Gender differences in basal protein kinetics in young adults

Gender affects energy expenditure and influences the relative utilization of carbohydrate and fat as fuels. However, little is known about the possible effects of gender on protein metabolism. Thus, we compared whole body and plasma (albumin and fibrinogen) protein kinetics in the basal postabsorptive state in young, untrained volunteers divided into two groups according to gender (women: n=17; age, 24+/-4 yr; men: n=17; age, 25+/-2 yr). The two groups were matched for body mass index. Protein kinetics were measured by means of L-[1-14C]leucine infusion. The leucine whole body rate of appearance, an index of proteolysis, and nonoxidative rate of disappearance, an index of protein synthesis, were similar in the two groups. However, the leucine oxidation rate was significantly lower in women compared to men (0.23+/-0.07 vs. 0.31+/-0.08 micromol/kg min; P=0.0062). Similar results were obtained when data were adjusted for estimated body composition. Albumin and fibrinogen fractional secretion rates were not different in the two groups. In conclusion, in the basal state leucine oxidation is lower in women than in men regardless of body composition. This could be one of the factors contributing to the lower metabolic rate in women.     

Endogenous loss of leucine and methionine in adult male rats

1. The fractional rate of loss of 14C and body-weight was measured in adult male rats after giving 14C-labelled methionine or leucine and maintaining rats for 30 d on a low-protein or a specific methionine+cystine-free diet: carcasses were then analysed for protein and fat 14C radioactivity. 2. The fractional loss of 14CO2 from [14C]methionine or [14C]leucine between day 20 and day 30 was always greater than the fractional loss of body-weight. 3. Carcass protein 14C radioactivity after giving [14C]leucine was higher than after giving [14C]methionine, but fat 14C radioactivity after either 14C-labelled amino acid was only a small proportion of the total body 14C radioactivity. 4. After correction of the fractional loss of 14CO2 for urinary 14C loss, but not body-weight loss, absolute amino acid loss was calculated using published values for methionine and leucine content of rats. 5. The best estimates of endogenous amino acid loss obtained using I-14C-labelled amino acids, expressed as mg/kg body-weight 0.75 per day were leucine 79, methionine 38.     

Protein metabolism in human obesity: relationship with glucose and lipid metabolism and with visceral adipose tissue

It is controversial whether metabolic disorders of human obesity include protein metabolism. Even less information is available concerning the effect of fat distribution on protein metabolism. Therefore, a comprehensive evaluation of glucose, lipid, and protein metabolism was performed in 11 obese nondiabetic and 9 normal women whose body composition and regional fat distribution were determined. [1-14C]Leucine and [3-3H]glucose were infused in the postabsorptive state and during an euglycemic hyperinsulinemic (35-40 microU/mL) clamp combined with indirect calorimetry for assessment of leucine flux, oxidation, and nonoxidative disposal, glucose turnover and oxidation, and lipid oxidation. Fat-free mass (FFM) was estimated by a bolus of 3H2O. Subcutaneous abdominal and visceral adipose tissues were determined by nuclear magnetic resonance imaging. During the clamp, obese women had lower glucose turnover (4.51 +/- 0.41 vs. 6.63 +/- 0.40 mg/min.kg FFM; P < 0.05), with a defect in both oxidation (3.27 +/- 0.22 vs. 3.89 +/- 0.21) and nonoxidative disposal (1.24 +/- 0.27 vs. 2.74 +/- 0.41; P < 0.005), whereas lipid oxidation was higher during the clamp (0.49 +/- 0.15 vs. 0.17 +/- 0.09 mg/min.kg FFM). There was no difference in leucine flux (basal, 2.23 +/- 0.17 vs. 2.30 +/- 0.29; clamp, 2.06 +/- 0.19 vs. 2.10 +/- 0.24 mumol/min.kg FFM), oxidation (basal, 0.37 +/- 0.04 vs. 0.36 +/- 0.05; clamp, 0.34 +/- 0.04 vs. 0.39 +/- 0.06) and nonoxidative leucine disposal (basal, 1.86 +/- 0.17 vs. 1.94 +/- 0.26; clamp, 1.72 +/- 0.20 vs. 1.71 +/- 0.19) in the two groups. In obese women, basal leucine oxidation was directly related with glucose oxidation and inversely to lipid oxidation (both P < 0.05), whereas visceral adipose tissue was inversely related to leucine flux both in the basal state and during the clamp (P < 0.05). In conclusion, in human obesity, 1) rates of protein metabolism in the basal state and in the range of insulin concentrations encountered after a meal are normal; 2) protein oxidation is positively related to glucose oxidation and negatively related to lipid oxidation; and 3) visceral adipose tissue is inversely related to all parameters of protein metabolism.     

Heritable disorder resembling neuronal storage disease in mice expressing prion protein with deletion of an alpha-helix

Ethanol abuse is frequently associated with protein malnutrition. To assess the acute effects of ethanol on whole-body protein metabolism, [1-13C]leucine kinetics were measured in eight postabsorptive normal male subjects three times, ie, during administration of two doses of ethanol (dose 1, 0.52 g/kg during 2 hours and 0.3 g/kg during 3 hours; dose 2, 0.69 g/kg during 2 hours and 0.3 g/kg during 3 hours) and during saline (controls). During the last 2 hours of the studies, glucose, insulin, and amino acids were infused to assess the effects of ethanol on protein kinetics under anabolic conditions (euglycemic clamp). The decreases in leucine flux (reflecting whole-body protein breakdown) and nonoxidative leucine disappearance (a parameter of protein synthesis) during saline infusion were abolished in both ethanol protocols (P < .05 or less v saline). The rate of leucine oxidation decreased during the higher dose of ethanol compared with saline (P < .005), indicating an anticatabolic effect. During anabolic conditions (clamp), leucine flux and nonoxidative leucine disappearance were significantly higher in both ethanol studies compared with saline (P < .05). Resting energy expenditure (REE) and oxygen consumption (VO2) during the euglycemic clamp increased to a greater degree during both ethanol studies than during saline (P < .05 or less). Thus, an elevation of blood ethanol concentrations to the levels observed in social drinking results in a net anticatabolic effect (diminished leucine oxidation) when ethanol is administered alone. However, during administration of other nutritional substrates, the anticatabolic effect was not detectable, possibly because ethanol enhanced nutrient-induced thermogenesis.     

Acute growth hormone effects on amino acid and lipid metabolism

The anabolic actions of GH are well known, although specific tissue responses and the mechanism of nitrogen conservation are less well understood. This study was designed to examine the acute metabolic effects of GH on whole body and regional protein metabolism, using an experimental protocol which controlled for confounding perturbations in other hormones by a simultaneous infusion of somatostatin. Control subjects received replacement doses of insulin, glucagon, and GH for the entire 7-h study period, whereas GH subjects received an identical protocol, except for an increased dose of GH sufficient to increase serum concentrations into the high-physiological range (12-20 ng/mL) for the final 3.5 h of the study (P < 0.001). Thirteen young, healthy male subjects were studied in the postabsorptive period; five served as control subjects and eight as treatment (GH) subjects. Each received continuous iv infusions of somatostatin, L-[13-C]leucine, and L-[2H5]phenylalanine throughout the study. Femoral arterial and venous sampling allowed for simultaneous measurements across the leg and in the whole body. C-Peptide levels were suppressed throughout the infusion; insulin, glucagon, insulin-like growth factor I, cortisol, epinephrine, norepinephrine, and glucose concentrations were not different between groups. Glycerol concentrations increased 3-fold in GH subjects during the final 3.5-h period (P = 0.04). Concentrations of several amino acids declined through the study, but no differences were observed between treatment groups. Leucine oxidation was reduced in GH compared to control subjects (P = 0.04). No changes in CO2 production or whole body leucine or phenylalanine flux were observed, whereas nonoxidative disposal of leucine was marginally higher in GH compared to control subjects (P = 0.07). By contrast, rates of appearance and disappearance of both leucine and phenylalanine across the leg all were relatively lower in GH compared to control subjects; leucine balance across the leg was reduced by GH (P = 0.03), whereas phenylalanine balance was not influenced by GH. Our data thus demonstrate an acute stimulatory effect of GH on lipolysis, a decrease in leucine oxidation, and no stimulation of muscle protein synthesis in spite of enhanced protein synthesis in nonmuscle tissue.     

Dietary protein restriction alters glucose but not protein metabolism in non-insulin-dependent diabetes mellitus

We determined whether a customary diet high or low in protein (1) influences postabsorptive amino acid catabolism, nitrogen (N) balance, and hepatic glucose output (HGO) in normal subjects or patients with non-insulin-dependent diabetes mellitus (NIDDM) or (2) alters blood glucose levels in NIDDM. Eight normal young adults and five obese middle-aged persons with NIDDM consumed low-protein (0.8 g/kg lean body mass [LBM]) or high-protein (3.0 g/kg LBM) diets at maintenance energy for consecutive 7-day periods. Fasting and average blood glucose and N balance were measured daily. The level of dietary protein had no effect on the basal plasma leucine rate of appearance (Ra) or urinary 3-methylhistidine excretion in either subject group. Basal leucine oxidation (and by inference, whole-body amino acid catabolism) was reduced on the low-protein diet but basal HGO was not, and although exogenous glucose effectively suppressed HGO, it did not reduce leucine oxidation with either diet. After adaptation to the low-protein diet, N balance in both the normal and NIDDM subjects was close to zero. The low-protein diet reduced the fasting and daily blood glucose of the diabetic subjects by approximately 2 mmol/L (P < .05). We conclude that physiologic variation in dietary protein does not affect basal whole-body protein turnover or HGO in either normal young adults or obese middle-aged NIDDM subjects. However, protein restriction to the level of the average daily requirement significantly reduces postabsorptive and average daily blood glucose concentrations in persons with NIDDM.     

Bed rails: a barrier to independence?

The influence of meal frequency on change of body weight and protein status, measured by level of amino acid oxidation (decarboxylation) in the postabsorptive state, was studied at a fixed daily protein intake. Growing rats (250g) were fed through gastric canula a feeding solution based on Nutrison Standard supplying 1.6g protein and 266kJ ME daily. This amount was given in either 2 large meals at the beginning and the end, or in 6 smaller meals, or by continuous infusion during entire dark period (10 hrs). After 3 weeks of feeding the mean growth rate of the rats fed continuously was nearly 20% higher than rats fed the same amount in 2 meals. The rats fed 6 meals a day had a growth rate rather similar to the rats fed continuously. The percentile recovery of label as 14CO2 in the breath after an intraperitoneal injection of [1-14C]leucine (4 hrs after last meal) was significantly higher (p.05) for the animals fed continuously (27% sd 2.6) compared to the rats fed 2 meals (21.9% sd 4.0). The value for 6 meal group was intermediate (24.5 sd 1.8). The results indicate that the metabolic utilization of a fixed daily amount of protein is clearly influenced by the way of supply. With respect to the change of body weight and protein status, animals have more benefit of the same amount of protein if the supply is more equable. It is suggested that the difference is caused by metabolic restriction for an adequate utilisation of large meals. Therefore large meals are supposed to cause a waste of amino acids in the postprandial phase. As a consequence amino acid amount that will be stored in the body to be available in the postabsorptive phase will be less.     

Metabolism in the hypothermically perfused dog kidney. Incorporation rate of leucine and threonine into proteins

The incorporation of [14C]leucine and [14C]threonine into kidney cortex proteins was studied during 6 days' hypothermic perfusion of dog kidneys at 8-10 degrees C and during in vitro incubation of dog kidney cortex slices at 37 degrees C. Leucine carbon was incorporated into proteins at a higher rate than threonine carbon both during in vitro incubation of kidney cortex slices and during hypothermic kidney perfusion. The incorporation of leucine and threonine during hypothermic perfusion was linear for 6 days but 50-100 times lower than the incorporation of leucine and threonine in kidney cortex slices at 37 degrees C. During hypothermic perfusion there was a decrease in specific activity of leucine and threonine in the perfusate corresponding to a degradation of proteins which was greater than protein synthesis as calculated from the incorporation of label into proteins. Leucine carbon was recovered in CO2 during hypothermic perfusion and in vitro incubation of kidney cortex slices at 37 degrees C. The incorporation of threonine carbon into CO2 was about 10% of the corresponding value for leucine both during hypothermic kidney perfusion and during in vitro incubation of kidney cortex slices at 37 degrees C. It is concluded that there is a turnover of kidney proteins during hypothermic perfusion with a perfusate containing amino acids.     

Comparison of leucine kinetics in endurance-trained and sedentary humans

Whole body leucine kinetics was compared in endurance-trained athletes and sedentary controls matched for age, gender, and body weight. Kinetic studies were performed during 3 h of rest, 1 h of exercise (50% maximal oxygen consumption), and 2 h of recovery. When leucine kinetics were expressed both per unit of body weight and per unit of fat-free mass, both groups demonstrated an increase in leucine oxidation during exercise (P < 0.01). Trained athletes had a greater leucine rate of appearance during exercise and recovery compared with their sedentary counterparts (P < 0.05) and an increased leucine oxidation at all times on the basis of body weight (P < 0.05). However, all of these between-group differences were eliminated when leucine kinetics were corrected for fat-free tissue mass. Therefore, correction of leucine kinetics for fat-free mass may be important when cross-sectional investigations on humans are performed. Furthermore, leucine oxidation, when expressed relative to whole-body oxygen consumption during exercise, was similar between groups. It is concluded that there was no difference between endurance-trained and sedentary humans in whole body leucine kinetics during rest, exercise, or recovery when expressed per unit of fat-free tissue mass.     

Gender-dependent alterations in serum leptin in alcoholic cirrhosis

BACKGROUND & AIMS: Leptin is a peptide that decreases food intake and increases energy expenditure. It is produced in fat cells, is stimulated by cytokines, and its levels in serum are higher in females. Because anorexia, hypermetabolism, and elevated cytokine levels are frequently observed in cirrhosis, we hypothesized that the serum leptin level would be elevated in cirrhosis. The aim of this study was to investigate the relationship of serum leptin to gender, body composition, and tumor necrosis factor (TNF). METHODS: Male (n = 18) and female (n = 10) abstinent alcoholic cirrhotic patients were studied and compared with control subjects (15 male and 8 female). Fat mass, fat-free body mass, and body cell mass were calculated by using H2[18O] and bromide dilution methodology. Serum leptin and TNF concentrations were measured by immunoassays. RESULTS: Fat mass was decreased only in male cirrhotics (P < 0.05), whereas body cell mass was decreased in both male and female cirrhotics (P < 0.01). Leptin levels were elevated in female (P < 0. 001) but not male cirrhotics compared with controls. When expressed per kilogram of fat mass, leptin was elevated in both male (P < 0. 01) and female (P < 0.01) cirrhotics. Women in both cirrhotic and control groups had higher leptin levels than men. TNF was elevated in both male and female cirrhotics and did not correlate with leptin levels. CONCLUSIONS: Cirrhotics have elevated serum leptin levels, which are related to both gender- and gender-dependent alterations in body composition.     

Variation in the apparent sensitivity of the insulin-mediated inhibition of proteolysis to amino acid supply determines the efficiency of protein utilization

1. The variability between normal individuals in the efficiency of postprandial protein utilization (PPU), a determinant of the apparent protein requirement, was examined in relation to the relative responses of protein synthesis and proteolysis to protein feeding by means of [1-13C]leucine turnover and balance studies.2. Twenty-five healthy adults were infused intravenously with L-[1-13C]leucine continuously for 9 h. This was started in the postabsorptive state (PA, 3 h) and followed by low-protein feeding (LP, 3 h), and then by isoenergetic high-protein feeding (HP, 3 h). This allowed protein intake to be varied against a constant postprandial insulin level so that the extent of any amino-acid-mediated responses which were additional to those exerted by insulin could be investigated. Leucine oxidation, O, and balance (intake-oxidation), protein synthesis, S, and degradation, D, were calculated from plasma [1-13C]alpha-ketoisocaproic acid enrichment and 13CO2 excretion.3.PPUprotein, calculated as change in leucine balance/change in intake (HP-LP), varied from 0.58 to 0.99 (mean=0. 81+/-0.10), independently of age or sex. PPUprotein varied directly with the inhibition of D and inversely with the increase in leucine concentration and stimulation of O and S.4. Efficient PPU, as demonstrated by the top quintile of individuals categorized in terms of PPUprotein, involves maximal inhibition of D by protein feeding with minimal increases in free amino acid concentrations, O and S. Lesser inhibition of D and greater stimulation of S and O characterized the lower, less efficient quintile. This indicates that the efficiency of protein utilization in individuals, and a component of their apparent protein requirement, is determined by the sensitivity of the insulin-mediated inhibition of proteolysis to amino acid supply.     

What makes a life good?

OBJECTIVE: The objective of this study was to evaluate the longitudinal changes in energy expenditure and body composition in relationship to alterations in carbohydrate metabolism in women with normal and abnormal glucose metabolism. We hypothesized that women with decreased insulin sensitivity before conception would have less fat accretion and smaller increases in energy expenditure. STUDY DESIGN: Six women with normal glucose tolerance and 10 women with abnormal glucose tolerance were evaluated before conception, and in early (12 to 14 weeks) and late (34 to 36 weeks) gestation. Body composition was estimated by hydrodensitometry, resting energy expenditure, and glucose and fat metabolism by indirect calorimetry, endogenous glucose production by infusion of [6-6 2H2] glucose, and insulin sensitivity using a hyperinsulinemic-euglycemic clamp (40 mU/m2/min). RESULTS: There was a smaller increase in fat mass (1.3 kg [P = .04]) in early pregnancy in women with abnormal glucose tolerance before pregnancy. Indirect calorimetry measured gestational age-related increases in basal oxygen utilization, with or without correction for fat-free mass (VO2, P = .002), resting energy expenditure (expressed in kilocalories, P = .0001), and carbohydrate oxidation (P = .0003). The insulin-mediated elevation in VO2 increased in later gestation VO2 (P = .005), as did resting energy expenditure (P = .0001) and fat oxidation (P = 0.0001). However, there was a decrease in respiratory quotient (P = .0001), carbohydrate oxidation (P = .002), and nonoxidative carbohydrate metabolism (P = .0001) with advancing gestation during insulin infusion. In early pregnancy, changes in fat mass correlated inversely with changes in insulin sensitivity (r= -0.52, P = .04) and changes in basal VO2 correlated inversely with decreases in basal endogenous glucose production (r = -0.74, P = .01). CONCLUSION: In early gestation, the changes in maternal fat mass and basal oxygen consumption are inversely related to the changes in insulin sensitivity. This response in lean women with decreased insulin sensitivity before conception may have survival value by providing a larger amount of available substrate to meet fetoplacental needs during gestation.     

Metabolism of [1-(14)C] and [2-(14)C] leucine in cultured skin fibroblasts from patients with isovaleric acidemia. Characterization of metabolic defects

Leucine metabolism in cultured skin fibroblasts from patients with isovaleric acidemia was compared with that in normal fibroblasts and in cells from patients with maple syrup urine disease using [1-(14)C] and [2-(14)C] leucine as substrates. Inhibitory effects of methylenecyclopropylacetic acid on leucine metabolism in normal cells were also investigated. Production of 14CO2 from [2-(14)C] leucine was very reduced (96-99%) in both types of mutant cells. Radioactive isovaleric acid accumulated in assay media with isovaleric acidemia cells but not in those with maple syrup urine disease cells. Unexpectedly, 14CO2 production from [1-(14)C] leucine was partially depressed (80%) in isovaleric acidemia cells whereas in maple syrup urine disease cells it was strongly depressed (99%) as expected. These two mutant cells were clearly distinguished by detection of 14C-isovaleric acid accumulation after incubation with [2-(14)C] leucine. A pattern of inhibition of leucine oxidation similar to that seen in isovaleric acidemia cells was induced in normal cells by the addition of 0.7 mM methylenecyclopropylacetic acid to the assay medium. The partial inhibition of [1-(14)C] leucine oxidation seen in isovaleric acidemia cells and also in normal cells in the presence of the inhibitor appears to be, at least in part, due to an accumulation of isovalerate in the cells. Isovaleric acid (5-10) mM) inhibited [1-(14)C] leucine oxidation 32-68% when added to the assay medium with normal cells. Addition of flavin adenine dinucleoside to culture medium or assay medium or both did not restore oxidation of either leucine substrate in isovaleric acidemia cells.     

Comparison of leucine, serine and glycine transport across the ovine placenta

To estimate the transport rate of maternal glycine across the placenta [1-13C]glycine and L-[1-13]serine were infused intravenously in pregnant sheep using both continuous and bolus infusions. Each tracer was infused together with L-[1-13C]leucine, to enable a comparison with the placental transport of an essential amino acid. At steady state, fetal plasma leucine enrichment was 40 per cent of maternal enrichment, indicating that approximately 60 per cent of the entry rate of leucine into fetal plasma is derived from protein breakdown in the placenta and fetus. Fetal plasma glycine enrichment was 11 per cent of maternal and there was no detectable fetal serine enrichment. The direct flux of maternal leucine into the fetal circulation was approximately 3.0 (bolus experiments) to 3.6 (continuous infusion experiments) mumol/min (kg fetus) and greater than the estimated 1.4 mumol/min (kg fetus) direct flux of maternal glycine, despite the fact that the net umbilical uptake of glycine exceeds that of leucine. This supports the conclusion that placental glycine production is a quantitatively important contribution to fetal glycine uptake via the umbilical circulation. The fetal glycine supply from the placenta is provided by a relatively small direct maternal glycine transplacental flux and a larger contribution derived from serine utilization within the placenta for glycine production.     

Albumin synthesis rates in cirrhosis: correlation with Child-Turcotte classification

Albumin-synthesis rates were measured in nine patients with stable cirrhosis and compared with those of eight healthy volunteers by means of a new technique using stable isotopes. Four grams of L-[1-13C]leucine was injected over 10 min, and blood samples were drawn at intervals. Serum free [13C]leucine enrichment, taken to be the precursor for albumin synthesis, and 13C enrichment of leucine in albumin, isolated with differential solubility in absolute ethanol from trichloracetic acid-precipitated serum proteins, were measured on mass spectrometry. Albumin synthesis, expressed as a fractional rate, was 7.9% +/- 0.3%/day in the controls and 7.9% +/- 1.1%/day in the cirrhotic patients. Albumin synthesis, expressed as an absolute rate, was lower in the cirrhotic group (cirrhotic, 119 +/- 17 mg/kg/day; controls, 146 +/- 8 mg/kg/day), but because of the relatively small number of patients the difference was not significant. However, the absolute rate of albumin synthesis significantly correlated with the Child-Turcotte score (p = 0.024) and its Pugh modification (p = 0.027). The rate of albumin synthesis also correlated with serum phenylalanine concentration but not with serum albumin concentration and intravascular albumin mass or with other clinical indexes of liver function or integrity when taken separately. However, the significant correlation between albumin synthesis and Child score suggests that albumin synthesis might be useful for the clinical judgment of patients with cirrhosis.     

Infusion of soy and casein protein meals affects interorgan amino acid metabolism and urea kinetics differently in pigs

For routine evaluation of the quality of dietary protein, amino acid scoring patterns were used. Evaluation of this pattern for soy and casein revealed that these proteins are of almost equal quality. However, in vivo studies showed a large difference. To study the biological effects of meals with casein and soy protein, the contributions of individual amino acids to net protein retention and amino acid kinetics in gut, liver and muscle in healthy pigs were investigated. Isonitrogenous enteral nutrition, infused at a rate of 10 mL. kg body wt-1. h-1 and consisting of maltodextrin (137 g/L) with added casein (53 g/L) or soy protein (68 g/L), was given to conscious, healthy female multicathetized pigs (20-22 kg, n = 12). A primed-constant infusion protocol with L-[ring-2,6-3H]phenylalanine, L-[3,4-3H]valine and [15N-15N]urea was used to measure amino acid and urea kinetics in gut, liver and muscle. Measurements were done postabsorptively and 2-6 h after initiation of the enteral nutrition. During the meal, appearance of amino acids into the portal vein and the uptake by the liver was lower with casein infusion. Muscle uptake did not differ. Gut protein synthesis tended to be lower with soy infusion (P = 0.1). Liver protein synthesis and degradation were higher with casein infusion (P < 0.05), while in muscle, soy infusion stimulated protein turnover (P < 0.05). In comparison to the postabsorptive condition, liver urea production was unchanged after casein infusion, while it was significantly increased after soy infusion. These results suggest that the quality of soy protein is inferior to that of casein protein.     

Nitrogen homeostasis in man: influence of protein intake on the amplitude of diurnal cycling of body nitrogen

1. The diurnal nature of nitrogen (N) homoeostasis was investigated in adults fed increasing protein intakes. N balance was estimated during a 48 h period of consecutive 12 h periods of feeding hourly meals and fasting, after 12 days of adaptation to diets containing 0.36 +/- 0.01, 0.77 +/- 0.03, 1.59 +/- 0.08 and 2.31 +/- 0.65 g of protein day-1 kg-1. N losses were determined from measured urinary N excretion corrected for changes in the body urea pool, and estimated faecal and miscellaneous losses. [13C]Leucine and [2H5]phenylalanine balances were measured during a primed, continuous infusion of the two amino acids during the fasting and feeding phase on the second day. 2. Increasing fasting N losses were observed (47 +/- 7, 60 +/- 6, 95 +/- 15 and 140 +/- 36 mg day-1 kg-1) on the four intakes, with corresponding increasing fed gains of 8.2 +/- 3.9, 40.2 +/- 7.1, 112 +/- 24 and 180 +/- 56 mg day-1 kg-1. 3. Increasing fed-state amino acid gains with increasing protein intake were observed with both [13C]leucine and [2H5]phenylalanine, whereas increasing fasting amino acid losses were confirmed with [13C]leucine. 4. The N equivalent of the leucine oxidation rate was mostly in the range of 10-50% lower than expected from the N excretion rates. This may reflect the timing of the amino acid balance measurements and non-uniform rates of gain and loss throughout the diurnal cycle. 5. We conclude on the basis of both N and amino acid balances that the amplitude of the diurnal cycling of body protein N in human adults increases with increasing dietary protein intake. Thus one component of the protein requirement for N balance reflects a demand for repletion of fasting losses which increases with increasing habitual protein intake.     

Free fatty acid metabolism in aerobically fit individuals

The impact of aerobic fitness level on the production and disposal of serum free fatty acids was investigated in 26 normal young volunteers. The fitness level was ascertained by history and confirmed by determination of maximal aerobic capacity. Energy expenditure and substrate oxidation at rest were measured with indirect calorimetry. Free fatty acid turnover was measured with an infusion of [14C]palmitic acid. All tests were done > or = 48 h after the last bout of exercise. The sedentary (SED) volunteers had higher rates of systemic delivery of fatty acids than aerobically fit (FIT) individuals (532 +/- 53.4 vs. 353 +/- 62.3 mumol/min; P = 0.05). This difference was accentuated when the values were normalized to fat-free mass (9.2 +/- 0.8 and 5.9 +/- 0.98 mumol.kg-1.min-1 for SED and FIT, respectively). Fatty acid oxidation was similar between FIT and SED volunteers in absolute numbers (209 +/- 25 vs. 202 +/- 21 mumol/min, respectively; NS) as well as when normalized to fat-free mass (3.8 +/- 0.9 vs. 3.6 +/- 1.4 mumol.kg-1.min-1, respectively; NS). In contrast, the nonoxidative disposal of serum fatty acids was higher in SED (330 +/- 46.1 mumol/min) than in FIT individuals (144 +/- 52 mumol/min; P = 0.026). Thus, the ratio of nonoxidative to oxidative disposal rates of fatty acids was higher in SED than in FIT individuals (1.65 +/- 0.29 vs. 0.75 +/- 0.17; P = 0.021). The data support the hypothesis that high aerobic fitness level is associated with a low rate of systemic delivery of fatty acids at rest. Nevertheless, subjects with high aerobic fitness levels have fat oxidation at the same rate as unfit individuals.     

A nitrogen-free hypocaloric diet and recombinant human growth hormone stimulate postoperative protein synthesis: fasted and fed leucine kinetics in the surgical patient

Twelve otherwise healthy patients undergoing elective surgery for resection of rectosigmoid adenocarcinoma were randomly allocated to two groups: one group receiving intravenous dextrose 5% 600 to 800 kcal.d-1 (DX, n = 6) and the other group receiving the same amount of dextrose intravenously plus recombinant human growth hormone (DX + rGH, n = 6). Supplementation with rGH started on the day of surgery and continued postoperatively for 5 days. No nitrogen was provided in the diet. This regimen was started 3 days before surgery and continued for 5 days after surgery. Protein kinetics were studied over a period of 8 hours in all patients. Following an overnight fast, a primed constant infusion of L-[1-13C]leucine was maintained for 4 hours (fasted state) and continued for a further 4 hours (fed state) during which 5% beet dextrose (low 13C content) with or without rGH was administered. The isotope studies were performed on the day before surgery and 6 days after surgery. Other measurements included urinary nitrogen excretion, gaseous exchange, and plasma concentrations of insulin, GH, and insulin-like growth factor-I (IGF-I). Addition of rGH to the dextrose diet had a significant positive effect on protein synthesis (P = .02). Surgery was responsible for a significant increase in postoperative whole-body protein breakdown and synthesis and leucine oxidation (P < .01), although lesser changes were observed in the DX group. An interaction between rGH and surgery was associated with a significant increase in protein synthesis (P = .009), but not with changes in either protein breakdown or leucine oxidation. Carbohydrate provision in the form of beet dextrose during the fed state of the isotopic study did not attenuate the significant decrease in protein synthesis (P = .01) or breakdown (P = .003) either before or after surgery, probably reflecting the absence of nitrogen in the diet. No significant interaction was found between rGH and feeding. These results of leucine kinetics indicate that addition of rGH to a low-dextrose intake in the absence of dietary nitrogen can actually promote protein synthesis. The low levels of leucine oxidation could be explained by the fact that amino acids resulting from protein degradation were directed preferentially toward resynthesis of new proteins rather than to oxidative pathways. There was a significant increase in plasma insulin and GH in the group receiving rGH (P < .05). The postoperative plasma concentration of IGF-I did not change in the latter group compared with the DX group, in which IGF-I concentration decreased significantly (P < .05) as part of the response to combined surgery and dietary restriction. Although both IGF-I and insulin are independently capable of stimulating protein synthesis, elevated levels of either hormone or GH itself may primarily modulate protein synthesis, even with a low intake of carbohydrates.     

Modulation of whole body protein metabolism, during and after exercise, by variation of dietary protein

The aim of this study was to investigate dietary protein-induced changes in whole body leucine turnover and oxidation and in skeletal muscle branched chain 2-oxo acid dehydrogenase (BCOADH) activity, at rest and during exercise. Postabsorptive subjects received a primed constant infusion of L-[1-13C,15N]leucine for 6 h, after previous consumption of a high- (HP; 1.8 g . kg-1 . day-1, n = 8) or a low-protein diet (LP; 0.7 g . kg-1 . day-1, n = 8) for 7 days. The subjects were studied at rest for 2 h, during 2-h exercise at 60% maximum oxygen consumption, then again for 2 h at rest. Exercise induced a doubling of both leucine oxidation from 20 micromol . kg-1 . h-1 and BCOADH percent activation from 7% in all subjects. Leucine oxidation was greater before (+46%) and during (+40%, P < 0.05) the first hour of exercise in subjects consuming the HP rather than the LP diet, but there was no additional change in muscle BCOADH activity. The results suggest that leucine oxidation was increased by previous ingestion of an HP diet, attributable to an increase in leucine availability rather than to a stimulation of the skeletal muscle BCOADH activity.