The use of low-calorie diets is a common strategy for body-weight reduction purposes, but the time-course of the metabolic changes induced by moderately energy-restricted, otherwise balanced, diets is still poorly known. The aim of this nutritional intervention design was to study in obese women the effect of a balanced low-calorie diet on the metabolic rate, and metabolic fuel utilization changes during the weight loss process through the application of breath tests with stable isotope-labeled tracers. Seven obese (body mass index >30 kg/m(2)) women were assigned to a 10-week dietary hypoenergetic intervention regime supplying 55% of energy as carbohydrate, 30% as fat and 15% as protein. Metabolic rate and substrate utilization were evaluated for 6 h in separate occasions during the weight loss program by indirect calorimetry and after 13C-labeled glucose, triolein and leucine administration. Body weight loss after 10 weeks was 4.2+/-1.1 kg, while the percent body fat decrease was about 5%. Slimming was accompanied by a marked decrease in fasting leptin (about 25%). Postprandial carbohydrate utilization after the administration of a test meal with the same macronutrient distribution as the experimental low-energy diet was decreased (24.1%, P<0.05) as a consequence of the dietary restriction, which was associated with lower insulin plasma levels (P<0.05). Although protein and lipid oxidation were not significantly different after weight reduction (day 1 versus day 70), the metabolic utilization of these substrates tended to increase. Moreover, marginally significant indications obtained on days 15 and 45 suggest that the weight and body composition changes are attributable to a shift in endogenous and exogenous glucose utilization in favor of lipid burning. The breath tests determinations, which were performed on different occasions along the experimental trial, confirmed that the cumulative 13C output decreased for labeled tracers with time, being only statistically significant for the glucose utilization between days 15 and 45. In summary, the weight and fat mass losses were associated with a lower carbohydrate oxidation, which were probably compensated by an increase in lipid oxidation without major changes in protein mobilization.