Varying the protein source in mixed meal modifies glucose, insulin and glucagon kinetics in healthy men, has weak effects on subjective satiety and fails to affect food intake.

OBJECTIVE: To evaluate the influence of three dietary protein types (casein, gelatin, soy protein) on satiety and food intake, at two levels of loading (total energy of test meals: 3.6 or 1.8 MJ). DESIGN: The study employed a repeated measures design. Test meals were controlled for energy, macronutrients, fiber and palatability, and contained about 23% energy as protein (of which about 65% was experimentally manipulated). Postprandial subjective satiety and hunger, plasma glucose, insulin and glucagon were assessed for 8 h, and energy and macronutrient intakes were monitored for 24 h. SUBJECTS: Nine healthy normal-weight men. RESULTS: No effect of the type of protein on 24 h energy and macronutrient intakes was observed despite a significant effect of protein source on the kinetics of peripheral metabolic responses (but only after 3.6 MJ lunches), and inconsistent effects on subjective hunger and satiety responses A casein-enriched lunch delayed glucose and insulin responses for 1.5 h, compared with soy protein, probably due to a lag in gastric emptying. CONCLUSION: Varying the protein source in a mixed meal modifies glucose, insulin and glucagon kinetics in healthy men, but these variations in satiety-implicated factors have inconsistent effects on subjective satiety and fail to affect food intake. SPONSORSHIP: Eridania Béghin-Say, Vilvoorde, Belgium and Association Nationale de la Recherche Technique, France (Convention CIFRE no 537/94).

Euglycemic hyperinsulinemic clamp to assess posthepatic glucose appearance after carbohydrate loading. 1. Validation in pigs.

BACKGROUND: Precise knowledge of the rate of glucose absorption after meal feeding requires invasive methods in humans. OBJECTIVE: This study aimed to validate in an animal model a technique combining the euglycemic hyperinsulinemic clamp and oral carbohydrate loading (OC-Clamp) as a noninvasive procedure to quantify the posthepatic appearance of glucose after oral carbohydrate loading. DESIGN: Twenty-one pigs were fitted with arterial, jugular, portal, and duodenal catheters and a portal blood flow probe. At glucose clamp steady state, duodenal glucose (0.9 g/kg; DG-Clamp) and oral carbohydrate (140 g corn or mung bean starch as part of a mixed meal; OC-Clamp) were administered while the glucose infusion was progressively reduced to compensate for the incremental posthepatic appearance of glucose. [3-3H]glucose was used to assess the glucose turnover rate. RESULTS: Hepatic glucose production was totally suppressed by insulin infusion, and the whole-body glucose turnover rate remained stable during glucose absorption. The incremental portal appearance of glucose after the DG load was not altered by hyperinsulinemia, and the cumulative posthepatic appearance of glucose was 63 +/- 3% (x +/- SEM) of the DG load. The net hepatic portal appearance of glucose remained constant during absorption (34 +/- 3% of the load). After the OC load, the respective portal appearance rates of glucose were significantly different between carbohydrate sources; however, the rates paralleled those of the posthepatic appearance of glucose. Again, net hepatic glucose uptake expressed as portal appearance was similar for both carbohydrates. CONCLUSIONS: The results validate the OC-Clamp method to monitor the posthepatic appearance of glucose after carbohydrate ingestion and to discriminate between different carbohydrate sources. The results suggest that the technique be used in humans.

Dietary (n-3) polyunsaturated fatty acids improve adipocyte insulin action and glucose metabolism in insulin-resistant rats: relation to membrane fatty acids.

To study the effects of dietary fish oil on insulin-stimulated glucose metabolism in adipocytes of insulin-resistant rats (rats fed 50% sucrose and 30% fat), eighteen 5-wk-old Sprague-Dawley rats were fed, for 6 wk, a diet containing 30% fat as either fish oil (FO) or a mixture of vegetable and animal oils [control oils (CO)]. A third reference group was fed a standard diet (62% corn starch and 13% fat). At the end of the 6-wk period, the two experimental groups had comparable plasma glucose concentrations that were higher than that found in the reference group. FO feeding corrected the hyperinsulinemia of the experimental rats (P < 0.05) to reach values in the reference group. Plasma triacylglycerol (P < 0.01) and cholesterol (P < 0.001) concentrations were also lower in rats fed FO than in those fed CO. The body weights of FO-fed rats were similar to that of CO-fed rats, but epididymal adipose tissue weight was lower (P < 0.01). Adipocytes of FO-fed rats, compared with those of CO-fed rats, had high insulin-stimulated glucose transport (P < 0.05), oxidation (P < 0.001) and incorporation into total lipids (P < 0.05). The incorporation of (n-3) polyunsaturated fatty acids in adipocyte membrane phospholipids was higher in FO-fed rats than in those fed CO (P < 0.0001). Insulin action was positively correlated with the fatty acid unsaturation index in membrane phospholipids. Thus dietary fish oil has beneficial effects on insulinemia, plasma lipids and insulin-stimulated glucose metabolism in insulin-resistant slightly diabetic rats.

Lack of effect of an acute ileal perfusion of short-chain fatty acids on glucose metabolism in healthy men.

Dietary fiber intake is associated with several beneficial effects on carbohydrate metabolism. Some authors have speculated that this improvement may be due to short-chain fatty acids (SCFA) produced by the colonic fermentation of dietary fibers. To test this hypothesis, six healthy men aged 26 +/- 2 (SE) yr with a body mass index of 20.9 +/- 0.7 received on three occasions an 18-h ileal perfusion infused at a flow rate of 3.3 ml/min, containing either 90 mmol/l of SCFA (60% acetate, 25% propionate, and 15% butyrate) (A), SCFA during the first 12 h and then a saline solution (A/S), or a saline solution (S). Basal hepatic glucose production (BHGP), insulin sensitivity (3-step euglycemic-hyperinsulinic clamp), and erythrocyte insulin binding (EIB) were studied 12 h after the beginning of the ileal perfusion. There was no change in BHGP or insulin sensitivity. However, maximal EIB was significantly different: 7.1 +/- 0.1 (A), 6.8 +/- 0.1 (A/S), vs. 6.5 +/- 0.1% (S) (P = 0.03). We conclude that acute administration of SCFA does not significantly alter glucose metabolism in healthy subjects.

Chronic consumption of short-chain fructooligosaccharides by healthy subjects decreased basal hepatic glucose production but had no effect on insulin-stimulated glucose metabolism.

We aimed to study the effects of chronic ingestion of short-chain fructooligosaccharides (FOS), an indigestible carbohydrate, on hepatic glucose production, insulin-mediated glucose metabolism, erythrocyte insulin binding, and blood lipids in healthy subjects. Twelve healthy volunteers received either 20 g FOS/d or sucrose for 4 wk in a double-blind crossover design. FOS did not modify fasting plasma glucose and insulin concentrations. Mean (+/- SEM) basal hepatic glucose production was lower after FOS than after sucrose consumption (2.18 +/- 0.10 compared with 2.32 +/- 0.09 mg.kg-1, min-1, respectively; P < 0.02, paired Student's t test). However, neither insulin suppression of hepatic glucose production nor insulin stimulation of glucose uptake measured by hyperinsulinemic clamp was significantly different between the two dietary periods. Erythrocyte insulin binding was also comparable. Serum triacylglycerols, total and high-density- lipoprotein cholesterol, apolipoproteins A-I and B, and lipoprotein(a) were not modified by FOS. To try to understand why FOS did not increase serum lipids, the in vitro production of short-chain fatty acids from FOS was evaluated by using human fecal inoculum and compared with that from lactulose, which was found to increase serum lipids. FOS produced an acetate-propionate ratio two times lower than that of lactulose. We conclude that 4 wk of 20 g FOS/d decreased basal hepatic glucose production but had no detectable effect on insulin-stimulated glucose metabolism in healthy subjects. The colonic fermentation pattern of undigestible carbohydrates may be relevant to predicting their metabolic effects.

Effects of dietary propionate on hepatic glucose production, whole-body glucose utilization, carbohydrate and lipid metabolism in normal rats.

Increased intake of dietary fibres is associated with several beneficial effects on carbohydrate and lipid metabolism. The colonic fermentation of dietary fibres produces short-chain fatty acids (SCFA; acetate, propionate and butyrate). Some authors have suggested that SCFA could be partly responsible for the effects of dietary fibres. The purpose of the present study was to test the effects of one of the SCFA, propionate. The effects of moderate amounts of dietary propionate on insulin sensitivity and hepatic glucose production were studied in male Sprague-Dawley rats. Two groups of twenty-one adult rats were fed for 3 weeks on a diet containing 78 g propionate/kg (P) or 78 g/kg of a poorly fermentable cellulose (control group; C). Feed intake, body weight, fasting plasma glucose, insulin, free fatty acids, alanine, lactate, glycerol and beta-hydroxybutyrate levels were measured weekly in anaesthetized rats. At the end of the feeding period basal hepatic glucose production (BHGP) was measured with a primed continuous infusion of [3-3H]glucose and the in vivo insulin sensitivity in rats was quantified by the euglycaemic-hyperinsulinaemic clamp technique (0.6 and 2 U/kg per h). At that time fasting plasma glucose measured in anaesthetized rats was significantly lower in group P than in group C: 7.7 (SE 0.2) v. 8.5 (SE 0.2) mmol/l respectively (P < 0.002); plasma insulin levels were not significantly different. Neither the BHGP (mg/min per kg; C 14.8 (SE 1.3), P 15.1 (SE 1.3); n 7, not significant) nor the basal metabolic clearance (ml/min per kg; 8.9 (SE 0.8) v. 9.9 (SE 1.1); not significant) were different between treatments. Hepatic glucose production and glucose utilization at the two insulin concentrations (approximately 500 and 1500 mU/l respectively, n 7) did not differ significantly between the two groups. These results show that dietary propionate chronically ingested by normal rats could decrease fasting glycaemia, but from our findings, no effect on hepatic glucose production and whole-body glucose utilization could be clearly demonstrated.

[Rapid assay of plasma acetate by gas chromatography: evaluation and comparison with two other methods]. / Dosage rapide de l'acétate plasmatique par chromatographie en phase gazeuse--technique espace de tête: évaluation et comparaison avec deux autres méthodes.

Human plasma acetate is derived from colonic fermentation of fiber and endogenous metabolism of dextrose and fatty acids. Acetate may have regulatory functions in hepatic carbohydrate metabolism. Intake of dietary fiber is associated with several beneficial effects on carbohydrates and lipids metabolisms. To study theses effects a valid and automated method for routine analysis of acetate in plasma is necessary. After oral administration of lactulose to healthy human volunteers, the concentration of plasma acetate was measured by head space gas chromatography (HS-GC), vacuum distillation gas chromatography (VD-GC) and enzymatic spectrometric method (ES). The method HS-GC was linear to 0.5 mmol.l-1 (n = 5, r = 0.998), the detection limit is 0.005 mmol.l-1. Within-day variation (CV) was 3.60% and day-to-day variation was 4.5% (0.1 mmol.l-1). The coefficients of correlation between CG-ET/CG-DsV and CG-ET/E-M are 0.903 (p = 0.0001) and 0.54 (p = 0.006) respectively, the mean square errors are respectively 0.118 and 0.138 mmol.l-1. The variation curves of plasma acetate measured by GC versus time show peak concentration of 0.323 to 0.380 mmol.l-1 at 120 min.

Neither dietary fructose, dextrose nor starch modifies in vitro glycerol release by adipocytes from streptozotocin-diabetic rats.

Because we found previously that fructose feeding could alter lipolytic responses to isoproterenol and insulin in normal rats, we studied the effects of the same diet in neonatal, streptozotocin-diabetic rats. Twenty-seven 5-wk-old diabetic Sprague-Dawley rats were fed a diet containing 57% carbohydrate as either fructose, dextrose or starch for 6 wk. At the end of the nutritional period, plasma glucose and insulin concentrations in fed rats were similar in the three diabetic groups. Plasma triacylglycerol concentrations were higher in the fructose-fed group than in the other two groups (P < 0.05). Neither the maximal adipocyte lipolytic response (fructose = 1147 +/- 165%, starch = 1823 +/- 329% and dextrose = 1287 +/- 239% of basal values) nor the sensitivity to isoproterenol (ED50) was changed by the dietary carbohydrate exchange. The maximal antilipolytic action of insulin (starch = 68 +/- 10%, dextrose = 41 +/- 13%, fructose = 95 +/- 29% of stimulated lipolysis values) was comparable in the three diet groups. Thus, 6 wk of fructose feeding in diabetic rats increased plasma triacylglycerol concentrations, but had no detectable effect on plasma glucose or insulin concentrations, isoproterenol-induced lipolysis or the antilipolytic action of insulin.

A new non-invasive method for treating insulin-reaction: intranasal lyophylized glucagon.

The main therapeutic indication for glucagon is the treatment of hypoglycaemia in insulin overdosed Type 1 (insulin-dependent) diabetic patients. We have previously shown that an intranasal spray of 7.5 mg glucagon with deoxycholic acid as surfactant was able to correct an i.v. insulin-induced hypoglycaemia in diabetic patients. However, bioavailability and stability needed to be improved before intranasal glucagon could be introduced into clinical practice. This has now been achieved with a freeze-dried mixture of glucagon (1 mg) and glycocholic acid (1 mg) as a surfactant. Kinetics and efficacy have been controlled by (1) comparing subcutaneous and intranasal glucagon in 12 healthy non-hypoglycaemic subjects; (2) testing intranasal glucagon in six Type 1 diabetic patients in whom hypoglycaemia was induced by an i.v. bolus of insulin and (3) comparing subcutaneous and intranasal glucagon in six Type 1 diabetic patients in whom hypoglycaemia was induced by adding extra subcutaneous regular insulin to their usual morning dosage. Our results show that 1 mg of intranasal glucagon is as effective as 1 mg of subcutaneous glucagon in terms of the rise in blood glucose. Differences in kinetics between the subcutaneous and the intranasal routes may be observed: intranasal glucagon initiates the blood glucose rise earlier than does the subcutaneous form but the effect of the latter is more sustained. Glycocholic acid appears to be a perfectly tolerated agent in acute conditions. The use of intranasal lyophylized glucagon, for the reversal of hypoglycaemia in Type 1 diabetes, seems to be a clinically relevant alternative to its parenteral equivalent and should now be ready to be introduced in the market.