Fuel homeostasis during physical inactivity induced by bed rest.

The consequences of physical inactivity on fuel homeostasis were evaluated during 7 days of head-down bed rest (HDBR), a model mimicking weightlessness. Eight men (32.4 +/- 1.9 yr; body mass index, 23.9 +/- 0.7 kg/m2) and eight women (27.9 +/- 0.9 yr; body mass index, 20.9 +/- 0.6 kg/m2) underwent an oral glucose tolerance test (OGTT; 1 g/kg) before and after HDBR. The glucose load was labeled with 13C and associated with D-[6,6-2H2] glucose infusion, indirect calorimetry, breath tests, and plasma measurements to determine the glucose turnover and biodisponibility, substrate oxidation, and endocrine responses. Body composition was assessed using H2(18)O dilution. In addition, hormones were measured in daily blood and 24-h urine samples. No change in body composition was noted. Daily fasting insulin increased during HDBR (men, 34%; women, 26%), as did the insulin to glucose ratio (men, 30%; women, 25%). The normetanephrine level dropped (men, 30%; women, 16%), but metanephrine was unchanged. During OGTTs, the insulin response was increased after HDBR (men, 47%; women, 67%), whereas plasma glucose levels were similar. Nonesterified fatty acids and beta-hydroxybutyrate levels were lower. Endogenous glucose production dropped (28%), and exogenous glucose oxidation increased (28%) only in men. Resting energy expenditure was unchanged, but nonproteic respiratory quotient increased (men, 10%; women, 14%). Basal levels of lipid oxidation dropped in both sexes (approximately 90%), but those of carbohydrate oxidation increased in men (40%); as did lipogenesis in women (570%). In response to OGTTs, lipid oxidation was 80% reduced in both sexes after HDBR, but carbohydrate oxidation increased (25%) in men. Lipogenesis occurred in men (304%) and women (74%), but the latter had higher absolute levels. Therefore, 7 days of HDBR resulted in 1) reduced sympathetic activity, 2) insulin resistance suggested at the muscle level in men and at both the muscle and liver levels in women, 3) no changes in glucose biodisponibility, suggesting no alterations in the gastrointestinal function, and 4) a shift toward carbohydrate oxidation in men and a net lipogenesis in women. Such results suggest gender differences in response to sedentary life style and warrant further analysis.

Energy and water metabolism, body composition, and hormonal changes induced by 42 days of enforced inactivity and simulated weightlessness.

Inactivity causes profound deleterious changes. We investigated in eight healthy men the impact of a 42-day head-down bed rest (HDBR) on energy and water metabolism and their interrelationships with body composition (BC) and catabolic and anabolic hormones. Total energy expenditure (TEE), total body water, water turnover, and metabolic water formation were assessed by the doubly labeled water method 15 days before and for the last 15 days of HDBR. Resting energy expenditure was determined by indirect calorimetry, and BC was determined by dual energy x-ray absorptiometry. Urinary excretion of cortisol, GH, normetanephrine, metanephrine, urea, and creatinine were measured daily. HDBR resulted in significant reductions in body weight (2%), total body water (5%), metabolic water (17%), and lean body mass (LBM; 4%), but fat mass and water turnover did not change. Segmental BC showed a decreased LBM in legs and trunk, whereas fat mass increased, no significant changes were noted in the arms. The hydration of LBM was unchanged. TEE and energy intake decreased significantly (20% and 13%), whereas resting energy expenditure was maintained. Expenditure for physical activity dropped by 39%. Subjects were in energy balance during HDBR, whereas it was negative during the control period (-1.5 MJ/day). There were decreases in urinary normetanephrine (23%) and metanephrine (23%), but urinary cortisol (28%; weeks 2 and 3), GH (75%; weeks 2-4), and urea (15%; weeks 3 and 4) increased. It was concluded that during prolonged HDBR no relevant modifications in water metabolism were triggered. BC changes occurred in the nonexercised body segments, and the reduction in TEE was due to inactivity, not to LBM loss. Moreover, body weight alone does not accurately reflect the subject's energy state, and energy balance alone could not explain the body weight loss, which involves a transient metabolic stress.

13C appearance in plasma glucose and breath CO2 during feeding with naturally 13C-enriched starchy food in normal humans.

We have used a recently developed technique (isotope-ratio mass spectrometer) to measure 13C appearance in plasma glucose and breath CO2 in eight normal subjects during feeding with naturally 13C-enriched starch. 13C in CO2 and plasma glucose, metabolites and insulin concentrations, carbohydrates, and lipid oxidation were measured after ingestion of 76 g glucose equivalent of crackers, pasta, or polenta. 13C in plasma glucose displays a very different pattern from plasma glucose concentration. It increases steadily for 90 min before reaching a plateau for approximately 2 h and slowly declines during the last 2 h of the study. No significant difference was observed with the three different starchy foods tested although plasma glucose tended to be higher during feeding with polenta. In summary measurement of 13C in plasma glucose during feeding with naturally 13C-labeled carbohydrates yields new insight in the study of carbohydrate bioavailability in humans.

Retroconversion and metabolism of [13C]22:6n-3 in humans and rats after intake of a single dose of [13C]22:6n-3-triacylglycerols.

The apparent retroconversion of docosahexaenoic acid (22:6n-3) to eicosapentaenoic acid (20:5n-3) and docosapentaenoic acid (22:5n-3) was studied in vivo, in rats and humans, after they ingested a single dose of triacylglycerols containing [13C]22:6n-3 ([13C]22:6-triacylglycerol), without 22:6n-3 dietary supplementation. The amount of apparent retroconversion and the distribution of the three n-3 polyunsaturated fatty acids (PUFAs) in plasma lipid classes were followed as a function of time by measuring the appearance of 13C in these PUFAs with gas-chromatography combustion-isotope ratio mass spectrometry. This [13C]22:6n-3 retroconversion, calculated by summing the amounts of [13C]22:5n-3 and [13C]20:5n-3 in plasma lipids, was lower in humans than in rats, reaching a maximum of approximately 9% of the total plasma [13C]22:6n-3 in rats, but only 1.4% in humans. The incorporation of [13C]22:6n-3 and [13C]22:5n-3 in lipid classes followed their endogenous distribution with a maximal accumulation in phospholipids, but a low incorporation into cholesterol esters (CEs), whereas [13C]20:5n-3 was equally present in phospholipids and CEs. The ratio of the amount of HDL-CE to HDL-phosphatidylcholine for [13C]20:5n-3 was higher than for [13C]22:6n-3, indicating a selectivity of the lecithin-cholesterol acyltransferase enzyme with regard to these PUFAs, which may be related to the differences in their biological properties after fish oil feeding. The occurrence of a weak basal 22:6n-3 retroconversion in humans supports feeding this pure PUFA in cases in which 20:5n-3 presents undesirable side effects and when specific alterations of blood lipids are expected.

Recovery of [13C]bicarbonate as respiratory 13CO2 in mechanically ventilated patients.

Measurement of the nutrient oxidation rate with 13C as a tracer requires knowledge of the value of its coefficient of fractional recovery in the expired gas (FR). We measured FR in nine intensive care patients who were mechanically ventilated and received total parenteral nutrition. NaH13CO3 was administered at a priming dose (3.75 mumol.kg-1.min-1) followed by a continuous infusion (0.05 mumol.kg-1.min-1). Metabolic rate and pulmonary carbon dioxide elimination (VCO2) were measured by using a mass-spectrometer system. The 13C-12C ratio was measured in the expired gas with an isotopic-ratio mass spectrometer and FR was calculated by using standard equations. The average value of FR was 0.899 +/- 0.026 (means +/- SE) and remained stable for each patient on 2 consecutive days. Between patients, the coefficient of variation of FR was 8.6%. Metabolic rate was the only physiological factor found to affect the FR value.

Influence of human obesity on the metabolic fate of dietary long- and medium-chain triacylglycerols.

The metabolic fate of an oral long-chain-triacylglycerol (LCT) load and of a mixed oral LCT and medium-chain-triacylglycerol (MCT) load was followed for 6 h in eight control and eight obese subjects with normal postabsorptive triacylglycerol concentrations. Labeled triacylglycerol and indirect calorimetry were used. Results showed that LCTs were less oxidized in obese than in control subjects (3.2+/-0.5 compared with 6.0+/-0.4 g, P < 0.01). Moreover, the amount of LCT oxidized was negatively correlated with fat mass (r = -0.77, P < 0.01). Appearance in plasma of dietary triacyglycerol-derived long-chain fatty acids was blunted in obese subjects and it was negatively related to fat mass (r = -0.84, P < 0.01) and positively to LCT oxidation (r = 0.70, P < 0.01). On the contrary, MCT oxidation was not altered in obese subjects compared with control subjects. Furthermore, the proportion of MCTs oxidized was higher in both groups compared with LCTs (x+/-SEM: 57.5+/-2.6% compared with 15.2+/-1.6%, P < 0.01, n = 16). Our conclusion is that obesity is associated with a defect in the oxidation of dietary LCTs probably related to an excessive uptake by the adipose tissue of meal-derived long-chain fatty acids. MCTs, the oxidation of which is not altered in obesity, could therefore be of interest in the dietary treatment of obesity.

Simultaneous validation of ten physical activity questionnaires in older men: a doubly labeled water study.

OBJECTIVE: The purpose of this study was to simultaneously validate 10 physical activity (PA) questionnaires in a homogenous population of healthy elderly men against the reference method: doubly labeled water (DLW). DESIGN: Cross-sectional study. SETTING: Community-based sample from Lyon, France. PARTICIPANTS: Nineteen healthy old men (age 73.4 +/- 4.1 years), recruited from various associations for elderly people in Lyon, agreed to participate in the study. MEASUREMENTS: The questionnaire-derived measures (scores) were compared with two validation measures: DLW and maximal oxygen uptake (VO2max). With the DLW method three parameters were calculated: (1) total energy expenditure (TEE), (2) physical activity level (PAL), i.e., the ratio of TEE to resting metabolic rate, (3) energy expenditure of PA. RESULTS: Relative validity. Correlation between the questionnaires and TEE ranged from 0.11 for the Yale Physical Activity Survey (YPAS) total index to 0.63 for the Stanford usual activity questionnaire. This questionnaire also gave the best correlation coefficients with PAL (0.75), and with VO2max (0.62). Significant results with TEE measured by the DLW method were also obtained for college alumni sports score, Seven Day Recall moderate activity, and Questionnaire d'Activité Physique Saint-Etienne sports activity (r = 0.54, r = 0.52, and r = 0.54, respectively). Absolute validity. No difference was found between PA measured by the Seven Day Recall or by the YPAS and DLW, on a group basis. The limits of agreement were wide for all the questionnaires. CONCLUSIONS: Only a few questionnaires demonstrated a reasonable degree of reliability and could be used to rank healthy older men according to PA. Correlation coefficients were best when the Stanford Usual Activity Questionnaire was compared with all the validation measures. The two questionnaires reporting recent PA, the Seven Day Recall, and YPAS accurately assessed energy expenditure for the group. The individual variability was high for all the questionnaires, suggesting that their use as a proxy measure of individual energy expenditure may be limited.

In vivo measurement of plasma cholesterol and fatty acid synthesis with deuterated water: determination of the average number of deuterium atoms incorporated.

Fractional lipid synthesis can be measured using the incorporation of deuterium from deuterated water. The calculations require knowledge of the maximum incorporation number (N) of deuterium atoms in the molecules synthesized. For both tissue palmitate and cholesterol, N values have been found to be higher during in vivo versus in vitro experiments. We determined the N values to be used for measuring the fractional synthesis of plasma cholesterol and of palmitate triglycerides (TG). Rats were given drinking water enriched (7% to 10%) with deuterated water, and N was determined from the mass isotopomer distributions of plasma cholesterol and plasma TG palmitate and the deuterium enrichment of plasma water. We found N to be 21 for palmitate and 27 for cholesterol. These values agree with those reported for tissue palmitate and cholesterol in vivo, and are higher than values found in vitro. We also found large deuterium enrichments in plasma glucose and in liver lactate and pyruvate. We suggest that, compared with in vitro studies, in vivo metabolism of these compounds leads to an additional pathway of incorporation of deuterium into lipids through deuterium-labeled acetyl coenzyme A (CoA). This could explain why N values are higher in vivo than in vitro.

Dietary specific sugars for serum protein enzymatic glycosylation in man.

All glycoprotein sugars can theoretically derive from glucose. However, dietary specific sugars could represent preferential substrates or have regulatory roles in enzymatic glycosylation. This hypothesis was tested in man using stable isotopes. Healthy subjects ingested different amounts (150, 300, or 550 mg) of artificially 13C-enriched sugar (galactose, mannose, or glucose) diluted in 200 mL water containing 50 g 13C-poor sucrose. 13C enrichment of expired CO2 was monitored for 8 hours during indirect calorimetry. Serum glycoproteins were precipitated and delipidated at various intervals. Glycoprotein neutral sugars were obtained by acidic hydrolysis, purified by ion-exchange chromatography, derivatized to alditol acetates, and analyzed by gas chromatography-isotope ratio mass spectrometry. The oxidation rate for galactose and mannose was slower than the rate for glucose. Total oxidation over the 8-hour period was less than 10% of the ingested amount of galactose or mannose. Galactose and mannose were readily incorporated into glycoprotein glycans, in the native form or after interconversion, despite ingestion of a large excess of sucrose: glycoprotein sugar 13C enrichment was strongly higher after 13C-galactose or 13C-mannose than after 13C-glucose. Thus, the metabolism of these three sugars appears to be different. Specific dietary sugars could represent a new class of non essential nutrients displaying interesting metabolic roles. This could have practical consequences especially in parenteral nutrition, where glucose is currently the only sugar available for metabolism.

Thermogenic effect of slight hyperglycemia during a lipid infusion.

Resistance to the glucoregulatory action of insulin is a common finding in obesity and may affect thermogenesis. In 13 healthy subjects, we studied the influence of acute insulin resistance induced by a lipid infusion on thermogenesis without any glucose load (n = 4) or during a euglycemic-hyperinsulinemic clamp (n = 5) and an oral glucose tolerance test (OGTT, n = 8). When substrates were not administered at the same time, the energy cost of storage was significantly (P < .05) lower for lipids (3.9%+/-0.9%) than for glucose (11.9%+/-0.5% during the clamp and 14.9%+/-4.0% during the OGTT, NS). The lipid infusion decreased glucose storage during the clamp (control, 3.99+/-0.40 mg x kg(-1) x min(-1); lipid infusion, 0.92+/-0.39; P < .05) but increased it during the OGTT (control, 1.76+/-0.22 mg x kg(-1) x min(-1); lipid infusion, 2.94+/-0.27; P < .05). Infused lipids were stored more (clamp, 3.31+/-0.16; OGTT, 2.65+/-0.11 mg x kg(-1) x min(-1); P < .01) and oxidized less (clamp, 0.64+/-0.21; OGTT, 1.02+/-0.09 mg x kg(-1) x min(-1); P < .05) during the clamp than during the OGTT. When lipids were infused, the energy cost of substrate storage was lower during the clamp versus the OGTT (clamp, 3.2%+/-0.8%; OGTT, 7.3%+/-1.0%; P < .05). This effect was attributed to a lipid-induced impairment of glucose tolerance, which overcomes the inhibitory effect of lipid infusion on glucose storage observed in euglycemia. A slight elevation of plasma glucose in response to a lipid infusion impairs thermogenesis by redirecting the storage of substrates from lipids to glucose, which has a higher energy cost.

Effects of glucose and amino acid infusion on glucose turnover in insulin-resistant obese and type II diabetic patients.

Glucose turnover was assessed from [6,6-2H]glucose and [U-13C]glucose dilution analysis in six lean nondiabetic subjects, six obese patients with normal glucose tolerance, and six obese patients with non-insulin-dependent diabetes mellitus (NIDDM) during sequential infusions of glucose (13.9 mumol/kg fat-free mass [FFM]/min) and glucose+amino acid (4.2 mg/kg FFM/min). Cori cycle activity was assessed from the difference between glucose turnover obtained from [6,6-2H]glucose and [U-13C]glucose. During infusion of glucose alone, total glucose turnover was increased by 70% in obese NIDDM patients. Amino acid infusion decreased glucose concentrations by 0.8, 0.5, and 1.8 mmol/L in controls, obese patients, and NIDDM patients, respectively. This decrease in glycemia occurred despite an increase in glucose turnover in lean and obese nondiabetic subjects, and was due to an increased metabolic clearance rate (MCR) of glucose. In NIDDM patients the MCR of glucose was unchanged, and the decrease in glycemia was explained by a diminution in hepatic glucose output. Glucose turnover obtained by [6.6-2H] dilution analysis exceeded significantly the values obtained by dilution analysis in obese subjects and obese NIDDM patients, but not in controls. This indicates an increased Cori cycle activity in these patients.

Fructose utilization during exercise in men: rapid conversion of ingested fructose to circulating glucose.

The aim of the present study was to compare the metabolic fate of repeated doses of fructose or glucose ingested every 30 min during long-duration moderate-intensity exercise in men. Healthy volunteers exercised for 3 h on a treadmill at 45% of their maximal oxygen consumption rate. "Naturally labeled" [13C]glucose or [13C]fructose was given orally at 25-g doses every 30 min (total feeding: 150 g; n = 6 in each group). Substrate utilization was evaluated by indirect calorimetry, and exogenous sugar oxidation was measured by isotope ratio mass spectrometry on expired CO2. Results were corrected for baseline drift in 13C/12C ratio in expired air due to exercise alone. Fructose conversion to plasma glucose was measured combining gas chromatography and isotope ratio mass spectrometry. Most of the ingested glucose was oxidized: 81 +/- 4 vs. 57 +/- 2 g/3 h for fructose (2P < 0.005). Exogenous glucose covered 20.8 +/- 1.4% of the total energy need (+/- 6.7 MJ) compared with 14.0 +/- 0.6% for fructose (2P < 0.005). The contribution of total carbohydrates was significantly higher and that of lipids significantly lower with glucose than with fructose. The blood glucose response was similar in both protocols. From 90 to 180 min, 55-60% of circulating glucose was derived from ingested fructose. In conclusion, when ingested repeatedly during moderate-intensity prolonged exercise, fructose is metabolically less available than glucose, despite a high rate of conversion to circulating glucose.

Measuring lipogenesis and cholesterol synthesis in humans with deuterated water: use of simple gas chromatographic/mass spectrometric techniques.

Lipogenesis and cholesterol synthesis can be studied by measuring the incorporation into fatty acids and cholesterol of deuterium from deuterated water. This has been previously achieved in human subjects using low levels of deuterium enrichment in plasma water, and thus in fatty acids and cholesterol. For the measurement of enrichment in lipids, this required the use of isotope ratio mass spectrometry, a tedious and time-consuming technique. It is shown that these measurements can be performed using the much simpler gas chromatography/mass spectrometry if higher, but always safe, deuterium enrichments in plasma water are obtained. Normal subjects ingested deuterated water in order to obtain stable enrichment in plasma water of 0.3% during a 60 h period. Enrichment in palmitate of plasma triglycerides (TG) plateaued (0.6-0.76%) whereas plasma cholesterol enrichment increased progressively [0.32 +/- 0.08% (12 h) to 0.78 +/- 0.18% (60 h)]. Endogenous synthesis was estimated to contribute, in post-absorptive subjects, 8-10% of the plasma TG pool and 3-5% of plasma free cholesterol pool. These data agree with results obtained previously using isotope ratio mass spectrometry. The present method will be useful for studies of normal and abnormal lipid metabolism in humans.

Total-body-water measurement with (18)O-labeled water in short-bowel patients with an ileostomy.

The aim of this study was to evaluate the use of H(2)(18)O for total-body-water (TBW) determination in patients with short bowel because ileostomy losses of labeled water can falsify TBW. Thirteen adult short-bowel patients were studied after an overnight fast. Saliva, blood, urine and ileostomy output were collected before and 30, 60, 90, 120, 240, and 360 min after oral ingestion of 0.88 g/kg of 2.5% H(2)(18)O to measure (18)O abundance by isotope-ratio mass spectrometry. TBW was calculated from the dilution of the isotopic water in saliva and plasma. The quantity of labeled water lost in the ileostomy was calculated from the isotopic enrichment of the ileostomy output from T0 to T360. The values obtained from saliva or plasma with (corrected) and without (uncorrected) considering H(2)(18)O lost in the ileostomy output were compared with a paired t test. Agreement was evaluated using the Bland-Altman method. From T0 to T360, the ileostomy output and the percentage of lost labeled water were 490 +/- 314 mL and 6.38 +/- 8.52%, respectively. TBW calculated from plasma or saliva isotopic enrichment was different, and a significant difference was also observed between corrected and uncorrected TBW values (saliva: uncorrected TBW = 32.35 +/- 7.52 L, corrected TBW = 30.29 +/- 6.09 L; plasma; uncorrected TBW = 30.80 +/- 7.29 L, corrected TBW = 28.79 +/- 5.79 L). The agreement between the values obtained from the two dilution spaces or between the two calculation methods was poor. Because of the large discrepancies between calculation methods, determination of TBW from oral ingestion of (18)O-labeled water in patients with short bowel should be calculated only from plasma (18)O space dilution and should consider ileostomy losses.

Use of compounds naturally labeled with stable isotopes for the study of the metabolism of glycoprotein neutral sugars by gas-liquid chromatography-isotope-ratio mass spectrometry. Technical validation in the rat.

In order to develop an alternative method to radioactive labeling for the study of the glycoprotein sugar metabolism in man, the possible use of stable isotopes provided by naturally, 13C-enriched dietary compounds has been explored in rat intestine and serum. Rats were fed a semisynthetic diet containing 67% wheat starch (containing 1.08692 13C atom/100 carbon atoms) for a week, and then the same diet containing corn starch (1.10042% 13C) for a week. Neutral sugars were prepared from delipidated, trichloroacetic acid-precipitable material from 200-400 mg of intestinal mucosa or 1 mL of serum, separated, and analyzed as alditol acetates by gas-liquid chromatography coupled to isotope-ratio mass spectrometry. This technique allowed the determination, in a single experiment, of the amount and 13C abundance of six neutral sugars (including xylose used as internal standard). Despite the low abundance of 13C in natural products, the sensitivity of the technique was found to be sufficient to detect isotopic enrichment as low as 0.001% with good accuracy and reproducibility in 2 micrograms of each glycoprotein neutral sugar. As an example, the pattern of labeling by dietary D-glucose from corn starch appears to be very different for fucose, ribose, mannose, galactose, and glucose of intestine or serum macromolecules.

Use of gas chromatography/isotope ratio-mass spectrometry to study triglyceride metabolism in humans.

The study of triglyceride (TG) metabolism using stable isotope tracers would be facilitated by being able to detect low 13C enrichment. To meet this goal, we developed a gas chromatography/isotope ratio-mass spectrometry technique to measure the enrichment of palmitate in nonesterified fatty acids (NEFA) and TG as its methyl derivative. This method allows accurate and reproducible measurements of enrichment as low as 0.009 mole percent excess (MPE), in a range between 0-0.65 MPE. The usefulness of this method is shown by two studies of lipid metabolism in human beings. First, we studied the metabolic fate of an oral TG load labeled with [1,1,1-13C3]tripalmitin. Labeled palmitate appeared concurrently in plasma NEFA and TG, and four hours after the load, the labeling was higher in NEFA than in TG (MPE NEFA: 1.53 +/- 0.31 vs. MPE TG: 0.78 +/- 0.06, P < 0.05). In a second study, the hepatic reesterification of NEFA was estimated by measuring the appearance of infused [1-13C]palmitate in circulating TG. The estimated contribution of plasma NEFA to circulating TG increased to a maximum of 22%. Thus, gas chromatography/isotope ratio-mass spectrometry appears to be a useful tool for future studies of lipid metabolism in humans.

In vivo compartmental metabolism of 13C-docosahexaenoic acid, studied by gas chromatography-combustion isotope ratio mass spectrometry.

The exchange of docosahexaenoic acid (22:6n-3) within lipid pools in rat and human has been followed as a function of time after the ingestion of triglycerides (TG) containing 22:6n-3 labeled with 13C(13C 22:6n-3). The 13C abundance in the fatty acid was measured by gas-chromatography-combustion isotope ratio mass spectrometry which allowed the detection of 0.001 atom 13C percent 12C. The 13C 22:6n-3 appearance was rapid in the TG of very low density lipoprotein plus chylomicron fraction, in which the maximal labeling was observed at 3 and 2 h after ingestion in rat and human, respectively. Concomitant with the TG utilization of this fraction by lipoprotein lipase from tissues, unesterified 13C 22:6n-3 appeared in the plasma albumin. 13C 22:6n-3 bound to albumin was mostly present in unesterified form before 12 h post-ingestion while after that period, lysophosphatidylcholine (lysoPC) bound to albumin carried higher 13C 22:6n-3 concentrations. These lyso-PC were mostly from hepatic origin and might represent a potential source of 22:6n-3 redistribution to tissues. The 13C 22:6n-3 uptake into rat brain PC and phosphatidylethanolamine was still increasing when the concentration of plasma unesterified 13C 22:6n-3 had already dropped to a minimal plateau value and during the period of maximal plasma circulation of 13C 22:6n-3-lysoPC bound to albumin. In contrast, the uptake of 13C 22:6n-3 into blood platelet PC occurred during the phase of important circulation of 13C-22:6n-3 bound to albumin, suggesting the in vivo efficiency of the Lands pathway for this fatty acid. It is concluded that 13C 22:6n-3 esterified in TG is rapidly absorbed and redistributed within plasma lipoproteins and that its redistribution within the two lipid species bound to albumin might influence its uptake by platelets and rat brain.

[New techniques for investigation of human nutrition]. / Techniques nouvelles d'exploration en nutrition humaine.

Accurate determination of energy requirement and nutrients metabolism is essential to improve physiological knowledge and for physiopathological purpose in human nutrition. This evaluation is an absolute necessity for food industry. Energy expenditure could be precisely determined by indirect calorimetry or doubly labeled water technic. Nutrients metabolism (kinetics?) and substrates turn-over studies are now accessible without health hazard using tracers labelled with stable isotopes. These three methods are the basic tools for the new Research Nutrition Center which are now in progress.

Whole body protein turnover measured with 13C-leucine and energy expenditure in preterm infants.

Our study was undertaken in preterm infants to examine the relationship of whole body protein kinetics with protein intake and energy expenditure. Leucine kinetics were determined in seven low birth wt preterm infants fed human milk or human milk enriched with protein (2.5 to 4.3 g protein/kg.d). The infants received a short (4-h) constant infusion of L-[1-13C]leucine and leucine turnover and oxidation were calculated from 13C-plasma leucine and expired 13CO2 enrichments measured by mass spectrometry. Energy expenditure was measured by indirect calorimetry. Nonoxidative leucine disposal (an estimate of protein synthesis) and leucine derived from protein (an estimate of protein breakdown) were, respectively, 2.98 +/- 0.82 and 2.06 +/- 0.74 mumol/kg.min. Whole body protein turnover and deposition, derived from leucine kinetics, were 8.22 +/- 2.31 and 2.17 +/- 0.50 g/kg.d, whereas energy expenditure was 56.3 kcal/kg.day. Protein turnover was correlated with protein intake but not with protein deposition. Energy expenditure was correlated with protein turnover, synthesis, and breakdown but not with protein deposition. These data are in agreement with the fact that protein deposition depends upon protein intake, but they also suggest that an elevated protein deposition is not necessarily the result of a rapid protein turnover or associated with an elevated energy expenditure.