To Assess the Influence of Aronox on Parameters Associated With Recovery Following Muscle Damaging Exercise
Condition: Muscle Damage
Study type: Interventional

Summary: 
      A randomized, double blind, counterbalanced, placebo controlled independent groups design.
      Prior to the visit, participants will be given a participant information sheet to inform of
      the procedure and requirements and undergo initial screening via email or telephone to
      ascertain suitability to participate. Stature, body mass, blood pressure and heart rate will
      be assessed. Participants will then be familiarized with the performance tests (MVC, vertical
      jump and sprint performance) and randomized to an investigational product group (2 groups:
      Aronox vs placebo; 1:1 allocation). The first investigational dose will be administered in
      the laboratory and participants will be given a 4-week supply of the investigational product
      to take in the morning with breakfast. Participants will also be asked to keep a food and
      activity diary for the 3 days preceding the baseline visit and for the duration of the
      damaging-recovery protocol.

      Following this supplementation period (28 days), participants will be asked to return to the
      lab in a fed (not less than 2 hours prior to the visit) and hydrated state. Participants will
      also be asked to abstain from strenuous exercise and caffeine for 24 h prior to each lab
      visit. Stature, body mass, blood pressure and heart rate will be assessed. This will be
      followed by baseline assessment of muscle damage which will consist of visual analogue scales
      to assess lower limb muscle soreness (DOMS); pain pressure threshold and baseline measures of
      functional performance (maximal voluntary contraction, vertical jump performance and sprint
      performance) and limb girth. Furthermore, a blood sample will be taken to analyze creatine
      kinase (index of muscle damage). This will be followed by a strenuous bout of exercise
      designed to cause muscle damage comprising of 100 drop jumps from a 0.6 m platform at a rate
      of 1 jump every 10 seconds. A short rest will be provided after every 20 jumps. Each jump is
      performed by the participant stepping from the platform and landing two-footed on the floor
      and descending quickly to ~90° and 'explosively jumping upward with maximum effort. This
      model for muscle damage has been used on numerous occasions in the literature and has been
      used with great success in our own laboratory. Participants will then return to the lab at
      24, 48 and 72 h post damaging protocol where muscle damage measures will be repeated to
      assess the level of recovery between the groups.
    

Detailed description: 
      A randomized, double blind, counterbalanced, placebo controlled independent groups design.
      Prior to the visit, participants will be given a participant information sheet to inform of
      the procedure and requirements and undergo initial screening via email or telephone to
      ascertain suitability to participate. If interested parties wish to participate, then they
      will be invited for a familiarisation visit, where they will:

        -  Be provided with an overview of the protocol and given the opportunity to ask questions

        -  Complete a health and physical activity questionnaire to ascertain study suitability

        -  Complete a training status questionnaire

        -  Assessment of ability to swallow capsules

        -  Complete an informed consent to participate in the study (assuming they meet the
           criteria and are willing to participate)

      Stature, body mass, blood pressure and heart rate will be assessed. Participants will then be
      familiarized with the performance tests (MVC, vertical jump and sprint performance) and
      randomized to an investigational product group (2 groups: Aronox vs placebo; 1:1 allocation).
      The first investigational dose will be administered in the laboratory and participants will
      be given a 4-week supply of the investigational product to take in the morning with
      breakfast. Participants will also be asked to keep a food and activity diary for the 3 days
      preceding the baseline visit and for the duration of the damaging-recovery protocol.

      Following this supplementation period (28 days), participants will be asked to return to the
      lab in a fed (not less than 2 hours prior to the visit) and hydrated state. Participants will
      also be asked to abstain from strenuous exercise and caffeine for 24 h prior to each lab
      visit. Stature, body mass, blood pressure and heart rate will be assessed. This will be
      followed by baseline assessment of muscle damage which will consist of visual analogue scales
      to assess lower limb muscle soreness (DOMS); pain pressure threshold and baseline measures of
      functional performance (maximal voluntary contraction, vertical jump performance and sprint
      performance) and limb girth. Furthermore, a blood sample will be taken to analyze creatine
      kinase (index of muscle damage). This will be followed by a strenuous bout of exercise
      designed to cause muscle damage comprising of 100 drop jumps from a 0.6 m platform at a rate
      of 1 jump every 10 seconds. A short rest will be provided after every 20 jumps. Each jump is
      performed by the participant stepping from the platform and landing two-footed on the floor
      and descending quickly to ~90° and 'explosively jumping upward with maximum effort. This
      model for muscle damage has been used on numerous occasions in the literature and has been
      used with great success in our own laboratory. Participants will then return to the lab at
      24, 48 and 72 h post damaging protocol where muscle damage measures will be repeated to
      assess the level of recovery between the groups.

      Exercise protocol details Muscle damage will be induced using a drop jump protocol our
      laboratory has used previously. It consists of 100 drop jumps performed 10 seconds apart with
      a 2-minute rest period provided every 20 jumps. Each jump will be conducted from a 0.6 m high
      box; upon landing, participants descend to a ~90º knee angle before performing a maximal
      effort vertical jump.

      Passive muscle soreness will be reported on a 200 mm visual analogue scale and muscle
      site-specific soreness will be assessed as pressure pain threshold (PPT) with a handheld
      algometer (N2). Measurements were taken with participants lying supine; pressure was applied
      continuously at a rate of ~10 N cm-2∙s-1 until a pressure of ~40 N cm-2∙s-1 is achieved on
      the muscle belly; whereupon the participant will report muscle soreness on the aforementioned
      visual analogue scale.

      Limb girth will be measured at the mid-thigh to examine limb swelling. This will be
      determined by identifying the midpoint between the inguinal crease and the superior border of
      the patella the right leg whilst the participant remained standing in anatomical zero. The
      location will be marked with permanent marker to ensure consistency on subsequent days.

      A venous blood sample will then taken to examine creatine kinase (CK) concentrations to
      provide evidence that muscle damage has occurred.

      Maximal isometric voluntary contractions (MIVCs) performed using a portable strain gauge (MIE
      Medical Research Ltd., Leeds, UK), will be used to examine strength loss and recovery. The
      peak value from 3 maximal contractions (separated by 60 seconds) will be used for analysis.
      In addition, counter movement jumps (CMJ) will be examined with an optical system to measure
      jump height in cm. With hands on their hips, participants will descend into a ~90° squat and
      jump vertically with maximal effort. The average of 3 maximal jumps (separated by 30 seconds)
      will be used for analysis. Participants will complete a single maximal effort 30 m sprint
      where sprint time will be recorded. The sprint will be initiated from a line 30 cm behind the
      start line in order to prevent false triggering of the timing gates (Brower, Utah, USA). In
      addition to the aforementioned measures that will be repeated at 24, 48 and 72 h, a perceived
      recovery questionnaire to assess more qualitative aspects of the intervention on recovery
      from the damaging protocol will be completed.

      Supplementation and dietary control Following group allocation, participants will be provided
      with aronia (Aronox® ) or placebo (PLA) supplementation and instructed to consume 500mg of
      the investigational product. The PLA will consist of 95% maltodextrin with 5% artificial dyes
      (red lake and blue lake). The 500mg of Aronox is fortified with maltodextrin in order to
      provide 200 mg of polyphenol in total. Aronia consumption has shown no safety issues when
      used as an ingredient in dietary supplements or as a study drug in clinical trials by adults
      at dose levels ranging from 150 to 300 mg of extract per day for 14 days up to 2 months
      (Broncel et al., 2010; Naruszewicz et al., 2007).

      Participants will be instructed not to consume the supplement on the day of initial
      assessment and consume the supplement after each laboratory visit as previous research
      (Bitsch et al. 2004; Kurilich et al. 2005) has demonstrated that systemic anthocyanin
      bioavailability increases to a peak between 1-2 hours post-ingestion. In addition, training
      logs will be kept to examine differences in training volume between groups and also general
      health log to record illness or consumption of any pharma during the supplementation period.

      Data analysis

      All data will be expressed as means ± SD, and statistical significance will be set at P <
      0.05 prior to analyses. Differences in participant group characteristics, training history,
      and dietary intake were analysed using Student t-tests. Nutritics dietary analysis software
      (Nutritics LTD, Dublin, Ireland) will be used to analyse participant's food diaries.
      Dependent variables (MIVC, CMJ, VAS, and all blood indices) will be analysed using a mixed
      model ANOVA with 2 independent group levels (Aronex vs. PLA) and 4 repeated measures time
      points (pre, 24,48,72 h). If the ANOVA indicates a significant interaction effect (drink ×
      time), Fisher LSD post hoc analysis will be performed to locate the significant differences.
      Homogeneity of variance will be checked with Mauchly's test of sphericity, and in the event
      of a significant result, GreenhouseGeisser adjustments will be used. All data will be
      analysed using IBM SPSS Statistics 22 for Windows (Surrey, UK).
    

Intervention name(s): 500mg of Aronox® >40% polyphenol aronia extract; Placebo
Intervention type(s): Dietary Supplement; Other
Intervention description(s): Name: Aronia PE 40% polyphenols Description: Powdered extract obtained from aronia berries (Aronia melanocarpa) Dosage form: Capsule (opaque, beige, size 00) Strength: 500 mg aronia extract Regimen 1 capsule, once a day with breakfast (except on days when attending the laboratory) Batch number: As per label Expiry Date: As per label Manufacturer: NATUREX / Virage sante; Name: Placebo Description: Identical formulation as the treatment consisting of colored maltodextrin using artificial colors Dosage form: Capsule (opaque, beige, size 00) Strength: 500 mg placebo Regimen 1 capsule, once a day with breakfast (except on days when attending the laboratory) Batch number: As per label Expiry Date: As per label Manufacturer: NATUREX (Advance nutraceutical) / Virage sante
Allocation: Randomized
Intervention model: Parallel Assignment
Masking: Triple (Participant, Care Provider, Investigator)
Primary purpose: Prevention