We investigated the use and possible mechanisms mediating the increased energy expenditure (EE) previously described for rats subjected to total or paradoxical sleep deprivation. Bomb calorimetry of wastes showed that during deprivation the efficiency of energy utilization was not reduced. Estimates of CO2 production by the doubly labelled water method of indirect calorimetry correlated with EE estimated from the caloric value of food, weight change, and wastes and confirmed an increase in EE during deprivation. Core temperatures decreased during the later stages of deprivation, suggesting the hypothesis that excessive heat loss may have required increased EE to protect body temperature. The increased EE could not be explained by the metabolic cost of increase wakefulness, water exposure, or motor activity; an increase in resting EE was indicated. The contribution of the hypothalamic-pituitary-adrenal axis, thyroid gland, and sympathoadrenal system to the mediation of the EE increases was evaluated by measuring the plasma levels of their hormones. Results appear to rule out the first as a mediator. Evidence for the other two was equivocal.

The accuracy and precision of the doubly labeled water method for measuring energy expenditure are influenced by isotope fractionation during evaporative water loss and CO2 excretion. To characterize in vivo isotope fractionation, we collected and isotopically analyzed physiological fluids and gases. Breath and transcutaneous water vapor were isotopically fractionated. The degree of fractionation indicated that the former was fractionated under equilibrium control at 37 degrees C, and the latter was kinetically fractionated. Sweat and urine were unfractionated. By use of isotopic balance models, the fraction of water lost via fractionating routes was estimated from the isotopic abundances of body water, local drinking water, and dietary solids. Fractionated water loss averaged 23% (SD = 10%) of water turnover, which agreed with our previous estimates based on metabolic rate, but there was a systematic difference between the results based on O2 and hydrogen. Corrections for isotopic fractionation of water lost in breath and (nonsweat) transcutaneous loss should be made when using labeled water to measure water turnover or CO2 production.

Human cholesterogenesis is measurable as the rate of Incorporation of deuterium derived from deuterium oxide (D 2 O) within the body water pool into plasma or erythrocyte cholesterol pools. Oral D 2 O equilibrates across body water, thus enabling extracellular sampling of pools (such as urine) to serve as accurate indicators of intracellular deuterium enrichments at the point of synthesis. Required doses of D 2 O fall below the threshold associated with negative side effects. Deuterium/carbon incorporation ratios into cholesterol during biosynthesis have been established that are applicable in humans. Models using unconstrained and constrained curve fitting permit improved flexibility in interpretation of deuteriumuptake kinetics. However, sample-size restrictions presently limit the ability of the technique to examine the kinetics within individual lipoprotein species. Correction of enrichment data for proton exchange during the combustion and reduction phases of sample preparation is an additional important procedural concern. In summary, the deuterated-water procedure is a useful tool in studies of human cholesterol synthesis that offers the advantages of short measurement interval, relative noninvasiveness, and provision of a direct index of synthesis in comparison with other available techniques. ( 910 Steady-state requirements vary across methods. With input-output analysis or kinetic approaches, cholesterol pool steadystate conditions must exist for the measurements to be valid. With other methods, cholesterogenesis can be measured without steady-state cholesterol pools. Moreover, the time window of measurement varies from 7 days or more for input-output analysis to a few hours for sterol precursors. Although wide-ranging, such methods are limited in three ways: they either are accurate but require extended measurement periods, are immediate but overly invasive, or measure synthesis indirectly.

The bioelectrical-impedance-analysis (BIA) method accurately measures body composition in weight-stable subjects. This study validates the use of BIA to measure change in body composition. Twelve obese females underwent weight loss at a mean rate of 1.16 kg/wk. Body composition was measured by deuterium oxide dilution (D2O), BIA, and skinfold anthropometry (SFA) at baseline and at 5% decrements in weight. Highly significant correlations were obtained between D2O and BIA (r = 0.971) and between D2O and SFA (r = 0.932). Overall, BIA predicted change in fat-free mass with greater accuracy (to 0.4 kg) and precision (+/- 1.28 kg) than did anthropometry (to 0.8 kg and +/- 2.58 kg, respectively). We conclude that BIA is a useful clinical method for measuring change in body composition.

To examine meal-frequency effects on circulating hormone concentrations and cholesterol synthesis, male subjects consumed liquid diets given as either six evenly spaced (ES) or three diurnal (DI) meals over 3 d. Deuterium oxide was given on day 2 and blood sampled every 4 h over days 2 and 3 to measure plasma cholesterol, glucose, insulin, and glucose-dependent insulinotropic polypeptide (GIP) concentrations and cholesterol synthesis. Cholesterol synthesis was determined from deuterium incorporation into plasma free cholesterol by using constrained and unconstrained curve-fit models. Plasma total cholesterol concentrations decreased in both ES and DI groups (P < 0.05). The ES group had lower insulin (P < 0.05) and GIP (P < 0.001) concentrations compared with the DI group. Cholesterol synthesis was reduced (P < 0.01) in the ES vs the DI group when determined by using constrained (0.050 +/- 0.002 vs 0.075 +/- 0.005 pools/d, respectively) and unconstrained (0.072 +/- 0.005 vs 0.119 +/- 0.011 pools/d, respectively) models. These data suggest meal frequency-dependent control of cholesterogenesis via hormonally mediated mechanisms.