Cross-validation of cut-points in preschool children using different accelerometer placements and data axes

The present study cross-validated various cut-points to assess physical activity and sedentary behaviour in preschoolers, using hip- and wrist-worn accelerometers and both vertical axis and vector magnitude data. Secondly, we examined the influence of epoch length on time estimates of physical activity and sedentary behaviour. Sixty-four preschoolers (34 girls) wore two accelerometers, on their right hip and dominant wrist, during 1 hour of free play. Preschoolers’ activities were observed by two trained researchers. Area under the curve (AUC) was calculated for the receiving operating characteristic (ROC) curves as a measure of precision. AUC ranges were 0.603–0.723 for sedentary behaviour, 0.472–0.545 for light physical activity and 0.503–0.661 for moderate-to-vigorous physical activity (MVPA), indicating poor to fair precision. Percentage of time classified as sedentary behaviour, light or MVPA according to observation and accelerometer data varied largely between cut-points, accelerometer placements and axes. The influence of epoch length on time estimates was minimal across cut-points, except for one hip-based vector magnitude cut-point. Across all accelerometer placements and data axes, no set of cut-points demonstrated adequate precision for sedentary behaviour, light physical activity and MVPA. The highly variable and omnidirectional activity pattern of preschoolers may explain the lack of adequate cut-points

Rapid genotyping of the OATP1B1 polymorphisms A388G and T521C with real-time PCR FRET assays

The polymorphisms (OATP)1B1 A388G and T521C of the solute carrier organic anion-transporter family member 1B1 gene (SLCO1B1), previously known as OATP-C, have potential impacts on drug metabolism. In order to establish a fast and consistent assay for these polymorphisms, rapid speed polymerase chain reaction (PCR) fluorescence resonance energy transfer (FRET) assays on the LightCycler(R) were developed for both OATP1B1 polymorphisms. A locked nucleic acid (LNA) on the polymorphic location within the sensor probe was necessary to discriminate both alleles of the OATP1B1 T521C polymorphism. To confirm the reliability of both real-time PCR FRET assays, these new methods were validated by genotyping 120 samples using a PCR restriction fragment length polymorphism (RFLP) assay and an allele-specific PCR. The results of the real-time PCR FRET assays were completely in line with conventional PCR methods, indicating that the real-time PCR FRET assays are appropriate for clinical settings