18 × weight), which has been shown to be the best equation for normal-weighted women undergoing energy restriction [12, 13]. Energy consumed in physical activity was estimated selleck chemicals llc using the internet application EnergyNet (University of Kuopio). The total energy need was 2340 ± 170 kcal for 1 KG and 2290 ± 120 kcal for 0.5 KG. Energy deficit and diet for each subject during the weight reduction period was then evaluated. The group 1 KG (energy deficit 1100 kcal/day, protein at least 1.4 g/kg/day) was supervised to reduce body weight by 1 kg per week and the group

0.5 KG (energy deficit 550 kcal/day, protein goal at least 1.4 g/kg/day) by 0.5 kg per week during the next four weeks, respectively. The subjects kept food diaries for two days each week and the researchers could then, with the diaries and with morning scale body weights, supervise that body weight was reducing as planned. All subjects were advised to continue their normal recreational find more resistance training and aerobic training during the weight reduction period which was also controlled each week. Measurements Body composition Body scale weight RG-7388 cost was determined in the familiarization

session, in the before and after measurements and in every week control with the same electric digital scale. Total body composition was determined using a dual-energy X-ray absorptiometry device (DXA; Lunar Prodigy Densitometer, GE Lunar Corporation, Madison, WI, USA). This method can differentiate bone mineral density (BMD), total percentage fat, total body tissue mass, fat mass, lean

mass, bone mineral content (BMC), and total bone calcium with precision errors of 0.62, 1.89, 0.63, 2.0, 1.11, 1.10, and 1.09%, respectively [14]. Blood sampling and hormone analysis Blood samples were drawn from the antecubital vein for analyze of hemoglobin, serum total testosterone, sex-hormone-binding globulin (SHBG), cortisol and dehydro-epiandrosterone sulfate (DHEAS) and pH were drawn on the morning of both measurement days after a 12 h fast. The intervention time interval was exactly 4 weeks for everyone so the menstrual cycle was in the same phase. The samples were taken in the sitting position two times with 30 minutes in between Immune system measurements. Serum samples were kept frozen at -80°C until assayed. Two milliliters of blood was taken in K2 EDTA tubes (Terumo Medical Co., Leuven, Belgium) for measurements of hemoglobin concentration with a Sysmex KX 21N Analyzer (Sysmex Co., Kobe, Japan). The intra-assay coefficient of variation (CV) is 1.5% for hemoglobin. For the determination of serum hormone concentrations five milliliters of blood was taken and the concentrations were analyzed by an immunometric chemiluminescence method with Immulite® 1000 (DPC, Los Angeles, USA). The sensitivity of the assay for serum testosterone is 0.5 nmol/l, for SHBG 0.2 nmol/l, for cortisol 5.5 nmol/l and for DHEAS 0.08 μmol/l. Coefficient of variations are 8.