Mangoes and Exercise Inflammation
Condition: Inflammatory Response; Muscular Injury
MeSH terms: Inflammation
Study type: Interventional

Summary: 
      Vigorous exercise bouts result in significant inflammation and muscle soreness. Our research
      group has published several papers showing that ingestion of various types of fruits enhances
      inflammation resolution after exercise. Mangoes have a unique nutrient profile (carotenoids,
      polyphenols, sugars, vitamins) that we hypothesize will mitigate post-exercise inflammation
      and muscle soreness. This study will examine the effect of 2 cups/day frozen mango ingestion
      (2 weeks, 330 g/day) in moderating exercise-induced (2.25 h cycling) inflammation and muscle
      soreness in a randomized crossover trial. Participants will include 20 male and female young
      adult cyclists (ages 18-60 years) who are capable of cycling 2.25 h in the laboratory on
      trainers. In random order, the cyclists will supplement their diets with 2 cups/day mangoes
      with 1 cup water or 1 cup water alone for two weeks, followed by the 2.25-h exercise
      challenge. Blood and urine samples will be collected pre- and post-2 weeks supplementation.
      Additional blood samples will be collected immediately post-exercise, and then 1.5-h, 3-h,
      and 24-h post-exercise. Urine samples will be analyzed for mango-related metabolites to
      confirm compliance to the supplementation regimen and to establish statistical relationships
      with inflammation-related outcomes. The blood samples will be analyzed for novel,
      cutting-edge outcomes related to inflammation including 70 oxylipins, inflammasome
      activation, and pro-inflammatory cytokines that we have previously shown are sensitive to
      exercise and nutrition-based interventions. If the data support our hypothesis, mango
      ingestion will be viewed as a nutritional strategy to counter exercise-induced inflammation
      by fitness enthusiasts and athletes who exercise vigorously on a regular basis.
    

Detailed description: 
      Subjects will come to the lab for orientation/baseline testing, pre-and post-supplementation
      (2 weeks mango ingestion compared to water alone) blood and urine sample collections with 3-d
      food records, two 2.25 h cycling sessions, and additional lab visits to provide 1-day 7:00 am
      recovery blood samples) (thus 7 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 6-week period).

      A. Orientation/Baseline Testing (Visit #1):

      After voluntarily signed IRB-approved consent forms, study participants will be tested for
      maximal aerobic capacity (VO2max) during a graded, cycling test with continuous metabolic
      monitoring 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. A
      symptom inventory will be conducted using a 2-week retrospective questionnaire. 3-day food
      records and 24-h urine collection kits will be supplied with thorough instructions.

      B. Pre-Supplementation Lab Visit #2 and 2-Week Mango Supplementation Protocol:

      Two weeks prior to the first running 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. Urine samples and 3-d food records will be turned in after review. A 2-week supply of
      frozen mangoes in date/time labeled freezer bags (1 cup or 165-gram amounts) will be given to
      the participants randomized to the mango trial, with instructions to consume 1 cup each with
      the first and last meals of the day (for two weeks). Participants will be given the frozen
      mangoes in cold containers and then instructed to store these in their home freezers.
      Subjects will be instructed to consume the mangoes in a variety of ways including in
      smoothies, with yogurt, or as is. Bottled water will be supplied to both groups with
      instructions to consume 1 cup each with the first and last meals of the day. Subjects will be
      instructed to store and then bring the freezer bags to their next lab visit. To assess
      potential adverse effects, a symptom inventory will be conducted using a 2-week retrospective
      questionnaire (before and after the 2-week supplementation period). Rationale for the mango
      dosing regimen: The amounts of carotenoids and polyphenols in 2 cups/d mangoes are similar to
      amounts we have used successfully in prior studies to mitigate post-exercise inflammation and
      muscle soreness. We recommend a 2-week supplementation period because our experience is that
      this is a sufficient time period for bioactive effects to emerge as the gut microbiome and
      tissues adapt. We recommend using water as the control condition to allow the full array of
      nutrient components found in mangoes (carbohydrate, carotenoids, polyphenols, vitamins C and
      A) to be tested when compared to just water.

      C. 2.25 h Cycling Session (Lab Visit #3):

      During the 3-day period prior to the 2.25-h cycling session, 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). We will also measure
      total flavonoid intake (and 6 flavonoid subgroups) using the USDA flavonoid database that we
      have incorporated into the Food Processor database. Study participants will report to the
      Human Performance Lab in an overnight fasted state and provide a blood sample, ingest 1 cup
      mango with 1 cup of water, or 1 cup of water alone, and then cycle 2.25 h at high intensity
      (70% VO2max) while ingesting water alone (3 ml/kg every 15 minutes). Blood samples will be
      collected at 0 h, 1.5 h, 3.0 h, and 24 h post-exercise.

      Testing protocol during the lab sessions with the 2.25-h cycling session:

        1. 7:00 am: Provide blood sample, delayed onset of muscle soreness (DOMS) rating (1-10
           scale), profile of mood states ratings (POMS), and turn in the 24-h urine sample and
           3-day food record. Fill in the 2-week retrospective symptom log.

        2. 7:10 am: Ingest 1 cup mangoes with 1 cup water or 1 cup water alone.

        3. 7:30 am: Start the 2.25 h cycling session. Subjects will cycle on trainers in the lab at
           70% VO2max (~race pace) for 2.25 hours. Oxygen consumption, carbon dioxide production,
           respiratory exchange ratio, and ventilation will be measured using the Cosmed Quark CPET
           metabolic cart after 15 minutes and then every 30 minutes. Subjects will consume 3 ml/kg
           water every 15 min. No other beverage or food containing energy or nutrients will be
           allowed during the 2.25-h cycling sessions.

        4. ~10:00 am to 1:00 pm: Blood samples will be taken via venipuncture immediately after
           completing the cycling session, and then 1.5-h and 3.0-h post-exercise. Subjects will be
           allowed to shower and change clothes. The DOMS and POMS questionnaires will be
           administered each time blood samples are collected. Subjects will ingest no food or
           beverage other than water (7 ml/kg during the 1.5 h post-exercise period). After the 1.5
           h post-exercise blood draw, subjects in the mango group will consume 1 cup mangoes with
           1 cup of water. Subjects in the water group will consume a calorie-matched volume of
           Gatorade (0.45 liters). Another blood sample will be collected 3-h post-exercise.
           Afterwards, subjects will be allowed to leave the lab with instructions to adhere to the
           food list requirements, and to keep exercise training moderate.

        5. 7:00 am, next morning: 24-h blood sample and DOMS and POMS ratings (Lab visit #4).

      D. Washout/Crossover/Repeat Subjects will engage in a 2-week washout period without the mango
      supplements, crossover to the opposite treatment arm, and then repeat all procedures (Lab
      visits #5,6,7).
    

Intervention name(s): Mangoes; Water
Intervention type(s): Dietary Supplement; Dietary Supplement
Intervention description(s): Frozen mangoes, 2 cups per day, 2 weeks; 2 cups/day bottled water, 2 weeks
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: Randomized treatment, crossover
Masking: None (Open Label)
Primary purpose: Prevention
Keywords: mangoes; exercise; inflammation; oxylipins; inflammasome