results showed that the changes of N 2 O, CO 2 emission rates, inorganic nitrogen (NO 3 - and NH 4 + ), total N and 15 N abundance in the soils which were affected by adding lettuce residue, paper wastes and mixture of both were investigated. Soil denitrification rate increased after lettuce residue was added into soil for 8 days. The maximum rate of N 2 O emission was 15 times higher than that in soil without any additive. However, paper wastes did not increase N 2 O emission in the first 8 days compared with other treatments, mixed residue and paper wastes could promote soil microbial activity, but N 2 O emission was lower than that in the soil with lettuce residue added and higher than that with paper wastes, indicating that mixture of residue and paper wastes was benefit to soil nitrogen immobilisation. CO 2 emission in all the treatments were declined to the same level on the 107 th day. In the treatment added mixed residues and paper wastes, the released CO 2 quantities were higher than those in other treatments every day. Effect of different bulk density on N 2 O and CO 2 emission were response to the change of bulk density, it seems that N 2 O and CO 2 emission increased with bulk density. High bulk density could affect decomposition of paper wastes and NO 3 - , NH 4 + concentration. (30 ref., 10 tabs.)

15N tracer kinetic studies on the validity of various 15N tracer substances for determining whole-body protein parameters in very small preterm infants

International Nuclear Information System (INIS)

Plath, C.; Heine, W.; Wutzke, K.D.; Krienke, L.; Toewe, J.M.; Massute, G.; Windischmann, C.

1987-01-01

Reliable 15 N tracer substances for tracer kinetic determination of whole-body protein parameters in very small preterm infants are still a matter of intensive research, especially after some doubts have been raised about the validity of [ 15 N]glycine, a commonly used 15 N tracer. Protein turnover, synthesis, breakdown, and further protein metabolism data were determined by a paired comparison in four preterm infants. Their post-conceptual age was 32.2 +/- 0.8 weeks, and their body weight was 1670 +/- 181 g. Tracer substances applied in this study were a [ 15 N]amino acid mixture (Ia) and [ 15 N]glycine (Ib). In a second group of three infants with a post conceptual age of 15 N-labeled 32.0 +/- 1.0 weeks and a body weight of 1,907 +/- 137 g, yeast protein hydrolysate (II) was used as a tracer substance. A three-pool model was employed for the analysis of the data. This model takes into account renal and fecal 15 N losses after a single 15 N pulse. Protein turnovers were as follows: 11.9 +/- 3.1 g kg-1 d-1 (Ia), 16.2 +/- 2.5 g kg-1 d-1 (Ib), and 10.8 +/- 3.0 g kg-1 d-1 (II). We were able to demonstrate an overestimation of the protein turnover when Ib was used. There was an expected correspondence in the results obtained from Ia and II. The 15 N-labeled yeast protein hydrolysate is a relatively cheap tracer that allows reliable determination of whole-body protein parameters in very small preterm infants

Endogenous and exogenous control of ecosystem function: N cycling in headwater streams

OpenAIRE

Valett, H. M.; Thomas, S. A.; Mulholland, P. J.; Webster, J. R.; Dahm, C. N.; Fellows, C. S.; Crenshaw, C. L.; Peterson, C. G.

2008-01-01

Allochthonous inputs act as resource subsidies to many ecosystems, where they exert strong influences on metabolism and material cycling. At the same time, metabolic theory proposes endogenous thermal control independent of resource supply. To address the relative importance of exogenous and endogenous influences, we quantified spatial and temporal variation in ecosystem metabolism and nitrogen (N) uptake using seasonal releases of (15)N as nitrate in six streams differing in riparian-stream ...

Application time of nitrogen fertilizer 15N by a potato crop (Solanum Tuberosum L.)

International Nuclear Information System (INIS)

Bastidas, O.G.; Urquiaga, S.

1987-01-01

This study was performed at the ''San Jorge'' experimental farm of the Instituto Colombiano Agropecuario (ICA), Bogota, Colombia. The study was performed to investigate the effect of timing of application of nitrogen fertilizer on the productivity of, and the efficiency of utilization of 15 N-labelled fertilizer by, a potato crop (Solanum tuberosum L.), cv. Tequendama. The crop was fertilized with 100, 200 and 100 Kg/ha -1 of N, P 2 O 5 and K 2 O respectively. The N fertilizers were either added as 15 N labelled urea (2.955 at.% 15 N excess) or as labelled ammonium sulphate (2.071 at.% 15 N excess). In all treatments with nitrogen, a total of 100 Kg N ha -1 was added, but the nitrogen was added either in two or three split doses (only one dose being labelled with 15 N) at the following times: at planting, 35 days after emergence (DAE) and/or 60 DAE. It was found that: a) Nitrogen fertilization increased tuber production from 24 to 43 t/ha -1 ; b) The tubers constituted approximately 80% of total plant dry matter and 70% of the total nitrogen and fertilizer N accumulated by the plant; c) The fertilizer use efficiency varied between 49 and 68%, and the highest efficiency occurred when the nitrogen was split in three doses; d) The urea and ammonium sulphate gave similar results in all parameters evaluated; e) When the total nitrogen difference method was applied to interpretation of the results the fertilizer use efficiency was overestimated by 15 to 30%

A system for oxygen-15 labeled blood for medical applications

International Nuclear Information System (INIS)

Subramanyam, R.; Bucelewicz, W.M.; Hoop, B. Jr.; Jones, S.C.

1977-01-01

Oxygen-15 labeled compounds in blood have been used successfully for cerebral circulation and cerebral oxygen metabolism measurements. The present paper describes a system for the rapid sequential production of 15 O-HgB, C 15 O-Hgb and H 2 15 O in blood under sterile and pyrogen-free conditions. A tonometer has been adopted for labeling blood without hemolysis and foam production. (author)

Crop uptake and leaching losses of 15N labelled fertilizer nitrogen in relation to waterlogging of clay and sandy loam soils

International Nuclear Information System (INIS)

Webster, C.P.; Belford, R.K.; Cannell, R.Q.

1986-01-01

Ammonium nitrate fertilizer, labelled with 15 N, was applied in spring to winter wheat growing in undisturbed monoliths of clay and sandy loam soil in lysimeters; the rates of application were respectively 95 and 102 kg N ha -1 in the spring of 1976 and 1975. Crops of winter wheat, oilseed rape, peas and barley grown in the following 5 or 6 years were treated with unlabelled nitrogen fertilizer at rates recommended for maximum yields. During each year of the experiments the lysimeters were divided into treatments which were either freely drained or subjected to periods of waterlogging. Another labelled nitrogen application was made in 1980 to a separate group of lysimeters with a clay soil and a winter wheat crop to study further the uptake of nitrogen fertilizer in relation to waterlogging. In the first growing season, shoots of the winter wheater at harvest contained 46 and 58% of the fertilizer nitrogen applied to the clay and sandy loam soils respectively. In the following year the crops contained a further 1-2% of the labelled fertilizer, and after 5 and 6 years the total recoveries of labelled fertilizer in the crops were 49 and 62% on the clay and sandy loam soils respectively. In the first winter after the labelled fertilizer was applied, less than 1% of the fertilizer was lost in the drainage water, and only about 2% of the total nitrogen (mainly nitrate) in the drainage water from both soils was derived from the fertilizer

Liver function tests using the stable istope 15N

International Nuclear Information System (INIS)

Faust, H.; Jung, K.; Hirschberg, K.; Krumbiegel, P.; Junghans, P.; Reinhardt, R.; Teichmann, B.

1988-01-01

Several liver function tests using oral application of a nitrogen compound labelled with 15 N and the subsequent determination of 15 N in a certain fraction of urine or in the total urine by emission spectrometry are described. Because of the key function of the liver in the metabolism of nitrogen compounds, the results of these tests allow conclusions concerning some disturbances of liver functions. (author)

Ecosystem partitioning of 15N-glycine after long-term climate and nutrient manipulations, plant clipping and addition of labile carbon in a subarctic heath tundra

DEFF Research Database (Denmark)

Sørensen, Pernille Lærkedal; Michelsen, Anders; Jonasson, Sven Evert

2008-01-01

of nitrogen (N). Here, we studied 15N label incorporation into microbes, plants and soil N pools after both long-term (12 years) climate manipulation and nutrient addition, plant clipping and a pulse-addition of labile C to the soil, in order to gain information on interactions among soil N and C pools...... addition. However, plants exerted control on the soil inorganic N concentrations and recovery of total dissolved 15N (TD15N), and likewise the microbes reduced these soil pools, but only when fed with labile C. Soil microbes in clipped plots were primarily C limited, and the findings of reduced N...... availability, both in the presence of plants and with the combined treatment of plant clipping and addition of sugar, suggest that the plant control of soil N pools was not solely due to plant uptake of soil N, but also partially caused by plants feeding labile C to the soil microbes, which enhanced...

Methods for sequential resonance assignment in solid, uniformly 13C, 15N labelled peptides: Quantification and application to antamanide

International Nuclear Information System (INIS)

Detken, Andreas; Hardy, Edme H.; Ernst, Matthias; Kainosho, Masatsune; Kawakami, Toru; Aimoto, Saburo; Meier, Beat H.

2001-01-01

The application of adiabatic polarization-transfer experiments to resonance assignment in solid, uniformly 13 C- 15 N-labelled polypeptides is demonstrated for the cyclic decapeptide antamanide. A homonuclear correlation experiment employing the DREAM sequence for adiabatic dipolar transfer yields a complete assignment of the C α and aliphatic side-chain 13 C resonances to amino acid types. The same information can be obtained from a TOBSY experiment using the recently introduced P9 1 12 TOBSY sequence, which employs the J couplings as a transfer mechanism. A comparison of the two methods is presented. Except for some aromatic phenylalanine resonances, a complete sequence-specific assignment of the 13 C and 15 N resonances in antamanide is achieved by a series of selective or broadband adiabatic triple-resonance experiments. Heteronuclear transfer by adiabatic-passage Hartmann-Hahn cross polarization is combined with adiabatic homonuclear transfer by the DREAM and rotational-resonance tickling sequences into two- and three-dimensional experiments. The performance of these experiments is evaluated quantitatively

Use of Bio-Organic Fertilizers to Develop N Uptake Using 15N Technique

International Nuclear Information System (INIS)

Galal, Y.G.M.

2008-01-01

Experimental work either in field scale or in green house conditions were conducted using 15 N technique to evaluate the role of different bio fertilizers and different plant residues as organic amendments on enhancement of plant N nutrition. Nitrogen fixation by a symbiotic bacteria has been observed in greenhouse and field experiments under dry land cropping systems. Biological N 2 fixation associated with crop residues (legumes or cereals) was investigated in pot experiments with wheat and chickpea cultivars. In these experiments, labelled wheat and rice straw were used as organic N sources in comparison with either 15 N-labelled ammonium sulfate or ammonium nitrate as chemical nitrogen fertilizers. Rhizobium inoculation extended to be used with wheat gave the best results of N uptake and N 2 fixation when combined with Azospirillum brasilense as heterotrophic diazotrophs. The nitrogen uptake by wheat plants was significantly increased by application of soybean residues and inoculation with Azospirillum brasilense. From the field trial we can conclude that soybean residue as enriched N material, and Azospirillum brasilense inoculation enhanced N yields of wheat cultivars grown in poor fertile sandy soil

NMR experiments for resonance assignments of 13C, 15N doubly-labeled flexible polypeptides: Application to the human prion protein hPrP(23-230)

International Nuclear Information System (INIS)

Liu Aizhuo; Riek, Roland; Wider, Gerhard; Schroetter, Christine von; Zahn, Ralph; Wuethrich, Kurt

2000-01-01

A combination of three heteronuclear three-dimensional NMR experiments tailored for sequential resonance assignments in uniformly 15 N, 13 C-labeled flexible polypeptide chains is described. The 3D (H)N(CO-TOCSY)NH, 3D (H)CA(CO-TOCSY)NH and 3D (H)CBCA(CO-TOCSY)NH schemes make use of the favorable 15 N chemical shift dispersion in unfolded polypeptides, exploit the slow transverse 15 N relaxation rates of unfolded polypeptides in high resolution constant-time [ 1 H, 15 N]-correlation experiments, and use carbonyl carbon homonuclear isotropic mixing to transfer magnetization sequentially along the amino acid sequence. Practical applications are demonstrated with the 100-residue flexible tail of the recombinant human prion protein, making use of spectral resolution up to 0.6 Hz in the 15 N dimension, simultaneous correlation with the two adjacent amino acid residues to overcome problems associated with spectral overlap, and the potential of the presently described experiments to establish nearest-neighbor correlations across proline residues in the amino acid sequence

Estimate of production of gaseous nitrogen in the human body based on (15)N analysis of breath N2 after administration of [(15)N2]urea.

Science.gov (United States)

Junghans, Peter

2013-01-01

After oral administration of [(15)N2]urea (1.5 mmol, 95 atom% (15)N), we found that breath N2 was significantly (15)N-labelled. The result suggests that molecular nitrogen in breath must be partly produced endogenously. Based on a metabolic model, the endogenous N2 production was estimated to be 0.40±0.25 mmol kg(-1) d(-1) or 2.9±1.8 % of the total (urinary and faecal) N excretion in fasted healthy subjects (n=4). In patients infected with Helicobacter pylori (n=5), the endogenous N2 production was increased to 1.24±0.59 mmol kg(-1) d(-1) or 9.0±4.3 % of the total N excretion compared to the healthy controls (pexchange measurements may be affected by endogenously produced nitrogen, especially in metabolic situations with elevated nitrosation, for instance in oxidative and nitrosative stress-related diseases such as H. pylori infections.

Fate of 15N applied as ammonium sulphate to a bean crop

International Nuclear Information System (INIS)

Cervellini, A.; Ruschel, A.P.; Matsui, E.

1980-01-01

An instrumented 10 X 10-m site was used to study the fate of 15 N-labelled ammonium sulphate (3.289 atom 15 N % excess) applied at the rate of 100 kg N.ha -1 with P and K supplements. Data were collected for the year January through December, 1977. The first bean crop (Phaseolus vulgaris, L) was planted in March, followed by the fertilizer, and harvested three months later. The plot was left fallow for three months when a second bean crop was planted without further fertilizer. The data indicated that the total drainage (22 cm) represented approximately 16% of the total water input (precipitation plus irrigation). It was estimated that nitrate leaching below the 120-cm depth was equivalent to approximately 16 kg N.ha -1 of which less than 10% was derived from the labelled fertilizer. The low recovery of labelled fertilizer (30%) by the first total harvest of straw and grain, and the less than 2.5% further recovery of initial labelled fertilizer by the second crop, suggest high immobilization by the soil-plant residue system. (author)

Uniform {sup 15}N- and {sup 15}N/{sup 13}C-labeling of proteins in mammalian cells and solution structure of the amino terminal fragment of u-PA

Energy Technology Data Exchange (ETDEWEB)

Hansen, A.P.; Petros, A.M.; Meadows, R.P.; Mazar, A.P.; Nettesheim, D.G.; Pederson, T.M.; Fesik, S.W. [Abbott Laboratories, Abbott Park, IL (United States)

1994-12-01

Urokinase-type plasminogen activator (u-PA) is a 54-kDa glycoprotein that catalyzes the conversion of plasminogen to plasmin, a broad-specificity protease responsible for the degradation of fibrin clots and extracellular matrix components. The u-PA protein consists of three individual modules: a growth factor domain (GFD), a kringle, and a serine protease domain. The amino terminal fragment (ATF) includes the GFD-responsible for u-PA binding to its receptor-and the kringle domains. This protein was expressed and uniformly {sup 15}N-and {sup 15}N/{sup 13}C-labeled in mammalian cells by methods that will be described. In addition, we present the three-dimensional structure of ATF that was derived from 1299 NOE-derived distance restraints along with the {phi} angle and hydrogen bonding restraints. Although the individual domains in the structures were highly converged, the two domains are structurally independent. The overall structures of the individual domains are very similar to the structures of homologous proteins. However, important structural differences between the growth factor domain of u-PA and other homologous proteins were observed in the region that has been implicated in binding the urokinase receptor. These results may explain, in part, why other growth factors show no appreciable affinity for the urokinase receptor.

Estimating legume N-2 fixation in grass-clover mixtures of a grazed organic cropping system using two N-15 methods

DEFF Research Database (Denmark)

Vinther, F.P.; Jensen, E.S.

2000-01-01

The input of Nitrogen (N) through symbiotic N-2 fixation (SNF) in grass-clover mixtures was determined in an organic cropping. system for grazing during 3 years. The mixture of perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) was established by undersowing in spring...... barley (Hordeum vulgare L.) and maintained subsequently for two production years. Dinitrogen fixation was determined using the N-15 isotope dilution techniques and two labelling procedures. Using either pre-labelling of the soil with immobilisation of the N-15 by addition of a carbon source before...

Fate of leaf-litter N in forest and grassland along a pedo-climatic gradient in south-western Siberia: an in situ 15N-labelling experiment

Science.gov (United States)

Brédoire, Félix; Zeller, Bernd; Nikitich, Polina; Barsukov, Pavel A.; Rusalimova, Olga; Bakker, Mark R.; Legout, Arnaud; Bashuk, Alexander; Kayler, Zachary E.; Derrien, Delphine

2017-04-01

The suitability of Siberia for agriculture is expected to increase in the next decades due to strong and rapid climatic changes, but little is known on the environmental drivers of soil fertility there, especially nitrogen (N). Plant-available N is mainly derived from litter decomposition. South-western (SW) Siberia is located on the transition between several bioclimatic zones that are predicted to shift and extend along with climate change (steppe, forest-steppe, sub-taiga). The soils of this region are formed on a common loess deposit but they are submitted to different climatic conditions and vegetation cover. In the south of the region, typically in steppe/forest-steppe, soil freezes over winter because of a relatively shallow snow-pack, and water shortages are frequent in summer. In the north, typically in sub-taiga, the soil is barely frozen in winter due to a thick snow-pack and sufficient soil moisture in summer. In this study, we characterized the dynamics of leaf litter decomposition and the transfer of N from leaf litter to the soil and back to plants. Four sites were chosen along a climate gradient (temperature, precipitation and snow depth). At each site, we applied 15N-labelled leaf litter on the soil surface in experimental plots in an aspen (Populus tremula L.) forest and in a grassland. Twice a year during three years, we tracked the 15N derived from the decomposing labelled-litter in the organic layers, in the first 15 cm of the soil, and in above-ground vegetation. Soil temperature and moisture were monitored at a daily timestep over three years and soil water budgets were simulated (BILJOU model, Granier et al. 1999). We observed contrasting patterns in the fate of litter-derived 15N between bioclimatic zones. Over three years, along with faster decay rates, the release of leaf litter-N was faster in sub-taiga than in forest-steppe. As such, higher quantities of 15N were transferred into the soil in sub-taiga. The transfer was also deeper there

The fate of ¹⁵N-labelled organic nitrogen in sheep manure applied to soils of different texture under field conditions

DEFF Research Database (Denmark)

Sørensen, P.; Jensen, E.S.; Nielsen, N.E.

1994-01-01

The fate of nitrogen from N-15-labelled sheep manure and ammonium sulfate in small lysimeters and plots in the field was studied during two growth seasons. In April 1991, N-15-labelled sheep faeces (87 kg N ha(-1)) plus unlabelled (NH4)(2)SO4 (90 kg N ha(-1)), and ((NH4)-N-15)(2)SO4 (90 kg N ha(-1......-17% of the labelled manure N and 56% of the labelled (NH4)(2)SO4-N. After 18 months 30% of the labelled manure N and 65% of the labelled (NH4)(2)SO4-N were accumulated in barley, the succeeding ryegrass crop and in leachate collected below 45 cm of soil, irrespective of the soil-sand mixture. Calculating the barley...

Effect of the supply dose on the 15N enrichment level of cow's milk nitrogenous fractions

International Nuclear Information System (INIS)

Colin, O.; Laurent, F.; Vignon, B.; Antoine, J.M.

1994-01-01

Production of cow milk 15 N-labelled proteins is necessary for the study of their digestion by man. An adequate enrichment is required for compatibility with utilization constraints (application dose, studied fractions...). A test was conducted with five cows in order to optimize the utilization of labelled ammonium sulphate in the cow diet for 15 N enrichment of the milk nitrogenous matter. Doses and supply timing of labelled compounds are discussed. 3 figs., 3 refs

Production of mungbean under reclaimed sandy soil and irrigation levels using N-15 labeled

International Nuclear Information System (INIS)

Abdallah, A.A.G.; Thabet, E.M.A.

2000-01-01

Field experiment were performed at the Atomic Energy Authority, Experimental farm, Inshas, Egypt. During 1998 growing season. In tafla and sand mixture soil (1:7 Wt./Wt.). The treatments were laid out using a single line source sprinkler irrigation system which allows a gradual variation of irrigation from excess to little irrigation in which the calculated amount of irrigation water levels were 2241, 1562 and 1093 m 3 / feddan (W 1 , W 2 and W 3 ). The obtained results indicated that, there was a clear relationship between adequate amount of irrigation water and both total seed yield and total green pods/plot as well as there were significant increase in both characters due to irrigation W 1 and W 2 compared with W 3 . The results also indicated that W 2 irrigation level could be used in irrigation under the same conditions. Water use efficiency was significantly increased with middle irrigation level than with other two irrigation treatments. The result concerned fertilizer use efficiency using N 15 labeled fertilizers and total seed protein content were increased with decreasing irrigation level

NMR studies on 15N-labeled creatine (CR), creatinine (CRN), phosphocreatine (PCR), and phosphocreatinine (PCRN), and on barriers to rotation in creatine kinase-bound creatine in the enzymatic reaction

International Nuclear Information System (INIS)

Kenyon, G.L.; Reddick, R.E.

1986-01-01

Recently, the authors have synthesized 15 N-2-Cr, 15 N-3-Crn, 15 N-2-Crn, 15 N-3-PCrn, 15 N-3-PCr, and 15 N-2-PCr. 1 H, 15 N, 31 P NMR data show that Crn protonates exclusively at the non-methylated ring nitrogen, confirm that PCrn is phosphorylated at the exocyclic nitrogen, and demonstrate that the 31 P- 15 N one-bond coupling constant in 15 N-3-PCr is 18 Hz, not 3 Hz as previously reported by Brindle, K.M., Porteous, R. and Radda, G.K.. The authors have found that creatine kinase is capable of catalyzing the 14 N/ 15 N positional isotope exchange of 3- 15 N-PCr in the presence of MgADP, but not in its absence. Further, the exchange does not take place when labeled PCr is resynthesized exclusively from the ternary complex E X Cr X MgATP as opposed to either E X Cr or free Cr. This suggests that the enzyme both imparts an additional rotational barrier to creatine in the complex and catalyzes the transfer of phosphoryl group with essentially complete regiospecificity

Nitrogen dynamics in stream wood samples incubated with [14C]lignocellulose and potassium[15N]nitrate

International Nuclear Information System (INIS)

Aumen, N.G.; Bottomley, P.J.; Gregory, S.V.

1985-01-01

Surface wood samples obtained from a Douglas fir log incubated in vitro with [ 14 C]lignocellulose in a defined mineral salts medium supplemented with 10 mg of N liter -1 of 15 N-labeled NO 3 - (50 atom % 15 N). Evolution of 14 CO 2 , distribution and isotopic dilution of 15 N, filtrate N concentrations, and the rates of denitrification, N 2 fixation, and respiration were measured at 6, 12, and 18 days of incubation. The organic N content of the lignocellulose-wood sample mixture had increased from 132 μg of N to a maximum of 231 μg of N per treatment after 6 days of incubation. Rates of [ 14 C]lignocellulose decomposition were greatest during the first 6 days and then began to decline over the remaining 12 days. Total CO 2 evolution was also highest at day 6 and declined steadily over the remaining duration of the incubation. Filtrate NH 4 + -N increased from background levels to a final value of 57 μg of N per treatment. Filtrate NO 3 - N completely disappeared by day 6, and organic N showed a slight decline between days 12 and 18. The majority of the 15 N that could be recovered appeared in the particulate organic fraction by day 6 (41 μg of N), and the filtrate NH 4 + N fraction contained 11 μg of 15 N by day 18. The 15 N enrichment values of the filtrate NH 4 + and the inorganic N associated with the particulate fraction had increased to approximately 20 atom % 15 N by 18 days of incubation, whereas the particulate organic fraction reached its highest enrichment by day 6. Measurements of N 2 fixation and denitrification indicated an insignificant gain or loss of N from the experimental system by these processes. The data show that woody debris in stream ecosystems might function as a rapid and efficient sink for exogenous N, resulting in stimulation of wood decomposition and subsequent activation of other N cycling processes

Liver function tests using the stable isotope /sup 15/N

Energy Technology Data Exchange (ETDEWEB)

Faust, H; Jung, K; Hirschberg, K; Krumbiegel, P; Junghans, P; Reinhardt, R; Matkowitz, R; Teichmann, B

1988-01-01

Several liver function tests using oral application of a nitrogen compound labelled with /sup 15/N and the subsequent determination of /sup 15/N in a certain fraction of urine or in the total urine by emission spectrometry are described. Because of the key function of the liver in the metabolism of nitrogen compounds, the results of these tests allow conclusions concerning some disturbances of liver functions.

Production of granules of urea, urea-ammonium sulphate and urea-potassium chloride enriched with 15N

International Nuclear Information System (INIS)

Bendassolli, J.A.

1991-01-01

Using a pearling tower it was possible to produce granulated urea, and granulated mixtures of ammonium sulphate and urea, potassium chloride and urea, Labelled in 15 N. Granulated urea with 1, 2, 3 and 4 mm of diameter was obtained using a system with a heating controller. A low concentration of biuret was observed in the granules produced ( 15 N-Labelled ( 15 NH 4' 15 NH 2 ) with variable proportion of ammonium sulphate and urea. (author)

MASS LOSS AND NITROGEN DYNAMICS DURING THE DECOMPOSITION OF A N-LABELED N2-FIXING EPOPHYTIC LICHEN, LOBARIA OREGANA (TUCK.) MULL. ARG.

Science.gov (United States)

We studied mass loss and nitrogen dynamics during fall and spring initiated decomposition of an N2-fixing epiphytic lichen, Lobaria oregana (Tuck.) Mull. Arg. using 15N. We developed a method of labeling lichens with 15N that involved spraying lichen material with a nutrient sol...

Estimation of dinitrogen fixation by cowpea (Vigna unguiculata) using residual soil 15N in poppy (Papaver somniferum L) cowpea sequence

International Nuclear Information System (INIS)

Patra, D.D.; Chand, Sukhmal; Anwar, M.

1994-01-01

Estimation of dinitrogen fixation by cowpea was carried out under greenhouse conditions using pots each containing 12 kg soil. Different 15 N sources included residual soil 15 N where urea was applied to opium poppy before planting of cowpea as fixing and maize as non-fixing crop. Other N sources were labelled urea, 15 N labelled poppy straw, and labelled urea + unlabelled poppy straw. The amount of N 2 fixed varied with the source of 15 N in soil. Plant material treatment gave a higher estimate at 40 days, whereas the estimate was highest with residual 15 N at 75 days. Such variation is attributed to variation in 1 5N enrichment which can be reduced by utilizing the residual 15 N which gives a more stable enrichment of soil 15 N with time. It may also alleviate the errors resulting from the differential pattern of 15 N uptake by fixing and nonfixing plant due to temporal variation in 15 N enrichment in soil. (author). 8 refs., 3 tabs

The influence of tannin, pectin and polyethylene glycol on attachment of 15N-labelled rumen microorganisms to cellulose

International Nuclear Information System (INIS)

Bento, M.H.L.; Acamovic, T.; Makkar, H.P.S.

2005-01-01

The microbial attachment to and gas production from α-cellulose (Sigma; C-8002) without and with mimosa tannin (MT), pectin (P), polyethylene glycol (PEG), MT + P or MT + PEG, were investigated using the in vitro gas production system. Microbial attachment based on 15 N-labelled rumen microorganisms in the residual pellet after 24 h incubation was estimated, which varied from 113.7 to 161.3 μg 15 N per g residual pellet. C + MT had the lowest microbial attachment (P 2 = 0.84, P 15 N) in the residual pellet measured for C + MT (0.054) and C + MT + P (0.159), compared with the other treatments (0.32 for C; 0.34 for C + P; 0.33 for C + PEG; and 0.33 for C + MT + PEG). A MT concentration of 194 g/kg diet reduced microbial attachment and activity of rumen microorganisms in vitro. Polyethylene glycol counteracted the effect of MT on microbial attachment and activity. Pectin exerted a beneficial effect on attachment and fermentation in the initial hours of incubation. A ratio of pectin to MT of 1:1 improved microbial activity of C + MT but inhibition of microbial activity by MT remained at 24 h as indicated by the lower gas production of C + MT + P compared with the control. The results support the hypothesis that there is considerable interaction between tannins, microbes and non-starch-polysaccharides (NSP) in animal feeds and that these interactions may influence the functional ability of microbes in the gastrointestinal tract of animals. (author)

[13N]ammonia in organic solvents; a potent synthetic precursor for 13N-labeling

International Nuclear Information System (INIS)

Tominaga, Toshiyoshi; Hirobe, Masaaki

1987-01-01

13 NH 3 in an organic solvent was prepared and its utility as a labeling precursor was studied. [ 13 N]adenine ([ 13 N]ADN), [ 13 N]nicotinamide ([ 13 N]NAM), [ 13 N]p-nitrophenyl carbamate ([ 13 N]NPC), and [ 13 N]L-glutamine ([ 13 N]Gln) were labeled utilizing this precursor. [ 13 N]ADN and [ 13 N]NAM were labeled in much better yields than from an aqueous solution of 13 NH 3 . [ 13 N]NPC and [ 13 N]Gln, which could not be labeled in an aqueous solution, were labeled in high radiochemical yields. Thus, the advantages of this precursor are the improvement of the labeling yield and the feasibility of labeling compounds unstable in aqueous conditions. (author)

Nitrogen cycling in an extreme hyperarid environment inferred from δ15N analyses of plants, soils and herbivore diet

Science.gov (United States)

Díaz, Francisca P.; Frugone, Matías; Gutiérrez, Rodrigo A.; Latorre, Claudio

2016-03-01

Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ15N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ15N and δ13C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ15N values span the entire gradient, soil δ15N values show a positive correlation with aridity as expected. In contrast, foliar δ15N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ15N values.

Nitrogen cycling in an extreme hyperarid environment inferred from δ(15)N analyses of plants, soils and herbivore diet.

Science.gov (United States)

Díaz, Francisca P; Frugone, Matías; Gutiérrez, Rodrigo A; Latorre, Claudio

2016-03-09

Climate controls on the nitrogen cycle are suggested by the negative correlation between precipitation and δ(15)N values across different ecosystems. For arid ecosystems this is unclear, as water limitation among other factors can confound this relationship. We measured herbivore feces, foliar and soil δ(15)N and δ(13)C values and chemically characterized soils (pH and elemental composition) along an elevational/climatic gradient in the Atacama Desert, northern Chile. Although very positive δ(15)N values span the entire gradient, soil δ(15)N values show a positive correlation with aridity as expected. In contrast, foliar δ(15)N values and herbivore feces show a hump-shaped relationship with elevation, suggesting that plants are using a different N source, possibly of biotic origin. Thus at the extreme limits of plant life, biotic interactions may be just as important as abiotic processes, such as climate in explaining ecosystem δ(15)N values.

Selective detection of carbon-13, nitrogen-15, and deuterium labeled metabolites by capillary gas chromatography-chemical reaction interface/mass spectrometry

International Nuclear Information System (INIS)

Chace, D.H.; Abramson, F.P.

1989-01-01

We have applied a new chemical reaction interface/mass spectrometer technique (CRIMS) to the selective detection of 13C-, 15N-, and 2H-labeled phenytoin and its metabolites in urine following separation by capillary gas chromatography. The microwave-powered chemical reaction interface converts materials from their original forms into small molecules whose mass spectra serve to identify and quantify the nuclides that make up each analyte. The presence of each element is followed by monitoring the isotopic variants of CO2, NO, or H2 that are produced by the chemical reaction interface. Chromatograms showing only enriched 13C and 15N were produced by subtracting the abundance of naturally occurring isotopes from the observed M + 1 signal. A selective chromatogram of 2H (D) was obtained by measuring HD at m/z 3.0219 with a resolution of 2000. Metabolites representing less than 1.5% of the total labeled compounds could be identified in the chromatogram. Detection limits from urine of 380 pg/mL of a 15N-labeled metabolite, 7 ng/mL of a 13C-labeled metabolite, and 16 ng/mL of a deuterium labeled metabolite were determined at a signal to noise ratio of 2. Depending on the isotope examined, a linear dynamic range of 250-1000 was observed using CRIMS. To identify many of these labeled peaks (metabolites), the chromatographic analysis was repeated with the chemical reaction interface turned off and mass spectra obtained at the retention times found in the CRIMS experiment. CRIMS is a new analytical method that appears to be particularly useful for metabolism studies

RAPID AND AUTOMATED PROCESSING OF MALDI-FTICR/MS DATA FOR N-METABOLIC LABELING IN A SHOTGUN PROTEOMICS ANALYSIS.

Science.gov (United States)

Jing, Li; Amster, I Jonathan

2009-10-15

Offline high performance liquid chromatography combined with matrix assisted laser desorption and Fourier transform ion cyclotron resonance mass spectrometry (HPLC-MALDI-FTICR/MS) provides the means to rapidly analyze complex mixtures of peptides, such as those produced by proteolytic digestion of a proteome. This method is particularly useful for making quantitative measurements of changes in protein expression by using (15)N-metabolic labeling. Proteolytic digestion of combined labeled and unlabeled proteomes produces complex mixtures that with many mass overlaps when analyzed by HPLC-MALDI-FTICR/MS. A significant challenge to data analysis is the matching of pairs of peaks which represent an unlabeled peptide and its labeled counterpart. We have developed an algorithm and incorporated it into a compute program which significantly accelerates the interpretation of (15)N metabolic labeling data by automating the process of identifying unlabeled/labeled peak pairs. The algorithm takes advantage of the high resolution and mass accuracy of FTICR mass spectrometry. The algorithm is shown to be able to successfully identify the (15)N/(14)N peptide pairs and calculate peptide relative abundance ratios in highly complex mixtures from the proteolytic digest of a whole organism protein extract.

Synthesis of specifically 15N- and 13C-labeled antitumor (2-Haloethyl)nitrosoureas. The study of their conformations in solution by nitrogen-15 and carbon-13 nuclear magnetic resonance and evidence for stereoelectronic control in their aqueous decomposition

International Nuclear Information System (INIS)

Lown, J.W.; Chauhan, S.M.S.

1981-01-01

The synthesis of certain specifically 15 N, 13 C, and 2 H isotope labeled 1-(2-chloroethyl)-3-alkyl-1-nitrosoureas (CENUs) is described. Spectroscopic examination of CENUs and their isotope-labeled counterparts by 1 H, 15 N, and 13 C NMR and infrared spectra indicates that they adopt preferred conformations in nonpolar aprotic solvents in which the NO group is aligned toward the 2-chloroethyl group. The result is in accord with the conformation of MeCCNU in the crystalline state derived from X-ray diffraction. The chemical shifts and coupling constants in the CENUs change with both solvent polarity and basicity. In aqueous phosphate buffer there is evidence for the formation of a tetrahedral intermediate, the conformation of which alters according to the reaction conditions and ultimately controls the formation of the aqueous decomposition products of CENUs. This is revealed most clearly by 13 C NMR of carbonyl- 13 C- and nitroso- 15 N-labeled BCNU and CCNU where two distinct 15 N-coupled 13 C doublets with different chemical shifts are observed. The rate of conformational change is comparable with the rate of decomposition of CENUs (via the second conformer) and may therefore represent the critical initial step of the latter process in vivo. The intermediacy of the postulated tetrahedral intermediates for CENUs is supported by observed 18 O exchange into the carbonyl group in 18 O-enriched water. Consideration of the conformations of the intermediates and of the alignment of the heteroatom lone pairs provides a satisfactory interpretation of the reactions of CENUs in aqueous solution as well as their pH dependence in terms of strict steroelectronic control and accounts for the formation of the observed products

Removal of nitrite impurity from nitrate labeled with nitrogen-15

International Nuclear Information System (INIS)

Malone, J.P.; Stevens, R.J.

1998-01-01

Potassium nitrate labeled with 15 N is often used as a tracer in studies of N dynamics in soil and water systems. Typically, 0.8% NO 2 - impurity has been found in the batches of K 15 NO 3 enriched to 99 atom % excess 15 N that were purchased by our laboratory. Nitrite is an intermediate in several N cycling processes so its addition when adding NO 3 - could produce misleading results. We have developed a safe, simple, and inexpensive method to remove NO 2 - impurity from any NO 3 - solution in a water matrix. The principle is the oxidation of NO2- to NO 3 - by UV light in the presence of a heterogenous TiO 2 catalyst. A NO 2 - concentration of 0.2 mM in 100 mL of 0.2 M NO 3 - solution could be oxidized in 12 min using 0.5 g L -1 TiO 2 in a specially constructed photoreactor with a 75-W UV facial tanning lamp. For the routine removal of NO 2 - , use of the same TiO 2 concentration in a standard beaker worked equally well when the irradiation time was extended to 2.5 h. After irradiation, the TiO2 is easily and totally removed from the solution by membrane filtration. (author)

Biosynthetic preparation of L-[13C]- and [15N]glutamate by Brevibacterium flavum

International Nuclear Information System (INIS)

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

1987-01-01

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

16 CFR 1500.15 - Labeling of fire extinguishers.

Science.gov (United States)

2010-01-01

... 16 Commercial Practices 2 2010-01-01 2010-01-01 false Labeling of fire extinguishers. 1500.15 Section 1500.15 Commercial Practices CONSUMER PRODUCT SAFETY COMMISSION FEDERAL HAZARDOUS SUBSTANCES ACT...)(1), the signal word “Danger” and the statement of hazard “Poisonous gases formed when used to...

N isotopes and N cycle in the TieShanPing subtropical forest ecosystem, Southwestern China.

Science.gov (United States)

Jiang, Chun-lai; Zhang, Xiao-Shan

2009-07-01

Nitrogen is essential for forest growth and forest stand development. It is commonly a limited factor for forest productivity. We examined delta (15)N values in soils and plants by studying the sources of N used by vegetation and cycles of N in a 43-year-old plantation of the TieShanPing forest ecosystem in southwestern China, dominated by massone pine (Pinus massoniana). The N concentration of plant materials ranges from 1.1% to 2.2%. The nitrogen concentration of P. massoniana was 1.3% while soils showed the concentration of 0.04-0.15%. Regarding natural abundance of (15)N, large significant variation (-6.0 per thousand to -3.8 per thousand) in delta (15)N values was observed among shrub and tree leaves. delta (15)N values were also significantly varied from -4.7 per thousand to -3.8 per thousand among the pioneer species in the plantation. Soil delta (15)N values (3.1-6.3 per thousand) were significantly enriched compared to those values in plant samples. Despite the negative delta (15)N values of the vegetation cover, the high delta (15)N values in the topsoil indicate that return of N to soils by litter-fall is minimal on TieShanPing and the present forests do not change very much the soil (15)N signals at the surface layer. The positive delta (15)N values may also indicate large N losses from the soil system vial leaching, volatilization and plant uptake.

15N kinetic analysis of N2O production by Nitrosomonas europaea: an examination of nitrifier denitrification

International Nuclear Information System (INIS)

Poth, M.; Focht, D.D.

1985-01-01

A series of 15 N isotope tracer experiments showed that Nitrosomonas europaea produces nitrous oxide only under oxygen-limiting conditions and that the labeled N from nitrite, but not nitrate, is incorporated into nitrous oxide, indicating the presence of the denitrifying enzyme nitrite reductase. A kinetic analysis of the m/z 44, 45, and 46 nitrous oxide produced by washed cell suspensions of N. europaea when incubated with 4 mM ammonium (99% 14 N) and 0.4 mM nitrite (99% 15 N) was performed. No labeled nitirte was reduced to ammonium. All labeled material added was accounted for as either nitrite or nitrous oxide. The hypothesis that nitrous oxide is produced directly from nitrification was rejected since (i) it does not allow for the large amounts of double-labeled (m/z 46) nitrous oxide observed; (ii) the observed patterns of m/z 44, 45, 46 nitrous oxide were completely consistent with a kinetic analysis based on denitrification as the sole mechanism of nitrous oxide production but not with a kinetic analysis based on both mechanisms; (iii) the asymptotic ratio of m/z 45 to m/z 46 nitrous oxide was consistent with denitrification kinetics but inconsistent with nitrification kinetics, which predicted no limit to m/z 45 production. It is concluded that N. europaea is a denitrifier which, under conditions of oxygen stress, uses nitrite as a terminal electron acceptor and produces nitrous oxide

Effect of 15n-labeled hairy vetch and nitrogen fertilization on maize nutrition and yield under no-tillage

International Nuclear Information System (INIS)

Almeida Acosta, Jose Alan de; Amado, Telmo Jorge Carneiro; Silva, Leandro Souza da; Silveira Nicoloso, Rodrigo da

2011-01-01

This study evaluated the effect of hairy vetch (Vicia villosa Roth) as cover crop on maize nutrition and yield under no tillage using isotope techniques. For this purpose, three experiments were carried out: quantification of biological nitrogen fixation (BNF) in hairy vetch; estimation of the N release rate from hairy vetch residues on the soil surface; quantification of 15 N recovery by maize from labeled hairy vetch under three rates of mineral N fertilization. This two year field experiment was conducted on a sandy Acrisol (FAO soil classification) or Argissolo Vermelho distrofico arenico (Brazilian Soil Classification), at a mean annual temperature of 18 deg C and mean annual rainfall of 1686 mm. The experiment was arranged in a double split-plot factorial design with three replications. Two levels of hairy vetch residue (50 and 100 % of the aboveground biomass production) were distributed on the surface of the main plots (5 x 12 m). Maize in the sub-plots (5 x 4 m) was fertilized with three N rates (0, 60, and 120 kg ha -1 N), with urea as N source. The hairy vetch-derived N recovered by maize was evaluated in microplots (1.8 x 2.2 m). The BFN of hairy vetch was on average 72.4 %, which represents an annual input of 130 kg ha -1 of atmospheric N. The N release from hairy vetch residues was fast, with a release of about 90 % of total N within the first four weeks after cover crop management and soil residue application. The recovery of hairy vetch 15 N by maize was low, with an average of 12.3 % at harvest. Although hairy vetch was not directly the main source of maize N nutrition, the crop yield reached 8.2 Mg ha -1 , without mineral fertilization. There was an apparent synergism between hairy vetch residue application and the mineral N fertilization rate of 60 kg ha -1 , confirming the benefits of the combination of organic and inorganic N sources for maize under no tillage. (author)

Nitrogen cycling in a forest stream determined by a 15N tracer addition

Science.gov (United States)

Patrick J. Mullholland; Jennifer L. Tank; Diane M. Sanzone; Wilfred M. Wollheim; Bruce J. Peterson; Jackson R. Webster; Judy L. Meyer

2000-01-01

Nitrogen uptake and cycling was examined using a six-week tracer addition of 15N-labeled ammonium in early spring in Waer Branch, a first-order deciduous forest stream in eastern Tennessee. Prior to the 15N addition, standing stocks of N were determined for the major biomass compartments. During and after the addition,

True cooking aroma or artefact. 15N gives the answer

International Nuclear Information System (INIS)

Metro, F.; Boudaud, N.; Dumont, J.P.

1994-01-01

In order to determine the respective contributions of the various nitrous precursor families in aroma preparations, the usually added amino acids were substituted with 15 N isotope labelled homologous components. Results concerning isotope ratios for the volatile fraction nitrous components collected from poultry meat aromatic preparations, are presented. Terminal product labelling appears to allow for a better determination of the substrate and functional additive contributions. 4 figs., 6 refs

Comparison of three 15N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages.

Science.gov (United States)

Kamoun, M; Ammar, H; Théwis, A; Beckers, Y; France, J; López, S

2014-11-01

The use of stable (15)N as a marker to determine microbial contamination in nylon bag incubation residues to estimate protein degradability was investigated. Three methods using (15)N were compared: (15)N-labeled forage (dilution method, LF), (15)N enrichment of rumen solids-associated bacteria (SAB), and (15)N enrichment of rumen liquid-associated bacteria (LAB). Herbage from forages differing in protein and fiber contents (early-cut Italian ryegrass, late-cut Italian ryegrass, and red clover) were freeze-dried and ground and then incubated in situ in the rumen of 3 steers for 3, 6, 12, 24, and 48 h using the nylon bag technique. The (15)N-labeled forages were obtained by fertilizing the plots where herbage was grown with (15)NH4 (15)NO3. Unlabeled forages (obtained from plots fertilized with NH4NO3) were incubated at the same time that ((15)NH4)2SO4 was continuously infused into the rumen of the steers, and then pellets of labeled SAB and LAB were isolated by differential centrifugation of samples of ruminal contents. The proportion of bacterial N in the incubation residues increased from 0.09 and 0.45 g bacterial N/g total N at 3 h of incubation to 0.37 and 0.85 g bacterial N/g total N at 48 h of incubation for early-cut and late-cut ryegrass, respectively. There were differences (P forage (late-cut ryegrass) was 0.51, whereas the corrected values were 0.85, 0.84, and 0.77 for the LF, SAB, and LAB methods, respectively. With early-cut ryegrass and red clover, the differences between uncorrected and corrected values ranged between 6% and 13%, with small differences among the labeling methods. Generally, methods using labeled forage or labeled SAB and LAB provided similar corrected degradability values. The accuracy in estimating the extent of degradation of protein in the rumen from in situ disappearance curves is improved when values are corrected for microbial contamination of the bag residue.

Improved diffusion technique for 15N:14N analysis of ammonium and nitrate from aqueous samples by stable isotope spectrometry

International Nuclear Information System (INIS)

Goerges, T.; Dittert, K.

1998-01-01

Nitrogen (N) isotope ratio mass spectrometry (IRMS) by Dumas combustion and continuous flow mass spectrometry has become a wide-spread tool for the studies of N turnover. The speed and labor efficiency of 15N determinations from aqueous solutions such as soil solutions or soil extracts are often limited by sample preparation. Several procedures for the conversion of dissolved ammonium (NH4+) or nitrate NO3- to gaseous ammonia and its subsequent trapping in acidified traps have been elaborated in the last decades. They are based on the use of acidified filters kept either above the respective solution or in floating PTFE envelopes. In this paper, we present an improved diffusion method with a fixed PTFE trap. The diffusion containers are continuously kept in a vertical rotary shaker. Quantitative diffusion can thus be achieved in only three days. For solutions with NH4+ levels of only 1 mg N kg-1 and NO3- concentrations of 12 mg N kg-1, recovery rates of 98.8-102% were obtained. By addition of 15N labeled and non-labeled NH4+ and NO3- it was shown that no cross-contamination from NH4+ to NO3- or vice versa takes place even when one form is labeled to more than 1 at %15N while the other form has natural 15N content. The method requires no intermediate step of ammonia volatilization before NO3- conversion

Incorporation of 15N-inorganic nitrogen into free-amino acids in germinating corn

International Nuclear Information System (INIS)

Samukawa, Kisaburo; Yamaguchi, Masuro

1979-01-01

Incorporation of 15 N-labeled compounds, (K 15 NO 3 ) and ( 15 NH 4 ) 2 SO 4 , into free-amino acids was measured in germinating corn. Sterilized seeds of sweet corn (Choko No. 865) were sown on the filter papers soaked in 10 ml of the solution containing one of the labeled compounds (40 ppm N, 99 atom % excess) in petri dishes and germinated at 30 deg C. After 48 hours and 72 hours, 15 N-incorporation was measured in 5 seedlings selected owing to uniform growth. A GC-MS was used for measuring the ratio of 15 N isotopes present in free-amino acids. 15 N incorporation into free-amino acids hardly occurred when corn was germinated in the solution containing K 15 NO 3 , which suggested that endogenous nitrogen was used during the early germination stage of corn when nitrate is present. Incorporation into amino acids was greater when corn was germinated in the medium containing ( 15 NH 4 ) 2 SO 4 , than the case of the solution containing K 15 NO 3 . When corn was germinated in the solution containing ( 15 NH 4 ) 2 SO 4 , assimilation of 15 N into asparagine or aspartic acid was comparatively higher than that into the other amino acids, though the incorporation rate was low. Thus, in intact germinating corn, the hydrolyzed product of protein was utilized for germination with priority, and dependence on exogenous nitrogen was low. (Kaihara, S.)

15N Kinetic Analysis of N2O Production by Nitrosomonas europaea: an Examination of Nitrifier Denitrification

OpenAIRE

Poth, Mark; Focht, Dennis D.

1985-01-01

A series of 15N isotope tracer experiments showed that Nitrosomonas europaea produces nitrous oxide only under oxygen-limiting conditions and that the labeled N from nitrite, but not nitrate, is incorporated into nitrous oxide, indicating the presence of the “denitrifying enzyme” nitrite reductase. A kinetic analysis of the m/z 44, 45, and 46 nitrous oxide produced by washed cell suspensions of N. europaea when incubated with 4 mM ammonium (99% 14N) and 0.4 mM nitrite (99% 15N) was performed....

1H, 15N and 13C NMR Assignments of Mouse Methionine Sulfoxide Reductase B2

Science.gov (United States)

Breivik, Åshild S.; Aachmann, Finn L.; Sal, Lena S.; Kim, Hwa-Young; Del Conte, Rebecca; Gladyshev, Vadim N.; Dikiy, Alexander

2011-01-01

A recombinant mouse methionine-r-sulfoxide reductase 2 (MsrB2ΔS) isotopically labeled with 15N and 15N/13C was generated. We report here the 1H, 15N and 13C NMR assignments of the reduced form of this protein. PMID:19636904

Methods of 15N tracer research in biological systems

International Nuclear Information System (INIS)

Hirschberg, K.; Faust, H.

1985-01-01

The application of the stable isotope 15 N is of increasing importance in different scientific disciplines, especially in medicine, agriculture, and the biosciences. The close correlation between the growing interest and improvements of analytical procedures resulted in remarkable advances in the 15 N tracer technique. On the basis of the latest results of 15 N tracer research in life sciences and agriculture methods of 15 N tracer research in biological systems are compiled. The 15 N methodology is considered under three headings: Chemical analysis with a description of methods of sample preparation (including different separation and isolation methods for N-containing substances of biological and agricultural origin) and special procedures converting ammonia to molecular nitrogen. Isotopic analysis with a review on the most important methods of isotopic analysis of nitrogen: mass spectrometry (including the GC-MS technique), emission spectrometry, NMR spectroscopy, and other analytical procedures. 15 N-tracer techniques with a consideration of the role of the isotope dilution analysis as well as different labelling techniques and the mathematical interpretation of tracer data (modelling, N turnover experiments). In these chapters also sources of errors in chemical and isotopic analysis, the accuracy of the different methods and its importance on tracer experiments are discussed. Procedures for micro scale 15 N analysis and aspects of 15 N analysis on the level of natural abundance are considered. Furthermore some remarks on isotope effects in 15 N tracer experiments are made. (author)

Synthesis 1-(5-oxohexyl)-3,7-dimethyl-xanthyne labelled with tritium into 8 position from purinic ring

International Nuclear Information System (INIS)

Mihaila, V.; Corol, D.

1999-01-01

This paper presents the work on synthesis of 1-(5-oxohexyl)-3,7-dimethyl-xanthyne labelled with tritium into 8 position from purinic ring. The obtaining of tritium labelled compound is realized by initial labelling of theobromine with tritium into 8 position and by coupling the purinic derivative to 1-Br-5-hexanone. Theobromine-8- 3 H was obtained by the bromination of theobromine with elementary bromine and after that the bromine was substituted with tritium i.e.: C 7 H 8 O 2 N 4 theobromine Br 2 /(-HBr) C 7 H 7 O 2 N 4 Br (8-Br-theobromine) ( 3 H 2 /cat)/(-KOH) C 7 H 7 3 HO 2 N 4 (theobromine-8- 3 H). Theobromine-8- 3 H was purified by thin layer chromatography with a solvent system i.e. n-BuOH:AcOH:H 2 O (4:1:1, v/v/v) and characterized radiochemically. It was then diluted by unlabelled theobromine to specific activity of 50 mCi/g. After dilution, theobromine-8- 3 H was coupled to 1-Br-5-hexanone i.e.: C 7 H 7 3 HO 2 N 4 (theobromine-8- 3 H) + Br-(CH 2 ) 4 -CO-CH 3 (1-Br-5-hexanone) (NaOH)/(CH 3 OH) C 13 H 17 3 HO 3 N 4 (1-(5-oxohexyl)- 3,7-dimethyl-xanthine-8- 3 H). The raw compound was purified by recrystallization from 2-propanol and it was characterized radiochemically. (authors)

Effect of method of N-application and modified urea on N-15 recovery by rice

International Nuclear Information System (INIS)

Soliman, S.M.; Abdelmonem, M.A.S.

1995-01-01

Rice is a very responsive crop to nitrogen fertilizer, but the efficiency of the applied N-fertilizer is low. Greenhouse experiment conducted to evaluate several methods to improve fertilizer efficiency and reduce N-losses in rice fields. N-15 labelled urea was applied to 10 kg soils in pots, urea was applied alone, with addition of two urease inhibitors (NBPT and HQ), with addition of nitrification inhibitor (DCD),or with the combination of both inhibitors. The fertilizers were applied either broadcast on soil surface or at depth of 8 cm below the surface. At maturity, plants were separated into grain and straw, dried and weighted. Soil and plant samples were analyzed for total N and N-15 excess. Both fertilizer placement and inhibitor application significantly increased straw and grain yield, as well as N-uptake. Nitrogen derived from fertilizer (%Ndff) was more than doubled, when urea was applied deep and in combination with inhibitors. Plant recovery of N-15 labelled urea ranged from 17% to 75% according to treatment. Regardless of inhibitors application, plant recovery was increased from 39% to 65% when urea was applied at depth of 8 cm. Approximately 2/3 of the applied urea (64%)was lost when urea was applied alone. Those losses were reduced down to 12% with deep placement and inhibitor application. The two management practices show significant effect on minimizing N-losses and increasing plant recovery. 1 fig., 3 tabs

Effect of method of N-application and modified urea on N-15 recovery by rice

Energy Technology Data Exchange (ETDEWEB)

Soliman, S M; Abdelmonem, M A.S. [Soil and Water Dept., Atomic Energy Auth., Cairo, (Egypt)

1995-10-01

Rice is a very responsive crop to nitrogen fertilizer, but the efficiency of the applied N-fertilizer is low. Greenhouse experiment conducted to evaluate several methods to improve fertilizer efficiency and reduce N-losses in rice fields. N-15 labelled urea was applied to 10 kg soils in pots, urea was applied alone, with addition of two urease inhibitors (NBPT and HQ), with addition of nitrification inhibitor (DCD),or with the combination of both inhibitors. The fertilizers were applied either broadcast on soil surface or at depth of 8 cm below the surface. At maturity, plants were separated into grain and straw, dried and weighted. Soil and plant samples were analyzed for total N and N-15 excess. Both fertilizer placement and inhibitor application significantly increased straw and grain yield, as well as N-uptake. Nitrogen derived from fertilizer (%Ndff) was more than doubled, when urea was applied deep and in combination with inhibitors. Plant recovery of N-15 labelled urea ranged from 17% to 75% according to treatment. Regardless of inhibitors application, plant recovery was increased from 39% to 65% when urea was applied at depth of 8 cm. Approximately 2/3 of the applied urea (64%)was lost when urea was applied alone. Those losses were reduced down to 12% with deep placement and inhibitor application. The two management practices show significant effect on minimizing N-losses and increasing plant recovery. 1 fig., 3 tabs.

Studies on 14C labelled chlorpyrifos in model marine ecosystem

International Nuclear Information System (INIS)

Pandit, G.G.; Mohan Rao, A.M.; Kale, S.P.; Murthy, N.B.K.; Raghu, K.

1997-01-01

Chlorpyrifos is one of the widely used organophosphorus insecticides in tropical countries. Experiments were conducted with 14 C labelled chlorpyrifos to study the distribution of this compound in model marine ecosystem. Less than 50 per cent of the applied activity remained in water in 24 h. Major portion of the applied chlorpyrifos (about 4.2 % residue per g) accumulated into the clams with sediment containing a maximum of 5 to 6 per cent of applied compound. No degradation of chlorpyrifos was observed in water or sediment samples. However, metabolic products were formed in clams. (author). 4 refs., 3 tabs

Nitrogen-15 natural abundance of different soil N pools as a tool for assessing N transformation processes in alpine soils

Science.gov (United States)

Makarov, Mikhail; Malysheva, Tatiana; Tiunov, Alexei; Kadulin, Maxim; Maslov, Mikhail

2017-04-01

Nitrogen availability, net N mineralization, nitrification and 15N natural abundance of total soil N and small soil N pools (N-NH4+, N-NO3-, DON and microbial biomass N) were studied in a toposequence of alpine ecosystems in the Northern Caucasus. The toposequence was represented by (1) low productive alpine lichen heath (ALH) of the wind-exposed ridge and upper slope; (2) more productive Festuca varia grassland (FG) of the middle slope; (3) most productive Geranium gymnocaulon/Hedysarum caucasicum meadow (GHM) of the lower slope and (4) low productive snow bed community (SBC) of the slope bottom. Nitrogen transformation in the alpine soils produces distinct N pools with different 15N enrichment: DON/microbial biomass N > total N > N-NH4+ > N-NO3-. Grassland and meadow soils of the middle part of the toposequence are characterized by higher nitrogen transformation activities and higher δ15 values of total N and N-NH4+. Field incubation of alpine soils increased δ15N of N-NH4+ from -2.6 - +2.0‰ to +6.1 - +15.7‰. The N-NO3-produced in the incubation experiment had extremely low (negative) δ15N values (up to -14‰). We found a positive correlation between δ15N of different soil N pools (total N, N-NH4+ and N-NO3-) and net N mineralization and nitrification. Nitrification controls the formation of 15N enriched N-NH4+ pool while N mineralization probably had an important role in regulation of 15N enrichment of DON pool in alpine soils. Overall, our results support the hypothesis that 15N is more enriched in N-rich and more depleted in N-poor ecosystems. We conclude that δ15N values of different soil N pools could be a good indicator of microbial N transformation in alpine soils of the Northern Caucasus. Acknowledgement: This study was supported by Russian Science Foundation (16-14-10208).

Syntheses of 18F-labeled reduced haloperidol and 11C-labeled reduced 3-N-methylspiperone

International Nuclear Information System (INIS)

Ravert, H.T.; Dannals, R.F.; Wilson, A.A.; Wong, D.F.; Wagner, H.N. Jr.

1991-01-01

18 F-Labeled reduced haloperidol and 11 C-labeled reduced 3-N-methylspiperone were synthesized in a convenient and quantitative one step reduction from 18 F-labeled haloperidol and 11 C-labeled N-methylspiperone, respectively. Both products were purified by semipreparative HPLC and were obtained at high specific activity and radiochemical purity. (author)

Effect of {sup 15}n-labeled hairy vetch and nitrogen fertilization on maize nutrition and yield under no-tillage

Energy Technology Data Exchange (ETDEWEB)

Almeida Acosta, Jose Alan de [Drakkar Solos, Santa Maria, RS (Brazil); Amado, Telmo Jorge Carneiro; Silva, Leandro Souza da, E-mail: tamado@smail.ufsm.b, E-mail: leandro@smail.ufsm.b [Universidade Federal de Santa Maria (UFSM), RS (Brazil). Soil Dept.; Neergaard, Andreas de; Vinther, Mads, E-mail: adn@life.ku.d [University of Copenhagen (Denmark); Silveira Nicoloso, Rodrigo da, E-mail: rodrigo.nicoloso@cnpsa.embrapa.b [Embrapa Swine and Poultry, Concordia, SC (Brazil)

2011-07-15

This study evaluated the effect of hairy vetch (Vicia villosa Roth) as cover crop on maize nutrition and yield under no tillage using isotope techniques. For this purpose, three experiments were carried out: quantification of biological nitrogen fixation (BNF) in hairy vetch; estimation of the N release rate from hairy vetch residues on the soil surface; quantification of {sup 15}N recovery by maize from labeled hairy vetch under three rates of mineral N fertilization. This two year field experiment was conducted on a sandy Acrisol (FAO soil classification) or Argissolo Vermelho distrofico arenico (Brazilian Soil Classification), at a mean annual temperature of 18 deg C and mean annual rainfall of 1686 mm. The experiment was arranged in a double split-plot factorial design with three replications. Two levels of hairy vetch residue (50 and 100 % of the aboveground biomass production) were distributed on the surface of the main plots (5 x 12 m). Maize in the sub-plots (5 x 4 m) was fertilized with three N rates (0, 60, and 120 kg ha{sup -1} N), with urea as N source. The hairy vetch-derived N recovered by maize was evaluated in microplots (1.8 x 2.2 m). The BFN of hairy vetch was on average 72.4 %, which represents an annual input of 130 kg ha{sup -1} of atmospheric N. The N release from hairy vetch residues was fast, with a release of about 90 % of total N within the first four weeks after cover crop management and soil residue application. The recovery of hairy vetch {sup 15}N by maize was low, with an average of 12.3 % at harvest. Although hairy vetch was not directly the main source of maize N nutrition, the crop yield reached 8.2 Mg ha{sup -1}, without mineral fertilization. There was an apparent synergism between hairy vetch residue application and the mineral N fertilization rate of 60 kg ha{sup -1}, confirming the benefits of the combination of organic and inorganic N sources for maize under no tillage. (author)

Towards fully automated structure-based NMR resonance assignment of 15N-labeled proteins from automatically picked peaks

KAUST Repository

Jang, Richard; Gao, Xin; Li, Ming

2011-01-01

In NMR resonance assignment, an indispensable step in NMR protein studies, manually processed peaks from both N-labeled and C-labeled spectra are typically used as inputs. However, the use of homologous structures can allow one to use only N-labeled NMR data and avoid the added expense of using C-labeled data. We propose a novel integer programming framework for structure-based backbone resonance assignment using N-labeled data. The core consists of a pair of integer programming models: one for spin system forming and amino acid typing, and the other for backbone resonance assignment. The goal is to perform the assignment directly from spectra without any manual intervention via automatically picked peaks, which are much noisier than manually picked peaks, so methods must be error-tolerant. In the case of semi-automated/manually processed peak data, we compare our system with the Xiong-Pandurangan-Bailey- Kellogg's contact replacement (CR) method, which is the most error-tolerant method for structure-based resonance assignment. Our system, on average, reduces the error rate of the CR method by five folds on their data set. In addition, by using an iterative algorithm, our system has the added capability of using the NOESY data to correct assignment errors due to errors in predicting the amino acid and secondary structure type of each spin system. On a publicly available data set for human ubiquitin, where the typing accuracy is 83%, we achieve 91% accuracy, compared to the 59% accuracy obtained without correcting for such errors. In the case of automatically picked peaks, using assignment information from yeast ubiquitin, we achieve a fully automatic assignment with 97% accuracy. To our knowledge, this is the first system that can achieve fully automatic structure-based assignment directly from spectra. This has implications in NMR protein mutant studies, where the assignment step is repeated for each mutant. © Copyright 2011, Mary Ann Liebert, Inc.

Towards fully automated structure-based NMR resonance assignment of 15N-labeled proteins from automatically picked peaks

KAUST Repository

Jang, Richard

2011-03-01

In NMR resonance assignment, an indispensable step in NMR protein studies, manually processed peaks from both N-labeled and C-labeled spectra are typically used as inputs. However, the use of homologous structures can allow one to use only N-labeled NMR data and avoid the added expense of using C-labeled data. We propose a novel integer programming framework for structure-based backbone resonance assignment using N-labeled data. The core consists of a pair of integer programming models: one for spin system forming and amino acid typing, and the other for backbone resonance assignment. The goal is to perform the assignment directly from spectra without any manual intervention via automatically picked peaks, which are much noisier than manually picked peaks, so methods must be error-tolerant. In the case of semi-automated/manually processed peak data, we compare our system with the Xiong-Pandurangan-Bailey- Kellogg\\'s contact replacement (CR) method, which is the most error-tolerant method for structure-based resonance assignment. Our system, on average, reduces the error rate of the CR method by five folds on their data set. In addition, by using an iterative algorithm, our system has the added capability of using the NOESY data to correct assignment errors due to errors in predicting the amino acid and secondary structure type of each spin system. On a publicly available data set for human ubiquitin, where the typing accuracy is 83%, we achieve 91% accuracy, compared to the 59% accuracy obtained without correcting for such errors. In the case of automatically picked peaks, using assignment information from yeast ubiquitin, we achieve a fully automatic assignment with 97% accuracy. To our knowledge, this is the first system that can achieve fully automatic structure-based assignment directly from spectra. This has implications in NMR protein mutant studies, where the assignment step is repeated for each mutant. © Copyright 2011, Mary Ann Liebert, Inc.

δ15N in the turtle grass from the Mexican Caribbean

Science.gov (United States)

Talavera-Saenz, A.; Sanchez, A.; Ortiz-Hernandez, M.

2013-05-01

Nutrient inputs associated with population growth threaten the integrity of coastal ecosystems. To assess the rapid increase in tourism, we compared the δ15N from Thalassia testudinum collected at sites with different levels of tourism development and population to detect the N inputs of wastewater discharge (WD) along the coast of Quintana Roo. The contributions of nitrogen enriched in 15N are directly related to the increase of WD inputs in areas of high tourism development (Nichupte Lagoon in Cancun, >3 million tourists per year from 2007 to 2011 and 0.7 million of resident population) and decreased towards Bahia Akumal and Tulum (>3 million tourists per year from 2007 to 2011 and 0.15 million of resident population). The δ15N from T. testudinum was significantly lower at Mahahual and Puerto Morelos (about 0.4 million tourists per year in 2007 to 2011 and 0.25 million of resident population) than other the sites. In areas of the lowest development and with tourist activity restricted and small population, such as the Yum Balam Reserve and Sian Ka'an Biosphere Reserve, the δ15N values were in much higher enrichment that Mahahual and Puerto Morelos. Therefore is suggested that Mahahual and Puerto Morelos may be used for baseline isotopic monitoring, over environmental pressure on the reef lagoon ecosystem, where tourist activities and population are growing very slow rate. The anthropogenic N input has the potential to impact, both environmentally and economically, the seagrass meadows and the coral reefs along the coast of Quintana Roo and the Caribbean.

Tracing the cycling and fate of the explosive 2,4,6-trinitrotoluene in coastal marine systems with a stable isotopic tracer, 15N-[TNT

Science.gov (United States)

Smith, Richard W.; Vlahos, Penny; Böhlke, John Karl; Ariyarathna, Thivanka; Ballentine, Mark; Cooper, Christopher; Fallis, Stephen; Groshens, Thomas J.; Tobias, Craig

2015-01-01

2,4,6-Trinitrotoluene (TNT) has been used as a military explosive for over a hundred years. Contamination concerns have arisen as a result of manufacturing and use on a large scale; however, despite decades of work addressing TNT contamination in the environment, its fate in marine ecosystems is not fully resolved. Here we examine the cycling and fate of TNT in the coastal marine systems by spiking a marine mesocosm containing seawater, sediments, and macrobiota with isotopically labeled TNT (15N-[TNT]), simultaneously monitoring removal, transformation, mineralization, sorption, and biological uptake over a period of 16 days. TNT degradation was rapid, and we observed accumulation of reduced transformation products dissolved in the water column and in pore waters, sorbed to sediments and suspended particulate matter (SPM), and in the tissues of macrobiota. Bulk δ15N analysis of sediments, SPM, and tissues revealed large quantities of 15N beyond that accounted for in identifiable derivatives. TNT-derived N was also found in the dissolved inorganic N (DIN) pool. Using multivariate statistical analysis and a 15N mass balance approach, we identify the major transformation pathways of TNT, including the deamination of reduced TNT derivatives, potentially promoted by sorption to SPM and oxic surface sediments.

Mineralization of nitrogen from nitrogen-15 labeled crop residues and utilization by rice

International Nuclear Information System (INIS)

Norman, R.J.; Gilmour, J.T.; Wells, B.R.

1990-01-01

The availability of N from the residues of the previous crop to the subsequent rice (Oryza sativa L.) crop is largely unknown. The objectives of this study were to (1) measure the mineralization of N from 15 N-labeled rice, soybean (Glycine max L.), and wheat (Triticum aestivum L.) residues and the uptake by a subsequent rice crop; and (2) compare the 15 N tracer method with the standard fertilizer-N response method used in field studies to quantify the N contribution from the crop residue to the next crop. Nitrogen mineralization from decomposing crop residues was measured by soil sampling prior to seeding the rice crop and after seeding by plant sampling the rice at maturity. The minimum estimate of the amount of residue N mineralized from the time of residue incorporation until rice harvest was 9, 52, and 38% of the rice, soybean, and wheat residue N, respectively. The amount of residue N recovered in the rice crop was 3, 11, and 37% of the rice, soybean, and wheat residue N, respectively. The lower the C/N ratio and the higher the amount of N in the residue, the lower was the amount of residue N recovered in the soil organic fraction at harvest and the higher was the amount of residue N mineralized. The 15 N tracer method compared favorably with the fertilizer N response method when the uptake efficiency of the fertilizer N was taken into account

Use of /sup 15/N in following organic matter turnover, with specific reference to rotation systems

Energy Technology Data Exchange (ETDEWEB)

Ladd, J N [Commonwealth Scientific and Industrial Research Organization, Glen Osmond (Australia). Div. of Soils

1981-01-01

The results of this experiment indicate that the use of the technique described, (based on the degree of /sup 15/N-labelling of an N/sub 2/ fixer and a non-fixer), may be of value in assessing N/sub 2/ fixation in the field by legumes, but it is apparent that there are some problems to be overcome. Analyses of the whole plant are necessary, since the proportions of legume N due to N/sub 2/ fixation vary with the plant part. The extent to which legumes take up available N from the soil obviously will vary with soil profile and plant properties; and they will be affected by sward density and competition from other plants. These latter factors will increase the difficulty of using this method for assessing N/sub 2/ fixation by legumes in grazed pastures, but probably they would not be big problems when applying the method to grain legume crops. It is important that, in comparing the extent of labelling of the N of fixing and non-fixing plants, both types of plants should have access to soil inorganic-N of the same enrichment. This will be difficult to achieve under field conditions. However soils which contain relatively stable /sup 15/N-labelled organic residues may yield NO/sub 3/-N of tolerably constant enrichments. An experiment is in progress at Avon in which soils, amended 15 months previously with /sup 15/N-labelled legume residues and then cropped to wheat, will remain in situ and will be sown with fixing and non-fixing plants during the 1980 and 1981 seasons. These soils may prove to be suitable for measuring N/sub 2/ fixation in the field.

Endogenous and exogenous control of ecosystem function: N cycling in headwater streams

Energy Technology Data Exchange (ETDEWEB)

Mulholland, Patrick J [ORNL; Valett, H. Maurice [Virginia Polytechnic Institute and State University (Virginia Tech); Thomas, Steve [University of Nebraska; Webster, Jackson [Virginia Polytechnic Institute and State University (Virginia Tech); Dahm, Cliff [University of New Mexico, Albuquerque; Fellows, Christine [Griffith University, Nathan, Queensland, Australia; Crenshaw, Chelsea [University of New Mexico, Albuquerque; Peterson, Chris G. [Loyola University

2008-01-01

Allochthonous inputs act as resource subsidies to many ecosystems, where they exert strong influences on metabolism and material cycling. At the same time, metabolic theory proposes endogenous thermal control independent of resource supply. To address the relative importance of exogenous and endogenous influences, we quantified spatial and temporal variation in ecosystem metabolism and nitrogen (N) uptake using seasonal releases of {sup 15}N as nitrate in six streams differing in riparian-stream interaction and metabolic character. Nitrate removal was quantified using a nutrient spiraling approach based on measurements of downstream decline in {sup 15}N flux. Respiration (R) and gross primary production (GPP) were measured with whole-stream diel oxygen budgets. Uptake and metabolism metrics were addressed as z scores relative to site means to assess temporal variation. In open-canopied streams, areal uptake (U; {micro}g N {center_dot} m{sup -2} {center_dot} s{sup -1}) was closely related to GPP, metabolic rates increased with temperature, and R was accurately predicted by metabolic scaling relationships. In forested streams, N spiraling was not related to GPP; instead, uptake velocity (v{sub f}; mm/s) was closely related to R. In contrast to open-canopied streams, N uptake and metabolic activity were negatively correlated to temperature and poorly described by scaling laws. We contend that streams differ along a gradient of exogenous and endogenous control that relates to the relative influences of resource subsidies and in-stream energetics as determinants of seasonal patterns of metabolism and N cycling. Our research suggests that temporal variation in the propagation of ecological influence between adjacent systems generates phases when ecosystems are alternatively characterized as endogenously and exogenously controlled.

Endogenous and exogenous control of ecosystem function: N cycling in headwater streams.

Science.gov (United States)

Valett, H M; Thomas, S A; Mulholland, P J; Webster, J R; Dahm, C N; Fellows, C S; Crenshaw, C L; Peterson, C G

2008-12-01

Allochthonous inputs act as resource subsidies to many ecosystems, where they exert strong influences on metabolism and material cycling. At the same time, metabolic theory proposes endogenous thermal control independent of resource supply. To address the relative importance of exogenous and endogenous influences, we quantified spatial and temporal variation in ecosystem metabolism and nitrogen (N) uptake using seasonal releases of 15N as nitrate in six streams differing in riparian-stream interaction and metabolic character. Nitrate removal was quantified using a nutrient spiraling approach based on measurements of downstream decline in 15N flux. Respiration (R) and gross primary production (GPP) were measured with whole-stream diel oxygen budgets. Uptake and metabolism metrics were addressed as z scores relative to site means to assess temporal variation. In open-canopied streams, areal uptake (U; microg N x m(-2) x s(-1)) was closely related to GPP, metabolic rates increased with temperature, and R was accurately predicted by metabolic scaling relationships. In forested streams, N spiraling was not related to GPP; instead, uptake velocity (v(f); mm/s) was closely related to R. In contrast to open-canopied streams, N uptake and metabolic activity were negatively correlated to temperature and poorly described by scaling laws. We contend that streams differ along a gradient of exogenous and endogenous control that relates to the relative influences of resource subsidies and in-stream energetics as determinants of seasonal patterns of metabolism and N cycling. Our research suggests that temporal variation in the propagation of ecological influence between adjacent systems generates phases when ecosystems are alternatively characterized as endogenously and exogenously controlled.

Leaching and mass balance of 15N-labeled urea applied to a Kentucky bluegrass turf

International Nuclear Information System (INIS)

Miltner, E.D.; Branham, B.E.; Paul, E.A.; Rieke, P.E.

1996-01-01

The fate of urea applied to Kentucky bluegrass (Poa pratensis L.) turf was studied over a 2-yr period using a combination of intact monolith lysimeters and small plots. Soil type was a Marlette fine sandy loam (fine-loamy mixed mesic Glossoboric Hapludalfs). Urea was applied at a rate of 196 kg N ha-1 yr-1 in five equal applications of 39.2 kg N ha-1, using two application schedules. Treatments were fertilized at approximately 38-d intervals with the 'Spring' treatment fertilized from late April through late September and the 'Fall' treatment from early June through early November. In 1991 only, the April and November applications used 15N-labeled urea (LFN). For the Spring treatment, 31% of LFN was recovered from thatch at 18 DAT. This value remained constant for the next year, then gradually declined to 20% after 2 yr. Only 8% of the LFN was recovered from soil at 18 DAT and increased to only 20% 2 yr after application. Approximately 35% of the LFN was harvested in clippings over 2 yr. Through May 1993 (748 DAT), LFN in leachate totaled 0.18% of the amount applied. For the Fall treatment, 62% of the LFN was recovered from thatch d 18 DAT. This value declined to 35% by the following June. LFN in soil increased from 12% to 25% over 2 yr. Approximately 38% of the LFN was harvested in clippings over 2 yr. Total leachate LFN recovery was 0.23% over the 2-yr period. Total recovery of LFN was 64 and 81% for the Spring and Fall treatments, respectively, suggesting volatile losses of N. Whether the N was applied in the spring or late fall, rapid uptake and immobilization of the LFN resulted. A maximum of 25% of applied LFN was recovered in the soil from either application timing at any time over the 2 yr of the experiment. A well-maintained turf intercepts and immobilizes LFN quickly making leaching an unlikely avenue of N loss from a turf system

(/sup 13/N)ammonia in organic solvents; a potent synthetic precursor for /sup 13/N-labeling

Energy Technology Data Exchange (ETDEWEB)

Tominaga, Toshiyoshi; Hirobe, Masaaki; Suzuki, Kazutoshi; Inoue, Osamu; Irie, Toshiaki; Yamasaki, Toshio

1987-01-01

/sup 13/NH/sub 3/ in an organic solvent was prepared and its utility as a labeling precursor was studied. (/sup 13/N)adenine ((/sup 13/N)ADN), (/sup 13/N)nicotinamide ((/sup 13/N)NAM), (/sup 13/N)p-nitrophenyl carbamate ((/sup 13/N)NPC), and (/sup 13/N)L-glutamine ((/sup 13/N)Gln) were labeled utilizing this precursor. (/sup 13/N)ADN and (/sup 13/N)NAM were labeled in much better yields than from an aqueous solution of /sup 13/NH/sub 3/. (/sup 13/N)NPC and (/sup 13/N)Gln, which could not be labeled in an aqueous solution, were labeled in high radiochemical yields. Thus, the advantages of this precursor are the improvement of the labeling yield and the feasibility of labeling compounds unstable in aqueous conditions.

Simple and Robust N-Glycan Analysis Based on Improved 2-Aminobenzoic Acid Labeling for Recombinant Therapeutic Glycoproteins.

Science.gov (United States)

Jeong, Yeong Ran; Kim, Sun Young; Park, Young Sam; Lee, Gyun Min

2018-03-21

N-glycans of therapeutic glycoproteins are critical quality attributes that should be monitored throughout all stages of biopharmaceutical development. To reduce both the time for sample preparation and the variations in analytical results, we have developed an N-glycan analysis method that includes improved 2-aminobenzoic acid (2-AA) labeling to easily remove deglycosylated proteins. Using this analytical method, 15 major 2-AA-labeled N-glycans of Enbrel ® were separated into single peaks in hydrophilic interaction chromatography mode and therefore could be quantitated. 2-AA-labeled N-glycans were also highly compatible with in-line quadrupole time-of-flight mass spectrometry (MS) for structural identification. The structures of 15 major and 18 minor N-glycans were identified from their mass values determined by quadrupole time-of-flight MS. Furthermore, the structures of 14 major N-glycans were confirmed by interpreting the MS/MS data of each N-glycan. This analytical method was also successfully applied to neutral N-glycans of Humira ® and highly sialylated N-glycans of NESP ® . Furthermore, the analysis data of Enbrel ® that were accumulated for 2.5 years demonstrated the high-level consistency of this analytical method. Taken together, the results show that a wide repertoire of N-glycans of therapeutic glycoproteins can be analyzed with high efficiency and consistency using the improved 2-AA labeling-based N-glycan analysis method. Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Use of 15N reverse gradient two-dimensional nuclear magnetic resonance spectroscopy to follow metabolic activity in Nicotiana plumbaginifolia cell-suspension cultures.

Science.gov (United States)

Mesnard, F; Azaroual, N; Marty, D; Fliniaux, M A; Robins, R J; Vermeersch, G; Monti, J P

2000-02-01

Nitrogen metabolism was monitored in suspension cultured cells of Nicotiana plumbaginifolia Viv. using nuclear magnetic resonance (NMR) spectroscopy following the feeding of (15NH4)2SO4 and K15NO3. By using two-dimensional 15N-1H NMR with heteronuclear single-quantum-coherence spectroscopy and heteronuclear multiple-bond-coherence spectroscopy sequences, an enhanced resolution of the incorporation of 15N label into a range of compounds could be detected. Thus, in addition to the amino acids normally observed in one-dimensional 15N NMR (glutamine, aspartate, alanine), several other amino acids could be resolved, notably serine, glycine and proline. Furthermore, it was found that the peak normally assigned to the non-protein amino-acid gamma-aminobutyric acid in the one-dimensional 15N NMR spectrum was resolved into a several components. A peak of N-acetylated compounds was resolved, probably composed of the intermediates in arginine biosynthesis, N-acetylglutamate and N-acetylornithine and, possibly, the intermediate of putrescine degradation into gamma-aminobutyric acid, N-acetylputrescine. The occurrence of 15N-label in agmatine and the low detection of labelled putrescine indicate that crucial intermediates of the pathway from glutamate to polyamines and/or the tobacco alkaloids could be monitored. For the first time, labelling of the peptide glutathione and of the nucleotide uridine could be seen.

Label Review Training: Module 1: Label Basics, Page 15

Science.gov (United States)

This module of the pesticide label review training provides basic information about pesticides, their labeling and regulation, and the core principles of pesticide label review. Learn about the consequences of improper labeling.

Synthesis and 125I labeling of N-succinimidyl-3-(tri-n-butylstannyl)benzoate

International Nuclear Information System (INIS)

Liu Zhenfeng; Wang Yongxian; Zhou Wei; Wang Lihua; Xia Jiaoyun; Yin Duanzhi

2005-01-01

N-succinimidyl-3-(tri-n-butylstannyl)benzoate (ATE) and N-succinimidyl-3-iodo-benzoate (SIB) is synthesized. The structures of ATE and SIB are confirmed with 1 HNMR, MS and IR. The yields of ATE and SIB are 45.4% and 71.4%, respectively. ATE is labeled with 125 I. The labeling field is 93.0% and radiochemical purity is over 98.0%. The synthesis and the labeling of ATE have a important value for indirect label of radiopharmaceuticals. (authors)

Effect of applying wheat stubble on preservation and utilization of n-fertilizer by 15N trace technique

International Nuclear Information System (INIS)

Xu Xinyu; Zhang Yumei; Xiang Hua; Hu Jisheng

1991-10-01

By using 15 N trace technique, the effect of applying wheat stubble on the preservation and utilization rate of 15 N- ammonium sulphate have been studied. The abundance of ( 15 NH 4 ) 2 SO 4 fertilizer was 8.92%. After three years pot test and field plot test, the results showed that the yields with ' 15 N+mulching' and ' 15 N+incorporating' treated were increased by 5.4∼30.0% for spring wheat and millet(pot test), and 18∼23% for winter wheat and summer corn(field plot test), as compared with only ' 15 N' treatment. The results of 15 N-fertilizer labelled tests showed that the utilization rates of 15 N-fertilizer treated by ' 15 N+mulching' for cropping seasons were 57.8%, 65.8%, 36.6% and 8.5% respectively. These were increased 3.7%, 10.2%, 21.5% and 2.8% as compared with only ' 15 N' treatment. Comparing with only ' 15 N'treatment, the N leached off by percolation water was decreasing 50%, the loss of N caused by volatilization was decreasing 30.3% and the N in humus was increasing 21.1%. All of these proved that the applying of wheat stubble in different mode would adjust and control the activation of microbe in the soil, and the preservation and utilization rate of fertilizer in the soul would be increased

Labeled Embedding Of (n, n-2-Graphs In Their Complements

Directory of Open Access Journals (Sweden)

Tahraoui M.-A.

2017-11-01

Full Text Available Graph packing generally deals with unlabeled graphs. In [4], the authors have introduced a new variant of the graph packing problem, called the labeled packing of a graph. This problem has recently been studied on trees [M.A. Tahraoui, E. Duchêne and H. Kheddouci, Labeled 2-packings of trees, Discrete Math. 338 (2015 816-824] and cycles [E. Duchˆene, H. Kheddouci, R.J. Nowakowski and M.A. Tahraoui, Labeled packing of graphs, Australas. J. Combin. 57 (2013 109-126]. In this note, we present a lower bound on the labeled packing number of any (n, n − 2-graph into Kn. This result improves the bound given by Woźniak in [Embedding graphs of small size, Discrete Appl. Math. 51 (1994 233-241].

Synthesis of tritium-labelled isopenicillin N, penicillin N and 6-aminopenicillanic acid

International Nuclear Information System (INIS)

Usher, J.J.; Loder, B.; Abraham, E.P.

1975-01-01

1. Phenoxymethylpenicillin sulphoxide 4-methoxybenzyl ester was labelled with 3 H in its 2-β-methyl group. Its specific radioactivity was 362mCi/mmol. 2. Removal of the side chain of this compound yielded the corresponding ester of 6-aminopenicillanic acid sulphoxide and coupling of the latter with the appropriate protected α-aminoadipic acid gave 4-methoxybenzyloxycarbonylisopenicillin N sulphoxide di-4-methoxybenzyl ester or the corresponding derivative of penicillin N. 3. Removal of the protective groups by hydrogenolysis and reduction of the sulphoxide group yielded 3 H-labelled isopenicillin N or penicillin N. 4. 3 H-labelled phenoxymethylpenicillin sulphoxide was obtained by hydrogenolysis from its 4-methoxybenzyl ester. Reducton of its sulphoxide group and subsequent removal of the side chain gave 3 H-labelled 6-aminopenicillanic acid. (author)

Absorption of ammonium sulphate 15N by coffee plants

International Nuclear Information System (INIS)

Fenilli, Tatiele Anete Bergamo; Reichardt, Klaus; Bacchi, Osny Oliveira Santos; Trivelin, Paulo Cesar Ocheuze; Dourado Neto, Durval

2005-01-01

The objective of this study was to quantify the absorption of ammonium sulphate 15 N by coffee plants. Treatments consisted of five sub-plots of 9 plants, of which the three central ones received 280 kg ha -1 of 15 N, applied at four times: 1/4 on 01 Set 03; 1/4 on 03 Nov 03; 1/4 on 15 Dec 03 and 1/4 on 30 Jan 04. The isotopic enrichment was 2,072 ± 0,001 atom % 15 N. The dry matter of the shoot was evaluated every 60 days, using one plant per replicate, collected outside the sub-plot. They were as similar as possible to the labeled plants, which were used only for isotopic and Total N analysis, after being dried at 65 deg C until constant weight. At harvest, plants had absorbed 42,88% of the fertilizer N. Leaves accumulated the largest amount of fertilizer N, and were also the compartments that received most N from other parts of the plant. The following partition of the fertilizer N was found at harvest: 23.01% in young leaves, 6.23% in old leaves, 4,46% in stem, 3.46% in fruits, 3.10% in young branches and 2.63% in old branches. (author)

/sup 15/N analysis in nutritional and metabolic research of infancy

Energy Technology Data Exchange (ETDEWEB)

Heine, W; Richter, I; Plath, C; Wutzke, K; Drescher, U [Rostock Univ. (German Democratic Republic). Bereich Medizin

1982-01-01

Investigation of protein metabolism in nutritional pediatric research by means of /sup 15/N tracer techniques has been relatively seldom used up to now. /sup 15/N-labelled compounds for these purposes are not injurious to health. The technique is based on oral or intravenous application of the tracer substances and on /sup 15/N analysis of the urine fractions. The subsequent calculation of protein synthesis and breakdown rate, turnover, and the reutilisation of amino acids from protein breakdown as well as the size of the metabolic pool offers detailed information of protein metabolism. Determination of these parameters were performed in infants on breast milk, formula feeding and on chemically defined diet. As an example of utilisation of D-amino acids for protein synthesis the /sup 15/N-D-phenylalanine retention of parenteral nutrition was found to be 33% of the applied doses at an average. An oral /sup 15/N-glycine loading test proved to be of value for the prediction of the therapeutic effect of human growth hormone. /sup 15/N tracer technique was also tested in utilizing /sup 15/N-urea for bacterial protein synthesis of the intestinal flora and by incorporation of /sup 15/N from /sup 15/N-glycine and /sup 15/N-lysine into the jejunal mucosa for measuring the enterocyte regeneration.

Metabolic regulation in Streptomyces parvulus during actinomycin D synthesis, studied with 13C- and 15N-labeled precursors by 13C and 15N nuclear magnetic resonance spectroscopy and by gas chromatography-mass spectrometry

International Nuclear Information System (INIS)

Inbar, L.; Lapidot, A.

1988-01-01

Recent studies have suggested that the onset of synthesis of actinomycin D in Streptomyces is due to a release from L-glutamate catabolic repression. In the present investigation we showed that S. parvulus has the capacity to maintain high levels of intracellular glutamate during the synthesis of actinomycin D. The results seem contradictory, since actinomycin D synthesis cannot start before a release from L-glutamate catabolic repression, but a relatively high intracellular pool of glutamate is needed for the synthesis of actinomycin D. Utilizing different labeled precursors, D-[U- 13 C]fructose and 13 C- and 15 N-labeled L-glutamate, and nuclear magnetic resonance techniques, we showed that carbon atoms of an intracellular glutamate pool of S. parvulus were not derived biosynthetically from the culture medium glutamte source but rather from D-fructose catabolism. A new intracellular pyrimidine derivative whose nitrogen and carbon skeletons were derived from exogenous L-glutamate was obtained as the main glutamate metabolite. Another new pyrimidine derivative that had a significantly reduced intracellular mobility and that was derived from D-fructose catabolism was identified in the cell extracts of S. parvulus during actinomycin D synthesis. These pyrimidine derivatives may serve as a nitrogen store for actinomycin D synthesis. In the present study, the N-trimethyl group of a choline derivative was observed by 13 C nuclear magnetic resonance spectroscopy in growing S. parvulus cells. The choline group, as well as the N-methyl groups of sarcosine, N-methyl-valine, and the methyl groups of an actinomycin D chromophore, arose from D-fructose catabolism. The 13 C enrichments found in the peptide moieties of actinomycin D were in accordance with a mechanism of actinomycin D synthesis from L-glutamate and D-fructose

Absorption of ammonium sulphate {sup 15}N by coffee plants; Recuperacao do {sup 15}N do sulfato de amonio por plantas de cafe

Energy Technology Data Exchange (ETDEWEB)

Fenilli, Tatiele Anete Bergamo; Reichardt, Klaus; Bacchi, Osny Oliveira Santos [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Fisica do Solo]. E-mail: tatiele@cena.usp.br; Trivelin, Paulo Cesar Ocheuze [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Isotopos Estaveis; Dourado Neto, Durval [Sao Paulo Univ., Piracicaba, SP (Brazil). Escola Superior de Agricultura Luiz de Queiroz. Dept. de Producao Vegetal

2005-07-01

The objective of this study was to quantify the absorption of ammonium sulphate {sup 15}N by coffee plants. Treatments consisted of five sub-plots of 9 plants, of which the three central ones received 280 kg ha{sup -1} of {sup 15}N, applied at four times: 1/4 on 01 Set 03; 1/4 on 03 Nov 03; 1/4 on 15 Dec 03 and 1/4 on 30 Jan 04. The isotopic enrichment was 2,072 {+-} 0,001 atom % {sup 15}N. The dry matter of the shoot was evaluated every 60 days, using one plant per replicate, collected outside the sub-plot. They were as similar as possible to the labeled plants, which were used only for isotopic and Total N analysis, after being dried at 65 deg C until constant weight. At harvest, plants had absorbed 42,88% of the fertilizer N. Leaves accumulated the largest amount of fertilizer N, and were also the compartments that received most N from other parts of the plant. The following partition of the fertilizer N was found at harvest: 23.01% in young leaves, 6.23% in old leaves, 4,46% in stem, 3.46% in fruits, 3.10% in young branches and 2.63% in old branches. (author)

Urinary excretion of 15N during intraportal infusion of 15N-ammonia in chickens fed low or high protein diet

International Nuclear Information System (INIS)

Karasawa, Yutaka; Koh, Katsuki; Takahashi, Akira; Sumiya, Ryuta

1985-01-01

The purpose of this study is to examine time courses of 15 N in urinary ammonia and total N when 15 N-labeled ammonium acetate was continuously infused for 1 hour into chickens fed a 5 or 20 % protein diet. 15 N-enrichment of urinary nitrogen in the two dietary groups increased sharply in ammonia for the first 20 minutes and to a less extent linearly in total N for the first 30 minutes, and then gradually in both ammonia and total N. Through the ammonia infusion, the 15 N-enrichment of urinary ammonia was higher in the chickens fed the low protein diet than in those fed the high protein diet; both of them were higher than 15 N-enrichments of urinary N, which were almost the same in the two dietary groups. The urinary total N from the infused ammonia rose linearly for the first 40 minutes but thereafter did not rise further in the two dietary groups, whereas the endogenous urinary total N tended to decrease a little in the chichens fed the high protein diet but unchanged in those fed the low protein diet. The urinary ammonia from the infused ammonia increased sharply for the first 20 minutes, then linearly but at a lower rate in the chickens fed the high protein diet, whereas that in the chickens fed the low protein diet rose linearly throughout ammonia infusion. In contrast, the endogenous urinary ammonia showed no change in the chickens fed the high protein diet while it showed a tendency to increase a little in these fed the low protein diet. These results indicate that the increased urinary ammonia and total N during ammonia infusion are derived mostly from the infused ammonia in chickens fed 5 and 20% protein diets. (author)

Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method

DEFF Research Database (Denmark)

Jørgensen, F.V.; Jensen, E.S.; Schjørring, J.K.

1999-01-01

Dinitrogen fixation in white clover (Trifolium repens L.) grown in pure stand and mixture with perennial ryegrass (Lolium perenne L.) was determined in the field using N-15 isotope dilution and harvest of the shoots. The apparent transfer of clover N to perennial ryegrass was simultaneously...... assessed. The soil was labelled either by immobilizing N-15 in organic matter prior to establishment of the sward or by using the conventional labelling procedure in which N-15 fertilizer is added after sward establishment. Immobilization of N-15 in the soil organic matter has not previously been used...

Fertilizer 15N balance and recovery of N from organic sources by rice in Typic Ustochrept

International Nuclear Information System (INIS)

Bhattacharyya, Ranjan; Sachdev, M.S.; Sachdev, P.; Kundu, S.; Sutradhar, G.

2002-01-01

To investigate the fertilizer-N balance and recovery of N from organics (as determined by A-value technique) by rice as affected by urea application alone or in combination with FYM or green manure, a field experiment was conducted in the khariff season if 1997 at IARI, New Delhi on a sandy loam soil (Typic Ustochrept). 15 N-labelled urea was applied at 0.90 and 120 kg N ha -1 levels alone and in combination with either FYM or green manure in 2:1 or 1:1 ratios. Organic sources were incorporated seven days before transplanting whereas, urea was applied in three equal splits at 15 DAT, 28 DAT and 42 DAT. The residual 15 N in soil was determined only in the surface soil layer (0-15 cm) of rice crop. The combined source helped in conserving more of urea-N in soil as residual (42-45%) than urea alone (23-27%) treatment due to the fact that the unaccounted fertilizer 15 N was more in urea alone treatment (43-45%) than combined sources (12-15%) at both the levels. The efficiency of uptake of organic N by rice as determined through A-value technique was similar or better than urea-N at both the levels. (author)

Absorption and translocation of 15N in Japonica (Hinohikari) and Indica (Hadsaduri) rice varieties

International Nuclear Information System (INIS)

Islam, N.; Inagaki, S.; Chishaki, N.; Horiguchi, T.

1997-01-01

The absorption and translocation of 15 N-labeled nitrogen (N) applied as three N levels of ammonium nitrate at the stages of panicle initiation (PI) and heading (HD) were compared between a japonica rice variety (var. Hinohikari) and a tall indica rice variety (var. Hadsaduri) by growing them hydroponically. With the supply of low N level, 15 N absorption by the japonica variety was larger, but at medium and high N levels, the tall indica variety absorbed larger amounts of 15 N at both stages. However, the amount of 15 N partitioned to the panicles at maturity was considerably smaller in the indica variety, since dry matter allocation to the panicles was also smaller in this variety. The tall indica variety showed a considerable loss of 15 N from heading to maturity at the high N-level unlike the japonica variety. (author)

Syntheses of sup 18 F-labeled reduced haloperidol and sup 11 C-labeled reduced 3-N-methylspiperone

Energy Technology Data Exchange (ETDEWEB)

Ravert, H T; Dannals, R F; Wilson, A A; Wong, D F; Wagner, Jr, H N [Johns Hopkins Medical Institutions, Baltimore, MD (USA)

1991-03-01

{sup 18}F-Labeled reduced haloperidol and {sup 11}C-labeled reduced 3-N-methylspiperone were synthesized in a convenient and quantitative one step reduction from {sup 18}F-labeled haloperidol and {sup 11}C-labeled N-methylspiperone, respectively. Both products were purified by semipreparative HPLC and were obtained at high specific activity and radiochemical purity. (author).

New series of Tc-99m-labeled hepatobiliary tracers: N'-acyl- and N'-sulfonyl ethylenediamine-N,N-diacetic acids

International Nuclear Information System (INIS)

Karube, Y.; Kono, A.; Maeda, T.; Ohya, M.; Matsushima, Y.

1981-01-01

Various Tc-99m-labeled N'-substituted derivatives of ethylenediamine-N,N-diacetic acid (EDDA) are evaluated as hepatobiliary imaging agents. N-substituted aromatic acyl and aromatic sulfonyl derivatives of EDDA, labeled with Tc-99m, were administered to rabbits and golden hamsters, and the distribution indicated clearance by the hepatobiliary system. N'-aromatic sulfonyl EDDAs were labeled with Tc-99m by the SnCl 2 method with more than 99% yield. Clearance of Tc-99m-p-toluenesulfonyl EDDA from the blood and the liver was as rapid as that of TC-99m N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid (Tc-99m benzenesulfonyl EDDA lowered urinary excretion. It is concluded that the sulfonyl EDDAs provide a fruitful source for Tc-99m-labeled hepatobiliary radiopharmaceuticals

15N and 13C abundances in marine environments with emphasis on biogeochemical structure of food networks

International Nuclear Information System (INIS)

Wada, E.

1987-01-01

Distributions of δ 15 N and δ 13 C for biogenic substances in the Antarctic Ocean and in the Otsuchi River estuary in Japan were investigated to construct isotope biogeochemical framework for assessing marine ecosystems. The isotopic compositions of phytoplankton were particularly low in the Antarctic Ocean. High nitrate and CO 2 concentrations in the surface sea waters, and the low light intensity seem to enhance the kinetic isotope fractionations that preferred the depletion of 15 N and 13 C in the algal body. A clear-cut linear relationship between animal δ 15 N and its trophic level was obtained in the Antarctic system. In the estuary, the variation of isotope ratios were principally governed by the mixing of land-derived organic matter, marine phytoplankton, and seagrasses. A food-chain effect of 15 N enrichment was also confirmed. An isotopically ordered structure was presented for a marine estuarine ecosystem. The isotopic abundances in a food network vary mainly because of the variation in 15 N and 13 C contents of primary producers grown under different environmental conditions and because of the enrichment of 15 N along food chains. (author)

Use of 15N in evaluating symbiotic N2 fixation of field-grown soybeans

International Nuclear Information System (INIS)

Ham, G.E.

1978-01-01

Various methods have been used to estimate N 2 fixation by legumes (i.e. Kjeldahl N and the acetylene-ethylene assay). Recently 'Asub(N)' values by the legume and a non-nodulating crop using 15 N-labelled N fertilizer were used to quantitatively estimate the amount of N 2 fixed by legume crops growing under field conditions. The objective of this research was to evaluate Kjeldahl N procedures, the acetylene-ethylene assay and the 'Asub(N)' technique as estimators of N 2 fixation by field-grown soybeans. The 'Asub(N)' value concept provided a reliable estimate of N 2 fixation by soybeans which agreed with acetylene-ethylene measurements made weekly and the values compared favourably with Kjeldahl N measurements. (author)

Study of the production yields of {sup 18}F, {sup 11}C, {sup 13}N and {sup 15}O positron emitters from plasma-laser proton sources at ELI-Beamlines for labeling of PET radiopharmaceuticals

Energy Technology Data Exchange (ETDEWEB)

Amato, Ernesto [Section of Radiological Sciences, Department of Biomedical and Dental Sciences and of Morphologic and Functional Imaging, University of Messina (Italy); Italiano, Antonio, E-mail: italianoa@unime.it [Istituto Nazionale di Fisica Nucleare, Gruppo Collegato di Messina (Italy); Margarone, Daniele [Institute of Physics ASCR, v.v.i. (FZU), ELI-Beamlines Project, 182 21 Prague (Czech Republic); Pagano, Benedetta [Nuclear Medicine Unit, University Hospital “G. Martino”, Messina (Italy); Baldari, Sergio [Section of Radiological Sciences, Department of Biomedical and Dental Sciences and of Morphologic and Functional Imaging, University of Messina (Italy); Nuclear Medicine Unit, University Hospital “G. Martino”, Messina (Italy); Korn, Georg [Institute of Physics ASCR, v.v.i. (FZU), ELI-Beamlines Project, 182 21 Prague (Czech Republic)

2016-03-01

The development of novel compact PET radionuclide production systems is of great interest to promote the diffusion of PET diagnostics, especially in view of the continuous development of microfluidics labeling approaches. We studied the feasibility to produce clinically-relevant amounts of PET isotopes by means of laser-accelerated proton sources such that expected at the ELI-Beamlines facility. {sup 18}F, {sup 11}C, {sup 13}N and {sup 15}O production yields were calculated through the TALYS software, by taking into account the broad proton spectra expected. With the hypothesized proton fluencies, clinically-relevant amounts of radionuclides can be obtained, suitable to prepare single doses of {sup 18}F-, {sup 11}C- and {sup 13}N-labeled radiopharmaceuticals exploiting fast and efficient microfluidic labeling systems.

Study of the production yields of "1"8F, "1"1C, "1"3N and "1"5O positron emitters from plasma-laser proton sources at ELI-Beamlines for labeling of PET radiopharmaceuticals

International Nuclear Information System (INIS)

Amato, Ernesto; Italiano, Antonio; Margarone, Daniele; Pagano, Benedetta; Baldari, Sergio; Korn, Georg

2016-01-01

The development of novel compact PET radionuclide production systems is of great interest to promote the diffusion of PET diagnostics, especially in view of the continuous development of microfluidics labeling approaches. We studied the feasibility to produce clinically-relevant amounts of PET isotopes by means of laser-accelerated proton sources such that expected at the ELI-Beamlines facility. "1"8F, "1"1C, "1"3N and "1"5O production yields were calculated through the TALYS software, by taking into account the broad proton spectra expected. With the hypothesized proton fluencies, clinically-relevant amounts of radionuclides can be obtained, suitable to prepare single doses of "1"8F-, "1"1C- and "1"3N-labeled radiopharmaceuticals exploiting fast and efficient microfluidic labeling systems.

Investigations on the production of labelled organic compounds by recoil labelling with gamma,n-produced 11-C-atoms

International Nuclear Information System (INIS)

Wagenbach, U.

1981-01-01

''Hot'' 11 C atoms are produced from 12 C(γ,n) 11 C nuclear reactions by bremsstrahlung at the 65 MeV electron linear accelerator in Giessen. The relative retention in various C-atoms of the amino acid, methionine, is determined by splitting of the terminal C-atoms of the molecule and by independent determination of the content of 11 C in the isolated and derived fragments. The terminal groups (thiomethyl or carboxyl groups) each carry approx. 25% of the total retained radioactivity, the remaining 50% being spread over the three inner carbon atoms. The activation of alkylamines, crystallised as hydrochlorides, hydrofluorides, oxalates and sulphates, leads to similar yields of direct labelling from 5 to 15%. Amines activated in the liquid state show a retention of less than 5%. The yields for labelled synthetic products are between 10 and 15% for amino acids and are often higher for crystallised amines. Amines activated in the liquid state produced greater yields of synthesis products but at the same time an increase in the product range. The labelled synthesis products can be separated faster by suitable methods such as preparative HPLC and are then available for carrier-free studies in the life sciences. (orig./EF) [de

15N-enrichments of ammonia and glutamine in blood after infusion of 15N-ammonia in chickens fed low or high protein diet

International Nuclear Information System (INIS)

Karasawa, Yutaka; Koh, Katsuki

1985-01-01

In this experment, the blood ammonia and glutamine amide came from infused ammonia were determined when N-15 labeled ammonium acetate was intraportally infused into the chickens fed 5 or 20 % protein diet. The data obtained indicated that the infused ammonia was taken into blood glutamine amide, and also accumulated in blood as it is, in both dietary groups. 10 to 12 months old White Leghorn male birds were used. The experimental diet was fed once a day for 5 days to the birds weighting about 1.2 kg by 35 g per kg body weight. The experimental diet was consumed within 40 min in all cases. Cardiac and portal catheterization were performed for blood collection and ammonia infusion, respectively. After finishing the infusion, blood samples were taken to analyze the ammonia and glutamine contents and their N-15 enrichment. Statistical difference was not observed in the appearance of N-15 in ammonia and glutamine amide between two dietary groups. The N-15 enrichment in blood ammonia and the amide of plasma glutamine, and the calculated exogenous nitrogen in the ammonia and glutamine amide tended to be more in the 5 % protein diet group than the other. (Kako, I.)

Does fertilizer (N15P15K15) amendment enhance phytoremediation of petroleum-polluted aquatic ecosystem in the presence of water hyacinth (Eichhornia crassipes [Mart.] Solms)?

Science.gov (United States)

Ndimele, Prince Emeka; Jenyo-Oni, Adetola; Chukwuka, Kanayo S; Ndimele, Chinatu Charity; Ayodele, Ibukunoluwa Augustine

2015-01-01

This study investigated the effects of inorganic fertilizer (N15P15K15) amendments on crude oil uptake by water hyacinth. Experimental units (water hyacinth grown in fresh water) were spiked with 0, 20, 40 and 60 mg/L crude oil. After 24 h, they were randomly assigned fertilizer (N15P15K15) at three different concentrations; 0, 6 and 10 mg/L. Crude oil degradation and absorption were determined by measuring total petroleum hydrocarbon (TPH) in the water column and water hyacinth, respectively. The measurements were taken monthly for six months (February-August 2010). The results showed that TPH concentration in the water column in the treatment amended at 6 mg/L (0.30 ± 0.01 mg/L) was significantly lower (p phytoremediation) absorbed significantly higher (p phytoremediation of crude oil by water hyacinth and biostimulation with fertilizer (N15P15K15) is possible.

New series of Tc-99m-labeled hepatobiliary tracers: N'-acyl- and N'-sulfonyl ethylenediamine-N,N-diacetic acids

International Nuclear Information System (INIS)

Karube, Y.; Kono, A.; Maeda, T.; Ohya, M.; Matsushima, Y.

1981-01-01

Various Tc-99m-labeled N'-substituted derivatives of ethylenediamine-N,N-diacetic acid (EDDA) are evaluated as hepatobiliary imaging agents. N'-substituted aromatic acyl and aromatic sulfonyl derivatives of EDDA, labeled with Tc-99m, were administered to rabbits and golden hamsters, and the distribution indicated clearance by the hepatobiliary system. N'-aromatic sulfonyl EDDAs were labeled with Tc-99m by the SnCl 2 method with more than 99% yield. Clearance of Tc-99m-p-toluenesulfonyl EDDA from the blood and the liver was as rapid as that of Tc-99m N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid (Tc-99m diethyl IDA). Substitution of a bulky group at the aromatic ring in Tc-99m benzene-sulfonyl EDDA lowered urinary excretion. It is concluded that the sulfonyl EDDAs provide a fruitful source for Tc-99m-labeled hepatobiliary radiopharmaceuticals

Imprint of oaks on nitrogen availability and δ15N in California grassland-savanna: A case of enhanced N inputs?

Science.gov (United States)

Perakis, S.S.; Kellogg, C.H.

2007-01-01

Woody vegetation is distributed patchily in many arid and semi-arid ecosystems, where it is often associated with elevated nitrogen (N) pools and availability in islands of fertility. We measured N availability and δ15N in paired blue-oak versus annual grass dominated patches to characterize the causes and consequences of spatial variation in N dynamics of grassland-savanna in Sequoia-Kings Canyon National Park. We found significantly greater surface soil N pools (0–20 cm) in oak patches compared to adjacent grass areas across a 700 m elevation gradient from foothills to the savanna-forest boundary. N accumulation under oaks was associated with a 0.6‰ depletion in soil δ15N relative to grass patches. Results from a simple δ15N mass balance simulation model, constrained by surface soil N and δ15N measured in the field, suggest that the development of islands of N fertility under oaks can be traced primarily to enhanced N inputs. Net N mineralization and percent nitrification in laboratory incubations were consistently higher under oaks across a range of experimental soil moisture regimes, suggesting a scenario whereby greater N inputs to oak patches result in net N accumulation and enhanced N cycling, with a potential for greater nitrate loss as well. N concentrations of three common herbaceous annual plants were nearly 50% greater under oak than in adjacent grass patches, with community composition shifted towards more N-demanding species under oaks. We find that oaks imprint distinct N-rich islands of fertility that foster local feedback between soil N cycling, plant N uptake, and herbaceous community composition. Such patch-scale differences in N inputs and plant–soil interactions increase biogeochemical heterogeneity in grassland-savanna ecosystems and may shape watershed-level responses to chronic N deposition.

Growth, development, and fertilizer-15N recovery by the coffee plant

International Nuclear Information System (INIS)

Fenilli, Tatiele Anete Bergamo; Reichardt, Klaus; Bacchi, Osny Oliveira Santos; Dourado-Neto, Durval; Favarin, Jose Laercio; Trivelim, Paulo Cesar Ocheuze; Costa, Flavio Murilo Pereira da

2007-01-01

The relationship between growth and fertilizer nitrogen recovery by perennial crops such as coffee is poorly understood and improved understanding of such relations is important for the establishment of rational crop management practices. In order to characterize the growth of a typical coffee crop in Brazil and quantify the recovery of 15 N labeled ammonium sulfate, and improve information for fertilizer management practices this study presents results for two consecutive cropping years, fertilized with 280 and 350 kg ha -1 of N, respectively, applied in four splittings, using five replicates. Shoot dry matter accumulation was evaluated every 60 days, separating plants into branches, leaves and fruits. Labeled sub-plots were used to evaluate N-total and 15 N abundance by mass spectrometry. During the first year the aerial part reached a recovery of 71% of the fertilizer N applied up to February, but this value was reduced to 34% at harvest and 19% at the beginning of the next flowering period due to leaf fall and fruit export. For the second year the aerial part absorbed 36% of the fertilizer N up to March, 47% up to harvest and 19% up to the beginning of the next flowering period. The splitting into four applications of the used fertilizer rates was adequate for the requirements of the crop at these growth stages of the coffee crop. (author)

Quantitative estimates of uptake and internal cycling of 15N-depleted fertilizer in mature walnut trees

International Nuclear Information System (INIS)

Weinbaum, S.; Kessel, C. van

1998-01-01

In mature fruit trees, internal recycling is an important source of N for the growth of new wood, leaves and fruits. Using 15 N-depleted fertilizer, i.e. 14 N-enriched, N-uptake efficiency and the magnitude of internal N cycling were studied in mature walnut trees. Two kg of 14 N-labelled ammonium sulfate N were applied per tree, and compartmentation of N was followed over a period of 6 years by analyzing catkins, pistillate flowers, leaves and fruits each year for total N content and isotopic composition. Subsequently, two of the six labelled trees were excavated and analyzed for labelled-N content. The data indicate that mature walnut uses most of the N accumulated from soil and fertilizer for storage purposes, to be remobilized for new growth within 2 years, and about half of the total-N pool in a mature tree is present as non-structural compounds, available for recycling. (author)

Infrared and Raman spectra of uric acid and its 15N and D labelled compounds

International Nuclear Information System (INIS)

Majoube, Michel

Infrared and Raman spectra of polycrystalline uric acid (2, 6, 8-trioxypurine) 1.3, 7 and 9- 15 N and deuterated analogues have been determined. Band shifts with 15 N substitution and with deuteration are discussed. An assignment of fundamental vibrations of uric acid is proposed from the comparison of the eight isotopically substituted analogues [fr

Reconstructing Century-Scale Changes in Nitrogen Cycling in Forests Throughout the United States using Tree-Ring δ15N Chronologies

Science.gov (United States)

Gerhart-Barley, L.; McLauchlan, K. K.; Battles, J. J.; Craine, J. M.; Higuera, P. E.; Mack, M. C.; McNeil, B. E.; Nelson, D. M.; Pederson, N.; Perakis, S. S.

2016-12-01

In recent decades, human perturbation of the global nitrogen (N) cycle has been immense with reactive nitrogen supply to ecosystems from anthropogenic sources now exceeding that of natural fixation. The impact of these perturbations on ecosystem nutrient cycling and plant communities is limited by the lack of long-term `baseline' assessments of N cycling prior to anthropogenic influences. Stable N isotope analysis (δ15N) of dendrochronological records have the potential to provide this baseline data, but to date have focused on short term, regional assessments. Here, we address this question with a data set incorporating 311 individual trees and 7,661 δ15N measurements from 50 sites throughout the contiguous United States. These sites represent the diversity of US forest types, climate conditions, N deposition, soil types, and disturbance histories. The chronologies span, on average, the last 162 calendar years, with the oldest chronology dating back to 1572 C.E. Consequently, this study is the first century- and continental-scale assessment of ecosystem N cycling using tree-ring chronologies. When aggregated, the chronologies show a consistent decline from 1825 C.E. to present, indicating declining N availability in US forests, despite global increases in N supply. Environmental factors such as mean annual precipitation (MAP), mean annual temperature (MAT), and mean annual nitrogen deposition (Ndep) did not contribute to average site δ15N values; however, MAP and MAT significantly affected temporal trajectories in tree-ring δ15N, with more negative slopes toward present occurring in regions with low MAT and high MAP. Quantity of atmospheric N deposition had no discernible impact on mean δ15N values or on the temporal slope. This lack of response is either because levels of N deposition are too low to produce a discernible response in any meaningful aspects of the N cycle, and/or the δ15N signature of depositional N is similar enough to ecosystem N pools that

Evaluation of automated analysis of 15N and total N in plant material and soil

DEFF Research Database (Denmark)

Jensen, E.S.

1991-01-01

Simultaneous determination of N-15 and total N using an automated nitrogen analyser interfaced to a continuous-flow isotope ratio mass spectrometer (ANA-MS method) was evaluated. The coefficient of variation (CV) of repeated analyses of homogeneous standards and samples at natural abundance...... was lower than 0.1%. The CV of repeated analyses of N-15-labelled plant material and soil samples varied between 0.3% and 1.1%. The reproducibility of repeated total N analyses using the automated method was comparable to results obtained with a semi-micro Kjeldahl procedure. However, the automated method...... analysis showed that the recovery of inorganic N in the NH3 trap was lower when the N was diffused from water than from 2 M KCl. The results also indicated that different proportions of the NO3- and the NH4+ in aqueous solution were recovered in the trap after combined diffusion. The method is most suited...

High retention of ¹⁵N-labeled nitrogen deposition in a nitrogen saturated old-growth tropical forest

DEFF Research Database (Denmark)

Gurmesa, Geshere Abdisa; Lu, Xiankai; Gundersen, Per

2016-01-01

ecosystem compartments were quantified 4 months after the last addition. Tracer recoveries in soil solution were monitored monthly to quantify leaching losses. Total tracer recovery in plant and soil (N retention) in the control plots was 72% and similar to those observed in temperate forests. The retention...

Measurement of undisturbed di-nitrogen emissions from aquatic ecosystems

Science.gov (United States)

Qin, Shuping, Clough, Timothy, Lou, Jiafa; Hu, Chunsheng; Oenema, Oene; Wrage-Mönnig, Nicole; Zhang, Yuming

2016-04-01

Increased production of reactive nitrogen (Nr) from atmospheric di-nitrogen (N2) during the last century has greatly contributed to increased food production1-4. However, enriching the biosphere with Nr through N fertilizer production, combustion, and biological N2 fixation has also caused a series of negative effects on global ecosystems 5,6, especially aquatic ecosystems7. The main pathway converting Nr back into the atmospheric N2 pool is the last step of the denitrification process, i.e., the reduction of nitrous oxide (N2O) into N2 by micro-organisms7,8. Despite several attempts9,10, there is not yet an accurate, fast and direct method for measuring undisturbed N2 fluxes from denitrification in aquatic sediments at the field scale11-14. Such a method is essential to study the feedback of aquatic ecosystems to Nr inputs1,2,7. Here we show that the measurement of both N2O emission and its isotope signature can be used to infer the undisturbed N2 fluxes from aquatic ecosystems. The microbial reduction of N2O increases the natural abundance of 15N-N2O relative to 14N-N2O (δ15N-N2O). We observed linear relationships between δ15N-N2O and the logarithmic transformed N2O/(N2+N2O) emission ratios. Through independent measurements, we verified that the undisturbed N2 flux from aquatic ecosystems can be inferred from measurements of N2O emissions and the δ15N-N2O signature. Our method allows the determination of field-scale N2 fluxes from undisturbed aquatic ecosystems, and thereby allows model predictions of denitrification rates to be tested. The undisturbed N2 fluxes observed are almost one order of magnitude higher than those estimated by the traditional method, where perturbation of the system occurs, indicating that the ability of aquatic ecosystems to remove Nr may have been severely underestimated.

Fate of 15N-labelled urea fertilizer under conditions of tropical flooded-rice culture

International Nuclear Information System (INIS)

Krishnappa, A.M.; Shinde, J.E.

1980-01-01

The fate of an initial pulse of 15 N urea (at the rate of 100 kg N.ha -1 ) was followed under conditions of tropical flooded-rice culture over a sequence of three crops and two intercrop fallows. The total crop recovery accounted for 24.3% of the added fertilizer nitrogen. Ammonia volatilization and leaching losses amounted to 9.7% and 7.5%, respectively. The major losses of the fertilizer nitrogen occurred during the crop season immediately following its application. At the end of the experiment, 26.5% of the fertilizer nitrogen was recovered in the root zone in the Kjeldahl fraction and 0.9% as clay-fixed, non-exchangeable ammonium-N. Total recovery thus amounted to about 69%. The maximum contribution of the 15 N pulse to the NO 3 -N content of the groundwater (about 2%) occurred in the first crop season. It had declined below 0.2% by the third crop season. Throughout the experimental period the total NO 3 -N concentration of the groundwater never exceeded 3.2 ppm. (author)

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

Science.gov (United States)

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

2007-10-01

Utilization of long-range (1)H--(15)N heteronuclear chemical shift correlation has continually grown in importance since the first applications were reported in 1995. More recently, indirect covariance NMR methods have been introduced followed by the development of unsymmetrical indirect covariance processing methods. The latter technique has been shown to allow the calculation of hyphenated 2D NMR data matrices from more readily acquired nonhyphenated 2D NMR spectra. We recently reported the use of unsymmetrical indirect covariance processing to combine (1)H--(13)C GHSQC and (1)H--(15)N GHMBC long-range spectra to yield a (13)C--(15)N HSQC-HMBC chemical shift correlation spectrum that could not be acquired in a reasonable period of time without resorting to (15)N-labeled molecules. We now report the unsymmetrical indirect covariance processing of (1)H--(13)C GHMBC and (1)H--(15)N IMPEACH spectra to afford a (13)C--(15)N HMBC-IMPEACH spectrum that has the potential to span as many as six to eight bonds. Correlations for carbon resonances long-range coupled to a protonated carbon in the (1)H--(13)C HMBC spectrum are transferred via the long-range (1)H--(15)N coupling pathway in the (1)H--(15)N IMPEACH spectrum to afford a much broader range of correlation possibilities in the (13)C--(15)N HMBC-IMPEACH correlation spectrum. The indole alkaloid vincamine is used as a model compound to illustrate the application of the method. (c) 2007 John Wiley & Sons, Ltd.

Protocol development for continuous nitrogen-15 measurement of N2O and its isotopomers for real-time greenhouse gas tracing