Blueberry-protein Supplementation and Exercise
Condition: Inflammatory Response
Study type: Interventional

Summary: 
      This study is the human clinical trial component of an SBIR grant with Sinnovatek (funded).
      Cyclists (n=20) will consume a blueberry-protein, blueberry, or placebo supplement for 2
      weeks prior to cycling for 2.5 h at high intensity. Washout periods (2 weeks) will separate
      the 3 trials, and the cyclists will crossover (randomized) to one of the two others
      supplements for 2 weeks, and then engage in additional 2.5 h cycling bouts. Blood and urine
      samples will be collected pre- and post-supplementation for each of the 3 trials. Blood
      samples will also be collected immediately after and 1.5h- and 24h-post-exercise. Blood
      samples will be analyzed for inflammation and oxidative stress outcomes. Urine samples will
      be analyzed for blueberry gut-derived phenolics.
    

Detailed description: 
      PURPOSE: To investigate the influence of 2-weeks blueberry, blueberry-protein, versus placebo
      gel supplementation (randomized, crossover) on oxinflammation markers and blueberry
      gut-derived phenolics after 2.5 h intensive cycling exercise in 20 subjects.

      HYPOTHESIS: We hypothesize that 2-weeks blueberry or blueberry-protein polyphenol (500
      mg/day) versus placebo gel supplementation will attenuate post-exercise increases in
      inflammation and oxidative stress biomarkers.

      RESEARCH PROCEDURES The research procedures will be conducted at the Human Performance
      Laboratory (Room 1201, Plants for Human Health Institute Building, 600 Laureate Way),
      operated by Appalachian State University at the North Carolina Research Campus (NCRC) in
      Kannapolis, NC. Subjects will come to the lab for orientation/baseline testing with
      pre-supplementation blood sample collections, three 2.5 h cycling sessions, and additional
      lab visits to provide 1-day 7:00 am recovery blood samples) (thus 9 total lab visits). The
      total amount of time subjects will be asked to volunteer for this study is about 20 hours at
      the Human Performance Laboratory (over a 10-week period).

      Orientation/Baseline Testing (Lab Visit #1) Two weeks prior to the 2.5 h cycling session,
      subjects will report to the Human Performance Laboratory at approximately 7:00-8:00 am. A
      blood sample will be collected in an overnight fasted state to coincide with the same time of
      the day for the post-supplementation blood draw. Study participants will be tested for
      maximal aerobic capacity (VO2max) during a graded, cycle ergometer test with the Cosmed CPET
      metabolic device (Cosmed, Rome, Italy). Body composition will be measured with the seca BIA
      and Bod Pod body composition analyzer (Life Measurement, Concord, CA). Demographic and
      training histories will be acquired with questionnaires. Gel blueberry, blueberry protein, or
      placebo supplements will be supplied. 3-day food records will be supplied with thorough
      instructions. Participants will be given two 24-h urine collection kits with thorough written
      and oral instruction on collection procedures. The first 24-h urine collection bottle will be
      collected during the 24-h period immediately after lab visit #1 and turned in the next
      morning. Supplementation will start right after the urine is turned in. The second 24-h urine
      collection bottle will be collected during the 24-h period immediately prior to lab visit #2.
      Subjects will collect every drop of urine in the collection bottle for 24 hours.

      2-Week Supplementation Periods Study participants will be randomized to Vitero
      (rice-pea-blueberry) gel, blueberry gel, and placebo gel trials, with supplements ingested
      daily for 2-weeks prior to participation in the first 2.5 h cycling session. After a 2-week
      washout period, participants will repeat all procedures two more times using the
      counterbalanced supplements.

      Vitero (500 mg total blueberry polyphenols per day) or placebo (containing all proximate
      matched ingredients except for the phytoactive polyphenolics) gels will be consumed with
      breakfast and lunch. The gels will look identical. The placebo has been formulated in house
      (NCSU at the Plants for Human Health Institute).

      2.5 h Cycling Session (Lab Visit #2) During the 3-day period prior to each of three the 2.5 h
      cycling sessions, subjects will taper exercise training and ingest a moderate-carbohydrate
      diet using a food list restricting high fat foods, visible fats, and polyphenols. Subjects
      will record all food and beverage intake during the 3-day period, with macro- and
      micro-nutrient intake assessed using the Food Processor dietary analysis software system
      (ESHA Research, Salem, OR).

      Study participants will report to the Human Performance Lab in an overnight fasted state and
      provide a blood sample, ingest the rice-pea-blueberry, blueberry, or placebo gel with water,
      and then cycle 2.5h at high intensity (60% watts max) while ingesting water alone (3 ml/kg
      every 15 minutes). Blood samples will be collected at 0 h, 1.5 h, and 21 h post-exercise.

      Testing protocol during the lab sessions with the 2.5 h cycling session:

        1. 7:00 am: Provide blood sample and delayed onset of muscle soreness (DOMS) rating.

        2. 7:10 am: Ingest supplements with water.

        3. 7:30 am: Start 2.5 h cycling session. Subjects are to complete the 2.5 h session at an
           average of 60% watts max. Subjects will use their own bicycles on Saris trainers with
           Zwift software. A course with moderate difficulty will be utilized using the Zwift
           software. Heart rate and rating of perceived exertion (RPE) will be recorded every 30
           minutes, and workload (watts) continuously monitored using the Zwift software system.
           Oxygen consumption, carbon dioxide production, respiratory exchange ratio, and
           ventilation will be measured using the Cosmed Quark CPET metabolic cart every 30
           minutes. The total distance cycled in 2.5 h will be recorded. Subjects will consume 3
           ml/kg water every 15 min. No other beverage or food containing energy or nutrients will
           be allowed during the cycling time sessions.

        4. ~10:00 am to 11:30 am: Immediate-post-exercise, and 1.5-h recovery blood samples. Blood
           samples will be taken via venipuncture immediately after completing the cycling session,
           and then 1.5-h post-exercise. Subjects will be allowed to shower and change clothes.
           DOMS questionnaire will be administered immediately post-exercise and 1.5-h
           post-exercise. Subjects will ingest no food or beverage other than water (7 ml/kg during
           the 1.5 h post-exercise period). Subjects will ingest the supplement gels, and then be
           allowed to leave the lab. Subjects will be instructed to adhere to the food list
           requirements, and to keep exercise training intensity moderate.

        5. 7:00 am, next morning: 21-h blood sample and DOMS rating (Lab visit #3).

      Washout/Crossover/Repeat Subjects will engage in a 2-week washout period without supplements,
      crossover, and then repeat all procedures (Lab visits #4,5,6, and then #7,8,9).
    

Intervention name(s): Blueberry-protein gel; Blueberry gel; Placebo
Intervention type(s): Dietary Supplement; Dietary Supplement; Dietary Supplement
Intervention description(s): Sport gel with blueberry extract and plant protein.; Sport gel with blueberry extract only.; Sport gel without the blueberry extract polyphenols and without protein.
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: Randomized treatment, double blinded procedures, crossover
Masking: Double (Participant, Investigator)
Primary purpose: Prevention
Keywords: blueberries; exercise; inflammation