Effect of 8-week Supplementation With Anthocyanin Compounds Contained in Black Chokeberry on Post-workout Recovery in Highly Trained Rowers

Study Overview
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Brief Summary
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Purpose: To analyze the effects of 8-week supplementation with anthocyanin compounds contained in black chokeberry (Aronia melanocarpa) on indicators of inflammation and oxidative stress. Hypothesis: Supplementation with anti-inflammatory and antioxidant anthocyanin compounds improves recovery from intense exercise by reducing inflammation and oxidative stress in highly trained rowers The study consisted of a nutritional intervention (supplementation) - with compounds of natural origin - chokeberry extract (capsules) - 18% standardized for anthocyanin content (dose 3 x 200 mg per day) - that is, 107 mg of pure anthocyanins per day, or a placebo product that was made from chokeberry fiber Study plan I term of the study Conduct a body composition analysis, 24 hour dietary recall Collection of blood samples before the exercise test for biochemical determinations. Exercise test of 2000m on a rowing ergometer. Collection of blood immediately after exercise and 1 hours after Collection of blood for biochemical determinations 24 hours after the end of the exercise test Supplementation for 8 weeks 2nd test date Repeat the measurements from the 1st test date. Participants: Youth National Rowing Team of Poland Experimental procedure: Observation of the effect of black chokeberry consumption on parameters of inflammation, oxidative stress and intestinal parameters during the immediate start preparation period in highly skilled rowers.

Detailed Description
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Participants Twenty male from the Youth National Rowing Team of Poland have been recruited for the study. In the end, sixteen rowers participated in the study (two did not show up for the first date, two more were absent for the second date). Before the first and second dates of the study, a body composition analysis was performed using the TANITA MC-780 with an accuracy of 0.05 kg. The results are shown in Table 1. The experiment was conducted in accordance with the Declaration of Helsinki. The study protocol was approved by the Ethical Committee of the Medical University of Poznan (resolution no. 390/22). All players were informed of the exact course of the study and gave written consent to participate. Youth National Rowing Team of Poland The athletes' diets were completely analyzed before each exercise test by a nutritionist. Participants, with the help of a dietician who was available during meals on test days, filled out questionnaires. The amount of energy, protein, carbohydrates and fats were then analyzed using a commercially available program. Experimental procedure The rowers were randomly divided into two groups-supplemented and control, it was a double-blind study. Athletes in the supplemented group (n=7) consumed capsules with highly concentrated 18% chokeberry extract. One capsule contained 200 mg. In addition to chokeberry extract, each capsule also contained chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydroxypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell color). The control group (n=8) consumed capsules that were made from chokeberry fiber. All additional substances the same as in capsules with chokeberry extract. The supplement and placebo product was manufactured by MBL BIOTRADE, Poznań. The rowers (both supplemented and control group) took 3 capsules a day (at breakfast, lunch and dinner) for 8 weeks. The athletes sipped each capsule with a glass of water. Exercise test On two testing dates (before and after supplementation), the athletes performed a 2,000-meter test on a rowing ergometer (Concept II, USA). Athletes had to complete the test in the shortest possible time overcoming the maximum load. The results of the test were taken into account in the selection for the championship team so the athletes performed the task with great motivation. Before the test, each competitor individually performed a 5-minute warm-up. Material collection and examination Samples were taken at three time points: pre (before the exercise test) post (immediately after the exercise test and 1 hour after) and recovery (24 hours after the exercise test) both before and after the supplementation period. Blood samples were collected from the elbow vein into 9-ml tubes (to obtain serum) and centrifuged at 3,000 rpm and into 2.7-ml tubes (to determine morphology). Morphology was determined on the same day the samples were collected, while serum was frozen and stored at -80oC until assays were performed. In addition, before and immediately after the exercise test, capillary blood was drawn from the earlobe to determine lactate levels. Measurements TAC (Total Antioxidant Capacity), TBARS (levels of lipid peroxidation products), IL-2, IL-6, IL-10, TNF-α and myoglobin levels were determined using ELISA kits (SunRed Biotechnology Company). Lactate levels were determined immediately after capillary blood sampling using a commercially available kit (Dr. Lange, Germany). The concentration of lactate was determined in mmol/l. intestinal parameters: LBP (lipopolysacharide-binding pro), Claudin3, I-FABP (intestinal fatty acid binding pro) Any data obtained during the research will be used only for the purposes of this research project. The participant has the right to access and correct his/her data, to obtain information regarding the processing of his/her personal data, to request the deletion or restriction of the processing of his/her data, the right to portability of the data provided, the right to object to the processing, and the right to withdraw consent to the processing of personal data at any time. Withdrawal of consent does not affect the lawfulness of processing carried out on the basis of consent before its withdrawal. The confidential nature of the research documentation will be respected throughout the study. Legal requirements for the protection of personal data will be observed. In the study documentation, participants will be identified by a nadem code at the beginning of the study. The results of this study will be published after pooled data analysis, thus the identity of the participants will remain confidential. The place of data storage is the university's branch office in Gorzow Wielkopolski. All parameters will be analyzed using Elisamtests and read on the ELISAMAR reader. In terms of statistical measures, the following will be performed: descriptive statistics, Shapiro-Wilk test to assess normality, and multivariate analysis of variance ANOVA test.

Official Title
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Effect of 8-week Supplementation With Anthocyanin Compounds Contained in Black Chokeberry on Inflammation and Oxidative Stress Induced by Very Intense Exercise in Highly Trained Rowers

Conditions
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Inflammation;Muscle(S), Oxidative Stress Injury, Permeability; Increased

Intervention / Treatment
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* Drug: black chokeberry (Aronia melanocarpa)
* Drug: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Training experience a minimum of five years Minimum training time per week 240 minute Membership in the Youth National Rowing Team of Poland Completion of the 2000m ergometer test on two test dates Exclusion Criteria: Taking other supplements containing anthocyanin compounds during the study period or a month earlier Antibiotic therapy Health problems

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 24 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Participants randomly divided into one of two groups. The supplemented group (n=7) receives chokeberry capsules, while the control group (n=8) receives a placebo product.
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Supplemented group<br>Athletes in the supplemented group (n=7) consumed capsules with highly concentrated 18% chokeberry extract. One capsule contained 200 mg. In addition to chokeberry extract, each capsule also contained chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydroxypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell color). The rowers (both supplemented and control group) took 3 capsules a day (at breakfast, lunch and dinner) for 8 weeks. The athletes sipped each capsule with a glass of water. | Drug: black chokeberry (Aronia melanocarpa)<br>* Black chokeberry capsules standardized for anthocyanin compounds<br>|
| Placebo Comparator: Placebo group<br>The control group (n=8) consumed capsules that were made from chokeberry fiber. The rowers (both supplemented and control group) took 3 capsules a day (at breakfast, lunch and dinner) for 8 weeks. The athletes sipped each capsule with a glass of water. | Drug: Placebo<br>* Chokeberry fiber placebo capsules<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Level of total antioxidants (TAC) | ELISA kits (SunRed Biotechnology Company) - Indicators of pro-oxidant-antioxidant balance | 8 weeks | 
| interleukins: IL-2, IL-6, IL-10 | ELISA kits (SunRed Biotechnology Company) - Immunological indicators | 8 weeks | 
| lactic acid | Diaglobal, Berlin, Germany) | 8 weeks | 
| LBP (lipopolysacharide-binding pro) | ELISA kits (SunRed Biotechnology Company) - Gut parameter | 8 weeks | 
| level of lipid peroxidation products (TBARS) | ELISA kits (SunRed Biotechnology Company) - Indicators of pro-oxidant-antioxidant balance | 8 weeks | 
| (NO) nitric oxide | obtained by indirect method Griessa - Indicators of pro-oxidant-antioxidant balance | 8 weeks | 
| comet assay (DNA damage) | comet assay BioTechne | 8 weeks | 
| tumor necrosis factor (TNF α) | ELISA kits (SunRed Biotechnology Company) - Immunological indicators | 8 weeks | 
| myoglobin | ELISA kits (SunRed Biotechnology Company) | 8 weeks | 
| I-FABP (intestinal fatty acid binding protein) | ELISA kits (SunRed Biotechnology Company) - Gut parameter | 8 weeks | 
| Claudin3 | ELISA kits (SunRed Biotechnology Company) - Gut parameter | 8 weeks |