Physical Activity Level Using Doubly-Labeled Water in Relation to Body Composition and Physical Fitness in Preschoolers.

Background and objectives: There is a lack of studies investigating associations of physical activity level (PAL) and activity energy expenditure (AEE) using the doubly-labeled water (DLW) method with body composition and physical fitness in young children. Thus, we aimed to examine cross-sectional associations of PAL and AEE with body composition indices and physical fitness components in Swedish preschool children. Materials and methods: PAL was calculated as total energy expenditure measured using DLW divided by the predicted basal metabolic rate in 40 children aged 5.5 (standard deviation 0.2) years. AEE was calculated as total energy expenditure minus basal metabolic rate and the thermic effect of food, and divided by fat-free mass. Body composition was assessed using the 3-component model by combining measurements based on isotope dilution and air-displacement plethysmography. Physical fitness (muscular strength, motor fitness, and cardiorespiratory fitness) was evaluated using the PREFIT test battery. Multiple linear regression models were conducted. Results: PAL and AEE were negatively associated with body mass index, percent body fat, and fat mass index (PAL: standardized ß -0.35, -0.41, and -0.45, all p < 0.036; AEE: standardized ß -0.44, -0.44, and -0.47, all p < 0.006, respectively). Furthermore, PAL and AEE were positively associated with the standing long jump test (PAL: standardized ß 0.37, p = 0.017; AEE: standardized ß 0.38, p = 0.014). There were no statistically significant associations found regarding PAL or AEE with fat-free mass index or any other physical fitness test. Conclusions: Greater PAL and AEE at the age 5.5 were significantly associated with body fatness and improved lower-body muscular strength. Therefore, increasing physical activity, and thus energy expenditure, at young ages may be beneficial for preventing overweight/obesity. However, further studies with larger sample sizes are needed to confirm the results.

Physical Activity Assessment with the ActiGraph GT3X and Doubly Labeled Water.

PURPOSE: To compare the degree to which four accelerometer metrics-total activity counts per day (TAC per day), steps per day (steps per day), physical activity energy expenditure (PAEE) (kcal·kg·d), and moderate- to vigorous-intensity physical activity (MVPA) (min·d)-were correlated with PAEE measured by doubly labeled water (DLW). Additionally, accelerometer metrics based on vertical axis counts and triaxial counts were compared. METHODS: This analysis included 684 women and 611 men age 43 to 83 yr. Participants wore the Actigraph GT3X on the hip for 7 d twice during the study and the average of the two measurements was used. Each participant also completed one DLW measurement, with a subset having a repeat. PAEE was estimated by subtracting resting metabolic rate and the thermic effect of food from total daily energy expenditure estimated by DLW. Partial Spearman correlations were used to estimate associations between PAEE and each accelerometer metric. RESULTS: Correlations between the accelerometer metrics and DLW-determined PAEE were higher for triaxial counts than vertical axis counts. After adjusting for weight, age, accelerometer wear time, and fat free mass, the correlation between TAC per day based on triaxial counts and DLW-determined PAEE was 0.44 in women and 0.41 in men. Correlations for steps per day and accelerometer-estimated PAEE with DLW-determined PAEE were similar. After adjustment for within-person variation in DLW-determined PAEE, the correlations for TAC per day increased to 0.61 and 0.49, respectively. Correlations between MVPA and DLW-determined PAEE were lower, particularly for modified bouts of ≥10 min. CONCLUSIONS: Accelerometer measures that represent total activity volume, including TAC per day, steps per day, and PAEE, were more highly correlated with DLW-determined PAEE than MVPA using traditional thresholds and should be considered by researchers seeking to reduce accelerometer data to a single metric.

Determination of urinary malondialdehyde by isotope dilution LC-MS/MS with automated solid-phase extraction: a cautionary note on derivatization optimization.

A highly sensitive quantitative LC-MS/MS method was developed for measuring urinary malondialdehyde (MDA). With the use of an isotope internal standard and online solid-phase extraction, urine samples can be directly analyzed within 10 min after 2,4-dinitrophenylhydrazine (DNPH) derivatization. The detection limit was estimated as 0.08 pmol. This method was further applied to assess the optimal addition of DNPH for derivatization and to measure urinary MDA in 80 coke oven emission (COE)-exposed and 67 nonexposed workers. Derivatization optimization revealed that to achieve complete derivatization reaction, an excess of DNPH is required (DNPH/MDA molar ratio: 893-8929) for urine samples that is about 100 times higher than that of MDA standard solutions (molar ratio: 10-80). Meanwhile, the mean urinary concentrations of MDA in COE-exposed workers were significantly higher than those in nonexposed workers (0.23±0.17 vs 0.14±0.05 µmol/mmol creatinine, P<0.005). Urinary MDA concentrations were also significantly associated with the COE (P<0.005) and smoking exposure (P<0.05). Taken together, this method is capable of routine high-throughput analysis and accurate quantification of MDA and would be useful for assessing the whole-body burden of oxidative stress. Our findings, however, raise the issue that derivatization optimization should be performed before it is put into routine biological analysis.

Quantification of deuterated bisphenol A in serum, tissues, and excreta from adult Sprague-Dawley rats using liquid chromatography with tandem mass spectrometry.

Bisphenol A (BPA) is an important industrial chemical used in the manufacture of polycarbonate plastic products, epoxy resin-based food can liners, and paper products. The presence of BPA in urine of >90% of Americans aged 6-60 suggests ubiquitous and frequent exposure and is problematic because of the potential for endocrine disruption. The ubiquity of environmental BPA in common laboratory supplies used for sample collection, storage, and analysis greatly increases the likelihood of false positive determinations, particularly at trace levels. The current study validated using liquid chromatography/tandem mass spectrometry (LC/MS/MS) in conjunction with deuterated BPA as the dosing material to circumvent contamination for high sensitivity quantifications in rat serum, tissues, urine, and feces. The methods described provided measurements of both estrogen receptor-active aglycone and metabolically deactivated conjugated forms of BPA, a distinction that is critical to assessing toxicological potential. The adequacy of the described methodology was substantiated by its utility in analyzing samples from rats treated orally with a 100 µg/kg body weight dose of d6-BPA. These results emphasize the challenges inherent in measuring BPA in biological samples and how employing stable isotope labeled dosing can facilitate pharmacokinetic studies needed to understand BPA metabolism and disposition. Such studies conducted in experimental animal models, in conjunction with properly validated human biomonitoring data, will be the basis for PBPK modeling of BPA in environmentally exposed humans.

Simplified method for microlitre deuterium measurements in water and urine by gas chromatography-high-temperature conversion-isotope ratio mass spectrometry.

Deuterium (2H) in water and urine can be measured by off-line and, more recently, on-line techniques using isotope ratio mass spectrometry (IRMS). We describe a new simple on-line pyrolysis method for the analysis of 2H/1H in water and urine samples by continuous flow IRMS, normally used for 2H/1H measurements in organic compounds. A deactivated column connected the split injector to a high-temperature conversion reactor (TC HD), and 0.5 microL of sample was injected. Accuracy and precision were determined with Vienna Standard Mean Ocean Water (VSMOW), Standard Light Antarctic Precipitation (SLAP), and Greenland Ice Sheet Precipitation (GISP). The range of linearity was measured with a calibration curve of enriched water from 0 up to 0.1 atom percent excess (APE) (i.e. -72 up to 6323 delta per mil (deltaD per thousand)) with a precision of <5 per thousand and accuracy ranging between 1 and 55 per thousand. Blinded reanalysis of urine samples by an equilibration device (Gas Bench) and by a dedicated pyrolysis system (TC/EA) was performed and results compared by the Bland-Altman test. Enrichments ranged between 600 and 2400 per thousand deltaD(VSMOW) with a precision of +/-5 per thousand. Urine enrichments described by our method were strongly correlated with values obtained by Gas Bench and TC/EA (p < 0.0001). There was a significant memory effect that was reduced by injecting the sample 15 times and discarding the first 10 injections, together with accurate furnace conditioning and appropriate cleaning of the syringe. Data indicate that the method is accurate, and that it can be used for water and urine deuterium determination when a Gas Bench or TC/EA instrument is not available and the amount of sample is limited.

Stable-isotope dilution gas chromatography-mass spectrometric measurement of 3-hydroxyglutaric acid, glutaric acid and related metabolites in body fluids of patients with glutaric aciduria type 1 found in newborn screening.

We developed a simple and sensitive stable-isotope dilution method for the quantification of 3-hydroxyglutaric acid (3HGA) and glutaric acid (GA) in body fluids. In our method, tert-butyldimethylsilyl (tBDMS) derivatives of 3HGA and GA were measured with a conventional electron-impact ionization (EI) mode in gas chromatography-mass spectrometry (GC-MS). The control values for 3HGA in nmol/ml were 0.15+/-0.08 (serum; n=10) and 0.07+/-0.03 (CSF; n=10). In addition, glutarylcarnitine and free carnitine were quantified by electrospray tandem mass spectrometry. Using these methods, we monitored 3HGA, GA, and glutarylcarnitine in the body fluids of three patients with glutaric aciduria type 1 found during newborn screening. None of the patients had experienced neurological strokes, which are possibly caused by the accumulation of 3HGA, at 15-24 months of age under a disease-specific treatment, including carnitine supplementation. Our data showed that 3HGA levels were relatively high in some serum samples with lower glutarylcarnitine and carnitine levels, suggesting that carnitine supplementation may play a role in preventing the accumulation of 3HGA in patients with this disease.

Relation between physical activity and energy expenditure in a representative sample of young children.

BACKGROUND: Strategies for the prevention and treatment of childhood obesity require a better understanding of the relation between the pattern of free-living physical activity and total energy expenditure (TEE). OBJECTIVE: We assessed the relations between TEE and physical activity level (PAL) during engagement in different intensities of physical activity. DESIGN: We used a cross-sectional study of 104 children (median age: 5.4 y) in Scotland. TEE was measured with use of doubly labeled water (DLW), and resting energy expenditure was predicted to determine PAL. Time spent sedentary and in light-intensity activity and in moderate- and vigorous-intensity physical activity (MVPA) was assessed by accelerometry concurrent with DLW measurements. Correlation and regression were used to assess the relations between measures of sedentary behavior, intensities of activity, and PAL as the dependent variable. RESULTS: Time spent sedentary was negatively correlated with PAL (r = -0.33, P < 0.01), and time spent in light-intensity activity was positively correlated with PAL (r = 0.31, P < 0.01). In multiple regression analyses, both time spent sedentary and in light-intensity activities were significantly associated with PAL. Time spent in MVPA was not associated with PAL; engagement in MVPA was limited in this sample (median: 3% of waking hours; range: 0-14%). PAL was significantly higher in boys than in girls. CONCLUSION: In this sample and setting, PAL was not influenced by engagement in MVPA but was influenced by time spent sedentary and in light-intensity activities. This study suggests that in young children, MVPA could make only a minor contribution to free-living TEE and PAL.

Habitual physical activity and physical activity intensity: their relation to body composition in 5.0-10.5-y-old children.

BACKGROUND: Concerns of a decrease in physical activity levels (PALs) of children and a concurrent increase in childhood obesity exist worldwide. The exact relation between these two parameters however has as yet to be fully defined in children. OBJECTIVE: This study examined the relation in 47 children, aged 5-10.5 y (mean age 8.4+/-0.9 y) between habitual physical activity, minutes spent in moderate, vigorous and hard intensity activity and body composition parameters. DESIGN: Total energy expenditure (TEE) was calculated using the doubly labelled water technique and basal metabolic rate (BMR) was predicted from Schofield's equations. PAL was determined by PAL=TEE/BMR. Time spent in moderate, vigorous and hard intensity activity was determined by accelerometry, using the Tritrac-R3D. Body fatness and body mass index (BMI) were used as the two measures of body composition. RESULTS: Body fat and BMI were significantly inversely correlated with PAL (r=-0.43, P=0.002 and r=-0.45, P=0.001). Times spent in vigorous activity and hard activity were significantly correlated to percentage body fat (r=-0.44, P=0.004 and r=-0.39, P=0.014), but not BMI. Children who were in the top tertiles for both vigorous activity and hard activity had significantly lower body fat percentages than those in the middle and lowest tertiles. Moderate intensity activity was not correlated with measures of body composition. CONCLUSIONS: As well as showing a significant relation between PAL and body composition, these data intimate that there may be a threshold of intensity of physical activity that is influential on body fatness. In light of world trends showing increasing childhood obesity, this study supports the need to further investigate the importance of physical activity for children.

Noninvasive evaluation of liver metabolism by 2H and 13C NMR isotopomer analysis of human urine.

Mammalian liver disposes of acetaminophen and other ingested xenobiotics by forming soluble glucuronides that are subsequently removed via renal filtration. When given in combination with the stable isotopes 2H and 13C, the glucuronide of acetaminophen isolated from urine provides a convenient "chemical biopsy" for evaluating intermediary metabolism in the liver. Here, we describe isolation and purification of urinary acetaminophen glucuronide and its conversion to monoacetone glucose (MAG). Subsequent 2H and 13C NMR analysis of MAG from normal volunteers after ingestion of 2H2O and [U-13C3]propionate allowed a noninvasive profiling of hepatic gluconeogenic pathways. The method should find use in metabolic studies of infants and other populations where blood sampling is either limited or problematic.

Energy expenditure and free-living physical activity in black and white women: comparison before and after weight loss.

BACKGROUND: The prevalence of obesity is higher in black than in white women. Differences in energy economy and physical activity may contribute to this difference. OBJECTIVE: The objective of this study was to compare free-living energy expenditure and physical activity in black and white women before and after weight loss. DESIGN: Participants were 18 white and 14 black women with body mass indexes (in kg/m(2)) between 27 and 30. Diet, without exercise, was used to achieve a weight loss of >/=10 kg and a body mass index <25. After 4 wk of energy balance in overweight and normal-weight states, body composition was assessed by using a 4-compartment model, sleeping and resting energy expenditures were assessed by using a chamber calorimeter, physiologic stress of exercise and exercise economy were measured by using standardized exercise tasks, and daily energy expenditure was assessed by using doubly labeled water. RESULTS: Weight loss averaged 12.8 kg. Sleeping and resting energy expenditures decreased in proportion to changes in body composition. Weight reduction significantly improved physiologic capacity for exercise in both groups of women, making it easier for them to be physically active. Black women had lower body composition-adjusted energy requirements than did white women-both before and after weight loss-during sleep (9% lower, 519 kJ/d; P < 0.001), at rest (14% lower, 879 kJ/d; P < 0.001), during exercise (6% lower; P < 0. 05), and as a daily total (9% lower, 862 kJ/d; P < 0.06). By contrast, free-living physical activity was similar between the groups. CONCLUSIONS: Weight-reduced women had metabolic rates appropriate for their body sizes. Black women had lower resting and nonresting energy requirements in both overweight and normal-weight states than did white women and did not compensate with greater physical activity, potentially predisposing them to greater weight regain.

Total body water in children with congenital heart disease, before and after cardiac surgery.

The aim of this study was to measure total body water in children with congenital heart disease before and after cardiac surgery and to compare the results of deuterium and 18oxygen dilution methods. Seventeen children (aged 4 to 33 months) were given aliquots of isotopically labeled water 1 week before and 6 hours after cardiac surgery. Isotope equilibration and analysis of the declining enrichment of daily urine samples allowed calculation of the total body water content. Before operation, total body water was significantly elevated (p < 0.001, Wilcoxon test); after operation it fell to approximately normal values. This finding is in contrast to those of previous reports, but may be explained in that the method used for calculation depended on measurements taken over a 7-day period rather than on a single measurement of isotope dilution as used elsewhere. Nevertheless, these results do suggest that surgery can correct the preoperative fluid overload. Comparison of deuterium and 18oxygen dilution methods showed a 2% to 2.5% overestimation of the total body water content with deuterium sampling.