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PubMed1980

Nitrogen Nutrition and Xylem Transport of Nitrogen in Ureide-producing Grain Legumes.

Xylem sap composition was examined in nodulated and nonnodulated cowpea (Vigna unguiculata [L.] Walp.) plants receiving a range of levels of NO(3) and in eight other ureide-forming legumes utilizing NO(3) or N(2) as sole source of nitrogen. A (15)N dilution technique determined the proportions of plant nitrogen derived from N(2) in the nodulated cowpeas fed NO(3). Xylem sap composition of NO(3)-fed, nodulated cowpea varied predictably with the relative extents to which N(2) and NO(3) were being utilized. The ratios of asparagine to glutamine (N/N) and of NO(3) to ureide (N/N) in xylem sap increased with increasing dependence on NO(3) whereas per cent of xylem nitrogen as ureide and the ratio of ureide plus glutamine to asparagine plus NO(3) (N/N) in xylem sap increased with increasing dependence on N(2) fixation. The amounts of NO(3) and ureides stored in leaflets, stems plus petioles, and roots of cowpea varied in a complex manner with level of NO(3) and the presence or absence of N(2) fixation. All species showed higher proportions of organic nitrogen as ureide and several-fold lower ratios of asparagine to glutamine in their xylem sap when relying on N(2) than when utilizing NO(3). In nodulated (minus nitrate) cowpea and mung bean (Vigna radiata [L.] Wilczek) the percentage of xylem nitrogen as ureide remained constant during growth but the ratio of asparagine to glutamine varied considerably. The biochemical significance of the above differences in xylem sap composition was discussed.

Journal Article↗
PubMed1995

Effect of nitrogen fixation, nitrogen fertilization and viral infection on yield, tannin and protein contents and in vitro protein digestibility of faba bean.

A field investigation of two faba bean cultivars (cv.), Agabat and Silaim, showed that bean yellow mosaic virus (BYMV) infection reduced (p < or = 0.001) yield (Kg/ha), protein content and in vitro protein digestibility (IVPD) but increased (p < or = 0.05) tannin content (mg/100 ml). Nitrogen fertilization with viral infection significantly reduced yield and IVPD for cv. Silaim and increased (p < or = 0.05) protein and tannin contents. Nitrogen fertilization alone was found to increase (p < or = 0.05) yield, protein and tannin contents but slightly reduced IVPD. Rhizobium inoculation with viral infection significantly decreased yield per unit area, protein content and IVPD, but increased (p < or = 0.05) tannin content. Rhizobium inoculation alone significantly increased (p < or = 0.001) yield and tannin content and slightly increased protein content but decreased IVPD. The results indicated that nitrogen fertilization or nitrogen fixation increased yield, protein and tannin contents and decreased IVPD. Viral infection had an adverse effect on yield, protein content and IVPD but had no effect on tannin content.

Digestion↗
PubMed2005

Sucrose synthase and RuBisCo expression is similarly regulated by the nitrogen source in the nitrogen-fixing cyanobacterium Anabaena sp.

In higher plants and cyanobacteria, sucrose (Suc) metabolism is carried out by a similar set of enzymes. The function and regulation of Suc metabolism in cyanobacteria has begun to be elucidated. In strains of Anabaena sp., filamentous nitrogen-fixing cyanobacteria, Suc synthase (SuS, EC 2.4.1.13) controls Suc cell level through the cleavage of the disaccharide. The present work shows that there are two sus genes in Anabaena (Nostoc) sp. that are co-regulated regarding the nitrogen source; however, only susA accounts for the extractable SuS activity and for the control of the Suc level. Primer extension analysis has uncovered the sequence of the Anabaena susA and susB ammonium-activated putative promoters, which share a high sequence similarity with that of rbcLS encoding ribulose bisphosphate carboxylase/oxygenase (EC 4.1.1.39) and other ammonium up-regulated genes. Moreover, susA and rbcLS expression is developmentally co-localized to the vegetative cells of the nitrogen-fixing cyanobacterial filaments. Our results strongly suggest the existence of a regulatory network that would coordinate the expression of key genes for Suc and nitrogen metabolism, carbon fixation, and development in Anabaena sp.

Anabaena↗
PubMed2005

Association between blood plasma urea nitrogen levels and reproductive fluid urea nitrogen and ammonia concentrations in early lactation dairy cows.

Two experiments were conducted to study the relationship of blood plasma urea nitrogen (PUN) concentrations with NH3, urea nitrogen, K, Mg, P, Ca, and Na concentrations in fluid of preovulatory follicles (experiment 1) and the relationships of PUN concentration and stage of estrus cycle with ammonia and urea nitrogen concentrations in uterine fluids (experiment 2) in early lactation dairy cows. Mean PUN levels were used to distribute cows into two groups: cows with PUN>or=20 mg/dl (HPUN), and cows with PUN<20 mg/dl (LPUN). In experiment 1, blood and follicular fluids from preovulatory follicles of 38 early lactation dairy cows were collected on the day of estrus (day 0) 4h after feed was offered. Follicular fluid NH3 was higher (P<0.01) in HPUN cows (339.0 micromol/L+/-72.2) compared to LPUN cows (93.9 micromol/L+/-13.1). Follicular fluid urea N was higher (P<0.001) in HPUN cows (22.4 mg/dl+/-0.4) compared to LPUN cows (17.0 mg/dl+/-0.3). PUN and follicular fluid urea N were correlated (r2=0.86) within cows. In experiment 2, blood and uterine fluids were collected from 30 cows on day 0 and on day 7. Uterine fluid NH3 was higher (P=0.05) in HPUN cows (1562 micromol/L+/-202) than in LPUN cows (1082 micromol/L+/-202) on day 7, but not on day 0. Uterine fluid urea N was higher (P<0.001) in HPUN cows than in LPUN cows on day 0 (26.9 mg/dl+/-1.3 and 20.4 mg/dl+/-0.7) and day 7 (26.5 mg/dl+/-1.1 and 21.4 mg/dl+/-1.1). There was a correlation (r2=0.17) between PUN and uterine fluid urea N within cows. The results of this study indicate that high PUN concentrations were associated with elevated NH3 and urea N concentrations in the preovulatory follicular fluids on the day of estrus and in the uterine fluid during the luteal phase of the estrous cycle in early lactation dairy cows. Elevated NH3 or urea N concentrations in the reproductive fluids may contribute to reproductive inefficiency in dairy cows with elevated plasma urea nitrogen due to embryo toxicity.

Ammonia↗
PubMed2005

Nitrogen assimilation and nitrogen control in cyanobacteria.

Nitrogen sources commonly used by cyanobacteria include ammonium, nitrate, nitrite, urea and atmospheric N(2), and some cyanobacteria can also assimilate arginine or glutamine. ABC (ATP-binding cassette)-type permeases are involved in the uptake of nitrate/nitrite, urea and most amino acids, whereas secondary transporters take up ammonium and, in some strains, nitrate/nitrite. In cyanobacteria, nitrate and nitrite reductases are ferredoxin-dependent enzymes, arginine is catabolized by a combination of the urea cycle and arginase pathway, and urea is degraded by a Ni(2+)-dependent urease. These pathways provide ammonium that is incorporated into carbon skeletons through the glutamine synthetase-glutamate synthase cycle, in which 2-oxoglutarate is the final nitrogen acceptor. The expression of many nitrogen assimilation genes is subjected to regulation being activated by the nitrogen-control transcription factor NtcA, which is autoregulatory and whose activity appears to be influenced by 2-oxoglutarate and the signal transduction protein P(II). In some filamentous cyanobacteria, N(2) fixation takes place in specialized cells called heterocysts that differentiate from vegetative cells in a process strictly controlled by NtcA.

Cyanobacteria↗
PubMed1993

Exogenous and endogenous contributions to nitrogen fluxes in the digestive tract of pigs fed a casein diet. III. Recycling of endogenous nitrogen.

The aim of the present study was to measure the incorporation of infused 15N in blood fractions, urine, digesta, faeces and in the exocrine pancreatic and biliary secretions, in order to estimate the endogenous part of nitrogen in the ileal digesta and in the faeces of pigs fed a casein diet and to calculate the total endogenous nitrogen secretion as well as its recycling in the digestive tract. For 8 d 11 Large White female pigs (50.1 +/- 1.8 kg) received a continuous infusion of L-[15N]leucine via a catheter in the jugular vein. The 15N-enrichment was measured in several fractions. The 15N-level of the pancreatic juice was higher than that in the biliary secretion, TCA-blood fractions, and urine during the whole experimental period. Using the 15N-isotope dilution method it was found that casein was completely digested up to the terminal ileum and that all the nitrogen in the ileal digesta was of endogenous origin. The total endogenous secretion was estimated at approximately 11 g N/d. The reabsorption of endogenous nitrogen amounted to 79% up to the end of the small intestine and 88% over the whole digestive tract.

Absorption↗
PubMed2003

Nitrogen comes down to earth: report from the 5th European Nitrogen Fixation Conference.

For four days and four nights, with almost 50 presentations and more than 175 posters, the 5th European Nitrogen Fixation Conference continued a tradition of excellence, bringing scientists from diverse fields such as microbiology, biochemistry, computational genomics, and plant physiology together to address the complex problems associated with biological nitrogen fixation (BNF). The conference was hosted by the John Innes Center and the University of East Anglia in Norwich, England and took place from September 6 through 10, 2002. A diverse range of topics was presented, from the evolution of rhizobial genomes to the plant genes involved in bacterial and fungal symbiosis, to the structure of nitrogenase, and to the means by which nitrogen is shuttled between the symbiotic bacteria and the plant. Additionally, sessions involving broader issues, such as nitrogen fertilizer use and work being done in developing countries, brought home the importance of the research being carried out in BNF around the world.

Nitrogen↗
PubMed1999

The global nitrogen regulator NtcA regulates transcription of the signal transducer PII (GlnB) and influences its phosphorylation level in response to nitrogen and carbon supplies in the Cyanobacterium synechococcus sp. strain PCC 7942.

The PII protein is encoded by a unique glnB gene in Synechococcus sp. strain PCC 7942. Its expression has been analyzed in the wild type and in NtcA-null mutant cells grown under different conditions of nitrogen and carbon supply. RNA-DNA hybridization experiments revealed the presence of one transcript species 680 nucleotides long, whatever the nutrient conditions tested. A second transcript species, 620 nucleotides long, absent in the NtcA null mutant, was observed in wild-type cells that were nitrogen starved for 2 h under both high and low CO2 and in the presence of nitrate under a high CO2 concentration. Primer extension analysis indicated that the two transcript species are generated from two tandem promoters, a sigma70 Escherichia coli-type promoter and an NtcA-dependent promoter, located 120 and 53 nucleotides, respectively, from the glnB initiation codon. The NtcA-dependent promoter is up-regulated under the conditions mentioned above, while the sigma70 E. coli-type promoter displays constitutive levels of transcripts in the NtcA null mutant and slightly different levels in the wild-type cells, depending on the nitrogen and carbon supplies. In general, a good correlation between the amounts of the two transcript species and that of the PII protein was observed, as revealed by immunodetection with specific antibodies. The phosphorylation level of PII in the wild type is inversely correlated with nitrogen availability and directly correlated with higher CO2 concentration. This regulation is correspondingly less stringent in the NtcA null mutant cells. In contrast, the dephosphorylation of PII is NtcA independent.

Bacterial Proteins↗
PubMed2004

Hemocompatibility of nitrogen-doped, hydrogen-free diamond-like carbon prepared by nitrogen plasma immersion ion implantation-deposition.

Amorphous hydrogenated carbon (a-C:H) has been shown to be a potential material in biomedical devices such as artificial heart valves, bone implants, and so on because of its chemical inertness, low coefficient of friction, high wear resistance, and good biocompatibility. However, the biomedical characteristics such as blood compatibility of doped hydrogen-free diamond-like carbon (DLC) have not been investigated in details. We recently began to investigate the potential use of nitrogen-doped, hydrogen-free DLC in artificial heart valves. In our experiments, a series of hydrogen-free DLC films doped with nitrogen were synthesized by plasma immersion ion implantation-deposition (PIII-D) utilizing a pulsed vacuum arc plasma source and different N to Ar (FN/FAr) gas mixtures in the plasma chamber. The structures and properties of the film were evaluated by Raman spectroscopy, Rutherford backscattering spectrometry (RBS), and X-ray photoelectron spectroscopy (XPS). To assess the blood compatibility of the films and the impact on the blood compatibility by the presence of nitrogen, platelet adhesion tests were conducted. Our results indicate that the blood compatibility of both hydrogen-free carbon films (a-C) and amorphous carbon nitride films are better than that of low-temperature isotropic pyrolytic carbon (LTIC). The experimental results are consistent with the relative theory of interfacial energy and surface tension including both dispersion and polar components. Our results also indicate that an optimal fraction of sp2 bonding is desirable, but an excessively high nitrogen concentration degrades the properties to an extent that the biocompatibility can be worse than that of LTIC.

Albumins↗
PubMed1988

Strict insulin therapy normalises organ nitrogen contents and the capacity of urea nitrogen synthesis in experimental diabetes in rats.

Rats with experimental diabetes due to streptozotocin (75 mg/kg body weight) and free access to food were divided into two groups. One group (n = 9) was optimally treated with insulin (glucosuria less than 4.0 mmol/24 h), using heat treated very long-acting ultralente insulin. The other group (n = 10) was poorly treated with insulin (glucosuria 20-30 mmol/24 h). The nitrogen balance and energy balance of optimally treated diabetic rats was positive and not different from the control group (n = 6). In the poorly treated diabetic rats the nitrogen balance was reduced whereas the energy balance was not different from that of control rats. After 4 weeks the fasting glucagon was: 50 +/- 21 ng/l (mean +/- SEM) in control rats, 62 +/- 18 ng/l in optimally treated diabetic rats and 249 +/- 58 ng/l in poorly treated diabetic rats (p less than 0.01). The capacity of urea nitrogen synthesis determined during alanine loading was: 9.6 +/- 1.0 mumol/(min 100 g body weight) in control rats, 10.6 +/- 1.7 mumol/(min 100 g body weight) in optimally treated diabetic rats and 17.3 +/- 1.3 mumol/(min 100 g body weight) in poorly treated diabetic rats (p less than 0.01). Nitrogen contents of carcass, heart, intestines, liver, and kidneys as determined by Kjeldahl analyses were identical in control rats and optimally treated diabetic rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Alanine↗
PubMed2005

Effects of carbon and nitrogen sources, carbon-to-nitrogen ratio, and initial pH on the growth of nematophagous fungus Pochonia chlamydosporia in liquid culture.

The effects of carbon and nitrogen sources, carbon-to-nitrogen ratio (C:N) and initial pH value on the growth and sporulation of the nematophagous fungus Pochonia chlamydosporia in liquid culture were examined. Among the 21 carbon sources and 15 nitrogen compounds tested, the optimal carbon and nitrogen sources for mycelial growth were sweet potato and L: -tyrosine, and for sporulation were sweet potato and casein peptone. A C:N ratio of 10:1 at pH 3.7 gave the maximum yield of conidia and a C:N ratio of 40:1 at pH 6.8 gave the maximum biomass. The initial pH value had a significant effect on mycelial growth and conidial production, with the optimal ranges being 3.5-4.5 for sporulation and 5-6 for growth. Maximum conidial production was obtained at an initial pH of 4.0 and the maximum biomass at pH 6.0. The results also showed that the final pH after 7 days cultivation was always higher than the initial value. The variability in growth and sporulation of seven strains of P. chlamydosporia in liquid culture was also compared and discussed.

Biomass↗
PubMed2005

Predicting total urinary nitrogen excretion from urinary urea nitrogen excretion in multiple-trauma patients receiving specialized nutritional support.

OBJECTIVE: We investigated the accuracy of methods to estimate total urinary nitrogen (TUN) excretion from urinary urea nitrogen (UUN) excretion for patients who have multiple trauma and receive specialized nutritional support. METHODS: Fifty-five critically ill, adult patients who had multiple trauma and were receiving specialized nutritional support were evaluated. A 24-h urine collection for urea nitrogen and total nitrogen was performed 4.4 +/- 2.6 d after admission to the trauma intensive care unit. Patients with significant renal impairment, liver dysfunction, or obesity (>150% of ideal body weight) were excluded from study entry. Eight publications that examined the relation between TUN and UUN were evaluated for bias and precision in estimating TUN from UUN. RESULTS: TUN was 20.8 +/- 10.8 g/d with an average difference of 3.8 +/- 2.8 g/d between TUN and UUN. Linear regression analysis comparing TUN with UUN indicated a significant correlative relation (TUN = 1.1 x UUN + 2; r = 0.958, P < 0.001). The difference between TUN and UUN varied based on UUN: for UUN lower than 10 g/d, TUN minus UUN was 1.5 +/- 1.0 g/d; for UUN 10 to 20 g/d, TUN minus UUN was 4.1 +/- 3.2 g/d; and for UUN higher than 20 g/d, TUN minus UUN was 5.3 +/- 1.9 g/d (P < 0.001). Six methods were biased toward underpredicting TUN, one method was unbiased, and one was biased toward overpredicting TUN. A practical method for estimating TUN from UUN was developed: TUN = UUN + 2 for those with UUN lower than 10 g/d and TUN = 1.1 x UUN + 2 for those with UUN of at least 10 g/d. CONCLUSIONS: Our method, the modified Velasco method, UUN/0.84, and UUN/0.85 provided reasonable estimates of TUN from UUN in critically ill, adult patients who had multiple trauma and were receiving specialized nutritional support; however, our method requires further validation.

Adult↗
PubMed2002

Low nitrogen losses with a new source of nitrogen for cultivation of conifer seedlings.

Losses of nitrogen (N) when cultivating plants may cause a number of adverse environmental effects. N losses from conifer nurseries, for instance, may have a considerable impact on the local environment, and studies indicate that the bulk of added N is not recovered in the cultivated plants. This study was conducted to obtain insight into the causes of the low recovery and to test an alternative N fertilizer. Hence, growth of the economically important Scots pine (Pinus sylvestris (L).) seedlings and the recovery of different forms of added nitrogen (N) were investigated in a greenhouse experiment. Containerized seedlings were grown in peat for one summer, with two different N fertilizers, one organic (arginine) and one inorganic (a commercial fertilizer (CF) containing a mixture of ammonium and nitrate) each at an N concentration of 3 mM. At the end of the growth period, some seedlings were labeled with solutions containing either U-[13C6], [15N4]-arginine, (15NH4)2SO4, or K15NO3 supplied to the growth substrate. Labeled seedlings were harvested 1 h, 5 days, and 19 days after tracer addition, and the recovery of each added nitrogen source in both the seedlings and the growth substrate was measured. The retention of the three N forms during discharge of solutions from the growth substrate, peat, was tested in a separate experiment. Arginine-fed seedlings grew better and had higher needle N concentrations than the CF-fed seedlings. Isotopic data showed that the arginine treatment gave significantly higher N recoveries (80%) compared to the CF treatment (50%). The low recovery of N in the CF treatment was largely due to very low recovery (30%) of NO3- -N. The retention of the different N forms during discharge of solutions from the growth substrate was highest for arginine, somewhat lower for NH44+, and very low for NO3-. The high rate of seedling growth and the small nitrogen losses observed when using arginine suggest that this amino acid may be an efficient and environmentally favorable N source for cultivating conifer seedlings.

Arginine↗
PubMed2005

Nitrogen isotope relationships between crops and fertilizer: implications for using nitrogen isotope analysis as an indicator of agricultural regime.

The effect of fertilizer type, amount of fertilizer applied, growing medium, and water type on the nitrogen-15 content of carrots, tomatoes, and lettuces has been investigated. Crops grown using synthetic nitrogen fertilizer were isotopically lighter than those grown using pelleted chicken manure. For example, for equivalent amounts of nitrogen applied, carrots grown with ammonium nitrate fertilizer had delta15N values between 3 per thousand and 4 per thousand lower than those grown using pelleted chicken manure. Plants grown in peat-based compost were generally found to be isotopically lighter than those grown in composted bark based compost. Results suggest that nitrate content and the delta15N of the nitrate in irrigation water may also influence crop delta15N. Wider implications of using crop delta15N more generally as an indicator of whether synthetic nitrogen fertilizers have been applied to a crop and the possible application and limitations of using crop delta15N as an indicator of agricultural regime (organic/conventional) are discussed.

Crops, Agricultural↗
PubMed1999

Nitration of unsaturated fatty acids by nitric oxide-derived reactive nitrogen species peroxynitrite, nitrous acid, nitrogen dioxide, and nitronium ion.

Reactive nitrogen species derived from nitric oxide are potent oxidants formed during inflammation that can oxidize membrane and lipoprotein lipids in vivo. Herein, it is demonstrated that several of these species react with unsaturated fatty acid to yield nitrated oxidation products. Using HPLC coupled with both UV detection and electrospray ionization mass spectrometry, products of reaction of ONOO- with linoleic acid displayed mass/charge (m/z) characteristics of LNO2 (at least three products at m/z 324, negative ion mode). Further analysis by MS/MS gave a major fragment at m/z 46. Addition of a NO2 group was confirmed using [15N]ONOO- which gave a product at m/z 325, fragmenting to form a daughter ion at m/z 47. Formation of nitrated lipids was inhibited by bicarbonate, superoxide dismutase (SOD), and Fe3+-EDTA, while the yield of oxidation products was decreased by bicarbonate and SOD, but not by Fe3+-EDTA. Reaction of linoleic acid with both nitrogen dioxide (*NO2) or nitronium tetrafluoroborate (NO2BF4) also yielded nitrated lipid products (m/z 324), with HPLC retention times and MS/MS fragmentation patterns identical to the m/z 324 species formed by reaction of ONOO- with linoleic acid. Finally, reaction of HPODE, but not linoleate, with nitrous acid (HONO) or isobutyl nitrite (BuiONO) yielded a product at m/z 340, or 341 upon reacting with [15N]HONO. MS/MS analysis gave an NO2- fragment, and 15N NMR indicated that the product contained a nitro (RNO2) functional group, suggesting that the product was nitroepoxylinoleic acid [L(O)NO2]. This species could form via homolytic dissociation of LOONO to LO* and *NO2 and rearrangement of LO* to an epoxyallylic radical L(O)* followed by recombination of L(O)* with *NO2. Since unsaturated lipids of membranes and lipoproteins are critical targets of reactive oxygen and nitrogen species, these pathways lend insight into mechanisms for the formation of novel nitrogen-containing lipid products in vivo and provide synthetic strategies for further structural and functional studies.

Chromatography, High Pressure Liquid↗
PubMed2002

Nitrogen cycle: what governs nitrogen loss from forest soils?

Nitrogen is lost as dissolved organic compounds in stream waters from unpolluted South American forests, but it is lost mainly as inorganic nitrate in streams flowing from North American forests that suffer nitrogen deposition from the atmosphere. From this it has been inferred that the standard thinking about how nature deals with nitrogen in soils and waters needs to be re-evaluated and that the conventional wisdom of how nitrogen is absorbed and released by plants must be wrong. We disagree, however, on the grounds that there are other, more likely interpretations of the new results.

Models, Biological↗
PubMed1995

Utilization of aspartate as a nitrogen source in Escherichia coli. Analysis of nitrogen flow and characterization of the products of aspartate catabolism.

15N NMR, reverse-phase high performance liquid chromatography and gas chromatography/mass spectrometry have been used to follow nitrogen metabolism in Escherichia coli labeled on medium containing L-[15N]aspartate. The flow of 15N through various nitrogen-containing metabolites was followed over the course of the labeling period. For wild-type E. coli labeled on L-[15N]aspartate as sole source of nitrogen, significant 15N labeling was detected only in the intracellular L-glutamate, L-alanine, L-aspartate, and putrescine pools. Intracellular concentrations of L-aspartate and L-glutamate differed significantly in extracts of an arginine auxotroph (argG-), which is deficient in a potential aspartate-assimilating reaction. When the L-[15N]aspartate containing labeling medium was supplemented with unlabeled arginine, extracts of wild-type E. coli were shown to contain significant amounts of unlabeled ammonia and putrescine. There was substantial dilution of 15N in the glutamate pool. The observation that glutamate, aspartate, and alanine are 15N-labeled, but ammonia is not, suggests that these amino acids are not the immediate source of ammonia. Our results suggest that arginine may be an intermediate for the degradation of some of the aspartate and that arginine may be an intermediate for ammonia production during nitrogen-limited growth. Our results also strongly suggest the presence of a previously uncharacterized pathway of arginine degradation.

Arginine↗