Assessment of usnic acid toxicity in rat primary hepatocytes using ¹³C isotopomer distribution analysis of lactate, glutamate and glucose.

The lichen metabolite usnic acid (UA) has been promoted as a dietary supplement for weight loss, although cases of hepatotoxicity have been reported. Here we evaluated UA-associated hepatotoxicity in vitro using isolated rat hepatocytes. We measured cell viability and ATP content to evaluate UA induced cytotoxicity and applied (13)C isotopomer distribution measuring techniques to gain a better understanding of glucose metabolism during cytotoxicity. The cells were exposed to 0, 1, 5 or 10 µM UA concentrations for 2, 6 or 24h. Aliquots of media were collected at the end of these time periods and the (13)C mass isotopomer distribution determined for CO(2), lactate, glucose and glutamate. The 1 µM UA exposure did not appear to cause significant change in cell viability compared to controls. However, the 5 and 10 µM UA concentrations significantly reduced cell viability as exposure time increased. Similar results were obtained for ATP depletion experiments. The 1 and 5 µM UA doses suggest increased oxidative phosphorylation. Conversely, oxidative phosphorylation and gluconeogenesis were dramatically inhibited by 10 µM UA. Augmented oxidative phosphorylation at the lower UA concentrations may be an adaptive response by the cells to compensate for diminished mitochondrial function.

Ingested fat oxidation contributes 8% of 24-h total energy expenditure in moderately obese subjects.

The role of ingested fat in the etiology of obesity is controversial. The aims of this study were to determine the contributions of ingested fat oxidation to: 1) 24-h total energy expenditure (TEE), and 2) substrate oxidation during acute stationary cycle exercises in adult humans. Healthy, moderately obese (n = 18; BMI = 31 +/- 1 kg/m2) subjects (8 men; 10 women) were each studied in a whole-room calorimeter for 24 h. They were fed mixed meals (55, 30, and 15% as energy from carbohydrate, fat and protein, respectively) to maintain energy balance. Each subject performed 1255-kJ cycle exercises at 50% VO2max in the calorimeter. Study test meal fat was labeled with carbon-13 (13C). Ingested fat oxidation was estimated from breath 13CO2 excretion and the subject's chamber CO2 production. Total fat and carbohydrate oxidations were estimated from nonprotein respiratory quotient (NP-RQ) values. Endogenous fat oxidation was estimated as the difference between total fat and ingested fat oxidations. TEE was estimated from gas exchanges; 28 +/- 3% of ingested fat was oxidized and it provided 8 +/- 1% of 24-h TEE. During cycle exercises, ingested fat provided 50% of total fat oxidized and 13.0 +/- 2% of energy expended. Endogenous fat oxidation contributed 10.4 +/- 3% of energy expenditure during cycle exercises. This study extended to 24-h observations of previous studies that lasted 6-9 h on ingested fat oxidation in humans. Understanding the factors that promote ingested fat oxidation could lead to more effective obesity intervention programs.

Multi-point estimation of total energy expenditure: a comparison between zinc-reduction and platinum-equilibration methodologies.

Reducing water to hydrogen gas by zinc or uranium metal for determining D/H ratio is both tedious and time consuming. This has forced most energy metabolism investigators to use the "two-point" technique instead of the "Multi-point" technique for estimating total energy expenditure (TEE). Recently, we purchased a new platinum (Pt)-equilibration system that significantly reduces both time and labor required for D/H ratio determination. In this study, we compared TEE obtained from nine overweight but healthy subjects, estimated using the traditional Zn-reduction method to that obtained from the new Pt-equilibration system. Rate constants, pool spaces, and CO2 production rates obtained from use of the two methodologies were not significantly different. Correlation analysis demonstrated that TEEs estimated using the two methods were significantly correlated (r=0.925, p=0.0001). Sample equilibration time was reduced by 66% compared to those of similar methods. The data demonstrated that the Zn-reduction method could be replaced by the Pt-equilibration method when TEE was estimated using the "Multi-Point" technique. Furthermore, D equilibration time was significantly reduced.

The validity of the Stanford Seven-Day Physical Activity Recall in young adults.

PURPOSE: To evaluate the criterion validity of the 7-Day Physical Activity Recall (7D-PAR) and factors associated with reporting error, in a sample of moderately overweight, young adult men and women. METHODS: Average total daily energy expenditure (TDEE) and physical activity energy expenditure (PAEE) from the 7D-PAR were compared with the same parameters assessed by doubly labeled water in 17 men, age = 23.9 +/- 3.8 yr, and 29 women, age = 23.3 +/- 4.6 yr, who volunteered to participate in a 16-month supervised aerobic exercise trial. PAEE was estimated from the 7D-PAR and from DLW [0.9 * TDEE -resting metabolic rate (RMR) (indirect calorimetry)]. In addition, peak oxygen uptake and percent body fat were obtained. RESULTS: No significant differences in TDEE (kJ.d-1) were noted between the 7D-PAR (11825 +/- 1779) and DLW (11922 +/- 2516) for the complete sample (N = 46) or for men (7D-PAR = 13198 +/- 1638, DLW = 13885 +/- 2754) or women (7D-PAR = 11018 +/- 1323, DLW = 10771 +/- 1457. The mean PAEE from the 7D-PAR was not different from DLW in the total sample (7D-PAR = 3286 +/- 502, DLW = 3508 +/- 1863) as well as in men (7D-PAR = 3650 +/- 490, DLW = 3989 +/- 2461) and women (3073 +/- 377, DLW = 3223 +/- 1360). In a regression model, PAEE, peak oxygen uptake, gender and percent fat accounted for 86% of the reporting error in total daily energy expenditure when using the 7D-PAR. CONCLUSION: The 7D-PAR provided a reasonable estimate of both the mean TDEE and PAEE in this sample; however, estimates of energy expenditure on an individual basis using the PAR were subject to considerable error.