Dc 320

All the subjects visited the laboratory prior to the experiment to become accustomed to the experimental instruments and the cognitive function tests and to reduce learning effects. At Pre, Post, and FU, the subjects visited the laboratory between 12:30 and 16:30, after having a prescribed lunch (200–400 kcal Calorie Mate, Otsuka Pharmaceutical, Japan) and bottle of noncaffeinated barley tea (350 mL). Subjects did not eat chocolate for at least 20 h prior. After urinating, the subjects’ weight and body fat were measured by bioelectrical impedance analysis (DC-320; Tanita, Tokyo, Japan). The subjects then entered an artificial climate chamber that was sound attenuated and controlled to an environmental temperature of 25 °C and a relative humidity of 50%. Subjects rested quietly in a relaxing sitting position for 30 min while the experimental instruments were attached. We measured the systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) using an automatic digital blood pressure monitor (HEM-907; Omron, Kyoto, Japan). Next, the subjects underwent the cognitive function tests (modified SCWT and D-CAT), and their PFCBF responses were measured during the cognitive function tests. The cognitive function tests and PFCBF measurements were independently performed for individuals in the same experimental conditions and were measured by same examiners.

The participants’ height (determined with the DST-210N; Muratec-KDS Corp., Kyoto, Japan) and weight, body fat percentage (determined with the DC-320; Tanita Corp., Tokyo, Japan) were measured after shoes were removed, and then we calculated BMI. A Smedley type dynamometer was used to measure grip strength, which represented the muscle strength^{25 (link),26 (link)}. All measurements were performed twice for each hand, and the highest result for each hand was used to calculate the mean grip strength.

Body height (BH) and mass (BM) were measured using a stadiometer and a leg-to-leg bioelectrical impedance analyzer with a computer-programmed athletic mode (DC-320, TANITA, Japan) to the nearest 0.1 cm and 0.1 kg, respectively. Participants were instructed to restrain from alcohol intake for 24 h prior to the experiment and from having a meal 2 h prior to the measurement. Waist circumference (W) was measured to the nearest 0.1 cm with a cloth tape (Rotary measure, Sodeyama Co., Ltd. Japan) immediately above the iliac crest [15 (link)]. To confirm the reproducibility of the waist circumference measurement, we measured twice at least three days apart for nine male athletes. The intra-class correlation coefficient (ICC) for the measurement was 0.97. The measurement error between 1st and 2nd measurements was 0.0 ± 1.1 cm, and the coefficient of variance (CV) was 0.9 ± 0.6%.

Height, body weight, body fat ratio (dc-320 Tanita, Tokyo, Japan), systolic blood pressure, diastolic blood pressure and pulse rate (H55 TERUMO CORPORATION, Tokyo, Japan) were measured at the screening test. BMI was calculated from height and body weight.

Body weight and %FM were determined using a bioelectrical impedance analyzer (BIA) (DC-320, Tanita Corp., Tokyo, Japan) [11 (link)]. Subjects wore indoor clothing with no shoes. Half a kilogram (0.5 kg) was deducted from the weight of each participant to account for the weight of clothing. The %FM was calculated as fat mass/weight × 100, and BMI was calculated as the weight (kg)/height (m)². The study defined +1 kg difference from the pre-pregnancy weight as a return to the pre-pregnancy weight. Subjects who returned to the pre-pregnancy weight within a year of delivery were categorized as the weight loss group, and those who did not were categorized as the no weight loss group.
NWO was defined as a BMI of 18.5–25 kg/m² and a %FM of ≥30 %. We defined “pre-NWO” as a BMI of 18.5–25 kg/m² and a %FM of 25–30 %.

The speed of sound (SOS), broadband ultrasound attenuation (BUA), and stiffness index of the calcaneus (SI) were measured using an ultrasound bone densitometer (Achilles InSight; GE Healthcare, Little Chalfont, UK). I used an ultrasound bone densitometer because it has no side effects and correlates well with the dual-energy X-ray absorptiometry (DXA)-measured bone mineral density (BMD) [19 –24 ] or bone mineral contents (BMC) [24 ] as well as quantitative computed tomography (QCT) [25 (link)]. The densitometer was operated by a researcher experienced with taking measurements, and the final intra-sample mean coefficient of variation for the stiffness index was 0.7%.
The subjects’ height (determined with a DST-210 N; Muratec-KDS Corp., Kyoto, Japan) and weight, body fat percentage, and muscle mass (determined with a DC-320; Tanita Corp., Tokyo, Japan) were measured after shoes had been removed, and then the body mass index (BMI) was calculated.

The questionnaire of the OHA-Q ver. 2 was completed by participants in private, to avoid any influence by physicians and medical care providers. The indices of body composition were measured using Tanita DC320 (Tokyo, Japan). The biochemical markers were measured using fasting blood samples and urine samples collected in hospital at baseline and at week 14.