Rate Of Amino Acid Oxidation Research Articles

The anabolic response to protein ingestion during recovery from exercise has no upper limit in magnitude and duration in vivo in humans

The belief that the anabolic response to feeding during postexercise recovery is transient and has an upper limit and that excess amino acids are being oxidized lacks scientific proof. Using a comprehensive quadruple isotope tracer feeding-infusion approach, we show that the ingestion of 100g protein results in a greater and more prolonged (>12 h) anabolic response when compared to the ingestion of 25g protein. We demonstrate a dose-response increase in dietary-protein-derived plasma amino acid availability and subsequent incorporation into muscle protein. Ingestion of a large bolus of protein further increases whole-body protein net balance, mixed-muscle, myofibrillar, muscle connective, and plasma protein synthesis rates. Protein ingestion has a negligible impact on whole-body protein breakdown rates or amino acid oxidation rates. These findings demonstrate that the magnitude and duration of the anabolic response to protein ingestion is not restricted and has previously been underestimated invivo in humans.

Leucine Is More Readily Oxidized When Ingested as an Isolated Nutrient versus Incorporated in Its Whole-Food Matrix

Abstract Objectives The ingestion of free amino acids, or isolated sources of protein, results in faster postprandial release of dietary amino acids into circulation, which stimulates muscle protein synthesis rates. However, indirect evidence suggests that this rapid release of dietary amino acids after the ingestion of free amino acids is coupled with higher amino acid oxidation rates when compared to a whole-food source. Whole food protein results in a reduced peak amplitude and prolonged postprandial aminoacidemia. This study aimed to assess the effects of eating a whole food source of protein on the stimulation of whole-body leucine oxidation rates versus eating these same nutrients in isolated form in healthy young adults. Methods In a crossover design, 10 recreationally active adults (24 ± 4 y; 5 M, 5 F) performed an acute bout of resistance exercise followed by the ingestion of salmon (SAL) (20.5 g protein and 7.5 g fat) or its matched constituents as crystalline amino acids and fish oil (ISO). Participants received priming doses of NaH13CO2 and L-[1–13C]leucine before initiating a constant L-[1–13C]leucine infusion. Blood and breath samples were collected at rest and after resistance exercise at regular intervals for the measurement of whole-body leucine oxidation rates and plasma leucine profiles. Data were tested using linear fixed effects models with time and group as fixed factors with Bonferroni's post hoc test. Results Postprandial plasma leucine concentrations did not differ between the SAL and ISO conditions (P > 0.05). Time to peak plasma leucine concentrations was faster in ISO (50 ± 27 min) vs. SAL (114 ± 64 min, P = 0.022) condition. Postprandial leucine oxidation rates were elevated from baseline at t = 30 min to t = 120 min in ISO and t = 60 min to t = 180 min in SAL (P < 0.001), with no total differences between group (P = 0.129). Time to peak leucine oxidation occurred sooner in ISO (66 ± 22 min; 1.358 ± 0.699 nmol · kg−1 · min−1) when compared to the SAL condition (105 ± 20 min; 1.067 ± 0.3076 nmol · kg−1 · min−1; P = 0.002). Conclusions We show that the ingestion of a whole-food source of protein resulted in a delayed stimulation of leucine oxidation when compared to free amino acid ingestion, but a similar net increase in oxidation during the 5 h postprandial period in healthy young adults. Funding Sources USDA National Institute of Food and Agriculture Hatch project 1017928

Oxidative removal of aromatic amino acids by manganese dioxide: kinetic modeling and effect of co-solutes

ABSTRACTThe degradation of aromatic amino acids, including L-tryptophan, L-tyrosine, and L-phenylalanine, by δ-MnO2 was investigated. L-tryptophan and L-tyrosine underwent rapid oxidative degradation in the δ-MnO2 suspension solution. In contrast, L-phenylalanine was not oxidized by δ-MnO2. The degradation of L-tryptophan was faster than that of L-tyrosine. The oxidation rates of amino acids increased with increasing concentration of δ-MnO2. The increase of concentrations of substrates and pH disfavored the degradation. The kinetics modeling showed that the reaction orders of amino acids, MnO2, and H+ were 0.67, 1.24, 0.72 and 0.32, 1.20, 1.05 for L-tryptophan and L-tyrosine, respectively. The coexistence of metal ions markedly inhibited the degradation in the order of Al3+ > Mn2+ > Zn2+ > Ca2+ > Mg2+. The inhibition effect of metal ions was related to the stability constant and hydration radius of metal ions. The addition of fulvic acid to the reaction solutions did not affect the degradation of substrate. This finding suggests that δ-MnO2 selectively oxidize aromatic amino acids and it can be used as an efficient oxidant for the removal of certain aromatic acid pollutants.

Short-term amino acid infusion improves protein balance in critically ill patients.

IntroductionEvidence behind the recommendations for protein feeding during critical illness is weak. Mechanistic studies are needed to elucidate the effects of amino acid and/or protein supplementation on protein metabolism before larger clinical trials with higher levels of protein feeding are initiated.MethodsWe studied the effects of parenteral amino acid supplementation (equivalent to 1 g/kg/day) over the course of 3 hours on whole-body protein turnover in critically ill patients in the intensive care unit (ICU) during the first week after admission. Patients were studied at baseline during ongoing nutrition and during extra amino acid supplementation. If the patient was still in the ICU 2 to 4 days later, these measurements were repeated. Protein kinetics were measured using continuous stable isotope-labeled phenylalanine and tyrosine infusions.ResultsThirteen patients were studied on the first study occasion only, and seven were studied twice. Parenteral amino acid supplementation significantly improved protein balance on both occasions, from a median of −4 to +7 μmol phenylalanine/kg/hr (P =0.001) on the first study day and from a median of 0 to +12 μmol phenylalanine/kg/hr (P =0.018) on the second study day. The more positive protein balance was attributed to an increased protein synthesis rate, which reached statistical significance during the first measurement (from 58 to 65 μmol phenylalanine/kg/hr; n =13; P =0.007), but not during the second measurement (from 58 to 69 μmol phenylalanine/kg/hr; n =7; P =0.09). Amino acid oxidation rates, estimated by phenylalanine hydroxylation, did not increase during the 3-hour amino acid infusion. A positive correlation (r =0.80; P <0.0001) was observed between total amino acids and/or protein given to the patient and whole-body protein balance.ConclusionExtra parenteral amino acids infused over a 3-hour period improved whole-body protein balance and did not increase amino acid oxidation rates in critically ill patients during the early phase (first week) of critical illness.Electronic supplementary materialThe online version of this article (doi:10.1186/s13054-015-0844-6) contains supplementary material, which is available to authorized users.

Metabolic Heat Stress Adaption in Transition Cows: Differences in Macronutrient Oxidation between Late-Gestating and Early-Lactating German Holstein Dairy Cows.

High ambient temperatures have severe adverse effects on biological functions of high-yielding dairy cows. The metabolic adaption to heat stress was examined in 14 German Holsteins transition cows assigned to two groups, one heat-stressed (HS) and one pair-fed (PF) at the level of HS. After 6 days of thermoneutrality and ad libitum feeding (P1), cows were challenged for 6 days (P2) by heat stress (temperature humidity index (THI) = 76) or thermoneutral pair-feeding in climatic chambers 3 weeks ante partum and again 3 weeks post-partum. On the sixth day of each period P1 or P2, oxidative metabolism was analyzed for 24 hours in open circuit respiration chambers. Water and feed intake, vital parameters and milk yield were recorded. Daily blood samples were analyzed for glucose, β-hydroxybutyric acid, non-esterified fatty acids, urea, creatinine, methyl histidine, adrenaline and noradrenaline. In general, heat stress caused marked effects on water homeorhesis with impairments of renal function and a strong adrenergic response accompanied with a prevalence of carbohydrate oxidation over fat catabolism. Heat-stressed cows extensively degraded tissue protein as reflected by the increase of plasma urea, creatinine and methyl histidine concentrations. However, the acute metabolic heat stress response in dry cows differed from early-lactating cows as the prepartal adipose tissue was not refractory to lipolytic, adrenergic stimuli, and the rate of amino acid oxidation was lower than in the postpartal stage. Together with the lower endogenous metabolic heat load, metabolic adaption in dry cows is indicative for a higher heat tolerance and the prioritization of the nutritional requirements of the fast-growing near-term fetus. These findings indicate that the development of future nutritional strategies for attenuating impairments of health and performance due to ambient heat requires the consideration of the physiological stage of dairy cows.

Myofibrillar protein synthesis following ingestion of soy protein isolate at rest and after resistance exercise in elderly men

BackgroundIncreased amino acid availability stimulates muscle protein synthesis, however, aged muscle appears less responsive to the anabolic effects of amino acids when compared to the young. We aimed to compare changes in myofibrillar protein synthesis (MPS) in elderly men at rest and after resistance exercise following ingestion of different doses of soy protein and compare the responses to those we previously observed with ingestion of whey protein isolate.MethodsThirty elderly men (age 71 ± 5 y) completed a bout of unilateral knee-extensor resistance exercise prior to ingesting no protein (0 g), or either 20 g or 40 g of soy protein isolate (0, S20, and S40 respectively). We compared these responses to previous responses from similar aged men who had ingested 20 g and 40 g of whey protein isolate (W20 and W40). A primed constant infusion of L-[1-13 C]leucine and L-[ring-13 C6]phenylalanine and skeletal muscle biopsies were used to measure whole-body leucine oxidation and MPS over 4 h post-protein consumption in both exercised and non-exercised legs.ResultsWhole-body leucine oxidation increased with protein ingestion and was significantly greater for S20 vs. W20 (P = 0.003). Rates of MPS for S20 were less than W20 (P = 0.02) and not different from 0 g (P = 0.41) in both exercised and non-exercised leg muscles. For S40, MPS was also reduced compared with W40 under both rested and post-exercise conditions (both P < 0.005); however S40 increased MPS greater than 0 g under post-exercise conditions (P = 0.04).ConclusionsThe relationship between protein intake and MPS is both dose and protein source-dependent, with isolated soy showing a reduced ability, as compared to isolated whey protein, to stimulate MPS under both rested and post-exercise conditions. These differences may relate to the lower postprandial leucinemia and greater rates of amino acid oxidation following ingestion of soy versus whey protein.

Metal-catalyzed degradation and racemization of amino acids in iron sulfate brines under simulated martian surface conditions

Recent lines of evidence indicate that Mars may have been substantially warmer and wetter, raising questions about whether the planet’s surface was once more conducive to the possibility of life. To better constrain the extent of preservation of ancient amino acid biosignatures, the effects of metal ions and UV radiation were investigated with respect to the rates of diagenesis of amino acids in metal-rich sulfate brines. Individual experimental results indicate that the oxidation rates of amino acids loosely followed trends characteristic of a radiolytic oxidation mechanism that is dependent on variations in side-chain constituents. Relative rates of racemization only show a limited dependence on the presence of metals; overall, rates of racemization in the presence of metals in solution are an order of magnitude slower than oxidation and both are several orders of magnitude faster than previously reported. This increase in rate constant is indicative of an extremely efficient catalytic enhancement of this destructive diagenetic pathway. These findings imply that chiral life-detection strategies should focus on specific geochemical environments that exhibit relatively slow rates of both degradation and racemization. Episodic liquid water during periods of increased water activity on the martian surface may have significant implications for the oxidation of organic matter through secondary diagenetic reactions and may act as a leveling mechanism for organic compounds.

Effect of dietary protein level on whole body protein turnover in the adult cat (Felis catus)

The domestic cat is an obligate carnivore and as such has a high protein requirement. However, little is known about how protein turnover is affected by dietary protein intake in the cat. In the current study, kinetics of [1‐13C]Leu metabolism were used to quantify rates of whole body protein synthesis, degradation, and oxidation in adult cats (n = 6) fed diets containing 15, 40, or 65% of the ME intake as CP. Cats were fed the test diets to maintain BW for a minimum of 3 wk before the day of [1‐13C]Leu infusion. They were fed hourly on day of infusion. Values for Leu flux, non‐oxidative Leu disposal (NOLD, an indication of protein synthesis), Leu rate of appearance into the plasma (Ra, an indication of protein degradation), and Leu oxidation on different diets were as follows:As indicated by rate of whole body Leu flux, cats fed a diet that supplied 65% of ME intake as CP had elevated rates of whole body protein turnover via chiefly increased rates of amino acid oxidation. This study showed that during a period of marginal protein intake, cats modulated protein turnover to conserve body protein. Research supported by the Massey University Research Fund.

Extracellular enzyme activity and uptake of carbon and nitrogen along an estuarine salinity and nutrient gradient

Amino acid oxidation (AAO) and peptide hydrolysis (PH) are processes affecting the recycling of organic material and nutrients. We compared extracellular AAO and PH rates to C and N uptake rates along estuarine gradients of salinity, nutrients and productivity in the Pocomoke River, a subestuary of the Chesapeake Bay. This estuary is seasonally depleted in inorganic N, and rich in dissolved organic material (DOM) throughout the year. AAO, PH, and N uptake rates measured in 1999 and 2000 were not limited to particular size fractions measured, or to auto- or heterotrophic groups of organisms. At a station near the turbidity maximum, where chlorophyll a biomass was highest, smaller ( 1.2 μm) size-fraction, except at the least saline station in August of both years. Rates of AAO and PH were not linearly correlated with each other seasonally or spatially. Uptake of NH 4 + dominated total N uptake (>50%) at all but the freshwater station, although uptake of organic compounds was measurable at all sites. Rates of dissolved free amino acid uptake, measured using dually labeled compounds, were substantial (up to 11% of the total N uptake) and contributed both C and N for growth. Dual labels unambiguously demonstrated that uptake rates of amino acid C and N were uncoupled; amino acid N was taken up preferentially to amino acid C even when rates were corrected for N uptake from AAO. Conceptual models of DOM cycling should include the realization that enzymatic processes and uptake of DOM occur in both 'microbial' and larger size fractions. Thus, competition between bacteria and phytoplankton mixotrophs may be an important factor determining the relative uptake of C and N from amino acids and other organic substrates.

Effects of birth weight and postnatal nutrition on neonatal sheep: III. Regulation of energy metabolism.

This study investigated effects of birth weight and postnatal nutrition on regulation of energy metabolism in the neonatal lamb. Low (mean +/- SD 2.289 +/- 0.341 kg, n = 28) and high (4.840 +/- 0.446 kg, n = 20) birth weight male Suffolk x (Finnsheep x Dorset) lambs were individually reared on a liquid diet to grow rapidly (ad libitum fed, ADG = 337 g, n = 20) or slowly (ADG = 150 g, n = 20) from birth to live weights (LW) up to approximately 20 kg. At birth, small newborns had higher plasma concentrations of urea nitrogen (mean +/- SEM 8.31 +/- 0.25 vs 6.39 +/- 0.32 mM, P = 0.002) and somatotropin (ST, 49.1 +/- 17.0 vs 10.8 +/- 4.3 ng/mL, P = .045) and lower IGF-I (36.1 +/- 6.8 vs 157.7 +/- 21.8 ng/mL, P < 0.001) than large newborns. Plasma glucose (1.42 +/- 0.23 vs 2.63 +/- 0.95 mM, P = 0.147) and insulin (0.09 +/- 0.02 vs 0.13 +/- 0.06 ng/mL, P = 0.264) concentrations did not differ. Urea nitrogen concentration in plasma peaked and then declined rapidly in all lambs during the first week postpartum, and plasma ST declined on a body-weight-related basis from birth. During rearing to 20 kg LW, plasma insulin was higher in low- vs high-birth-weight lambs. Lambs fed ad libitum had greater plasma concentrations of glucose, urea nitrogen, insulin, and IGF-I compared to those fed a restricted diet (ADG = 150 g). The results suggest that during the early postpartum period, newborn lambs exhibit the fetal characteristic of high rates of amino acid oxidation. The results also support the notion that, at birth, low-birth-weight lambs are less mature than high-birth-weight lambs in aspects of metabolic and endocrine development, which may enhance their capacity to utilize amino acids for energy production and to support gluconeogenesis during the immediate postpartum period. Being small at birth also resulted in elevated plasma insulin concentrations when adequate nutriment to support moderate or rapid growth was provided postpartum, although it remains to be elucidated whether this more chronic effect persists in the longer term.

Peptide hydrolysis, amino acid oxidation, and nitrogen uptake in communities seasonally dominated by Aureococcus anophagefferens

Elevated levels of dissolved organic nitrogen (DON) and dissolved inorganic nitrogen (DIN) are among the factors implicated in the initiation of algal blooms. However, the degree to which phytoplankton augment their autotrophic metabolism with heterotrophic uptake of organic carbon that is associated with DON is unknown. We evaluated the relative importance of peptide hydrolysis, amino acid oxidation, and amino acid uptake over a seasonal cycle in an embayment on Long Island, New York, that had high concentrations of dissolved organic matter (DOM) and a bloom of the brown tide pelagophyte, Aureococcus anophagefferens. Amino acids were a significant component (up to 50%) of the total N uptake, particularly during the late summer. About half of the associated amino acid C was also taken up. Amino acid oxidation rates were an order of magnitude lower than free amino acid uptake rates, but still supplied up to 32.5% of the NH taken up. Up to 75% of the amino acid oxidation was in the bacterial size fraction (‹1.2 μm), and rates were significantly correlated with bacterial densities. Peptide hydrolysis rates were high, and most (up to 72%) occurred in the brown tide size fraction (1.2–5 μm). The high rates of peptide hydrolysis and amino acid uptake measured in cultures of A. anophagefferens confirm that this species can readily hydrolyze peptides and take up N and C from amino acids. Laboratory findings and size‐fractionation studies in the field suggest that A. anophagefferens plays a major role in consumption of both C and N from DOM.

Modeling mammary amino acid metabolism

Milk pricing schemes place economic importance on milk components. Most current nutrient requirement models do not predict milk component yields accurately. Deaggregation of energy and protein terms in those models may improve prediction accuracy. Descriptions of energy metabolism by the major postabsorptive tissues have progressed over the last 20 years. More recent efforts have been directed at representing amino acid metabolism. Mammary amino acid metabolism appears to be a function of amino acid supply and regulatory elements. Regulation of uptake and blood flow occurs and is represented in some models. Intracellular metabolism of amino acids and possibly energy are determinants of removal. Both the rate of amino acid oxidation and use for protein synthesis appear to be functions of intracellular concentrations. Experimental observations suggest that representation of protein synthesis as a linear function of the first-limiting amino acid is inadequate. A multi-substrate Michaelis–Menten equation form is more consistent with experimental observations and appears to yield better predictions as compared to the single-limiting model. Consideration of energy supply as a driver of milk protein synthesis also appears to be warranted. Additional knowledge of the substrate response surface for protein synthesis and how it is regulated is needed.

A tracer investigation of obligatory oxidative amino acid losses in healthy, young adults

Estimation of the minimum requirement for indispensable amino acids (IAAs) has been attempted by assuming that obligatory oxidative losses (OOLs) of IAAs can be approximated from nitrogen losses and that the efficiency of utilization of IAAs at requirement intakes is approximately 70%. We wished to determine the rates of OOLs in healthy adults, using L-[1-(13)C]leucine and L-[1-(13)C, methyl-(2)H(3)]methio-nine as tracers, after adjustment to a protein-free diet and how these rates compare with those when either sulfur amino acids (SAAs: methionine and cyst(e)ine) or leucine were removed from an otherwise adequate diet. Eleven subjects were randomly assigned to a 5-d protein-free diet or a 5-d diet providing adequate nitrogen and amino acids except for the SAAs or leucine. A 24-h constant intravenous infusion of [(15)N, (15)N]urea and L-[1-(13)C]leucine (Leu group; n = 5) or L-[1-(13)C, methyl-(2)H(3)]methionine (Met group; n = 6 ) began at 1800 on day 5 and rates of amino acid oxidation were determined. Mean (+/-SD) oxidation rates (mg kg(-)(1) d(-)(1)) of methionine and leucine were 6.4 +/- 1.4 and 24.7 +/- 3.6, respectively, with the protein-free diet; rates were significantly lower (3.9 +/- 2.2 and 7. 2 +/- 3.4, respectively) after the SAA- and leucine-free diets. Urea production was significantly lower (P < 0.01) with the protein-free than with the SAA- or leucine-free diet. Isotopically determined OOLs for methionine and leucine are consistent with losses predicted from nitrogen excretion, and consistent with our previous measurements of cysteine oxidation as an index of total SAA losses. The data further support our earlier conclusions regarding methionine sparing by cysteine and tentative recommended SAA requirements in adults.

Rates of Lysine Catabolism Are Inversely Related to Rates of Protein Synthesis When Measured Concurrently in Adult Female Rats Induced to Grow at Different Rates

To test the effect of changes in the rate of protein synthesis on amino acid oxidation, both were studied concurrently in individual 200-g female Sprague-Dawley rats. In a growth trial (Experiment 1), recombinant bovine somatotropin (rbST) was injected subcutaneously (0, 2 or 12 mg/d) over 6 d (n = 4 rats per rbST level). Weight gain increased with rbST level (P < 0.01); 1.96 ± 0.8, 4.24 ± 0.8 and 8.67 ± 0.8 g/d, respectively. After treatment with rbST (0 or 12 mg/d) for 4 d (Experiment 2), rats were injected via a tail vein catheter with valine (400 mmol, 4.07 mBq L-[3,4(n)-3H]valine) at 0, 4, 10, 13 or 16 h after the daily rbST injection and killed 20 min later. This flooding dose was 5 to 6 times, not 10 times, the free pool as hoped. Protein synthesis in rbST-treated rats increased 46% in muscle (P < 0.001) and 36% in liver (P < 0.01). The ks was unaltered with time after rbST injection (0–16 h, P > 0.05). When 600 mmol valine (4.4 mBq L-[3,4(n)-3H]valine) was used in Experiment 3, specific activity (SA) of free valine was constant over 20 min and was 94 ± 4% of that injected. Finally, in Experiment 4, protein synthesis and amino acid oxidation rates measured in the same rat revealed a 35% increase (P < 0.01) in protein synthesis in hind leg muscle and a 29% increase in liver (P < 0.05) from rbST-injected (12 mg/d) rats (n = 6). Lysine oxidation was estimated by continuous (12 h) infusion of L-[1-14C]lysine via the opposite tail vein catheter. Expired CO2 was collected over 20-min intervals and SA at plateau was estimated by fitting an exponential model. Lysine oxidation was reduced (P < 0.05) by 44% in rbST-treated rats. The idea that an increase in protein synthesis results in decreased amino acid oxidation remains tenable.

Whole-body protein kinetics in children with kwashiorkor and infection: a comparison of egg white and milk as dietary sources of protein

This study tested the hypothesis that during treatment of kwashiorkor (including marasmic kwashiorkor) with infection there is a lower rate of amino acid oxidation when the dietary intake of amino acids resembles the amino acid composition of acute phase proteins (APPs). Twenty-two children in Blantyre, Malawi, with kwashiorkor and acute infection were fed an isoenergetic, isonitrogenous diet with either egg white or milk as a protein source. The whole-body amino acid oxidation rate was measured after 24 h by determining the plasma urea rate of appearance, and whole-body protein breakdown and synthesis rates were determined from the plasma leucine rate of appearance. Plasma concentrations of C-reactive protein, alpha1-antitrypsin, tumor necrosis factor alpha (TNF-alpha), and interleukin 6 (IL-6) were determined on admission and at 24 and 48 h. The 11 children who received milk had a lower rate of amino acid oxidation than the children who received egg white (x +/- SD: 137 +/- 65 compared with 195 +/- 66 micromol urea x kg body wt(-1) x h(-1), P < 0.05). No significant differences were found between the two groups in the rate of whole-body protein breakdown or protein synthesis. The TNF-alpha concentration correlated inversely with whole-body protein breakdown and synthesis rates, and the IL-6 concentration correlated directly with C-reactive protein. We conclude that by making the amino acid composition of the diet resemble that of APPs in the treatment of acute kwashiorkor, the rate of amino acid oxidation can be decreased.

Substrate and acute temperature effects on turtle heart and liver mitochondria.

The oxidative properties of heart and liver mitochondria from the Western painted turtle (Chrysemys picta bellii) were characterized on the basis of substrate preferences and temperature sensitivity. Turtle heart and liver mitochondria oxidize most substrates at 2- and 10-fold less, respectively, than rates obtained from the corresponding rat mitochondria. Krebs cycle intermediates, ketone bodies, and glutamate were oxidized at similarly high rates by turtle heart mitochondria (70.0-121.2 nmol O.min-1.mg protein-1). Fatty acylcarnitines were oxidized at approximately one-half of the above rates, and rates of amino acid oxidation were either not detectable or very low. Heart mitochondria oxidize ketone bodies at rates as high as pyruvate plus malate. Liver mitochondria oxidized Krebs cycle intermediates, amino acids, and fatty acids at similarly low rates (3.0-8.0 nmol O.min-1.mg protein-1). Typically, succinate was oxidized at the highest rates, 20.6 +/- 4.0 and 121.2 +/- 1.2 nmol O.min-1.mg protein-1, for liver and heart, respectively. Values for the rate of change of oxidation with a 10 degrees C increase (Q10) from heart and liver mitochondria oxidizing glutamate were calculated over the range 25-5 degrees C in 5 degrees C intervals; over the range 10-5 degrees C, Q10 values were 3.2 and 19.8, respectively. Q10 values calculated over the higher temperature intervals were lower. It is suggested that the large difference in temperature sensitivity between mitochondria from these two tissues and the ability of the heart mitochondria to oxidize ketone bodies at high rates are adaptations to recover from the long anoxic overwintering bouts experienced by this species.

Effects of Glucose Infusion on Leucine Transamination and Oxidation in the Ovine Fetus

During fasting of the ewe, the rate of amino acid oxidation by the ovine fetus increases substantially. We hypothesized that the increase in amino acid oxidation derived mainly from reduced protein synthesis. We further hypothesized that fetal glucose supplementation would result in diminished amino acid oxidation. To test these hypotheses, nine ovine fetuses were infused with [15N,1-13C]leucine to determine the rates of leucine appearance and disposal. Simultaneously, the fetal uptake of leucine was determined. Animals were studied in the fed and fasted state. After baseline measurements, glucose was infused into the fetal inferior vena cava at a rate estimated to match the fetal glucose uptake. Results of these studies indicate that leucine nitrogen flux, leucine carbon flux and fetal leucine uptake were constant. Leucine oxidation was increased by 50% in the fasted state (6.3 versus 13.4 mumol/min); glucose infusion resulted in a 25% decline in oxidation (to 10.4 mumol/min) in the fasted state, but had no effect in the fed state. Mean leucine umbilical uptake during fasting was 9.3 mumol/min, 4.1 mumol/min less than leucine oxidation. These data suggest that leucine (and potentially other amino acids) may be in negative balance during maternal fasting, and can be spared by supplementation of the fetus with exogenous glucose.

Fenton chemistry. Amino acid oxidation.

The oxidation of amino acids by Fenton reagent (H2O2 + Fe(II] leads mainly to the formation of NH+4, alpha-ketoacids, CO2, oximes, and aldehydes or carboxylic acids containing one less carbon atom. Oxidation is almost completely dependent on the presence of bicarbonate ion and is stimulated by iron chelators at levels which are substoichiometric with respect to the iron concentration but is inhibited at higher concentrations. The stimulatory effect of chelators is not due merely to solubilization of catalytically inactive polymeric forms of Fe(OH)3 nor to the conversion of Fe(II) to complexes incapable of scavenging hydroxyl radicals. The results suggest that an iron chelate and another as yet unidentified form of iron are both required for maximal rates of amino acid oxidation. The metal ion-catalyzed oxidation of amino acids is likely a "caged" process, since the oxidation is not inhibited by hydroxyl radical scavengers, and the relative rates of oxidation of various amino acids by the Fenton system as well as the distribution of products formed (especially products of aromatic amino acids) are significantly different from those reported for amino acid oxidation by ionizing radiation. Several iron-binding proteins, peptides, and hemoglobin degradation products can replace Fe(II) or Fe(III) in the bicarbonate-dependent oxidation of amino acids. In view of their ability to sequester metal ions and their susceptibility to oxidation by H2O2 in the presence of physiological concentrations of bicarbonate, amino acids may serve an important role in antioxidant defense against tissue damage.

Protein-metabolism kinetics and energy-substrate utilization in infants fed parenteral solutions with different glucose-fat ratios

The relative effect of glucose and lipids on whole-body protein-metabolism kinetics was assessed in seven infants undergoing parenteral feeding. Protein intake was kept constant and nonprotein energy was either provided as glucose alone or as an isoenergetic glucose-lipid mixture according to a randomized crossover trial. Protein metabolism and energy-substrate utilization were assessed by a primed, constant L-[13C]leucine infusion, combined with indirect calorimetry. There was a significant difference in the pattern of energy-substrate utilization according to regime. Protein turnover (11.3 +/- 0.7 vs 9.8 +/- 0.4 g.kg-1.d-1; P less than 0.05), protein breakdown (8.4 +/- 0.6 vs 7.1 +/- 0.4 g.kg-1.d-1; P less than 0.05), and amino acid oxidation rates (2.7 +/- 0.4 vs 1.4 +/- 0.5 g.kg-1.d-1; P less than 0.05) were higher for the glucose than the glucose-lipid treatment, whereas protein-synthesis rates did not significantly differ. These results suggest that the nature of energy substrates delivered to parenterally fed infants may affect protein metabolism.

A theoretical basis for increasing current estimates of the amino acid requirements in adult man, with experimental support

Estimates of the minimum (obligatory) rates of loss of indispensable acids via irreversible oxidation in healthy young adults are predicted from published data for obligatory nitrogen losses and from published values for whole-body protein turnover and the efficiency of amino acid recycling. The estimates obtained by these two different approaches are consistent and equal to or in most cases considerably higher than current values for the upper range of the individual amino acid requirements in healthy adults. From these estimates of the obligatory, or minimum, rates of amino acid oxidation, the minimum requirements for indispensable amino acids are predicted. These are about two to three times higher than current requirement figures. Experimental support for these higher requirement figures is presented, based on published 13C-labeled amino acid tracer studies. When these revised estimates are expressed per unit of protein need, the amino acid pattern is similar to the 1985 Food and Agriculture Organization/World Health Organization/United Nations University amino acid scoring pattern for the preschool child (aged 2-5 y).