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1.
In experiments on 4 sheep fed on a low protein diet [6.2 g N/day] and given a single i.v. dose of 15N-labelled urea [15 mg 15N/kg body mass], the authors found that, from 0.5 to 6 h, mean 15N incorporation rose progressively in the total rumen fluid nitrogen from 0.23 to 0.44 at. % 15N and in the rumen bacterial nitrogen from 0.11 to 0.51 at. % 15N. Up to 3 h, total nitrogen enrichment was greater (0.5 at. % 15N) than enrichment of bacterial nitrogen (0.28 at. % 15N), but from 3 to 6 h there was little difference between them. The mean 15N values in the nucleic acids isolated from rumen fluid bacteria in samples collected 3 and 6 hours after injecting labelled urea into the blood were 0.15 and 0.19 at. % 15N respectively, in nucleic acids isolated from the liver 0.042 and 0.04 at. % 15N, in the total rumen bacterial nitrogen 0.28 and 0.51 at. % 15N and in the total liver nitrogen 0.11 and 0.11 at. % 15N. It is concluded from the results that blood urea nitrogen is utilized for synthesis of the total nitrogenous substances of the sheep's rumen bacteria and liver far more intensively than for synthesis of the nucleic acids isolated from them. At the same time, it is utilized more intensively for nucleic acid synthesis in the rumen bacteria than in the liver.  相似文献   

2.
The metabolic fate of nitrogen from 15N-labeled sodium nitrate has been investigated in four healthy Polish Merino ewes. 15N-labeled sodium nitrate was administered intravenously at the dosage of 400 micromol.kg(-1) body weight. Blood plasma and urine concentrations of nitrate, ammonia, and urea and 15N enrichment of ammonia and urea were estimated over a 50-h period following 15N-nitrate administration. Nitrate (NO3-) was slowly eliminated from the blood plasma, and the presence of NO3(-) in the blood plasma above the nitrate "background" was observed for 50 h. 15N enrichment of blood plasma urea already appeared at 15 min and reached the maximum 6 h after 15N-nitrate administration. The urinary excretion of nitrate occured during 50 h after 15N-nitrate injection; the total urine excretion of NO3(-) was 23.63+/-2.39% of the administered dose. The mean urinary recoveries of nitrogen as 15N-urea and 15N-ammonia were 14.76+/-1.32% and 0.096+/-0.015% of the administered 15N-nitrate dose, respectively. It should be pointed out that in total only 38.49% of the administered nitrate-N was excreted in urine (as nitrate, ammonia and urea nitrogen) during 50 h. The results obtained indicate that sheep are able to store nitrate nitrogen in their body. The fate of the remaining approximately 60% of the 15NO3(-) administered dose is unknown. The results obtained do not allow one to conclude what fraction of the unrecovered approximately 60% of the 15NO3(-) dose was utilized by gastrointestinal microorganisms, and (or) metabolized, or stored in sheep tissues.  相似文献   

3.
The retention and excretion of intrajejunally administered 15N-urea was studied in four experiments on two sheep with a permanently fistulated small intestine. In the first 7 days after the administration of 2 g 15N-urea, 18.26% was excreted in the faeces and 19% in the urine; 62.74% was retained in the organism. Urinary excretion took place mainly on the first day and from the 3rd to the 7th day no 15N was present in the urine. The rate of 15N excretion in the faeces was roughly the same for the first 4 days and then fell; on the 7th day there was no 15N in the faeces. The proportion of 15N-urea retained in the organism and excreted in the urine was 81% showing that urea in the ruminant gastrointestinal tract is largely linked up into metabolic circulation as part of the general exchange of nitrogenous compounds.  相似文献   

4.
The endogenous biosynthesis of nitrate in rats was investigated by using 15NH3 administered as a continuous intravenous infusion for as long as 96 h. A comparison of the enrichment of 15N in urinary nitrate after a 24 h infusion revealed that it was 36% of the enrichment of plasma NH3 and about 50% of the enrichment of plasma urea and urinary NH3. Continuous infusion of 15NH3 for 96 h showed that a plateau for the incorporation of NH3 into nitrate is reached by 24 h, whereas the enrichment of urinary NH3 and urea increase during the 96 h. After the infusion of progressively larger doses of 15NH3, the concentration of nitrate synthesized de novo increased. Although there was a significant correlation between plasma 15NH3 concentration and 15NO3- appearance, a given change in plasma NH3 concentration does not produce a direct proportional change in nitrate synthesis. Our findings indicate that NH3 is a quantitatively significant nitrogen precursor for nitrate, but that approx. 50% of nitrate nitrogen is derived from other, as yet unidentified, sources.  相似文献   

5.
1. 15N-Percentage of the amide of glutamine in total blood non-protein-15N was 42 and 48% in chickens fed 5 and 20% protein diets, respectively, when 15N-ammonia was intraportally-infused for 6 hr. 2. The infused ammonia-15N also appeared in the amide of free glutamine in the liver and kidney in large amounts at both levels of protein intake. 3. The 15N incorporated into glutamine-amide in the blood, liver and kidney and non-protein-15N in plasma were greater in chickens fed the low protein diet than in those fed the high protein diet (P less than 0.05 except kidney of P less than 0.01). 4. About 60% of the amide-N of the glutamine which was increased during 6 hr infusion of ammonia was derived from infused ammonia-N and the remainder from endogenous nitrogen, irrespective of protein intake. 5. These results suggest that glutamine is the most important intermediate in detoxication of intraportal ammonia in chickens.  相似文献   

6.
The effect of cyanide on ammonia and urea metabolism was studiedwith intact cells of Chlorella ellipsoidea Gerneck, a greenalga which apparently lacks urease. Ammonia uptake was inhibited more readily by cyanide than wasurea uptake. Urea uptake was stimulated by lower concentrationsof cyanide. The addition of cyanide caused the formation ofammonia from some cellular nitrogenous compounds. In the presenceof exogenously added urea, the molar ratio of ammonia accumulatedin the medium to urea taken up exceeded 2.0 as the cyanide concentrationincreased. However, the molar ratio of ammonia actually producedfrom urea nitrogen to urea taken up was less than 1.35 at anyconcentration of cyanide tested. In the presence of higher concentrationsof cyanide, the rate of incorporation of 15N into amino acidsfrom 15N-urea was higher than that from 15N-ammonium sulfate. The results suggest that Chlorella ellipsoidea possesses a pathwaythrough which urea nitrogen is assimilated directly withouta preliminary breakdown to ammonia. (Received October 18, 1976; )  相似文献   

7.
Urea can be one of the major sources of nitrogen for phytoplankton, but little is known about its importance for corals. Experiments were therefore designed to assess the uptake rates of urea by the scleractinian coral Stylophora pistillata; 15N-urea was used to follow the incorporation of nitrogen into the zooxanthellae and animal tissue. The uptake kinetics of urea in the tissue of S. pistillata showed that there is a concentration-dependent uptake of urea. The transport of urea was composed of a linear component (diffusion) at concentrations higher than 6 μmol N-urea l− 1 and an active carrier-mediated component, at lower concentrations. The value of the carrier affinity (Km = 1.05 μmol urea l− 1) indicates a good adaptation of the corals to low levels of urea in seawater. At the in situ concentration of ca. 0.2 μmol N-urea l− 1, the uptake rate was equal to 0.1 nmol N h− 1 cm− 2. Urea uptake was at least four times higher in the animal than in the algal fraction, and five times higher when corals were incubated in the light than in the dark. These results could be explained by the involvement of urea in the calcification process, which is also enhanced by light. Comparison of urea uptake rates with nitrate or ammonium uptake rates for the same S. pistillata species, at in situ concentrations, showed that urea is preferred to nitrate and may therefore be an important source of nitrogen for scleractinian corals.  相似文献   

8.
Effect of dietary energy intake on tubular reabsorption of urea in sheep   总被引:2,自引:0,他引:2  
The aim of the experiment was to determine the effect of dietary energy intake on renal urea excretion in sheep with different nitrogen intakes. The control sheep, with a high nitrogen and energy intake, were given a daily feed dose of 21.18 g N and 15.2 MJ digestible energy (DE). The two experimental groups, with an equal, low nitrogen intake, were given diets with a different energy content. The high energy diet contained 3.63 g N and 14.18 MJ DE, the low energy diet 3.4 g N and 6.44 MJ DE. After nine weeks' adaptation to the diets, renal functions were measured by a standard clearance technique. It was found that, under stable urine flow conditions, both groups given the low nitrogen diet had a significantly lower glomerular filtration rate, fractional urea excretion and total urea excretion. A reciprocal comparison of these two groups showed that fractional urea excretion by the sheep with a high energy intake was significantly lower than in the group with a low energy intake. There were no differences in the glomerular filtration rate. A raised dietary energy intake in the presence of a low nitrogen intake caused marked natriuresis and kaliuresis. The results indicate that a raised dietary energy intake can be a significant factor in potentiating the renal effect of urea retention in sheep with a low nitrogen intake.  相似文献   

9.
The renal response of sheep to a low dietary nitrogen intake   总被引:1,自引:0,他引:1  
Renal functions were tested in sheep fed on a low nitrogen diet (LN sheep), with a daily N intake of 4.7 g (gross energy 17.76 . 10(6) J). Sheep given a high nitrogen diet (HN sheep) with 21.2 g N (24.12 . 10(6) J) acted as the control. The functions of the left kidney were measured in anaesthetized animals by the standard clearance technique. A comparison of HN and LN sheep showed that a low nitrogen intake led to a drop in the plasma urea level (from 5.91 +/- 0.35 to 2.87 +/- 0.36 mmol.1-1, (P less than 0.001), the glomerular filtration rate (GFR, from 36.6 +/- 3.6 to 20.7 +/- 2.4 ml.min-1, P less than 0.005), amount of urea excreted (from 106.7 +/- 18.1 to 15.7 +/- 3.3 mumol.min-1, P less than 0.001), fractional urea excretion (from 51.0 +/- 3.0 to 24.6 +/- 3.1 %, P less than 0.001) and the absolute tubular reabsorption of urea (Reaburea/GFR (from 3.06 +/- 0.27 to 2.12 +/- 0.28 mumol.ml-1, P less than 0.05), without a significant change in the effective renal plasma flow (182.6 +/- 20.0 and 138.5 +/- 21.0 ml.min-1, non-significant - N.S.) and in sodium and potassium excretory function. Free water clearance rose in LN sheep (from -0.53 +/- 0.11 to -0.19 +/- 0.06 ml.min-1, P less than 0.05) owing to inhibited urea excretion. A regression analysis of the relationship of the tubular reabsorption of urea to the amount of filtered urea (both normalized to the GFR) showed that the urea transport capacity of the tubules of LN sheep was significantly higher.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

10.
15N kinetic labeling studies were performed on seedlings of Hordeum vulgare L. var. Golden Promise growing under steady state conditions. Patterns of label incorporation in the pools of nitrogen compounds of roots fed [15N]ammonium were compared with computer-simulated labeling curves. The data were found to be quantitatively consistent with a three-compartment model in which ammonium is assimilated solely into the amide-N of glutamine. Labeling data from roots fed [15N]nitrate were also found to be at least qualitatively consistent with the assimilation of ammonia into glutamine. Methionine sulfoximine almost completely blocked the incorporation of 15N label into the amino acid pools of barley roots fed [15N]nitrate. These observations suggest that ammonia assimilation occurs solely via the glutamine synthetase/glutamate synthase pathway in both nitrate- and ammonia-grown barley roots.  相似文献   

11.
A gas chromatographic-mass spectrometric method for analysis of L-[2-15N]- and L-[5-15N]glutamine is described. The method is based on direct acylation of glutamine with trifluoroacetic anhydride and the formation of the N,N-bis-trifluoroacetyl-L-glutamine derivative. This simple and sensitive method is capable of detecting approximately 0.5 atom% excess 15N in as little as 10 microliter of plasma with a mean coefficient of variance of 11.6%. The method was applied to determine the appearance of 15N enrichment in plasma amino-N and amide-N of glutamine in a healthy adult volunteer during a constant infusion of 15NH4Cl. A plateau level of 3.7 and 2.6 atom% excess was observed in amide-N and amino-N, respectively, at 1 and 2 h after 15NH4Cl infusion was started.  相似文献   

12.
Urea concentration and urease activity in the midgut content were compared between larvae of the silkworm, Bombyx mori fed an artificial diet and those fed fresh mulberry leaves. A considerable amount of urea was found in the midgut content of the both larvae, however it was significantly lower in the larvae fed fresh mulberry leaves than in the larvae fed the artificial diet; average urea concentrations in the midgut content of the larvae fed fresh mulberry leaves and the artificial diet were 2.9 and 4.6 &mgr;mol/g, respectively. Urea in the midgut content seems to be secreted from the insect itself since the amount of urea in both diets were negligibly small. Urease activity was detected only in the midgut content of the larvae fed fresh mulberry leaves but not in other tissues of the larvae. On the other hand, no urease activity was detected in the midgut content of the larvae fed the artificial diet. Subsequently, to elucidate the role of mulberry leaf urease in the midgut lumen, larvae that had been reared on the artificial diet were switched to fresh mulberry leaves. The diet switch caused a rapid decrease in urea concentration in the midgut content and an increase in ammonia concentration in the midgut content, suggesting that secreted urea could be hydrolyzed to ammonia by mulberry leaf urease in the midgut lumen. Furthermore, to investigate the physiological significance of mulberry leaf urease on urea metabolism of the silkworm, (15)N-urea was injected into the hemocoel, and after 12 h the larvae were dissected for (15)N analysis. A considerable amount of (15)N was found to be incorporated into the silk-protein of the larvae fed fresh mulberry leaves, but there was little incorporation of (15)N into the silk-protein of the larvae fed the artificial diet. These data indicate that urea is converted into ammonia by the action of mulberry leaf urease in the midgut lumen and used as a nitrogen source in larvae fed mulberry leaves.  相似文献   

13.
Splanchnic tissues are largely involved in the postprandial utilization of dietary amino acids, but little is yet known, particularly in humans, about the relative contributions of different splanchnic protein pools to splanchnic and total postprandial anabolism. Our aim was to develop a compartmental model that could distinguish dietary nitrogen (N) incorporation among splanchnic constitutive, plasma (splanchnic exported), and peripheral proteins after a mixed-protein meal in humans. Eight healthy subjects were fed a single mixed meal containing 15N-labeled soy protein, and dietary N postprandial kinetics were measured in plasma free amino acids, proteins, and urea and urinary urea and ammonia. These experimental data and others previously obtained for dietary N kinetics in ileal effluents under similar experimental conditions were used to develop the compartmental model. Six hours after the mixed-meal ingestion, 31.5, 7.5, and 21% of ingested N were predicted to be incorporated into splanchnic constitutive, splanchnic exported, and peripheral proteins, respectively. The contribution of splanchnic exported proteins to total splanchnic anabolism from dietary N was predicted to be approximately 19% and to remain steady throughout the simulation period. Model behavior and its predictions were strongly in line with current knowledge of the system and the scarce, specific data available in the literature. This model provides the first data concerning the anabolism of splanchnic constitutive proteins in the nonsteady postprandial state in humans. By use of only slightly invasive techniques, this model could help to assess how the splanchnic anabolism is modulated under different nutritional or pathophysiological conditions in humans.  相似文献   

14.
选用15N同位素标记的新型回收塑料包膜控释肥和大颗粒尿素,采用池栽试验研究夏玉米-冬小麦轮作体系中肥料氮的去向及利用率。结果表明,整个轮作体系中,控释肥处理(PCU)作物吸收的肥料氮为241.03 kg/hm,高于尿素处理(Urea)的211.02 kg/hm。控释肥处理施用的肥料氮主要残留在0~40 cm土层,而尿素处理则残留在0~60 cm土层,控释肥延缓了肥料氮向土壤深层迁移的趋势。在夏玉米和冬小麦轮作体系中,控释肥处理的氮肥利用率(32.86%,32.47%)高于尿素处理(28.23%,30.16%)。在冬小麦季,控释肥处理损失率相比尿素处理从36.07% 降至28.75%,而夏玉米季,控释肥处理损失率相比尿素处理从37.17%降至29.50%。玉米季控释肥处理与尿素处理差异不显著,但在冬小麦季控释肥处理的产量显著高于尿素处理。因此,在玉米和小麦整个生长季,新型回收塑料包膜控释肥的养分释放与作物养分需求吻合,既提高氮肥利用率,也降低了肥料氮的损失。  相似文献   

15.
Summary Nitrogen metabolism and urea kinetics were studied in rock hyraxes (Procavia habessinica) fed diets of different protein content.The maintenance nitrogen (N) requirement of the rock hyrax (311 mg·kg–0.75·24 h–1 of dietary N, or 209 mg·kg–0.75·24 h–1 of truly digestible N) is similar to that of several marsupial species, and thus lower than that of other eutherians.Urea recycled to the gut, measured with single injections of14C-urea, was 63% and 60% of urea entry rate on diets with 14.6% and 8.2% crude protein, respectively. Urea recycling increased to 70% when water intake was restricted, but decreased to 40% on a low (5.3%) protein diet, presumably because of a low energy intake.Urea utilization in the gut, measured with single injections of15N-urea, was 59% and 53% of urea degradation on the 14.6% and 8.2% protein diets, respectively. Urea utilization increased to 71% on the low protein diet, and increased to 98% with water restriction.The low maintenance nitrogen requirement appears to be the main physiological attribute of the rock hyrax enabling it to survive periods of low dietary protein availability. However, this low requirement can be related to the low basal metabolic rate of the Hyracoidea in general, and thus is not necessarily a primary adaptation to the environment.  相似文献   

16.
We investigated the effect of human milk feeding on the nitrogen metabolism of appropriate-for-gestational age infants of birth weight 1.5-2.0 kg. Eight infants received pooled mature human milk. The remaining 20 were divided into two equal groups, who received one of two low-protein, milk-based formulae. The formulae were identical in composition except for the protein source, which was either casein- or whey-predominant. The three diet groups received similar total nitrogen (390 mg N.kg-1.d-1) and energy (500 kJ.kg-1.d-1) intakes. The human-milk-fed group, however, received a significantly higher intake of nonprotein and urea nitrogen and a significantly lower true protein nitrogen. Nitrogen metabolism was studied using a modified constant infusion of [15N]glycine, mixed with the feeding every 2-3 h. Urine was collected in approximately 3-h aliquots and analysed for total ammonia and urea nitrogen. Excretion of the 15N label was measured in urinary urea and ammonia. No differences were seen between the three diet groups in total [15N]urea or [15N]ammonia urinary excretion. However, the concentration of 15N in urinary urea in the human-milk-fed group was lower than in the two formula-fed groups. This reduction in concentration appeared due to a higher dietary intake of urea among the human-milk-fed group, and the consequent dilution of the label in the urine. As a result, protein turnover rates calculated from the [15N]urea end product were artificially raised in the milk-fed group, and were significantly higher than those in the formula groups.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

17.
The derivation of alanine in fibroin was investigated using NMR and selective isotopic labelling. 2H2O infused orally into 5th instar larvae was incorporated into the proton of the methyl group of alanine in fibroin. Proton exchange among alanine, glycine and serine was also found. Incorporation of 13C from [2-(13)C]acetate into alanine C2 and C3 and glycine C2 in fibroin, and also C4 of free glutamine plus glutamate was observed in vivo. Hemolymph contained a peak for C4 of glutamate plus glutamine, and an alanine C3 peak appeared transiently. Thus, it is suggested that the C-skeleton of alanine formed was derived from L-malate via the TCA-cycle, and that this alanine is utilized in part for fibroin synthesis. Spectra of the hemolymph extract of larvae infused orally with [15N2]urea showed no 15N-compounds, whereas those of larvae injected subcutaneously showed only one peak of urea, whose intensity decreased with time, as shown in the in vivo spectra of a living larva infused with [15N2]urea. The solution NMR spectrum of fibroin showed no 15N-labelled compounds. Temporal changes in the peak intensities of six compounds in the spectra of a living larva infused with [15N]ammonium demonstrated a process in which 15N was incorporated into fibroin containing 15N-alanine through the amide group of glutamine and the amino group of glutamate. Thus, alanine biosynthesis from the TCA-cycle originates mainly from water, L-malate and ammonium. The fact that no 15N-urea was detected in the hemolymph extract of larvae infused with [15N]ammonium suggests that 15N-urea found in the above in vivo spectra may be that accumulated in the hindgut. Thus, excess ammonium in the body causes the production of urea by the urea-cycle. In Samia larvae, urea was not reutilized but excreted. The metabolic relationships between the assimilation of ammonium and the function of the urea-cycle are discussed.  相似文献   

18.
We measured the incorporation of recycled urea-nitrogen (N) by ruminal microbes, using five ruminally and duodenally fistulated steers (237 kg) fed low-quality grass hay (47 g crude protein/kg dry matter (DM)). Three received 1 kg/day of soybean meal (SBM) and two received no supplemental protein (control). The experiment was 15 days long. Background enrichments of 15N were measured on day 9 and continuous jugular infusion of 0.12 g/day [15N15N]urea began on day 10. Daily samples of urine, feces, ruminal bacteria and duodenal digesta from days 10 through 14 were used to determine plateaus in 15N enrichment. Duodenal and bacterial samples collected on day 15 were used to measure duodenal N flows. Bacterial N flow was calculated as duodenal N flow multiplied by duodenal 15N enrichment divided by bacterial 15N enrichment. Bacterial N from recycled urea-N was calculated as bacterial N flow multiplied by bacterial 15N enrichment divided by urinary urea 15N enrichment. Urinary enrichment of [15N15N]urea plateaued within 24 h, whereas 14N15N urea plateaued within 48 h of [15N15N]urea infusion. Bacteria reached a plateau in 15N enrichment within 24 h and duodenal samples within 48 h. Urea production was 17.6 g of urea-N/day for control and 78.0 g/day for SBM. Gut entry was 0.99 g of urea-N/g of urea-N produced for control and 0.87 g/g for SBM. Incorporation of recycled N into microbial N was 9.0 g of N/day for control and 23.0 g/day for SBM. Recycled urea-N accounted for 0.33 g of N/g of microbial N at the duodenum for control and 0.27 g/g for SBM. Our methods allowed measurement of incorporation of recycled urea-N into ruminal microbial N.  相似文献   

19.
用渗漏池模拟洞庭湖区2种主要稻田土壤(河沙泥和紫潮泥),研究了施用尿素(CF)和控释氮肥(CRNF)对双季稻田表面水pH、电导率(EC)、全氮(TN)、铵态氮(NH4+-N)和硝态氮(NO3--N)浓度变化规律及TN径流损失的影响.结果表明,双季稻田施用尿素后,表面水TN、NH4+-N浓度分别在第1、3天达到高峰,然后迅速下降;NO3--N浓度普遍很低;早稻表面水pH在施用尿素后15 d内(晚稻3 d)逐渐升高;EC与NH4+动态变化一致.与尿素相比,施用CRNF能显著降低双季稻田表面水pH、EC、TN和NH4+-N浓度,70% N控释氮肥的控制效果最显著;但后期NO3--N浓度略有升高.径流监测结果表明,洞庭湖区种植双季稻期间施用尿素的TN径流损失为7.70 kg·hm-2,占施氮量的2.57%;施肥后20 d内发生的径流事件对双季稻田TN径流损失的贡献极为显著;与施用尿素相比,施用控释氮肥显著降低了施肥后10 d内发生的第1次径流液中的TN浓度,施用CRNF和70%N CRNF的氮素径流损失分别降低24.5%和27.2%.  相似文献   

20.
Rates of whole body amino nitrogen flux were measured in 16 obese adolescents undergoing weight reduction with a high protein low energy diet. The subjects received approximately 2.5 g of animal protein per day per kilogram ideal body weight and maintained nitrogen balance throughout the 18 days on the diet. Flux rates were calculated separately from the cumulative excretion of 15N in urinary ammonia and urea following the administration of a single dose of [15N]glycine. The pattern of 15N label appearance in urinary ammonia and urea nitrogen was followed for 72 h after the administration of [15N]glycine. Significant amounts of label continued to be excreted in both urinary ammonia and nitrogen for 36-48 h after label administration. The weight-reducing diet accelerated 15N cumulative excretion in urinary urea, but not in ammonia nitrogen compared with the control diet. Whole body nitrogen flux rates increased rapidly and significantly on the diet. Using the urea end product, this increase was evident on the 4th diet day, but not by the 7th or subsequent days. On the other hand, using the ammonia end product, flux rate increased markedly (p less than 0.0001) and remained elevated throughout the whole study. Our results demonstrate adaptive changes in whole body amino-nitrogen metabolism in response to the reducing diet. Different patterns of change are seen depending upon whether an ammonia or a urea end product is used. Our data thus add to the evidence for compartmentation of the body's amino-nitrogen pools.  相似文献   

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