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.

The 24-h kinetics of leucine oxidation in healthy adults receiving a generous leucine intake via three discrete meals.

The significance of meal size and frequency for the 24-h leucine tracer-balance technique was examined. Continuous measurements of leucine oxidation throughout a 24-h d were performed in six healthy, young adults who were given a weight-maintaining diet (188 kJ.kg-1.d-1; 1 g protein.kg-1.d-1) for 6 d followed by primed, continuous intravenous infusions of L-[1-13C]leucine and [15N-15N]urea. The 24-h study was started at 1800 on day 6 and three equal discrete meals were given at 2000, 0600, and 1200. Leucine oxidation was assessed from plasma [13C]alpha-ketoisocaproate enrichment and 13CO2 excretion. The mean (+/- SD) leucine oxidation after each meal (over 6 h) was not significantly different (P > 0.5) among the three discrete meals: 20.0 +/- 3.9, 20.2 +/- 1.9, and 20.3 +/- 2.4 mg.kg-1.d-1 for the meals given at 2000, 0600, and 1200, respectively. Twenty-four-hour leucine oxidation was 75.0 +/- 7.8 mg.kg-1.d-1 for a leucine dietary intake of 80 mg.kg-1.d-1 (and approximately 9.7 mg tracer.kg-1.d-1). The 24-h pattern in leucine oxidation was paralleled by plasma leucine concentrations. Further, leucine oxidation and urea excretion predicted relatively similar values for 24-h protein oxidation. These data are compared with results from our similar previous studies using a multiple-small-meal feeding protocol.

The 24-h pattern and rate of leucine oxidation, with particular reference to tracer estimates of leucine requirements in healthy adults.

Daily leucine oxidation and derived values for whole-body leucine balance, obtained by continuous measurement throughout a 24-h period, were compared with those predicted from short-term measurements during fasted and fed states in five healthy adults studied during two 6-d experimental diet periods, each immediately followed by a 24-h continuous intravenous tracer infusion of L-[1-13C]leucine. Leucine intake was either 14 or 38.3 mg.kg-1.d-1. Mean measured daily leucine oxidation (mg leucine.kg-1.d-1) was 27.8 and 45.2 for the 14- and 38.3-mg intakes, respectively. Oxidation rates predicted by extrapolation of rates measured during the final hour of fasting (15 h after last meal) and the 5th h of feeding were approximately 12% higher (P < 0.01) than measured rates for both diets. For the prediction based on the 12th h of fasting and 5th h of feeding, it was 4% higher or 0.4% lower than measured rates for the 38.3- and 14- mg intakes, respectively. Hence, relatively small differences exist between measured vs predicted estimates of daily leucine oxidation and balance. These studies support previous conclusions that the current, international requirement value for leucine in healthy adults is far too low.

Validation of the tracer-balance concept with reference to leucine: 24-h intravenous tracer studies with L-[1-13C]leucine and [15N-15N]urea.

The validity of tracer-derived estimates of whole-body leucine balance was investigated. Seven healthy young adult subjects received an adequate protein diet for 6 d; at 1800 on the last day, L-[1-13C]leucine and [15N-15N]urea were given as primed, continuous intravenous infusions for 24 h. Subjects were in a fasting state for the first 12 h and at 0600 on day 7 they then received hourly 10 equal meals to achieve a fed state. Total leucine intake (diet plus tracer) was 89.4 mg.kg-1.d-1. Mean daily leucine oxidation was equivalent to 89.5 +/- 3.3 mg leucine/kg. The predicted daily oxidation rate, from measurements made during the last hour of the fast and the fifth hour of the fed period, was 91.2 +/- 5.8 mg/kg (P = 0.25 from measured). Measured and predicted whole-body leucine balances were 0.76 +/- 2.99 and -0.98 +/- 5.54 mg/kg, respectively (P = 0.25). Urea production exceeded urea excretion by 20%; daily protein oxidation was the same when estimated from leucine oxidation or nitrogen excretion. Thus, the tracer-balance concept is valid, and reliable predictions of total daily leucine oxidation and whole-body leucine balance can be obtained from short-term measurements of leucine oxidation during fasted and fed states.