BioSteel Supplementation and Performance

This study is expected to start in January 2014 and be completed by December 2014.

Ten exercise trained male athletes (18-35 years) will complete a repeated testing protocol under 2 experimental conditions in a systematically rotated blind crossover design: BSD and isoenergetic carbohydrate control, separated by 1 week. The testing protocol will consist of sprint interval exercise (4-repeated Wingate cycle tests, resistance at 10% body mass separated by 4 min recovery bouts, followed by 4 muscular fatigue and 2 cognitive function tests).

Approximately 1-2 weeks prior to testing, participants will complete two familiarization sessions in the laboratory. During the first session (~2h), participants will be asked to complete a physical activity readiness questionnaire and a participant information form for personal and familial health history to rule out any pathologies or health issues that may preclude them from participating in the present study. Additionally, their body composition will be determined via BodPod (air displacement chamber), and they will also be screened via 5 exercise tests used in National Hockey League Combine (150 lb bench press, 8 lb sitting medicine ball toss, standing long jump, vertical jump and maximal oxygen consumption test on a cycle ergometer). During the second session (~1h), they will familiarize themselves with the 2 cognitive tests. This testing is designed to eliminate any learning effect of repeat testing.

On test days (~5h), participants will arrive at the laboratory at ~0800h following a 12-h overnight fast, and with limited activity (drive/use of the elevator to get to the lab). Participants will be fed a standardized breakfast (7 kcal/kg body weight) and allowed 2 hours to digest prior to testing. Thirty minutes before exercise, participants will ingest either 500ml of BSD (0.18 calories/kg body weight) or an isoenergetic carbohydrate control of similar taste/appearance in a blind fashion. Participants will be given a 5-minute warm up, then complete a session of sprint interval exercise (4 repeated Wingate tests, separated by 4 minutes recovery). Following the exercise, 4 National Hockey League Combine tests will be completed to evaluate muscular fatigue (150 lb bench press, sitting medicine ball toss, standing long jump, and vertical jump), and 2 cognitive function tests. Participants will then receive a 15-minute non-exercise recovery where they will consume another 500ml of supplement (or placebo) and complete the testing protocol again. Ratings of perceived exertion (RPE) will be collected throughout the exercise and muscular fatigue tests.

Test day timeline:

0800h: Arrive at lab 0805h: Standard breakfast 1025h: Blood sample 1030h: Take supplement (double blind) 1100h: Blood sample 1105h: Warm up 1110h: Sprint interval exercise: post test RPE 1130h: Muscular fatigue tests: post test RPE 1150h: Cognitive function tests 1200h: Rest 1215h: Sprint interval exercise: post test RPE 1245h: Muscular fatigue tests: post test RPE 1305h: Cognitive function tests 1315h: Blood sample 1320h: Leave lab

During the experimental trial, participants will be able to drink water ad libidum (volume consumed will be quantified). The Gatorade condition will have a similar taste/appearance as BioSteel. Blood samples will be collected via venipuncture in the forearm at 3 time points: prior to supplement ingestion, 30 minutes post-ingestion/pre-exercise and immediately after the second testing protocol. Blood obtained will be analyzed for leucine, isoleucine, valine and tryptophan. All blood sampling will be done by a trained phlebotomist.

Eligibility Criteria: Individuals must be healthy, exercise-trained males aged 18-35 years. Participants will have never consumed BioSteel Sports Drink. To ensure participants are trained, they must achieve the 2013 National Hockey League Combine average scores or better on 5 exercise tests (150 lbs bench press, 4 kg sitting medicine ball toss, standing long jump, vertical jump and maximal oxygen consumption test on cycle ergometer). Women will be studied subsequently to asses whether gender differences exist.

Outcome Measures:

Blood Collection: Blood samples will be collected by venipuncture of an arm vein by a trained phlebotomist as with previous studies conducted in our lab. Briefly, participants will be sitting comfortably in a chair with an armrest for the specimen collection. In addition, subjects will be thoroughly briefed regarding the blood draws, and blankets, pillows, food and drink will be available in the event a subject reacts negatively. Standard sterile blood handling techniques will be used including disposable medical examination gloves, alcohol swabs, and needles. All blood contaminated materials will be disposed of in a hazardous material labeled disposal container, promptly after analysis. The specimens will be collected in EDTA-treated tubes, undergo treatment following ELISA kit protocols, centrifuged, and stored in a freezer at -70°C in the Exercise Nutrition Research Lab until hormone analysis in the same laboratory. The frozen specimens will remain stable for approximately 1 month. Therefore, the blood analyses will be completed within that time frame, after which they will be disposed of appropriately. The samples will be controlled and cared for by the principal investigator Dr. Peter Lemon and Kolten Abbott.

Body composition: Will be measured by densitometry using air displacement plethysmography via a BodPod® (involves sitting comfortably in a chamber for about five minutes while the space the body takes up is measured).

Aerobic Capacity: Will be quantified via an incremental exercise cycle ergometer test. This test is conducted routinely in our laboratory and will follow American College of Sport Medicine guidelines (Gold standard).

Wingate test: Will be conducted using a cycle ergometer which electronically measures power (average, peak and decline). Resistance will be equal to 10% of subject body mass in kg.

Muscular power and endurance: Four measurements will be conducted in accordance to the National Hockey League Combine guidelines (Gledhill \& Jamnik, 2007).

1. Bench press repetitions of 150 lb (including the barbell) will be performed at a rate of 25 repetitions per minute, the number of consecutive repetitions until the individual can not keep up the cadence will be recorded.
2. Sitting medicine ball toss using a standard 8 lb medicine ball. Subject will sit on the floor with their back flat against the wall and legs straight out (hips flexed and the knees extended). The ball will start at the chest and be pushed out. When the ball first lands will be measured as the total distance (recorded in cm.)
3. Standing long jump will be conducted from a standing start with feet together. The distance from the jumping line to the individuals heels will recorded as the total distance (recorded in cm.)
4. Vertical jump will be conducted from a standing start. Fingers outstretched on the dominant arm will reach as a high as possible deflecting a plastic marker. Without a pre-step individual will jump as a high as possible. Measurement is to the nearest cm.

Cognitive function: Two tests will be conducted on a computer located within the Exercise Nutrition Research Laboratory. Subjects will receive 5 minutes to complete each cognitive test.

1. Modified Stroop test- a measurement of attentional fatigue and executive function, will be scored based on error rate and mean response time (Kane \& Kay, 1992).
2. Trail Making Test Part B- a measurement of visual attention and speed of processing, will be scored on total time to completion (Sanchez-Cubillo et al., 2009).

Ratings of perceived exertion: Will be assessed after each sprint interval trial and at the conclusion of muscular endurance and power testing via a Borg Scale (Noble et al., 1983).