The Response to Exercise with Constant Energy Intake in Identical Twins

Seven pairs of young adult male identical twins completed a negative energy balance protocol during which they exercised on cycle ergometers twice a day, 9 out of 10 days, over a period of 93 days while being kept on a constant daily energy and nutrient intake. The total energy deficit caused by exercise above the estimated energy cost of body weight maintenance reached 244 ± 9.8 MJ (Mean ± SEM). Baseline energy intake was estimated over a period of 17 days preceding the negative energy balance protocol. Mean body weight loss was 5.0 kg (SEM = 0.6) (p <0.001) and it was entirely accounted for by the loss of fat mass (p <0.001). Fat-free mass was unchanged. Body energy losses reached 191 MJ (SEM = 24) (p <0.001) which represented about 78% of the estimated energy deficit. Subcutaneous fat loss was slightly more pronounced on the trunk than on the limbs as estimated from skinfolds, circumferences, and computed tomography (CT). The reduction in CT-assessed abdominal visceral fat was quite striking, from 81 cm² (SEM = 5) to 52 cm² (SEM = 6) (p <0.001). At the same submaximal power output level, subjects oxidized more lipids than carbohydrates after the program as indicated by the changes in the respiratory exchange ratio (p <0.05). Intrapair resemblance was observed for the changes in body weight (p <0.05), fat mass (P <0.01), percent fat (p <0.01), body energy content (p <0.01), sum of 10 skinfolds (p <0.01), abdominal visceral fat (p <0.01), fasting plasma triglycerides (p <0.05) and cholesterol (p <0.05), maximal oxygen uptake (p <0.05), and respiratory exchange ratio during submaximal work (p <0.01). We conclude that even though there were large individual differences in response to the negative energy balance and exercise protocol, subjects with the same genotype were more alike in responses than subjects with different genotypes particularly for body fat, body energy, and abdominal visceral fat changes. High lipid oxidizers and low lipid oxidizers during sub-maximal exercise were also seen despite the fact that all subjects had experienced the same exercise and nutritional conditions for about three months.     

Variation of bone collagen amino acid δ13C values in archaeological humans and fauna with different dietary regimes: developing frameworks of dietary discrimination

We present bone collagen amino acid (AA) δ(13)C values for a range of archaeological samples representing four \"benchmark\" human diet groups (high marine protein consumers, high freshwater protein consumers, terrestrial C(3) consumers, and terrestrial C(4) consumers), a human population with an \"unknown\" diet, and ruminants. The aim is to establish an interpretive palaeodietary framework for bone collagen AA δ(13)C values, and to assess the extent to which AA δ(13)C values can provide additional dietary information to bulk collagen stable isotope analysis. Results are analyzed to determine the ability of those AAs for which we have a complete set, to discriminate between the diet groups. We show that very strong statistical discrimination is obtained for all interdiet group comparisons. This is often obvious from suitably chosen bivariate plots using δ(113)C values that have been normalized to compensate for interdiet group differences in bulk δ(13)C values. Bi-plots of non-normalized phenylalanine and valine δ(13)C values are useful for distinguishing aquatic diets (marine and freshwater) from terrestrial diets. Our interpretive framework uses multivariate statistics (e.g., discriminant analysis) to optimize the separation of the AA δ(13)C values of the \"benchmark\"' diet groups, and is capable of accurately assigning external samples to their expected diet groups. With a growing body of AA δ(13)C values, this method is likely to enhance palaeodietary research by allowing the \"unknown\" diets of populations under investigation to be statistically defined relative to the well-characterized or \"known\" diets of previously investigated populations.     

Rapid Carbon Turnover During Growth of Atlantic Salmon (Salmo salar) Smolts in Sea Water, and Evidence for Reduced Food Consumption by Growth-Stunts

Wild Atlantic salmon smolts were captured during spring out-migration in the Northwest Miramichi River, New Brunswick, Canada, and placed on an isotopically distinct hatchery diet to determine the relative contributions of growth and metabolic turnover to isotopic change. As expected for an ectothermic species, growth explained a large amount of isotopic variation in changing stable carbon ratios of muscle tissue (average r²= 0.46 ) for the 3 years of study. Turnover rates of muscle carbon in all 3 years in growing fish (0.24–0.66 month^–1) were higher than previously reported values for other ectothermic species, but there was little evidence for isotopic change in non-growers (average r²= 0.041, p > 0.1). It is unlikely that non-growers had consumed any of the hatchery diet over a 2-month period, thus preventing them from acquiring the new carbon isotopic signature. This period of food deprivation resulted in nitrogen-15 enrichment in liver relative to muscle (p= 0.003). It is advised that future isotope studies of metabolic turnover rates in ectotherms be conducted on slow-growing animals over a long time period. This would serve to avoid the obscuring effects of growth on isotopic change, and provide stronger comparisons to endothermic tissue turnover rates.     

Indole-3-acetic acid biosynthesis in Lemna gibba studied using stable isotope labeled anthranilate and tryptophan

IAA biosynthesis in many plants, including Lemna gibba, has been shown to involve at least two different pathways, one from tryptophan and a tryptophan-independent route. To study the kinetics of IAA biosynthesis in Lemna, we simultaneously measured the incorporation of label from [15N]-anthranilate and [2H5]-tryptophan into IAA by Lemna plants in short term feeding studies. The data show that label from anthranilate rapidly goes into IAA and tryptophan. Labeling of the IAA pool by [15N]-anthranilate slightly precedes labeling of the tryptophan pool, confirming that more than one route to IAA exists in these plants. Longer term feeding studies (5–25 h) suggest that exogenous tryptophan is used preferentially to label IAA as compared to tryptophan made by the plant. This is indicated by the fact that the IAA pool was more enriched than the tryptophan pool in [2H5]-label, but less enriched than the tryptophan pool in [15N] (which comes about by de novo synthesis of tryptophan from [15N]-anthranilate by the plant).     

An LC/MS method for d8-β-carotene and d4-retinyl esters: β-carotene absorption and its conversion to vitamin A in humans

The intestinal absorption and metabolism of β-carotene is of vital importance in humans, especially in populations that obtain the majority of their vitamin A from provitamin A carotenoids. MS has provided a better understanding of the absorption of β-carotene, the most potent provitamin A carotenoid, through the use of stable isotopes of β-carotene. We report here an HPLC-MS method that eliminates the need for complicated sample preparation and allows us to detect and quantify newly absorbed d8-β-carotene as well as its d4-retinyl ester metabolites in human plasma and chylomicron fractions. Both retinoids and β-carotene were recovered in a single simple extraction that did not involve saponification, thus allowing subsequent quantitation of individual fatty acyl esters of retinol. Separation of d8-β-carotene and its d4-retinyl ester metabolites was achieved using the same C30 reversed-phase liquid chromatography followed by mass spectrometry in selected ion monitoring and negative atmospheric pressure chemical ionization modes, respectively. Total time for the two successive runs was 30 min. This HPLC-MS method allowed us to quantify the absorption of intact d8-β-carotene as well as its extent of conversion to d4-retinyl esters in humans after consumption of a single 5 mg dose of d8-β-carotene.     

Evaluation of regression models in metabolic physiology: predicting fluxes from isotopic data without knowledge of the pathway

This study explores the ability of regression models, with no knowledge of the underlying physiology, to estimate physiological parameters relevant for metabolism and endocrinology. Four regression models were compared: multiple linear regression (MLR), principal component regression (PCR), partial least-squares regression (PLS) and regression using artificial neural networks (ANN). The pathway of mammalian gluconeogenesis was analyzed using [U−¹³C]glucose as tracer. A set of data was simulated by randomly selecting physiologically appropriate metabolic fluxes for the 9 steps of this pathway as independent variables. The isotope labeling patterns of key intermediates in the pathway were then calculated for each set of fluxes, yielding 29 dependent variables. Two thousand sets were created, allowing independent training and test data. Regression models were asked to predict the nine fluxes, given only the 29 isotopomers. For large training sets (>50) the artificial neural network model was superior, capturing 95% of the variability in the gluconeogenic flux, whereas the three linear models captured only 75%. This reflects the ability of neural networks to capture the inherent non-linearities of the metabolic system. The effect of error in the variables and the addition of random variables to the data set was considered. Model sensitivities were used to find the isotopomers that most influenced the predicted flux values. These studies provide the first test of multivariate regression models for the analysis of isotopomer flux data. They provide insight for metabolomics and the future of isotopic tracers in metabolic research where the underlying physiology is complex or unknown.We acknowledge the support of NIH Grant DK58533 and the DuPont-MIT Alliance.     

Abnormalities of Energy Expenditure and the Development of Obesity

The role of energy expenditure in the development of obesity remains unclear. This issue is examined using data from prospective studies of energy expenditure and obesity, the effects of overfeeding and diet composition on energy expenditure, and studies of the relationship between energy expenditure for physical activity and body composition. The combined results from these investigations strongly support the view that low energy expenditure can facilitate rapid weight gain in susceptible individuals. It is speculated that, in susceptible individuals, low energy expenditure for resting energy expenditure as well as physical activity are part of a range of mechanisms available for providing surplus energy for rapid weight gain. In addition, both cross-sectional and intervention studies indicate that there is an equilibration between the level of energy expenditure for physical activity and body fat content. While genetic and other factors clearly play an important role in this relationship, it appears that a modest reduction in body fat content can be achieved by increasing energy expenditure for physical activity in physical exercise programs.