The D-BIICEP Study

During this study the subjects will be asked to visit the labs on 4 different occasions. There will be two occasions, at the beginning of the study for baseline measurements, and again on two occasions at the end of the study.

Data collection will take place across two testing sessions (separated by ~24 - 72 hrs) in the Autumn and Spring period.

Session 1

• Peripheral Quantitative Computed Tomography (pQCT) scan of the bone mineral composition of the tibia.
• Total and percentage lean and fat mass and total bone mineral density via Dual Energy X-ray Absorptiometry (DEXA).
• Sport performance: Vertical jump height and maximal oxygen uptake (VO2 max) test.

Session 2

• Fasting serum 25(OH)D levels, lipid profile, glucose, insulin, serum calcium, albumin, parathyroid hormone, C- terminal telopeptide (CTX), full blood count, kidney, thyroid and liver function (≈15ml of whole blood will be collected for these measurements).
• Collection of self-reported food diaries to assess dietary intake
• Sport performance: muscular strength

Aerobic fitness will be tested using a VO2 max exercise protocol using a stationary cycle ergometer, the participants will be expected to perform this to maximal effort or exhaustion. The test will consist of progressive increments in cycling workload (power output, W) until volitional fatigue. The test will take about 8 - 12 minutes to complete and is dependent upon the fitness of the participants. This will take place in the Sport \& Exercise Science laboratories based in the Clinical Investigation Unit (22AX00). All subjects will be supervised during exercise at all times and will implement an adequate warm-up and cool-down to minimize the risk of injury. Staff will be on hand throughout the testing trained adequately in basic life support (including defibrillation).

Muscular strength assessments will be performed at the Surrey Human Performance Institute. Muscular strength of the knee extensor and handgrip muscles will be determined using an Isokinetic Dynamometer and a handgrip dynamometer respectively. For each assessment three maximal effort isometric contractions will be performed and peak torque (nM) and strength (kg) will be recorded. Muscular power will be assessed through performance of a countermovement jump (CMJ).

Body composition (absolute and relative amount of lean and fat mass), whole body bone mineral density and hip and femur bone mineral density will be measured with the use of DEXA (located in the Sport \& Exercise Science laboratories based in the Clinical Investigation Unit). Two scans will be performed: one for the assessment of whole body bone mineral density and body composition, and the other scan is performed to specifically assess fracture risk by scanning the hip and femoral head. Effective exposure doses for these scans are ~8uSv and ~4uSv respectively. The total effective dose is depending on the size ('thickness' \& height) of the volunteer and is approximately equivalent to the cosmic radiation exposure received in 45min of a transatlantic flight, or the radiation dose received on eating 80g of Brazil nuts (Health Protection Agency).

Bone mineral content of the tibia will be assessed using pQCT and this will be correlated to bone mineral density data obtained from DEXA. One scan will be performed and effective exposure doses will be between 1.5-1.8uSv which is comparable to 10 min on a transatlantic flight or one handful of Brazil nuts. The estimated total exposure dose for a single subject during the entire study is ~28uSv which is the same level of exposure as an average UK citizen is exposed to over the course of 4 days via background radiation.

Results of the body composition, bone mineral density, jump height, muscular strength, aerobic fitness (maximal oxygen uptake: VO2max) and dietary intake from the self-reported food diaries will be made available to the subjects upon request. The results from the blood analysis will be reported to the subject if there are any health concerns raised. If the results are within healthy ranges the participants will not be contacted unless specifically requested for this information. The investigators will not be contacting their GPs if there are any concerns raised in the study however it will be stressed that participants should contact a GP to discuss the results found.

Upon commencing the study, the participants will be required to track their incidence of illness and training habits during the 20-week experimental period between baseline and conclusion measurements. Throughout this period the incidence of illness and days trained during the months must be recorded in a check-list booklet, which will be provided. An email reminder will be sent to prompt all participants to complete these booklets on a daily basis during the experimental period, this should only take 1-2 minutes every evening. Throughout the duration of the trial, the participants will be contacted via telephone/ email on a monthly basis to discuss any issues and maintain good communication.