Catherine Luengo

Exogenous and endogenous nitrogen flow rates and level of protein hydrolysis in the human jejunum after [15N]milk and [15N]yoghurt ingestion.

Milk and yoghurt proteins were 15N-labelled in order to measure the flow rate of exogenous N during digestion in the human intestine. After fasting overnight, sixteen healthy volunteers, each with a naso-jejunal tube, ingested either [15N]milk (n 7) or [15N]yoghurt (n 9). Jejunal samples were collected every 20 min for 4 h. A significant stimulation of endogenous N secretion was observed during the 20-60 min period after yoghurt ingestion and the 20-40 min period after milk ingestion. The endogenous N flows over a 4 h period did not differ between the groups (44.3(SEM 6.5) mmol for milk and 63.5(SEM 5.9) mmol for yoghurt). The flow rates of exogenous N indicated a delayed gastric emptying of the yoghurt N compared with N from milk. The jejunal non-protein N (NPN) flow rate increased significantly after milk and yoghurt ingestion due to an increase in the exogenous NPN flow rate. The NPN fraction of exogenous N ranged between 40 and 80%. The net gastro-jejunal absorption of exogenous N did not differ significantly between milk (56.7(SEM 8.5)%) and yoghurt (50.9(SEM 7)%). The high level of exogenous N hydrolysis is in accordance with the good digestibility of milk products. Fermentation modifies only the gastric emptying rate of N and does not affect the level of diet hydrolysis, the endogenous N stimulation or the digestibility rate.

Metabolism of enterostatin in rat intestine, brain membranes, and serum: differential involvement of proline-specific peptidases.

The degradation of enterostatin (VPDPR), a potent inhibitor of food intake, by intestinal brush-border membranes, brain membranes, and rat serum has been investigated in the presence of specific inhibitors. Hydrolysis by intestinal membranes was found to be 10 and 100 times faster than in serum and brain membranes, respectively. Enterostatin hydrolysis by intestinal and brain membranes involves the removal of C-terminal arginine by carboxypeptidase P, leading to the production of des-Arg-enterostatin, and the splitting of the Pro2-Asp3 bond by dipeptidyl aminopeptidase IV (DPP IV). A small amount of the potent anorectic peptide Pro2-Asp3-Pro4 was released during hydrolysis of des-Arg-enterostatin by brain membranes and rat serum. In rat serum, enterostatin degradation was mainly due to DPP IV.

High-protein diets differentially modulate protein content and protein synthesis in visceral and peripheral tissues in rats

OBJECTIVE High-protein diets give rise to increased amplitude in the diurnal cycling of protein gains and losses at the whole-body level, but the tissue localization and mechanisms underlying these metabolic adaptations remain unclear. We investigated tissue-specific responses to increasing protein intakes in rats. METHODS Protein synthesis rates (flooding dose with (13)C-valine) and accretion were assessed in individual tissues of fasted or fed rats (n = 32) after a 2-wk adaptation to a normal- or high-protein (HP) diet. RESULTS In livers of HP rats, a strong inhibition of protein synthesis rates (-34%) occurred in the fasted and fed states, whereas a higher protein content (+10%) was observed. In the kidneys, a slight inhibition of synthesis rates after the HP diet was also observed but remained without effect on kidney protein pool size. Stomach and skin protein synthesis rates were significantly increased under HP conditions, whereas protein anabolism in skeletal muscle remained insensitive to the dietary protein level. This was also true for specific muscle protein fractions: myosin, mitochondrial, or sarcoplasmic protein synthesis rates were influenced by neither the dietary protein level nor the nutritional status. CONCLUSION Modulation of protein kinetics and accretion by the HP diet is tissue-specific and the liver plays a critical role in such adaptations in a unique situation associating an inhibition of protein synthesis and protein pool expansion. The mechanisms underlying these changes and their physiologic incidence remain to be elucidated.

Dietary Proteins Contribute Little to Glucose Production Even Under Optimal Gluconeogenic Conditions in Healthy Humans

Dietary proteins are believed to participate significantly in maintaining blood glucose levels, but their contribution to endogenous glucose production (EGP) remains unclear. We investigated this question using multiple stable isotopes. After overnight fasting, eight healthy volunteers received an intravenous infusion of [6,6-2H2]-glucose. Two hours later, they ingested four eggs containing 23 g of intrinsically, uniformly, and doubly [15N]-[13C]–labeled proteins. Gas exchanges, expired CO2, blood, and urine were collected over the 8 h following egg ingestion. The cumulative amount of dietary amino acids (AAs) deaminated over this 8-h period was 18.1 ± 3.5%, 17.5% of them being oxidized. The EGP remained stable for 6 h but fell thereafter, concomitantly with blood glucose levels. During the 8 h after egg ingestion, 50.4 ± 7.7 g of glucose was produced, but only 3.9 ± 0.7 g originated from dietary AA. Our results show that the total postprandial contribution of dietary AA to EGP was small in humans habituated to a diet medium-rich in proteins, even after an overnight fast and in the absence of carbohydrates from the meal. These findings question the respective roles of dietary proteins and endogenous sources in generating significant amounts of glucose in order to maintain blood glucose levels in healthy subjects.

[13C] GC–C-IRMS analysis of methylboronic acid derivatives of glucose from liver glycogen after the ingestion of [13C] labeled tracers in rats

We developed a complete method to measure low [(13)C] enrichments in glycogen. Fourteen rats were fed a control diet. Six of them also ingested either [U-(13)C] glucose (n=2) or a mixture of 20 [U-(13)C] amino acids (n=4). Hepatic glycogen was extracted, digested to glucose and purified on anion-cation exchange resins. After the optimization of methylboronic acid derivatization using GC-MS, [(13)C] enrichment of extracted glucose was measured by GC-C-IRMS. The accuracy was addressed by measuring the enrichment excess of a calibration curve, which observed values were in good agreement with the expected values (R=0.9979). Corrected delta values were -15.6+/-1.6 delta(13)C (per thousand) for control rats (n=8) and increased to -5 to 8 delta(13)C (per thousand) per thousand and 12-14 delta(13)C (per thousand) per thousand after the ingestion of [U-(13)C] amino acids or [U-(13)C] glucose as oral tracers, respectively. The method enabled the determination of dietary substrate transfer into glycogen. The sequestration of dietary glucose in liver glycogen 4 h after the meal was 35% of the ingested dose whereas the transfer of carbon skeletons from amino acids was only 0.25 to 1%.

Whole-body protein turnover in humans fed a soy protein-rich vegetable diet

Objectives: This study was designed to compare the whole-body protein turnover in humans after the ingestion of a soy protein-rich vegetable diet with that of a control group fed a western animal protein-rich diet. Subjects: Twelve male volunteers were divided into two groups of six subjects who were given for two weeks either a 85% vegetable protein diet (diet VP) or a control western animal protein-rich diet (diet AP). Interventions: Whole-body protein turnover was estimated at the end of the two-week controlled diet period using the [15N]-glycine end-product method. Nitrogen flux rates were determined in the fed state (1.3 g protein/kg) over a 9 h period after the dose of [15N]-glycine was given. Results: After the 9 h of the test, the urinary ammonia excretion was significantly higher in the group receiving the diet AP than that in the group receiving the diet VP (P<0.05). In contrast, there was no significant difference for both total nitrogen and urea nitrogen excretions. Both the protein synthesis and the protein breakdown were similar in both groups. In the same way, the net protein deposition measured in the fed state during 9 h was similar for both diets at 0.07 g/kg/h. Conclusions: Young adults fed 1.3 g/kg/d of either meat or vegetable protein-rich diet for two weeks did not show a different protein turnover. Sponsorship: This work was supported in part by grant from Eridania Béghin-Say Compagny.