Leptin and body composition of Nigerians, Jamaicans, and US blacks.

The role of leptin in humans remains controversial. Leptin concentrations are highly correlated with body fat stores. We tested whether or not this relation was consistent across the range of body composition encompassing the lean as well as the obese. Individuals participating in community-based comparative research in Nigeria (n = 363), Jamaica (n = 372), and the United States (Maywood, IL; n = 699) had their plasma leptin concentrations and body compositions (with bioelectrical impedance analysis) measured. All participants identified themselves as being black. Body mass index (in kg/m2) ranged from 14 to 62. Large differences in mean plasma leptin were noted across populations for both men and women in Nigeria, Jamaica, and the United States, respectively (men: 2.8, 3.9, and 6.8 microg/L; women: 10.3, 18.6, and 27.7 microg/L). An exponential function fit the relation between percentage body fat or total fat mass and leptin for men and women at each site. For women and men the exponential function with either percentage body fat or total fat mass was of the same shape, but increased by a constant in women, yielding higher leptin concentrations than in men at every level of body fat. On the basis of this broad distribution of body composition, the data suggest an exponential response of leptin to increases in body fat stores, consistent with the development of leptin resistance in individuals developing obesity. These findings likewise confirm that men and women exhibit different set points in terms of leptin production.

Rapid 18O analysis of CO2 samples by continuous-flow isotope ratio mass spectrometry.

Continuous-flow isotope ratio mass spectrometry (CF-IRMS) has been shown to minimize sample preparation and reduce the sample size requirements and time for the determination of carbon and nitrogen. An inexpensive CF inlet was developed for the determination of the 18O isotopic abundance in CO2. Initial efforts were hampered by significant exchangeable oxygen within the inlet system, which was minimized by design changes and post-analysis mathematical correction. The system was tested for accuracy in the measurement of 18O abundance in standards and was accurate to 0.04/1000 with a precision of 0.03/1000. Additional tests were performed using urinary water for the measurement of energy expenditure by the doubly labeled water method and the results agreed to within 1% with those from traditional off-line, dual-inlet IRMS. The CF-IRMS reduced the analysis time to 2 min per sample compared with 15 min for traditional duel-inlet IRMS.

Compound-specific carbon isotope ratio determination of enriched cholesterol.

On-line gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) was evaluated for use in stable isotope studies of human cholesterol metabolism. Calibration curves were constructed by analyzing underivatized and TMS-cholesterol over a range of 13C abundances from natural abundance to 0.05 mol of [3,4-13C]cholesterol per mole of unlabeled cholesterol (delta 13C -24 to 27/1000). The calibration curves were highly linear (r2 = 0.99) with slopes of 0.98 and 0.88 for underivatized and TMS-cholesterol, respectively. Higher precision of 13C/12C ratio determination was obtained using TMS-cholesterol, with a detection limit (2 SD) of 0.3/1000 or 0.004 mol% of [3,4-13C]cholesterol. By comparison standard GC/MS had a detection limit of 0.06 mol% for [3,4-13C]cholesterol. A 57-day study of cholesterol elimination was conducted in one human subject with oral administration of 50 mg of [3,4-13C]cholesterol. Peak enrichment was 9/1000 above baseline (40 times the detection limit) and could be followed for 40-50 days. The results indicate that GC/C/IRMS provides 15-fold lower detection limits for [3,4-13C]cholesterol than GC/MS and is useful for human studies of cholesterol metabolism.

Relative dilution spaces of 2H- and 18O-labeled water in humans.

The doubly labeled water method for measuring energy expenditure can be very sensitive to small differences in the ratio of the 2H to 18O isotope dilution spaces. Recently it has been suggested that the average ratio is higher than the 1.03 we previously recommended. We therefore combined the data from 99 recently studied subjects. Subjects (85 females and 14 males) were between the ages of 4 and 78 yr (mean = 34 yr) and between 10 and 52% (mean = 35%) fat. The average 2H-to-18O dilution space ratio was 1.034 +/- 0.014, which was very similar to the original assumption. As in recent reports, we did find that most of the variance (60%) was due to random analytic error and that there was no correlation between the dilution space ratio and age or body fat. However, in contrast to recent reports we found no evidence of a gender difference. Use of the constant dilution space ratio of 1.034 to recalculate CO2 product in published validation studies demonstrated improved accuracy, and thus the value of 1.034 is suggested for use in future studies.