Safety assessment of gamma-cyclodextrin.

Gamma-cyclodextrin (gamma-CD) is a cyclic alpha-(1,4)-linked oligosaccharide consisting of eight glucose molecules. Like other cyclodextrins, gamma-CD can form inclusion complexes with a variety of organic molecules because the inner side of the torus-like molecule is less polar than the outer side. In foods, gamma-CD may be used as a carrier for flavors, vitamins, polyunsaturated fatty acids, and other ingredients. It also has useful properties as a stabilizer in different food systems. The daily intake from all its intended uses in food at highest feasible concentrations has been estimated at 4.1g/person/day for consumers of gamma-CD containing foods. The present review summarizes the safety data of gamma-CD. The toxicity studies consist of standard genotoxicity tests, subchronic rat studies with oral and intravenous administration of gamma-CD for up to 3 months, a subchronic (3-month) toxicity study in dogs, a (1-year) oral toxicity study in rats, and embryotoxicity/teratogenicity studies in rats and rabbits. In the studies with oral administration, gamma-CD was given at dietary concentrations of up to 20%. All these studies demonstrated that gamma-CD is well tolerated and elicits no toxicological effects. Metabolic studies in rats showed that gamma-CD is rapidly and essentially completely digested by salivary and pancreatic amylase. Therefore, the metabolism of gamma-CD closely resembles that of starch and linear dextrins. A human study with ingestion of single doses of 8 g gamma-CD or 8 g maltodextrin did not reveal a difference in gastrointestinal tolerance of these two products. An interaction of ingested gamma-CD with the absorption of fat-soluble vitamins or other lipophilic nutrients is not to be expected because the formation of inclusion complexes is a reversible process, gamma-CD is readily digested in the small intestine, and studies with beta-CD, a non-digestible cyclodextrin, have shown that the bioavailability of vitamins (A, D, and E) is not impaired. On basis of these studies it is concluded that gamma-CD is generally recognized as safe (GRAS) for its intended uses in food.

L-arginine transport in the human coronary and peripheral circulation.

BACKGROUND: Nitric oxide synthase (NOS) uses arginine for the production of nitric oxide (NO). High intracellular concentrations of arginine suggest that NOS activity should be independent of plasma arginine supply. However, under certain conditions, increased plasma arginine concentrations appear to be associated with increased NOS activity. The purpose of this study was to explore arginine transport within the human coronary and peripheral circulation METHODS AND RESULTS: Mass-labeled 15N2-arginine was infused to steady state before cardiac catheterization in 31 patients. After diagnostic angiography, a catheter was placed in the coronary sinus. The transcardiac concentration gradient (aorta-coronary sinus) of 15N2-arginine was used as a measure of arginine transport at baseline and during infusions of acetylcholine and N(G)-monomethyl-L-arginine (L-NMMA). No gradient was detected at rest. During the infusion of acetylcholine, a significant gradient was detected (2.5+/-1.2 micromol/L, P=0.01) corresponding to a fractional extraction of 11.7+/-7.5%. This is consistent with in vitro studies that suggest that stimulation of NOS induces arginine transport. During the infusion of L-NMMA, the concentration of 15N2-arginine increased in the coronary sinus, producing a gradient of -3.9+/-1.3 micromol/L (P=0.0002), corresponding to a fractional production of 20.5+/-5.0%. This is consistent with in vitro studies that suggest that L-NMMA induces the efflux of arginine from the cell to the extracellular space via transporter-mediated transstimulation. CONCLUSIONS: The use of steady-state 15N2-arginine to examine transorgan L-arginine gradients represents a novel tool for the study of L-arginine transport and the mechanisms of endothelial and NOS dysfunction.

Daily requirement for and splanchnic uptake of leucine in healthy adult Indians.

BACKGROUND: The 1985 FAO/WHO/UNU requirement for leucine is too low according to tracer-derived estimates of leucine oxidation and balance in adults from developed regions. OBJECTIVE: The leucine requirement in populations in developing countries was assessed with use of the 24-h tracer balance method and on the basis of nitrogen balances. DESIGN: Twenty healthy Indian men were studied during their consumption for 6 d of 2 L-amino acid diets that supplied either 14 and 30 (n = 10) or 22 and 40 (n = 10) mg leucine x kg(-1) x d(-1) in random order. At 1800 on day 7, a 24-h constant intravenous [13C]leucine tracer-infusion protocol was conducted to determine leucine oxidation and daily leucine balance. During the intake of 40 mg leucine/d, [2H3]leucine was given orally to assess the splanchnic uptake of leucine. RESULTS: Mean 24-h leucine oxidation rates were 29.8, 30.6, 33.6, and 39.3 mg x kg(-1) x d(-1) at leucine intakes of 14, 22, 30, and 40 mg x kg(-1) x d(-1), respectively; daily leucine balances were -16.5, -9.0, -3.3, and 0.5 mg x kg(-1) x d(-1), respectively. Mixed-models linear regression of balance against leucine intake resulted in a zero balance at a leucine intake of 37.3 mg x kg(-1) x d(-1). Nitrogen balances were -12.7, -17.9, -3.9, and 1.0 mg x kg(-1) x d(-1) at leucine intakes of 14, 22, 30, and 40 mg x kg(-1) x d(-1). Regression of nitrogen balance against intake resulted in a zero balance at a leucine intake of 37.6 mg x kg(-1) x d(-1). The first-pass splanchnic uptake of leucine was 45.7% and 33.9% in the fasted and fed periods, respectively. CONCLUSION: A tentative mean leucine requirement of 40 mg x kg(-1) x d(-1) is proposed for healthy Indian adults, as it is for Western subjects.

Glutamine: the emperor or his clothes?

In this introduction to the Proceedings of the Symposium on Glutamine, we consider various lines of evidence that might potentially lead to an answer to the question posed in the title. We begin with a short summary of the multiple functions of glutamine, which are extensive and, superficially at least, equally as impressive as those of glutamate. However, each of these amino acids may serve an equivalent role in some of these functions due to their ready metabolic interconversion. We raise the question whether glutamine is of primordial or rudimentary significance or whether it is a product of somebody else's existence. Thus, there is a short account of the prebiotic events of evolution that led to the appearance of glutamine and life on Earth. In doing this, it then appears that glutamine is a rather schizophrenic molecule, stable and thermodynamically reliable in biochemical environments, but labile in chemical ones. We then turn to the involvement of glutamine in mammalian N (nitrogen) commerce, with initial emphasis on the nitrogen cycle on Earth, then N transport and N excretion, before assessing its contribution to carbon/energy or C/E commerce. We hypothesize that, in addition to its utilization in immune cell function and in normal intestinal tissues, glutamine is a particularly key anapleurotic and energy-yielding substrate in conditions of hypoxia, anoxia and dysoxia. It also serves as a quantitatively important gluconeogenic metabolite under normal postabsorptive conditions. We postulate that in certain conditions, this carbon-energy econometric function might be by-passed with ornithine. In conclusion, the answer to the question above depends on the context, and this point will receive elaboration in many of the individual contributions that collaborate to form these Proceedings.

Cysteine metabolism and whole blood glutathione synthesis in septic pediatric patients.

OBJECTIVE: To investigate whole body in vivo cysteine kinetics and its relationship to whole blood glutathione (GSH) synthesis rates in septic, critically ill pediatric patients and controls. DESIGN: Prospective cohort study. SETTING: Multidisciplinary intensive care unit and pediatric inpatient units at a children's hospital. PATIENTS: Ten septic pediatric patients and ten controls (children admitted to the hospital for elective surgery). INTERVENTIONS: Septic patients (age, 31 months to 17 yrs) and controls (age, 24 months to 21 yrs) received a 6-hr primed, constant, intravenous tracer infusion of l-[1-13C]cysteine. Blood samples were obtained to determine isotopic enrichment of plasma cysteine and whole blood [1-13C]cysteinyl-glutathione by gas-chromatography mass spectrometric techniques. The plasma flux and oxidation rate of cysteine and the fractional and absolute synthesis rates of GSH were determined. Septic patients received variable protein and energy intake, as per routine clinical management, and controls were studied in the early postabsorptive state. MEASUREMENTS AND MAIN RESULTS: Plasma cysteine fluxes were increased in the septic patients when compared with the controls (68.2 +/- 17.5 [sd] vs. 48.7 +/- 8.8 micromol x kg(-1) x hr(-1); p <.01), and the fraction of plasma cysteine flux associated with oxidative disposal was similar among the groups. The absolute rates of GSH synthesis in whole blood were decreased (p <.01) in the septic patients (368 +/- 156 vs. 909 +/- 272 micromol x L(-1) x day(-1)). The concentration of whole blood GSH also was decreased in the septic group (665.4 +/- 194 vs. 1059 +/- 334 microM; p <.01) CONCLUSIONS: Whole blood glutathione synthesis rates are decreased, by about 60%, in critically ill septic children receiving limited nutritional support. Plasma cysteine fluxes and concentration of cysteine were increased in the septic patients, suggesting a hypermetabolic state with increased protein breakdown. The mechanisms whereby GSH synthesis rates are decreased in these patients are probably multifactorial, presumably involving an inflammatory response in the presence of limited nutritional support. The role of nutritional modulation and the use of cysteine prodrugs in maintaining GSH concentration and synthesis remain to be established.

Lysine requirements of healthy adult Indian subjects, measured by an indicator amino acid balance technique.

BACKGROUND: In an earlier study, using a modification of the indicator amino acid oxidation approach, we concluded that the 1985 FAO/WHO/UNU-proposed lysine requirement of 12 mg x kg(-1) x d(-1) is likely inadequate to maintain body amino acid homeostasis in apparently healthy south Asian subjects and that our proposed requirement of 30 mg x kg(-1) x d(-1) is more appropriate. OBJECTIVE: We assessed the lysine requirement in a similar population by using 4 test lysine intakes (12, 20, 28, and 36 mg x kg(-1) x d(-1)) with an indicator amino acid balance approach. DESIGN: Sixteen healthy male Indians were studied during each of 2 randomly assigned 8-d L-amino acid diets that supplied either 12 and 28 or 20 and 36 mg lysine. At 1800 on day 8, a 24-h intravenous [(13)C]leucine tracer-infusion protocol was conducted to assess leucine oxidation and daily leucine balance at each lysine intake. RESULTS: Mean 24-h leucine oxidation rates decreased significantly (P = 0.005) across different lysine intakes and were 104.1, 97.8, 87.3, and 87.3 mg x kg(-1) x d(-1) at intakes of 12, 20, 28, and 36 mg x kg(-1) x d(-1), respectively; mean 24-h leucine balances were 3.3, 9.1, 19.7, and 20.7 mg x kg(-1) x d(-1), respectively (P = 0.015, mixed-model analysis of variance). Oxidation and balances differed significantly between the lower and higher lysine intakes but were not significantly different between the 12- and 20-mg and 28- and 36-mg test intakes. Two-phase regression analysis indicated a mean breakpoint at 29 mg lysine x kg(-1) x d(-1) in the relation between lysine intake and leucine oxidation or balance. CONCLUSION: We propose a mean lysine requirement of 30 mg x kg(-1) x d(-1) for healthy Indian adults, which is the same amount we proposed previously for Western populations.

Metabolism 2000: the emperor needs new clothes.

Metabolism is one of the corner stones of nutritional science. As biology enters the post-genomic era and with functional genomics beginning to takeoff, we anticipate that the study of metabolism will play an increasingly important role in helping to link advances made via the reductionist paradigm, that has been so successful in molecular and cellular biology, with those emerging from observational studies in animals and human subjects. A reconstructive metabolically-focused approach offers a timely paradigm for enhancing the elegance of nutritional science. Here we give particular attention to the use of tracers as phenotyping tools and discuss the application of our metaprobe concepts with respect to some novel features of metabolism, including 'underground metabolism', 'metabolic hijacking', 'catalytic promiscuity' and 'moonlighting proteins'. The opportunities for enhancing the study of metabolism by new and emerging technologies, and the importance of the interdisciplinary research enterprise are also touched upon. We conclude that: (1) the metaprobe concepts and approach, discussed herein, potentially yield a quantitative physiological (metabolic) phenotype against which to elaborate partial or focused genotypes; (2) physiological (metabolic) phenotypes which have a whole-body or kinetically-discernible inter-organ tissue-directed metabolic signature are an ideal target for this directed tracer-based definition of the 'functional' genotype; (3) metabolism, probed with tracer tool kits suitable for measuring rates of turnover, change and conversion, becomes in the current sociology of the 'Net', like AOL, Yahoo. Alta Vista, Lycos or Ask Jeeves, the portal for an exploration of the metabolic characteristics of the 'Genomics Internet'.

Arginine and ornithine kinetics in severely burned patients: increased rate of arginine disposal.

Arginine serves multiple roles in the pathophysiological response to burn injury. Our previous studies in burn patients demonstrated a limited net rate of arginine de novo synthesis despite a significantly increased arginine turnover (flux), suggesting that this amino acid is a conditionally indispensable amino acid after major burns. This study used [15N2-guanidino-5,5-2H2]arginine and [5-13C]ornithine as tracers to assess the rate of arginine disposal via its conversion to and subsequent oxidation of ornithine; [5,5-2H2]proline and [5,5,5-2H3]leucine were also used to assess proline and protein kinetics. Nine severely burned patients were studied during a protein-free fast ("basal" or fast) and total parenteral nutrition (TPN) feedings. Compared with values from healthy volunteers, burn injury significantly increased 1) fluxes of arginine, ornithine, leucine, and proline; 2) arginine-to-ornithine conversion; 3) ornithine oxidation; and 4) arginine oxidation. TPN increased arginine-to-ornithine conversion and proportionally increased irreversible arginine oxidation. The elevated arginine oxidation, with limited net de novo synthesis from its immediate precursors, further implies that arginine is a conditionally indispensable amino acid in severely burned patients receiving TPN.

Nitrogen and amino acid requirements: : the Massachusetts Institute of Technology amino acid requirement pattern.

We review the current international recommendations concerning the protein (nitrogen) and amino acid requirements of healthy individuals, from infancy to the later years of adult life and describe the changes in the recommendations for protein that have been made, since those issued in 1985 by Food and Agriculture Organization/World Health Organization/United Nations University (FAO/WHO/UNU), by the International Dietary Energy Consultative Group. The current international requirements for the specific indispensable amino acids are critiqued briefly, and the rationale and basis for the proposed Massachusetts Institute of Technology (MIT) amino acid requirement pattern are presented. The evidence is then summarized that supports its use in practical considerations of protein nutrition. It is suggested that this MIT amino acid requirement pattern provides the best current estimates of the minimum physiological requirements for the indispensable amino acids in children and adults. It is further concluded that it would be difficult to argue for the continued use of the amino acid requirement values proposed by FAO/WHO/UNU in 1985 in the planning and assessment of dietary protein intakes for population groups worldwide. The MIT amino acid requirement pattern supports and strengthens the relevance of dietary protein quality as an important factor in human protein and amino acid nutrition.

Twenty-four-hour oral tracer studies with L-[1-13C]lysine at a low (15 mg.kg (-1).d (-1) and intermediate (29 mg.kg (-1).d(-1)) lysine intake in healthy adults.

BACKGROUND: We proposed previously that the mean lysine requirement value is approximately 30 mg * kg(-)(1) * d(-)(1) rather than the proposed 1985 FAO/WHO/UNU estimate of the upper range of the requirement, which is 12 mg * kg(-)(1) * d(-)(1). OBJECTIVE: Our objective was to explore the 24-h pattern and rate of whole-body lysine [l-(13)C]oxidation and status of whole-body lysine balance in healthy, young adults given an L-amino acid diet supplying either a low lysine intake (14-15 mg * kg(-)(1) * d(-)(1)) or an intermediate lysine intake (29 mg * kg(-)(1) * d(-)(1)) for 6 d before a continuous tracer study with L-[1-(13)C]lysine. DESIGN: Five subjects received the low lysine intake, 6 subjects received the intermediate intake, and all were studied by using a standard 24-h oral tracer protocol that was described earlier for studies at a generous lysine intake. RESULTS: The rate of lysine oxidation was not significantly different between the 12-h fasted and 12-h fed states. The daily oxidation rate (f1.gif" BORDER="0"> +/- SD) was 27. 9 +/- 8.8 and 27.3 +/- 17.6 mg lysine * kg(-)(1) * d(-)(1) for the low- and intermediate-intake groups, respectively (NS). Daily lysine balance was -12.4 +/- 92 and 1.8 +/- 17.7 mg * kg(-)(1) * d(-)(1), respectively (P < 0.025), for the low and intermediate intakes. The balance was significantly less than zero (P < 0.001) for the low intake. CONCLUSION: The FAO/WHO/UNU lysine requirement value is not sufficient to maintain lysine homeostasis in healthy adults. From the results of this and tracer studies done by others, the mean lysine requirement of healthy adults was determined to be 30 mg * kg(-)(1) * d(-)(1).

Arginine, citrulline, and nitric oxide metabolism in end-stage renal disease patients.

The kidneys are thought to be a major site of net de novo arginine synthesis, but the quantitative status of arginine metabolism and its substrate precursor relationship to nitric oxide (NO) synthesis in end stage renal disease (ESRD) patients have not been characterized. We have investigated kinetic aspects of whole body arginine metabolism in six patients with ESRD. They received two pre- and two post-hemodialysis intravenous tracer infusion studies with L-[guanidino-(15)N(2)]arginine and L-[(13)C]leucine during the first study, and L-[5-(13)C]arginine and L-[5-(13)C-ureido,5,5, (2)H(2)]citrulline during the second study. Arginine homeostasis in ESRD patients was found to be associated with a lower rate of arginine oxidation, and despite the decrease in renal function, the rate of de novo arginine synthesis appeared to be preserved. Plasma citrulline concentrations and flux were also elevated in these subjects compared with healthy adults. The rate of whole body NO synthesis was increased in the ESRD patients, but apparently not different pre- and post-hemodialysis therapy. The anatomic site(s) responsible for the maintenance of net de novo arginine synthesis and for the elevated NO synthesis and its pathophysiological importance in ESRD remain to be established.

Blood glutathione synthesis rates in healthy adults receiving a sulfur amino acid-free diet.

The availability of cysteine is thought to be the rate limiting factor for synthesis of the tripeptide glutathione (GSH), based on studies in rodents. GSH status is compromised in various disease states and by certain medications leading to increased morbidity and poor survival. To determine the possible importance of dietary cyst(e)ine availability for whole blood glutathione synthesis in humans, we developed a convenient mass spectrometric method for measurement of the isotopic enrichment of intact GSH and then applied it in a controlled metabolic study. Seven healthy male subjects received during two separate 10-day periods an L-amino acid based diet supplying an adequate amino acid intake or a sulfur amino acid (SAA) (methionine and cysteine) free mixture (SAA-free). On day 10, L-[1-(13)C]cysteine was given as a primed, constant i.v. infusion (3 micromol x kg(-1) x h(-1)) for 6 h, and incorporation of label into whole blood GSH determined by GC/MS selected ion monitoring. The fractional synthesis rate (mean +/- SD; day(-1)) of whole blood GSH was 0.65 +/- 0.13 for the adequate diet and 0.49 +/- 0.13 for the SAA-free diet (P < 0.01). Whole blood GSH was 1,142 +/- 243 and 1,216 +/- 162 microM for the adequate and SAA-free periods (P > 0.05), and the absolute rate of GSH synthesis was 747 +/- 216 and 579 +/- 135 micromol x liter(-1) x day(-1), respectively (P < 0.05). Thus, a restricted dietary supply of SAA slows the rate of whole blood GSH synthesis and diminishes turnover, with maintenance of the GSH concentration in healthy subjects.

Kinetics of L-[1-(13)C]leucine when ingested with free amino acids, unlabeled or intrinsically labeled casein.

In two groups of five adults, each adapted to two different dietary regimens for 6 days, the metabolic fate of dietary [1-(13)C]leucine was examined when ingested either together with a mixture of free amino acids simulating casein (extrinsically labeled; condition A), along with the intact casein (extrinsically labeled; condition B), or bound to casein (intrinsically labeled; condition C). Fed state leucine oxidation (Ox), nonoxidative leucine disposal (NOLD), protein breakdown, and splanchnic uptake have been compared using an 8-h oral [1-(13)C]leucine and intravenous [(2)H(3)]leucine tracer protocol while giving eight equal hourly mixed meals. Lower leucine Ox, increased NOLD, and net protein synthesis were found with condition C compared with condition A (19.3 vs. 24.9; 77 vs. 55.8; 18.9 vs. 12.3 micromol. kg(-1). 30 min(-1); P < 0.05). Ox and NOLD did not differ between conditions B and C. Splanchnic leucine uptake calculated from [1-(13)C]- and [(2)H(3)]leucine plasma enrichments was between 24 and 35%. These findings indicate that the form in which leucine is consumed affects its immediate metabolic fate and retention by the body; the implications of these findings for the tracer balance technique and estimation of amino acid requirements are discussed.

Inverse relationship between protein intake and plasma free amino acids in healthy men at physical exercise.

The effect of a "normal" (n = 8) and "high" (n = 6) protein intake (1 and 2.5 g x kg(-1) x day(-1), respectively) and of exercise on plasma amino acid (AA) concentrations, insulin, and glucagon concentrations was followed throughout a continuous 24-h period in adult male subjects at energy balance after six days on a standardized diet and exercise program. Subjects were fasting from 2100 on day 6 to 1200 on day 7 and then fed 10 identical meals hourly until 2100. Physical exercise was performed (46% maximal oxygen uptake) between 0830 and 1000 (fasting) and in a fed state (1600-1730) on each day. The normal-protein group showed fasting plasma AA concentrations that were higher (P < 0.05) than those for the high-protein group, except for leucine, methionine, and tyrosine. Glutamine, glycine, alanine, taurine, and threonine concentrations were distinctly higher ( approximately 30% or greater) throughout the 24-h period in subjects consuming the normal- vs. the high-protein diets. Exercise appeared to increase, although not profoundly, the plasma concentrations of amino acids except for glutamate, histidine, ornithine, and tryptophan. The profound diet-related differences in plasma AA concentrations are only partially explained by differences in the renal clearance of the amino acids. We speculate on the possible metabolic basis for these findings.

Oxoproline kinetics and oxoproline urinary excretion during glycine- or sulfur amino acid-free diets in humans.

L-5-oxoproline (L-5-OP) is an intermediate in glutathione synthesis, possibly limited by cysteine availability. Urinary 5-OP excretion has been proposed as a measure of glycine availability. We investigated whether 5 days of dietary sulfur amino acid (SAA-free) or glycine (Gly-free) restriction affects plasma kinetics of 5-OP and urinary excretion of L- and D-5-OP in 6 healthy men. On day 6, L-5-[1-(13)C]oxoproline and [3,3-(2)H(2)]cysteine were infused intravenously for 8 h (3 h fast/5 h fed). In a control study (adequate amino acid mixture), plasma oxoproline fluxes were 37.8 +/- 13.8 (SD) and 38.4 +/- 14.8 micromol x kg(-1) x h(-1); oxidation accounted for 85% of flux. Cysteine flux was 47.9 +/- 8.5 and 43.2 +/- 8.5 micromol x kg(-1) x h(-1) for fast and fed phases, respectively. Urinary excretion of L- and D-5-OP was 70 +/- 34 and 31.1 +/- 13.3 micromol/mmol creatinine, respectively, during days 3-5, and 46.4 +/- 13.9 and 22.4 +/- 8.3 micromol/mmol over the 8-h tracer study. The 5-OP flux for the Gly-free diet was higher (P = 0. 018) and tended to be higher for the SAA-free diet (P = 0.057) when compared with the control diet. Oxidation rates were higher on the Gly-free (P = 0.005) and SAA-free (P = 0.03) diets. Cysteine fluxes were lower on the the Gly-free (P = 0.01) and the SAA-free diets (P = 0.001) compared with the control diet. Rates of L-5-OP excretion were unchanged by withdrawal of SAA or Gly for 5 days but increased on day 6 (P = 0.005 and P = 0.019, respectively). Thus acute changes in the dietary availability of SAA and Gly alter oxoproline kinetics and urinary 5-OP excretion.

Rates of urea production and hydrolysis and leucine oxidation change linearly over widely varying protein intakes in healthy adults.

The quantitative relationships between nitrogen (N) intake, urea production, excretion and amino acid oxidation are currently a matter of debate. Some investigators have proposed that urea production is essentially constant over a wide range of N intakes and that urea hydrolysis is regulated according to the N needs of the organism. We have assessed this proposal by compiling results from four separate experiments in healthy young adults (n = 34) carried out in our laboratories and all at the end of the respective diet periods using an identical 24-h continuous intravenous infusion of [(15)N, (15)N]urea and L-[1-(13)C]leucine. The N intakes were: expt. 1; protein-free diet for 5 d; expt. 2; N at 44 mg N. kg(-1). d(-1) from a balanced L-amino acid mixture for 13 d; expt. 3; N at 161 mg. kg(-1). d(-1) from egg protein for 6 d; expt. 4 -one group received 157 mg. kg(-1). d(-1) and the other 392 mg. kg(-1). d(-1) from milk-protein-based diets for 6 d. Urea production and excretion were linearly correlated with N intake (r = 0.98 and 0.94, respectively; P < 0.01). Urea hydrolysis increased linearly with N intake (r = 0.7; P < 0.05), with considerable variation in the rate among individuals, especially at the N intake of approximately 160 mg N. kg(-1)d(-1). These findings are consistent with the generally accepted view that a control of body N balance is via a regulation of urea production. They do not support the concept that urea hydrolysis is the more important site in the control of body N loss.

Glutamate: an amino acid of particular distinction.

In this introductory paper to the symposium, we consider why L-glutamate (GLU) is such an abundant biomolecule. We begin with a brief discussion of the prebiotic dawn of events and some evolutionary features of GLU in the biological and metabolic world. The properties of GLU are then examined with reference to its overall structural motif and to the reactivity of the molecule at the tautomeric 2 carbon and at the 4- and 5-C positions. This chemical viewpoint reveals that the GLU molecule offers a number of features/properties not shared by its homologs (amino adipic and aspartic acids). These properties make GLU a favorable choice for facilitating its involvement in multiple metabolic processes that play major roles in the nitrogen economy of the host, as well as serving as a nutrient, an energy-yielding substrate, a structural determinant and an excitatory molecule.

Cysteine kinetics and oxidation at different intakes of methionine and cystine in young adults.

BACKGROUND: We previously studied methionine kinetics and oxidation with the tracer L-[1-(13)C, methyl-(2)H(3)]methionine. OBJECTIVES: We sought to explore methionine-cysteine interrelations in adults by using L-[1-(13)C]cysteine under different dietary conditions. DESIGN: In experiment 1, 12 adults consumed a protein-free diet for 6 d. On day 7, methionine (n = 6) or cysteine (n = 6) oxidation rates were measured during an 8-h continuous infusion of L-[1-(13)C, methyl-(2)H(3)]methionine or L-[1-(13)C]cysteine, respectively. In experiment 2, 6 young men consumed 3 diets for 6 d each before a tracer study on day 7 with L-[1-(13)C]cysteine. The amounts (in mg*kg(-)(1)*d(-)(1)) of methionine and cysteine, respectively, were: high-methionine (HM) diet, 13 and 0; low-methionine (LM) diet, 6.5 and 0; and methionine-plus-cystine (MC) diet, 6.5 and 5.6. Cysteine flux and oxidation rates were determined and sulfur amino acid (SAA, methionine plus cysteine) balances were estimated. RESULTS: In experiment 1, rates of methionine and cysteine oxidation were similar to losses predicted from obligatory nitrogen losses. In experiment 2, SAA balance was less negative when subjects consumed the HM diet than the LM and MC diets (interaction, P = 0.034), largely because of a difference in fed-state balance (HM compared with LM, P < 0.01; HM compared with MC, P < 0.05). There was no evidence of a sparing effect of dietary cystine on the methionine requirement. CONCLUSION: These studies support use of [1-(13)C]cysteine for studying whole-body SAA oxidation and conclusions that maintenance of SAA balance is best achieved by supplying methionine at approximately the FAO/WHO/UNU recommendations for total SAA intake (13 mg*kg(-)(1)*d(-)(1)).