Effect of Resveratrol Supplementation on Exercise Performance and Recovery

This study will utilize a randomized counter balanced cross-over repeated measures design. Participants will complete a total of 13 visits to the Exercise Physiology Intervention and Collaboration (EPIC) lab.

6.2 Procedures of each individual visit: Visit 1 (V1): will consist of informed consent and eligibility screening. Individuals who have provided written informed consent to participate will be screened using the Physical Activity Readiness Questionnaire (PAR-Q+), Medical History Questionnaire (MHQ), and a Caffeine Consumption Questionnaire (CCQ) to determine eligibility to participate in the study. Eligible participants will also be examined by a certified phlebotomist to determine the likelihood of successful venipuncture. Eligible participants will subsequently be randomized in a counter balanced fashion into two treatment groups (n=10/group) corresponding to either Verisperse® Resveratrol (VRES) or Placebo (PLA) using an online randomization tool. Participants will also select their snack choice from a list of pre-approved options. Following this, participants will be scheduled for visit 2 (V2), which will occur at least 24 hours following V1.

Visit 2 (V2): Participants will arrive for V2 in a fasted state (≥8 hours) having abstained from alcohol for at least 12 hours prior. Upon arrival, participants will complete a hydration assessment by urinating into a provided sample cup no more than 4oz. Participants will then provide a venous blood sample to determine baseline systemic concentrations of creatine kinase and C-reactive protein. Next, participants will complete anthropometric assessments of height, weight, and body composition via bioelectrical impendence analysis (BIA). Participants will be asked to remove shoes and all jewelry for BIA. There are no risks or discomfort associated with the bioelectrical impendence analysis. Next, participants will be rested in a quiet, dimly lit laboratory in a semi-recumbent position for 15 minutes before completing the flow-mediated dilation (FMD) assessment. Participants will then be provided with their standardized snack of choice (energy bar) before undergoing familiarization with the active range of motion (AROM), pain pressure threshold (PPT), muscle soreness (MS) perceived recovery scale (PRS), maximal voluntary isometric contraction (MVIC), and vertical jump (VJ) assessments and the isoinertial squat protocol (ISP) (3 sets of 6 repetitions at a resistance of .067kg/mm2). Following the ISP, participants will be familiarized with the online MyFitnessPal application.

Visits 3-5 (V3-V5): Upon arrival for V3-V5, participants will undergo a standardized warm-up identical to V2 before completing additional familiarization with the ISP. All procedures for V3-V5 will be identical. V2-V5 will be separated by at least 24-hours but no more than 72 hours.

Visit 6 (V6): Upon arrival to the lab participants will fill out a sleep quality questionnaire (SQQ), provide a urine sample to assess hydration status, and provide a pre-exercise venous blood sample to establish baseline measures of creatine kinase (CK) and C-reactive protein (CRP). Next, participants will complete PRE ultrasound assessments of muscle thickness (MT) and echo intensity (EI) followed by the FMD assessment. Participants will then be provided with the standardized snack (energy bar) of their choice and complete a standardized warmup identical to V2, before completing PRE assessments of AROM, PPT, MS, PRS, MVIC, and VJ. Participants will then be fitted with a heart rate monitor and complete the ISP followed by a 3-minute cycle-based cool down period, a venous blood sample, and follow-up assessments of MT, EI, AROM, PPT, MS, PRS, MVIC, and VJ immediately post-exercise (IP). MVIC and VJ will be separated by 5 minutes to allow for adequate recovery.

Visits 7-9 (V7-V9): V7-V9 will be identical in nature and will occur 24, 48, and 72 hours following V6, respectively. Upon arrival to the lab for each experimental trial (V7-V9) participants will fill out a sleep quality questionnaire (SQQ), provide a urine sample to assess hydration status, and provide a pre-exercise venous blood sample to establish baseline measures of creatine kinase (CK) and C-reactive protein (CRP). Next, participants will complete ultrasound assessments of muscle thickness (MT) and echo intensity (EI) followed by the FMD assessment. Participants will then complete a standardized warmup identical to V2, before completing assessments of AROM, PPT, MS, PRS, MVIC, and VJ. MVIC and VJ will be separated by 5 minutes to allow for adequate recovery.

Visit 10 (V10): V10 will occur 32 days after V9 and be conducted in an identical fashion to V6.

Visits 11-13 (V11-V13): V11-V13 will occur 24, 48, and 72 hours following V10, respectively. V11-V13 will be conducted in an identical fashion to V7-V9)

Consent and Screening Prior to beginning the study, individuals interested in participating in the study will be asked to report to the EPIC lab to review and sign an informed consent document. Participants who provide consent to participate will then be screened for eligibility via the PAR-Q+, MHQ, and CCQ. Participants will also be asked to fill out a contact form that will record their preferred email and cell phone number.

Hydration Status Assessment Upon arriving to the lab, participants will be given a sample cup to take to the restroom and provide a urine sample, so that hydration may be assessed. After providing a urine sample, a member of the research team will measure urine specific gravity (USG) using a refractometer to ensure participants are euhydrated (1.002-1.025). If dehydrated (\>1.025), they will be asked to consume 16 oz of water and be retested after waiting 15 minutes post ingestion. If hyperhydrated (\<1.002) their visits will be rescheduled. Hydration status will be checked immediately upon arrival to the laboratory for V2 and each experimental trial (V6-V13). Urine samples will be disposed of according to UCF policy.

Venous Blood Sample Collection Blood samples at V2, PRE-, IP, 24H, 48H, and 72H will be obtained using a 21-gauge, 1¼ inch vacutainer blood collection needle or 23-gauge butterfly needle taken from a superficial forearm vein. A maximum of 2 unsuccessful attempts to insert a needle in one of the participant's arms will be made and, with their permission, a third and final attempt will be made in the opposite arm. Three unsuccessful venipuncture attempts will result in termination of the blood draw. Issues with blood collection devices that prevent the drawing of blood are not grounds for removal from the study and will not affect participants continuation with the remainder of the study. All blood draws will be conducted under aseptic conditions. To minimize the risks, the skin area where the needle is inserted will be cleaned and prepared with a disinfectant alcohol wipe before the needle is inserted. Following the blood draw, the puncture site will be covered with a bandage.

A total of 8mL (slightly less than 2 tsp) of blood will be collected at each time point (V2 \& V6-V13) in one 4mL BD Vacutainer™ Serum separation tube (SST) and one 4mL K2EDTA plasma tube (EDTA).

Blood samples collected in the 4mL SST tube will be used to determine serum concentrations of C-reactive protein (CRP) and creatine kinase (CK). Serum tubes will be inverted ten times and allowed to clot at room temperature for 30-minutes prior to centrifugation for 10-minutes at 4000rpm. After centrifugation, serum will be aliquoted and frozen at -80°C within one hour of collection.

Blood samples collected in EDTA tube will be gently inverted ten times and left at room temperature for same day analysis. EDTA whole blood samples will be analyzed directly in-house using an automated hematology analyzer (Sysmex XN-450 Automated Hematology Analyzer) located in the Cellular Exercise Physiology Laboratory (CEPL) in BIO 224, Building 20 (Biological Sciences). Sample analytes will include hematocrit, hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and total (white blood cell and red blood cell) and differential (lymphocyte, monocyte, and granulocyte, platelet) cell counts.

Standardized Warm-up Prior to all performance assessments, participants will be required to complete a dynamic warm-up. Participants will begin by pedaling on a cycle ergometer for 5-minutes at a self-selected pace at a set resistance of 75 watts. Participants will then complete ten body-weight squats, ten body-weight walking lunges, ten dynamic straight leg kicks, and ten dynamic walking quadricep stretches.

Sleep Quality Questionnaire Participants will be asked to fill out a brief 7 question sleep questionnaire consisting of questions pertaining to their sleep and sleep quality within the last 24 hours. Participants will complete the SQQ immediately upon arriving for each experimental trial (V6-V13) Diet and Dietary Log Participants will be asked to maintain two separate 5-day dietary logs using the "MyFitnessPal" application on their own cellular or computer device. Participants will be asked to begin each 5-day log 24 hours before ET1 and ET2 and to maintain the log by tracking all food and beverage consumption to the best of their ability for the 5 days until the completion of the 72H follow up visits (visits 9 \& 13, respectively). Additionally, participants will be asked to keep their food and beverage consumption during the two 5-day periods as consistent as possible and to match macronutrient (fats, carbohydrates, and proteins) consumption between the two periods to the best of their ability. Participants refusing to voluntarily report food and beverage consumption will be withdrawn from the study.

Assessments Anthropometric Assessments: During V2, participants will complete anthropometric assessments of height, weight, and body composition. Height and weight will be assessed using a stadiometer and scale (Health-o-meter Professional Patient Weighing Scale, Model 500 KL, Pelstar, Alsip, IL, USA). Body composition will be assessed using bioelectrical impedance analysis (BIA; InBody 770, Biospace Co, Ltd. Seoul, Korea) or bioelectrical impedance spectroscopy analyzer (Sozo; Carlsbad, CA, USA). Participants will be asked to void their bladder, and remove shoes, socks, and all jewelry for this assessment. They will also be asked to be eight hours fasted and well-hydrated as determined by USG analysis. There are no risks or discomforts associated with the BIA analysis.

Flow Mediated Dilation (FMD) Standardization of FMD procedures and protocol will be conducted following established guidelines (40). Prior to testing, subjects will be rested in a quiet, dimly lit laboratory in a semi-recumbent position for 15 minutes. Automated brachial artery sphygmomanometry (IntelliSense® Professional Monitor, OMRON Healthcare Inc., Lake Forest, IL) will be used to measure resting systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) on the left arm. Mean arterial pressure (MAP) will be calculated to determine peripheral vascular resistance using the following equation: {MAP=\[0.33×(SBP-DBP)\]+DBP}. Brachial artery diameter and blood velocity will be measured by duplex, high-resolution Doppler ultrasound (GE Logiq P9, GE Healthcare, Chicago, IL) using a (5)-MHz linear array transducer. An arm tourniquet cuff with rapid inflation system (Hokanson E20; D.E. Hokanson Inc., Bellevue, WA) will be placed around the right forearm, immediately distal to the antecubital fossa for assessment of reactive hyperemia and brachial artery FMD. Stable imaging of the brachial artery will be accomplished using foam positioning aids and transducer stabilizing clamp.

Brachial artery FMD will be continuously measured and recorded using a 10-minute protocol. Baseline diameter and blood velocity will be recorded for two minutes. The tourniquet cuff will then be infla

• Hydration Status [Срок оценки: Visit 2 & visits 6-13 approx=8 weeks]
• Sleep Quality Questionnaire [Срок оценки: Visit 2 & visits 6-13 approx=8 weeks]
• Diet and Dietary Log [Срок оценки: Visit 2 & visits 6-13 approx=8 weeks]
• Anthropometric Assessments: [Срок оценки: Visit 2]