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Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters.

In plants, many of the proteins involved in transport of nitrogenous compounds have not been identified so far. The use of heterologous complementation in yeast mutants has enabled the isolation of a gene family encoding amino acid permease (AAP). A highly sensitive selection procedure was used to identify other proteins capable of transporting amino acids. In addition to members of the AAP gene family, an integral membrane protein (NTR1) that shows significant similarities to the low affinity nitrate transporter from Arabidpsis and to peptide transporters from yeast and rabbit was identified. NTR1 seems to be involved in the supply of reproductive organs with nitrogen as it is expressed at low levels in leaves and highly in developing pods.

Amino Acid Sequence↗

Ultrasound-induced formation of S-nitrosoglutathione and S-nitrosocysteine in aerobic aqueous solutions of glutathione and cysteine.

S-Nitrosocompounds are formed when aqueous solutions of cysteine or glutathione are exposed to ultrasound (880 kHz) in air. The yield of the S-nitrosocompounds was as high as 10% for glutathione and 4% for cysteine of the initial thiol concentrations (from 0.1 to 10 mM) in the aqueous solutions. In addition to the formation of S-nitrosocompounds, thiol oxidation to disulfide forms was observed. After the oxidation of over 70% of the sulfhydryl groups, formation of peroxide compounds as well as cysteic acid derivatives was recorded. The formation of the peroxide compounds and peroxide radicals in the ultrasound field reduced the yield of S-nitrosocompounds. S-Nitrosocompounds were not formed when exposing low-molecular-weight thiols to ultrasound in atmospheres of N(2) or CO. In neutral solutions, ultrasound-exposed cysteine or glutathione released NO due to spontaneous degradation of the S-nitrosocompounds. N(2)O(3), produced due to the spontaneous degradation of the S-nitrosocompounds in air, nitrosylated sulfhydryl groups of glutathione manifested in the appearance of new absorption bands at 330 and 540 nm. The nitrogen compounds formed in an ultrasound field modified the sulfhydryl groups of apohemoglobin and serum albumin. The main target for ultrasound-generated oxygen free radicals were cystine residues oxidized to cysteic acid residues.

Air↗

Nickel as a micronutrient element for plants.

The detrimental effects of excessive Ni on plant growth have been well known for many years. More recent evidence indicates that Ni is required in small amounts for normal plant growth and development. Ni is an essential component of urease in plants and microorganisms. A deficiency of Ni in plants is reported to result in necrotic lesions in leaves in response to toxic accumulations of urea. Urease plays an essential role in mobilization of nitrogenous compounds in plants, a process that is especially important during seed germination and fruit formation when protein reserves are degraded into amino acids. Arginine, an abundant amino acid in plants, when degraded produces urea as a product and urease is needed for urea utilization. Theories of urea formation during allantoin degradation in Glycine max have been recently refuted. In G. max ureides apparently are metabolized via an amidohydrolase reaction with subsequent degradation of ureidoglycine, yielding glyoxylate, NH+4 and CO2. No evidence is available for the formation of urea in this pathway. Nitrogen-fixing symbionts, such as Rhizobium and Bradyrhizobium, contain two known Ni enzymes: urease and hydrogenase. Optimum growth of nodulated legumes and actinorhizal plants may depend on an adequate supply of Ni to meet the requirements of the Ni-requiring enzymes in host plants and endophytes. The seeds of severely Ni-deficient Hordeum are completely inviable, thus providing conclusive evidence for the essentiality of Ni for this species. The evidence indicates that Ni must be added to the list of micronutrient elements generally required by plants.

Nickel↗

Sulfur and nitrogen chemiluminescence detection in gas chromatographic analysis.

Sulfur and nitrogen compounds exist in a wide variety of analytical samples, from harmful contaminants in refinery streams to important ingredients in certain beverages, spices and condiments. Accurate determination of sulfur/nitrogen concentrations in these diverse samples is very important for process monitoring, quality control, product development, as well as basic research in these different industries. Due to the complex nature of the matrices, and the fact that the sulfur/nitrogen analytes usually exist at low concentrations, an element-selective detector for sulfur/nitrogen is indispensable to the GC analysis of these samples. To meet this challenge, sulfur and nitrogen chemiluminescence detectors for GC have become one of the most powerful tools available to analytical chemists, thanks to their high sensitivity, selectivity and equimolar detector response. In this brief review, an overview of the chemiluminescence detector operating principles, as well as the detector characteristics is presented. Examples of their applications in petroleum refinery, environmental analysis and food/flavor industry are presented. The two chemiluminescence detectors have also been combined into one convenient package and an example of applications of the simultaneous sulfur/nitrogen chemiluminescence detector is also provided.

Chromatography, Gas↗

Nitric oxide in star-forming regions: further evidence for interstellar N-O bonds.

Nitric oxide has been newly detected towards several star-forming clouds, including Orion-KL, Sgr B2(N), W33A, W51M, and DR21(OH) via its J = 3/2 --> 1/2 transitions near 150 GHz, using the FCRAO 14 m telescope. Both lambda-doubling components of NO were observed towards all sources. Column densities derived for nitric oxide in these clouds are N approximately 10(15)-10(16) cm-2, corresponding to fractional abundances of f approximately 0.5-1.0 x 10(-8), relative to H2. Towards Orion-KL, the NO line profile suggests that the species arises primarily from hot, dense gas. Nitric oxide may arise from warm material toward the other clouds as well. Nitric oxide in star-forming regions could be synthesized by high-temperature reactions, although the observed abundances do not disagree with values predicted from low-temperature, ion-molecule chemistry by more than one order of magnitude. The abundance of NO, unlike other simple interstellar nitrogen compounds, does appear to be reproduced by chemical models, at least to a good approximation. Regardless of the nature of formation of NO, it appears to be a common constituent of warm, dense molecular clouds. N-O bonds may therefore be more prevalent than previously thought.

Extraterrestrial Environment↗

[Change in biochemical composition of amaranth leaves as a result of selection for increased level of amaranthine pigment].

The composition and content of secondary compounds produced by the shikimate pathway and the contents of protein and cellulose were determined in leaves of amaranth (Amaranthus tricolor L.) K-99 and the cultivar Valentina raised from it by family selection and enriched in the pigment amaranthine. It was found that intense biosynthesis of amaranthine, tyrosine, and phenylalanine resulted in a decrease in the contents of lignin, protein, and cellulose in leaves of Valentina by comparison with K-99 and in changed the morphological traits: color deepening and a decrease in leaf density. It is concluded that amaranth biosynthesis is related to nitrogen metabolism and amaranthine is an intermediate involved in conversion of nitrogen compounds in the cell.

Amaranthus↗

The effect of nitrogen nutrition on cluster root formation and proton extrusion by Lupinus albus.

Nitrogen nutrition can influence cluster root formation in many wild species, but the effect of N form on cluster root formation and root exudation by white lupin is not known. In a solution culture study, we examined the effect of N nutrition (ammonium, nitrate, both or N2 fixation) on cluster root formation and H+ extrusion by white lupin plants under deficient and adequate P supply. The number of cluster roots increased greatly when plants were supplied with I microM P compared with 50 microM P, the increase being 7.8-fold for plants treated with (NH4)2SO4, 3-fold for plants treated with KNO3 and NH4NO3, and 2-4-fold for N2-fixing plants. Under P deficiency. NH4+-N supply resulted in production of a greater number and biomass of cluster roots than other N sources. Dry weight of cluster roots was 30 % higher than that of non-cluster roots in P-deficient plants treated with (NH4)2SO4 and NH4NO3. In plants treated with sufficient P (50 microM), the weight of non-cluster roots was approx. 90 % greater than that of cluster roots. Both total (micromol per plant h(-1)) and specific (micromol g(-1) root d. wt h(-1)) H+ extrusions were greatest from roots of plants supplied with (NH4)2SO4, followed by those supplied with NH4NO3 and N2 fixation, whereas plants receiving KNO3 had negative net H+ extrusion between the third and fifth week of growth (indicating uptake of protons or release of OH- ions). The rate of proton extrusion by NH4+-N-fed plants was similar under P-deficient and P-sufficient conditions. In contrast, proton exudation by N2-fixing plants and KNO3-treated plants was ten-fold greater under P deficiency than under P sufficiency. In comparison with P deficiency, plants treated with 50 microM P had a significantly higher concentration of P in roots, shoots and youngest expanded leaves (YEL). Compared with the N2 fixation and KNO3 treatments, total N concentration was highest in roots, shoots and YEL of plants supplied with (NH4)2SO4 and NH4NO3, regardless of P supply. Under P deficiency, K concentrations in roots decreased at all N supplies, especially in plants treated with (NH4)2SO4 and NH4NO3, which coincided with the greatest H+ extrusion at these P and N supplies. In conclusion, NH4-N nutrition stimulated cluster root formation and H+ extrusion by roots of P-deficient white lupin.

Biomass↗

Ammonia and urea in the maternal-fetal trophic relationship of the viviparous blenny (eelpout) Zoarces viviparus.

A high correlation was observed between the concentration of urea in the maternal plasma and the concentration of urea in the ovarian fluid during post-yolk sac growth of the embryos in the ovary of the viviparous blenny (eelpout) Zoarces viviparus. A high correlation was observed between maternal plasma and ovarian fluid ammonia as well. Ammonia but not urea was excreted to the external medium by the mother fish during pregnancy. Intraovarian loading with either urea or ammonia resulted in a steady decrease from initial high concentrations in the ovarian fluid and a concomitant increase in the maternal plasma levels of both nitrogenous compounds. Injected ammonia was eliminated much faster from the ovarian fluid than urea. No significant effect of any ammonia could be observed on urea excretion rates by embryos in vitro. Patterns of accumulation for urea and ammonia in the external medium were investigated by exposure of embryos in vitro to concentrations of urea and ammonia similar to those normally found in the ovarian fluid. No significant changes could be observed in the external urea or ammonia levels during the experiment. The results indicate that during post-yolk sac development of embryos in vivo, net catabolism of nitrogen-containing organics and formation of urea may be reduced by high ambient concentrations of urea in the ovarian fluid.

Ammonia↗

P(II) signal transduction proteins, pivotal players in microbial nitrogen control.

The P(II) family of signal transduction proteins are among the most widely distributed signal proteins in the bacterial world. First identified in 1969 as a component of the glutamine synthetase regulatory apparatus, P(II) proteins have since been recognized as playing a pivotal role in control of prokaryotic nitrogen metabolism. More recently, members of the family have been found in higher plants, where they also potentially play a role in nitrogen control. The P(II) proteins can function in the regulation of both gene transcription, by modulating the activity of regulatory proteins, and the catalytic activity of enzymes involved in nitrogen metabolism. There is also emerging evidence that they may regulate the activity of proteins required for transport of nitrogen compounds into the cell. In this review we discuss the history of the P(II) proteins, their structures and biochemistry, and their distribution and functions in prokaryotes. We survey data emerging from bacterial genome sequences and consider other likely or potential targets for control by P(II) proteins.

Bacteria↗

Performance evaluation of SBR treatment for nitrogen removal from tannery wastewater.

Performance of SBR treatment for nitrogen removal from tannery is evaluated for a wide range of wastewater temperature between 7 and 30 degrees C. A pilot-scale SBR unit fed with plain-settled wastewater is operated on site for this purpose. Effective nitrogen removal is sustained by adjustment of the sludge age from 28 to 5 days. Concentration profiles of nitrogen compounds within a selected complete SBR cycle during the steady state operation at different wastewater temperatures and sludge ages are evaluated by model simulation. System performance is also interpreted in terms of modeling and stoichiometric calculation. Additional nitrate loss was observed during aerobic period when the aeration intensity was reduced by the factor of 50%.

Industrial Waste↗

[Studies on the syntheses and reaction of nitrogen-containing heterocyclic compounds centered the naphthyridines].

1, X-Naphthyridines (X = 5, 6, and 8) (1-3) were synthesized in a high yield by the Skraup reactions of 4-, 3-, and 2-aminopyridines with glycerol, in the presence of ferrous sulfate and boric acid. The improved syntheses were applied to the syntheses of 1,5-, 1,6-, and 1,8-naphthyridines (4-28), pyridonaphthyridines (31, 32), benzonaphthyridines (38-42), naphthonaphthyridines (44, 48, 49, 59). New synthetic methods of naphtho[1,2-b or 2,1-b][1,8]naphthyridine (62, 63), benzo[b][1,8]naphthyridine (41) and benzo[g]quinoline (67) were developed and their compounds obtained conveniently. Reissert reaction of 1,6-naphthyridine (2) using triethylbenzylammonium chloride (TEBA) produced Reissert compounds and ring-opened compounds. Reissert reaction of 1,7-naphthyridine (29), 4,7- (87), 4,6-phenanthroline (38) gave the Reissert compounds, and the compounds (2, 38) gave their pseudo bases (79, 90). Reissert-type reactions of benzo[f]quinoline (68), and 1,7-phenanthroline (105) with acyl chloride and phosphite, gave alpha- and gamma-phosphonate, and 87 produced alpha,alpha'-(108a-c) and alpha,gamma'-diphosphonates (109a-c). The structure of 108c-1 was determined to be trans-type by an X-ray analysis. Compounds 1-3, 29, and benzo[f or h]quinoline were treated with dimetyl sulfoxide in the presence of sodium hydride to give mainly methylated compounds at the position para to the ring-nitrogen. When benzo[f or h]quinoline N-oxide was treated with methylsulfinylmethyl carbanion in the usual procedure, a new reaction took place to produce phenanthrene in an excellent yield. The same reaction of 1,10-, 1,7-, or 4,7-phenanthroline N-oxides (143-145) resulted in the liberation of the N-oxide group to form benzo[f or h]quinoline, but isoquinoline N-oxide afforded to benzazepine derivatives (161). Reaction of quinaldine with methylsulfinylmethyl carbanion gave novel tricyclic compound (121a). The oxidation of 41, 62, and 63 with peroxy acids afforded novel products such as seven-membered 1,4-oxazepine derivatives.

Chemistry, Organic↗

[The effect of exogenous factors on prececal nutrient and amino acid absorption, ascertained from swine with ileo-rectal anastomoses. 3. The effect of crude fiber-rich coarse meal supplements to a basic ration].

In digestion trials using pigs fitted with ileo-rectal anastomoses and parallel intact pigs the influence of wheat straw meal (WSM) or grass meal (GM) supplemented on two different levels to a basal diet was examined with regard to nutrient and amino acid digestibility resp. absorption. Both roughages reduced, obviously because of their high cell wall contents, nearly at equal amounts and partly significantly the precaecal as well as the total digestibility of dry matter, organic matter, carbohydrates (= crude fibre + NFE), crude fibre and hemicelluloses (arabinose, xylose). By the roughage supplementations the crude protein digestibility at the terminal ileum was less reduced than at the end of the total digestive tract, the starch digestibility was hardly influenced and that of ether extract mostly increased. In comparison with precaecal glucose and fructose digestibilities it could be shown that the anthrone method is not suitable for determinations of the precaecal digestibility of water soluble carbohydrates. Beside these compounds other substances in the ileum digesta must evidently be dyed by anthrone too. The crude protein digestibility and the amino acid absorption were precaecally hardly or not reduced by WSM supplementations, therefore it can be concluded that it is possible to dilute the energy concentration in diets (e.g. for sows) by addition of WSM without impairing protein digestion and amino acid absorption. The GM supplementations, however, impaired protein digestibility and amino acid absorption of the whole diet, probably caused by the encrusted, possibly heat damaged protein present in the GM itself. When the roughage supplemented diets were fed, the excretion of nitrogen compounds in the faeces was enlarged due to the more intensive bacterial activity in the hind gut and the additional sorption effects to cell wall substances, so that a too low apparent and also true digestibility is made believe. The applicability of the difference method to the calculation of protein and amino acid digestibilities in roughages is very questionable because of their low contents and the missing additivity. The crude fibre and hemicellulose digestibility values calculated by the difference method demonstrate for the both roughages--in spite of high standard deviations--that the precaecal digestibility of crude fibre is about zero and that of the hard lignified hemicellulose fraction in contrast to the predominantly endospermic hemicelluloses of the basal diet is very low. The faecal crude fibre and hemicellulose digestibilities of WSM--especially on the lower supplementation levels--are markedly worse than those of GM.

Amino Acids↗

Past and future exceedances of nitrogen critical loads in Europe.

Critical loads of acidity and nutrient nitrogen--simple measures of the sensitivity of ecosystems to deposition--have been widely used for setting emission reduction targets in Europe. In contrast to sulfur, the emissions of nitrogen compounds remain high in the future. This is also true for the exceedances of critical loads until 2010. Looking further into the future, climate change is likely to influence ecosystem sensitivity, and thus critical loads. It is shown that higher temperatures, changed precipitation patterns, and modified net primary production mainly increase critical loads, except in mountainous and arid regions. Using consistent scenarios of climate change and air pollution from a recently completed European study (AIR-CLIM), it is shown that the exceedances in 2100 of the critical loads are declining in comparison to 2010. However, exceedances of critical loads of nutrient nitrogen remain substantial, even under the most stringent scenario. This confirms the increasing role nitrogen plays in environmental problems in comparison to sulfur. Thus research should focus on the effects of nitrogen in the environment, especially under conditions of climate change, to support nitrogen-emission mitigating policies. This not only reduces acidification and eutrophication, but also helps curb the formation of tropospheric ozone.

Environmental Monitoring↗

Arginine catabolism in Aphanocapsa 6308.

The catabolic products of arginine metabolism were observed in Aphanocapsa 6308, a unicellular cyanobacterium, by thin layer chromatography of growth media, by limiting growth conditions, and by enzymatic analysis. Of the organic, nitrogenous compounds examined, only arginine supported growth in CO2-free media. The excretion of ornithine at a concentration level greater than citrulline suggested the existence in Aphanocapsa 6308 of the arginine dihydrolase pathway which produced ornithine, CO2,NH4,+ adenosine 5'-triphosphate. Its existence was confirmed by enzymatic analysis. Although cells could not grow on urea as a sole carbon source a very active urease and subsequently an arginase were also demonstrated, indicating that Aphanocapsa can metabolize arginine via the arginase pathway. The level of enzymes for both pathways indicates a lack of genetic control. It is suggested that the arginase pathway provides only nitrogen for the cells wheras the arginine dihydrolase pathway provides not only nitrogen, but also CO2 and adenosine 5'-triphosphate.

Adenosine Triphosphate↗

How partial nitrification could improve reclaimed wastewater transport in long pipes.

Reclaimed wastewater transport is studied in a concrete-lined cast iron pipe, where a nitrification-denitrification process occurs. The pipe is part of the Reuse System of Reclaimed Wastewater of South Tenerife (Spain), 0.6 m in diameter and 61 km long. In order to improve wastewater quality, at 10 km from the inlet there is injection of fresh water saturated in dissolved oxygen (DO), after which a fast nitrification process usually appears (less than two hours of space time). The amount of oxidized nitrogen compounds produced varies between 0.8 and 4.4 mg/l NOx(-)-N. When DO has disappeared, a denitrification process begins. The removal of nitrite is complete at the end of the pipe, whereas the nitrate does not disappear completely, leaving a concentration of about 0.4-0.5 mg/l. For a COD/NOx(-)-N ratio higher than 5, a first order nitrification rate in NOx(-)-N has resulted, with the constant k20 = 0.079 h-1, for a NOx(-)-N concentration range of 0.8-4.4 mg/l. Finally the following temperature dependency for the first order denitrification rate constant has been found: k = k20 x 1 x 15T-20. Although nitrogen could be used as nutrient in the agricultural reuse, its removal from reclaimed wastewater could be useful in order to diminish the chlorine needs for reclaimed wastewater disinfection.

Agriculture↗

Ammonia-specific regulation of Gln3 localization in Saccharomyces cerevisiae by protein kinase Npr1.

Events directly regulating Gln3 intracellular localization and nitrogen catabolite repression (NCR)-sensitive transcription in Saccharomyces cerevisiae are interconnected with many cellular processes that influence the utilization of environmental metabolites. Among them are intracellular trafficking of the permeases that transport nitrogenous compounds and their control by the Tor1,2 signal transduction pathway. Npr1 is a kinase that phosphorylates and thereby stabilizes NCR-sensitive permeases, e.g. Gap1 and Mep2. It is also a phosphoprotein for which phosphorylation and kinase activity are regulated by Tor1,2 via Tap42 and Sit4. Npr1 has been reported to negatively regulate nuclear localization of Gln3 in SD (ammonia)-grown cells. Thus we sought to distinguish whether Npr1: (i) functions directly as a component of NCR control; or (ii) influences Gln3 localization indirectly, possibly as a consequence of participating in protein trafficking. If Npr1 functions directly, then the ability of all good nitrogen sources to restrict Gln3 to the cytoplasm should be lost in an npr1Delta just as occurs when URE2 (encoding this well studied negative Gln3 regulator) is deleted. We show that nuclear localization of Gln3-Myc(13) in an npr1Delta occurred only with ammonia as the nitrogen source. Other good nitrogen sources, e.g. glutamine, serine, or asparagine, restricted Gln3-Myc(13) to the cytoplasm of both wild type and npr1Delta cells. In other words, the npr1Delta did not possess the uniform phenotype for all repressive nitrogen sources characteristic of ure2Delta. This suggests that the connection between Gln3 localization and Npr1 is indirect, arising from the influence of Npr1 on the ability of cells to utilize ammonia as a repressive nitrogen source.

Ammonia↗

Perturbation of conjugation in internally solvated allylic lithium compounds: variation of ligand structure. NMR and X-ray crystallography.

Several allyic lithium compounds were prepared with different potential ligands tethered at C2. These are with CH3OCH2CH2NCH3CH2-, 5 and 1-TMS 6, with (CH3)2NCH2CH2NCH3CH2-, 1-TMS 7, and with ((CH3)2NCH2CH2)2NCH2-, 8 and 1-TMS 9. In all these compounds Li is fully coordinated to the pendant ligand and is sited off the axis perpendicular to the allyl plane at one of the allyl termini as indicated by a combination of X-ray crystallography and NMR spectra. Compounds 5 and 8 are Li-bridged dimers as shown by X-ray crystallography and also dimeric in benzene solution as determined from freezing point determinations. Compounds 6, 7, and 9 are monomeric in THF-d8 or diethyl ether-d10 solution and exhibit one bond 13C1, 6Li scalar coupling at low temperature. Taken together the crystallographic and NMR data indicate that all of these compounds incorporate partially delocalized allylic moieties. Compounds 5 and 8 undergo fast 1,3-Li-sigmatropic shifts that are proposed to take place within low concentrations of monomers in fast equilibrium with prevalent dimers. Averaging with increasing temperature of the one-bond 13C, 6Li coupling constant in 6, 7, and 13 provided the dynamics of bimolecular C-Li exchange with Delta H++ values of 6.7, 12, and 13 kcal x mol(-1), respectively. Averaging of the diastereotopic N(CH3)2 13C resonances of 7 is indicative of fast transfer of coordinated ligand between faces of the allyl plane Delta H++ = 5.3 kcal x mol(-1) combined with slower inversion at nitrogen. Compound 8 exhibits similar effects. It is concluded that variation of the ligand structure changes dynamic behavior of the compounds but has little influence of their degrees of delocalization.

Journal Article↗