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Nitrogen metabolism and nitrogen control in corynebacteria: variations of a common theme.

The published genome sequences of Corynebacterium diphtheriae, Corynebacterium efficiens, Corynebacterium glutamicum and Corynebacterium jeikeium were screened for genes encoding central components of nitrogen source uptake, nitrogen assimilation and nitrogen control systems. Interestingly, the soil-living species C. efficiens and C. glutamicum exhibit a broader spectrum of genes for nitrogen transport and metabolism than the pathogenic species C. diphtheriae and C. jeikeium. The latter are characterized by gene decay and loss of functions like urea metabolism and nitrogen-dependent transcription control. The global regulator of nitrogen regulation AmtR and its DNA-binding motif are conserved in C. diphtheriae, C. efficiens and C. glutamicum, while in C. jeikeium, an AmtR-encoding gene as well as putative AmtR-binding motifs are missing.

Amino Acid Motifs↗

Energy, industry and nitrogen: strategies for decreasing reactive nitrogen emissions.

Nitrogen oxides are released during atmospheric combustion of fossil fuels and biomass, and during the production of certain chemicals and products. They can react with natural or man-made volatile organic compounds to produce smog, or else can be further oxidized to produce particulate haze, or acid rain that can eutrophy land and water. The reactive nitrogen that begins in the energy sector thus cascades through the atmosphere, the hydrosphere and soils before being eventually partially denitrifed to the global warming and stratospheric ozone-depleting gas nitrous oxide or molecular nitrogen. This paper will suggest how an economic analysis of the nitrogen cycle can identify the most cost-effective places to intervene. Nitrogen oxides released during fossil-fuel combustion in vehicles, power plants and heating boilers can either be controlled by add-on emission control technology, or can be eliminated by many of the same technical options that lead to carbon dioxide reduction. These integrated strategies also address sustainability, economic development and national security issues. Similarly in industrial production, it is more effective to focus on redesigning industrial processes rather than on nitrogen oxide pollution elimination from the current system. This paper will suggest which strategies might be utilized to address multiple benefits rather than focusing on single pollutants.

Acid Rain↗

Studies on biochemical effects of nitrogen dioxide: I. Lipid peroxidation as measured by ethane exhalation of rats exposed to nitrogen dioxide.

This research was in order to follow the periodic fluctuation of lipid peroxidation by a new method in rats exposed to nitrogen dioxide. Wistar male rats were examined for lipid peroxidation as demonstrated by ethane exhalation. In rats continuously exposed to 10 ppm nitrogen dioxide for 2 weeks, the amount of ethane exhaled fluctuated in a complex manner during the exposure. Ethane exhalation decreased slightly after the first day of exposure and then increased rapidly. The maximal values were observed after the fourth day of exposure and then decreased gradually to the initial level. Furthermore, the activity of glutathione peroxidase in lungs of rats exposed to 10 ppm nitrogen dioxide varied symmetrically against the change of ethane formation. Similar changes in ethane exhalation were observed in rats exposed to the lowest levels of nitrogen dioxide (0.4, 1.2 and 4.0 ppm) for 4 months. Compared to 10 ppm nitrogen dioxide exposure for 14 days, the characteristics in rats exposed to the low levels (0.4-4.0 ppm) of nitrogen dioxide were: the decline of ethane formation, the delay in alterations, and the tendency toward gradual increase during the longer period exposure.

Animals↗

Effect of ruthenium on nitrogen fixation by some nitrogen fixers.

The effect of ruthenium chloride in the culture media on the nitrogen-fixing ability of the three nitrogen fixers (unidentified species of Azotobacter, designated here as D3, B3, and B), isolated from Allahabad soil, was studied. It was observed that the nitrogen-fixing ability of the organisms is much increased in presence of 25-75 micro M concentration of ruthenium chloride in the culture media, while sugar consumption remains more or less steady. Also, if mg nitrogen fixed/g carbon consumed in two culture media with successive increasing concentrations of ruthenium chloride is compared by calculating the difference in increase of the amount of nitrogen fixed and carbon consumed in these two culture media, it was observed that high amounts of nitrogen are fixed by B3 and B between 25-50 micro M, and between 50-75 micro M concentration of ruthenium chloride by D3.

Azotobacter↗

Effects of inland nitrogen transport and attenuation modeling on coastal nitrogen load abatement.

Modeling of the spatial distribution of nitrogen transport and attenuation from various inland sources and along different hydrological pathways to coastal waters is needed for relevant decisions on effective allocation of measures for coastal nitrogen load abatement. We identify, classify, and quantify uncertainties associated with main discrepancies between spatial process representations in different catchment-scale nitrogen transport-attenuation models. The results show important model differences, indicating scientific disagreement on the realistic spatial process understanding, representation, and quantification in nitrogen transport-attenuation modeling. By further developing solutions for economic optimization of spatially differentiated nitrogen source abatement in coastal catchments, we find this disagreement to considerably affect the economic efficiency of coastal nitrogen load reduction. It may also lead to stakeholder mistrust and conflict and needs to be recognized and handled in environmental policy.

Algorithms↗

A study on the spatial distribution of nitrogen and phosphorus balance, and regional nitrogen flow through crop production.

The spatial distribution patterns of the nitrogen and phosphorus input/intake amounts in crop production within two small basins are examined, based upon a cropping unit distribution map that is obtained from remote sensing data analysis. Firstly, we examine the availability and suitability of approaches to the spatial distribution analysis of cultivation patterns classified from material flow characteristics of crop production using seasonal remote-sensing data. Secondly, material flow units in crop production are grouped according to the cultivation patterns obtained from the remote-sensing data analysis. Consequently, the spatial patterns of the amounts of both nitrogen and phosphorus inputs/intakes through crop production on farmland are examined and their spatial distribution maps are prepared according to the material flow units. In addition, we developed a nitrogen flow and runoff model and the model is simulated based on the examination of the results of spatial distribution patterns of the material flow units. The annual nitrogen runoff from small catchments, where various crops are cultivated, varies from 2.7 kg ha(-1) year(-1) to 108 kg ha(-1) year(-1) and the annual balanced losses of nitrogen in small catchments varied from -30 kg ha(-1) year(-1) to 101 kg ha(-1) year(-1). Also, the monthly changes in soil nitrogen of each material flow unit is estimated at -55 kg ha(-1) as a maximum decrease and 114 kg ha(-1) as a maximum increase. These results indicate that the spatial distribution patterns of nutrient input and intake through agricultural activities should be considered when analyzing the material flows and nutritient movement in soil-water systems in rural areas for watershed environmental control and regional agricultural management.

Agriculture↗

Metabolic engineering with Dof1 transcription factor in plants: Improved nitrogen assimilation and growth under low-nitrogen conditions.

Utilization of transcription factors might be a powerful approach to modification of metabolism for a generation of crops having superior characteristics because a single transcription factor frequently regulates coordinated expression of a set of key genes for respective pathways. Here, we apply the plant-specific Dof1 transcription factor to improve nitrogen assimilation, the essential metabolism including the primary assimilation of ammonia to carbon skeletons to biosynthesize amino acids and other organic compounds involving nitrogen in plants. Expressing Dof1 induced the up-regulation of genes encoding enzymes for carbon skeleton production, a marked increase of amino acid contents, and a reduction of the glucose level in transgenic Arabidopsis. The results suggest cooperative modification of carbon and nitrogen metabolisms on the basis of their intimate link. Furthermore, elementary analysis revealed that the nitrogen content increased in the Dof1 transgenic plants (approximately 30%), indicating promotion of net nitrogen assimilation. Most significantly, the Dof1 transgenic plants exhibit improved growth under low-nitrogen conditions, an agronomically important trait. These results highlight the great utility of transcription factors in engineering metabolism in plants.

Amino Acids↗

Saccharomyces cerevisiae Sit4 phosphatase is active irrespective of the nitrogen source provided, and Gln3 phosphorylation levels become nitrogen source-responsive in a sit4-deleted strain.

Tor1,2 control of type 2A-related phosphatase activities in Saccharomyces cerevisiae has been reported to be responsible for the regulation of Gln3 phosphorylation and intracellular localization in response to the nature of the nitrogen source available. According to the model, excess nitrogen stimulates Tor1,2 to phosphorylate Tip41 and/or Tap42. Tap42 then complexes with and inactivates Sit4 phosphatase, thereby preventing it from dephosphorylating Gln3. Phosphorylated Gln3 complexes with Ure2 and is sequestered in the cytoplasm. When Tor1,2 kinase activities are inhibited by limiting nitrogen, or rapamycin-treatment, Tap42 can no longer complex with Sit4. Active Sit4 dephosphorylates Gln3, which can then localize to the nucleus and activate transcription. The paucity of experimental data directly correlating active Sit4 and Pph3 with Gln3 regulation prompted us to assay Gln3-Myc(13) phosphorylation and intracellular localization in isogenic wild type, sit4, pph3, and sit4pph3 deletion strains. We found that Sit4 actively brought about Gln3-Myc(13) dephosphorylation in both good (glutamine or ammonia) and poor (proline) nitrogen sources. This Sit4 activity masked nitrogen source-dependent changes in Gln3-Myc(13) phosphorylation which were clearly visible when SIT4 was deleted. The extent of Sit4 requirement for Gln3 nuclear localization was both nitrogen source- and strain-dependent. In some strains, Sit4 was not even required for Gln3 nuclear localization in untreated or rapamycin-treated, proline-grown cells or Msx-treated, ammonia-grown cells.

Active Transport, Cell Nucleus↗

[Energy and nitrogen metabolism of pregnant and lactating sows and suckling piglets. 6. Comparative studies of energy and nitrogen metabolism of non-pregnant and pregnant sows].

Aim of the studies was to attain new scientific findings for the influence of the physiological state of sows (non-pregnant and pregnant respectively) on energy and nitrogen metabolism. The experiments were carried out according to a 3 x 3 factorial experimental plan with 3 variants of litter number (1, 2 and 4) and 3 variants of energy supply (120, 100 and 80%). Within the variants the non-pregnant and pregnant sows were fed equally related to the metabolic live weight. The metabolism measurements run from the 1st to the 115th experimental and gestation day respectively. The methods of indirect calorimetry and slaughtering technique were applied. The chemical composition of the whole bodies of the non-pregnant sows was like that of the maternal bodies of the pregnant sows (dry matter 410 g/kg, protein 160 g/kg and fat 210 g/kg). The physiological state for the sows did not influence the nutrients digestibility (OM 77.0 and 76.7%) and the energy metabolizability. About conformable intakes of metabolizable energy (601 and 586 kJ/kg LW0.75.d) and of digestible nitrogen (0.77 and 0.75 g/kg LW0.75.d) in non-pregnant and pregnant sows resulted in higher deposition of energy (64 and 79 kJ/kg LW0.75.d) and of nitrogen (0.15 and 0.19 g/kg LW0.75.d) in the pregnant sows due to energy and nitrogen deposition in the conception products and in the reproductive organs. The energy maintenance requirement and the partial efficiency of the energy utilization for the deposition as well as the nitrogen maintenance requirement and the partial efficiency of the nitrogen utilization for the deposition in the non-pregnant and the pregnant sows were determined.

Animals↗

Photosynthetic light acclimation in peach leaves: importance of changes in mass:area ratio, nitrogen concentration, and leaf nitrogen partitioning.

Photosynthetic light acclimation of leaves can result from (i) changes in mass-based leaf nitrogen concentration, Nm, (ii) changes in leaf mass:area ratio, Ma, and (iii) partitioning of total leaf nitrogen among different pools of the photosynthetic machinery. We studied variations in Nm and Ma within the crowns of two peach (Prunus persica L. Batsch) trees grown in an orchard in Portugal, and one peach tree grown in an orchard in France. Each crown was digitized and a 3-D radiation transfer model was used to quantify the intra-crown variations in time-integrated leaf irradiance, . Nitrogen concentration, leaf mass:area ratio, chlorophyll concentration, and photosynthetic capacity were also measured on leaves sampled on five additional peach trees in the orchard in Portugal. The data were used to compute the coefficients of leaf nitrogen partitioning among carboxylation, bioenergetics, and light harvesting pools. Leaf mass:area ratio and area-based leaf nitrogen concentration, Na, were nonlinearly related to , and photosynthetic capacity was linearly related to Na. Photosynthetic light acclimation resulted mainly from changes in Ma and leaf nitrogen partitioning, and to a lesser extent from changes in Nm. This behavior contrasts with photosynthetic light acclimation observed in other tree species like walnut (Juglans regia L.) in which acclimation results primarily from changes in Ma.

Chlorophyll↗

Utilization of ammonium as a nitrogen source: effects of ambient acidity on growth and nitrogen accumulation by soybean.

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

Biomass↗

Growth and metabolism of Saccharomyces cerevisiae in chemostat cultures under carbon-, nitrogen-, or carbon- and nitrogen-limiting conditions.

Aerobic chemostat cultures of Saccharomyces cerevisiae were performed under carbon-, nitrogen-, and dual carbon- and nitrogen-limiting conditions. The glucose concentration was kept constant, whereas the ammonium concentration was varied among different experiments and different dilution rates. It was found that both glucose and ammonium were consumed at the maximal possible rate, i.e., the feed rate, over a range of medium C/N ratios and dilution rates. To a small extent, this was due to a changing biomass composition, but much more important was the ability of uncoupling between anabolic biomass formation and catabolic energy substrate consumption. When ammonium started to limit the amount of biomass formed and hence the anabolic flow of glucose, this was totally or at least partly compensated for by an increased catabolic glucose consumption. The primary response when glucose was present in excess of the minimum requirements for biomass production was an increased rate of respiration. The calculated specific oxygen consumption rate, at D = 0.07 h-1, was more than doubled when an additional nitrogen limitation was imposed on the cells compared with that during single glucose limitation. However, the maximum respiratory capacity decreased with decreasing nitrogen concentration. The saturation level of the specific oxygen consumption rate decreased from 5.5 to 6.0 mmol/g/h under single glucose limitation to about 4.0 mmol/g/h at the lowest nitrogen concentration tested. The combined result of this was that the critical dilution rate, i.e., onset of fermentation, was as low as 0.10 h-1 during growth in a medium with a low nitrogen concentration compared with 0.20 h-1 obtained under single glucose limitation.

Ammonium Sulfate↗

Influence of energy and nitrogen contents of enteral diets on nitrogen balance: a double blind prospective controlled clinical trial.

Results of recent controlled studies show that because of difficulties in administering adequate quantities of enteral diet, positive nitrogen balance is not consistently achieved during enteral feeding. In order to determine whether nitrogen balance can be improved in routine clinical practice by prescribing enteral diets containing higher concentrations of nutrients, 118 patients with normal gastrointestinal function needing enteral nutrition were randomised to receive daily 21 of one of three polymeric diets: Standard diet (1.0 kcal/ml; 6.3 gN/l), Energy Dense diet (1.5 kcal/ml; 7.8 gN/l), and Energy-Nitrogen Dense diet (1.5 kcal/ml; 9.4 gN/l. The three diets, administered by continuous nasogastric infusion, were equally well tolerated. Results were analysed only for patients fed five or more days and who received at least 60% of prescribed enteral diet (n = 42). Positive nitrogen balance was achieved only in the patients receiving the Energy-Nitrogen Dense diet (n = 16; + 1.6 (SE) 0.6 gN/d, compared with the Standard diet (n = 12; -3.8 (1.1) gN/d; p less than 0.001), and the Energy Dense diet (m = 14; -1.9 (0.8) gN/d; p less than 0.005). As the findings of this prospective controlled trial show that positive nitrogen balance was not consistently achieved by administering 21 enteral diet containing up to 15.6 gN, consideration could, therefore, be given to routinely using enteral diets containing up to 9.4 gN/l.

Adolescent↗

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. I. Development of a model for predicting nonprotein nitrogen utilization by cattle.

The influence of ration composition on mean ruminal ammonia concentration was studied by collecting samples of ruminal ingesta from cattle fed rations varying in crude protein and total digestible nutrient content. A minimum of four sampling times distributed throughout the day permitted calculation of mean ruminal ammonia concentrations. Mean ruminal ammonia concentration was positively related to dietary crude protein concentration and negatively related to total digestible nutrient concentration. It is postulated that mean ruminal ammonia concentration may be a useful criterion for predicting efficacy of nonprotein nitrogen supplementation. A quantitative approach for evaluating nonprotein nitrogen supplementation based upon determination of the point at which ruminal ammonia exceeds the requirement (5 mg ammonia nitrogen/100 ml rumen fluid) of the ammonia-utilizing bacteria is proposed. Dietary conditions expected to result in excessive concentrations of ruminal ammonia are defined and recommended upper limits for nonprotein nitrogen supplementation are presented. Theoretical relationships between composition of the unsupplemented ration, amount of nonprotein nitrogen added, and efficiency of nonprotein nitrogen utilization are discussed. The practice of adding nonprotein nitrogen so as to exceed 12 to 13% crude protein in typical dairy or feedlot rations needs to be reevaluated.

Ammonia↗

Influence of nonprotein nitrogen and protein of low rumen degradability on nitrogen flow and utilization in lactating dairy cows.

Four lactating cows, each fitted with a rumen cannula and duodenal and ileal t-cannulae, were used to measure flow and digestion of nitrogenous compounds in the digestive tract. Dietary dry matter contained 17% crude protein and 50:50 forage:concentrate. Treatments were: 1) corn silage-soybean meal; 2) corn silage-heated soybean meal; 3) ammonia-treated corn silage-soybean meal; and 4) ammonia-treated corn silage-heated soybean meal. Flow of organic matter to the duodenum was overestimated when lanthanum or chromium ethylene-diaminetetraacetate was used as an indigestible marker. This resulted in low estimates of ruminal digestion of organic matter and high estimates of nitrogen flow to the duodenum. However, calculations using lanthanum or lignin as markers yielded similar organic matter flow to ileum and feces. With acid-detergent lignin as a marker, estimates of dietary nitrogen degraded in the rumen were: corn silage-heated soybean meal, 55.0% ammonia-treated corn silage-heated soybean meal, 58.8%; ammonia-treated corn silage-soybean meal, 63.3%; and corn silage-soybean meal, 66.0%. Digestion in the small intestine of nonammonia nitrogen was equal for all treatments suggesting that availability of heated soybean meal in the intestine was not different, although ruminal degradability tended to be lower. Feeding diets containing nonprotein nitrogen did not decrease available nitrogen at the duodenum.

Animals↗

Urea index and nitrogen balance in uremic patients on miminal nitrogen intakes.

Normal anduremic subjects were studied on different protein intakes. The uremic patients were also given minimal amounts of nitrogen in the form of 8 essential amino acids, with or without the addition of histidine. It was noticed that as little as 416 mg of essential amino acid nitrogen was sufficient to cover body needs if enough non-essential nitrogen was present. It was also demonstrated that on low-nitrogen intakes, the ratio of urinary urea to total urinary nitrogen varied with nitrogen balance.

Adolescent↗

Pyrochemiluminescence: real-time, cost-effective method for determining total urinary nitrogen in clinical nitrogen-balance studies.

Total urinary nitrogen (TUN) determinations for nitrogen-balance studies were traditionally performed by the Kjeldahl method, but this method is laborious, hazardous, prone to error, and no longer widely available in most clinical laboratories. During the last several decades, urinary urea nitrogen (UUN) determinations have replaced TUN as an index of urinary nitrogen excretion in many clinical laboratories, owing to its ease of determination, decreased cost, and wide availability. However, the validity of using UUN for estimating nitrogen loss has been questioned in many disease states, owing to wide variations in the proportional amount of urea found in TUN. Chemiluminescence has been proposed as an alternative to the Kjeldahl method for TUN. TUN values obtained from 24-h urine collections measured by both micro-Kjeldahl (x) and Pyrochemiluminescence (y) (Antek Instruments, Inc.) techniques were comparable by linear regression analysis: n = 97; r = 0.996; r2 = 0.992; y = 1.048x - 0.606; P less than 0.001. Automated induction of samples and calculation of results allows up to 42 samples to be run unattended. This dramatically reduces labor and overall costs for TUN determinations, while providing a more accurate and economical assessment of nitrogen excretion than UUN in a clinical setting.

Cost-Benefit Analysis↗

Nitrogen dioxide and respiratory illness in children. Part II: Assessment of exposure to nitrogen dioxide.

Repeated measurements of nitrogen dioxide were obtained from 1988 to 1991 in the homes of 1,205 infants living in Albuquerque, NM. Passive diffusion samplers were used to obtain a series of two-week integrated measurements from the home of each infant for use in a cohort study of the relation of residential exposure to nitrogen dioxide and respiratory illnesses. Information on stove use and time spent inside the residence was collected at two-week and two-month intervals, respectively. During the winter, in the bedrooms of homes with gas cooking stoves, mean nitrogen dioxide concentrations were 21 parts per billion (ppb); mean concentrations in the living room and kitchen were 29 ppb and 34 ppb, respectively. In homes with electric cooking stoves, the mean bedroom concentration was 7 ppb during the winter. Lower indoor concentrations were observed during the summer in homes with both gas and electric stoves. On average, infants spent approximately 12.3 hours per day in their bedrooms, 7.3 hours in the living rooms, 35 minutes in the kitchens, and 3.8 hours out of their homes. (As a condition of participation, none of the infants spent more than 20 hours per week in day care outside of their homes). The mean time infants spent in the kitchen during cooking was approximately nine minutes per day. We tested whether exposures of infants living in homes with gas stoves could be reasonably estimated by measurements in the bedroom in comparison with time-weighted average concentrations based on time-activity data and simultaneous nitrogen dioxide measurements in the kitchen, living room, and bedroom. In 1,937 two-week intervals from 587 infants, 90% of time-weighted exposure (on the three-level classification used in this study) estimates were in agreement with estimates based on bedroom concentrations alone. The agreement of the time-weighted nitrogen dioxide exposure estimates with the bedroom concentrations is attributed to limited amounts of cooking stove use (the mean was 29 minutes per day), small room-to-room differences in nitrogen dioxide concentrations (the mean kitchen-bedroom difference was 12 ppb), and the relatively large proportion of time that infants spent in their bedrooms.

Air Pollution, Indoor↗