2.4. Assessment Procedures
dehydration, hand dynamometer, dual-energy X-ray absorptiometry
DEHYDRATION, OXIDATIVE STRESS
All assessments were performed by a physician, radiologist, nurse, dietitian, and ten physical therapists. The assessments before and after the intervention lasted for approximately 60 min. They were conducted at the Mihara Campus of Prefectural University of Hiroshima using a standardized protocol by the same researcher.All assessments were conducted between 09:00 and 15:00 in the same laboratories. The participants were instructed to fast for 2 h before the assessments. The consumption of Japanese tea was allowed before and during the assessments to prevent dehydration. Upon arrival, the participants were instructed to sit. Next, their blood pressure and heart rate were evaluated and their health was examined by the physician. Their body mass was measured at increments of 0.1 kg using a calibrated digital scale and their height was evaluated at increments of 0.1 cm using a wall-mounted height meter. Body mass index (BMI) was calculated as weight (kg) divided by squared height (mWhile in the sitting position, the nurse collected blood samples from a superficial vein in the forearm of all patients via venipuncture. Approximately 4 mL blood was collected and placed into prechilled silicone-coated and lithium heparin-coated tubes. A portion of the blood sample was used to measure the level of diacron-reactive oxygen metabolites (d-ROMs), which is an indicator of blood oxidative stress, using the FREE Carrio Duo (WISMERLL Co., Tokyo, Japan) as per the manufacturer’s instructions. The level of d-ROMs was expressed in Carratelli units (U.CARR) [Body composition and bone mineral density (BMD) were assessed by a radiologist using Discovery (Hologic, Inc., Bedford, MA, USA) with dual-energy X-ray absorptiometry (DXA), which is a highly accurate method [To evaluate hand grip and knee extension strength, which are significantly correlated with the total body skeletal muscle mass, skeletal muscle function was assessed by physical therapists. The hand grip strength of the dominant hand was analyzed to the nearest 0.5 kg using a hydraulic hand dynamometer (T.K.K. 5401; Takei Scientific Instruments Co., Ltd., Niigata, Japan) while the participants remained in a standing position and the upper limbs remained in a drooping position. Evaluations were conducted twice at intervals of 1 min, and the best score was recorded as the hand grip strength. The knee extension strength of the dominant leg was evaluated using a seated measuring device (Isoforce GT-330, OG Giken, Okayama, Japan). The measurements were conducted while the participants were sitting in a chair with the backrest tilted back at 12° from upright position, with two belts on the thighs and one on the waist, and with the knee joint flexed at 90°. Measurements were conducted at intervals of 2 min, and the best score was recorded as the knee extension strength. During the measurements of hand grip and knee extension, the participants were instructed not to stop breathing to prevent the occurrence of the Valsalva effect.