This study was Momelotinib order conducted to determine whether betaine is a component selleckchem of sweat that may be lost from the body during exercise. Methods Subjects Eight trained female Scottish Highland dancers (10-17 yr) were recruited from the Stirling Highland Dance Company, Oakdale CT. The subjects trained regularly, and were actively competing in dance competitions. Subjects attended a briefing meeting before any experimentation
to ensure an understanding of the testing parameters and the benefits/risks of the study. The subjects and parents signed a written informed consent statement. The study was part of the Somers High School (SHS) Science Research Program and the protocol was approved by the SHS IRB. Experimental Protocol Sweat patches were prepared by placing two 2″” × 2″” gauze squares onto 4″” × 4.5″” adhesive film. Care was taken to minimize any cross-contamination. New disposable latex gloves were utilized for each subject. The
skin on the lower back of the subjects was cleaned with gauze and distilled water, dried, and two patches were placed on both sides of the spine. The dancers then conducted a 2 hour class. The sweat patches were removed, placed in plastic 6-ml centrifuge tubes and stored on ice prior to centrifugation. The tubes were spun for 2 min at 1315 g in a benchtop centrifuge (Model 0151; Clay Adams, Parsippany, NJ). The patches were removed from the tubes, and the sweat (1-2 ml) at the bottom of the tubes was recovered. Each subject had two tubes from the two patches. The EPZ015938 in vitro sweat from the two tubes was combined and stored frozen at -20°C prior to analysis. Measurements Betaine, choline, and choline metabolites were determined in duplicate by liquid chromatography/electrospray ionization-isotope dilution mass spectrometry [22]. Lactate and glucose were determined in duplicate by enzymatic techniques (YSI 2300 Stat Plus, Yellow Springs, OH). Sodium, potassium and chloride were measured in duplicate using ion selective electrodes (Medica Easy Electrolytes, Medica Corp., Bedford,
MA). Urea and ammonia were check details measured using a COBAS Mira Plus Analyzer (Roche Diagnostics, Indianapolis, IN) and Pointe Scientific (Canton, MI) reagent sets and standards. Instruments were calibrated using NIST certified standards. Statistics Grubbs’ test http://graphpad.com/quickcalcs/Grubbs1.cfm was used to determine outliers in data sets (alpha = 0.05). Pearson’s correlation test (SigmaPlot v11, Systat Software Inc, San Jose, CA) and Passing-Bablok regression analysis (MedCalc, Mariakerke, Belgium) were conducted to compare data sets. Results The measures of sweat composition are shown in Table 1. Phosphatidylcholine and sphingomyelin were also measured, but were not detected (data not shown). The mean betaine content was 232 ± 84 μmol·L-1. The other components of sweat were found at levels similar to that of previous studies [18, 19, 21].