In Vivo and in Vitro Anabolic Potential of Essential Amino Acids Following Resistance Exercise

Study Overview
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Brief Summary
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This study seeks to investigate the anabolic potential of a dileucine-enriched essential amino acid (EAA) formulation compared with a branched chain amino acid (BCAA) alternative and a collagen beverage on muscle protein anabolism and catabolism following a bout of resistance exercise training. To do this, investigators will employ a novel 'breath test' method developed in our laboratory as well as blood and urine sampling. The results of this study will allow us to better understand the anabolic potential of dileucine which could have implications for people engaging in regular resistance training (such as athletes) as well as people that need to preserve muscle mass (older people who are susceptible to anabolic resistance and sarcopenia, or muscle wasting).

Official Title
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In Vivo and in Vitro Anabolic Potential of Essential Amino Acids Following Resistance Exercise

Conditions
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Interventional

Intervention / Treatment
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* Dietary Supplement: Di-Leucine Supplement
* Dietary Supplement: BCAA Supplement
* Dietary Supplement: Collagen Supplement


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Trained individuals currently performing structured exercise (e.g., running, weightlifting, team-sport activity) at least 2 days per week for the previous 3 months Exclusion Criteria: Inability to perform physical activity as determined by the PAR-Q Inability to adhere to protocol guidelines (e.g., alcohol, habitual diet) Regular tobacco use Illicit drug use (e.g., growth hormone, testosterone, etc.) Diagnosed medical condition under the care of a physician (e.g. type 2 diabetes) Inability to abstain from supplements (e.g. protein, creatine, HMB, BCAA, phosphatidic acid, etc.) at least three weeks before the trial Individuals on any medications known to affect protein metabolism (e.g., corticosteroids, non-steroidal anti-inflammatories, or prescription-strength acne medications) Participants that are amenorrheic (females only) On medications that may interfere with protein metabolism (e.g. anti-inflammatory drugs, hormone therapy) Regular tobacco use (e.g. daily use of oral or inhaled tobacco) Illicit drug use (e.g. growth hormone, testosterone, etc.) Inability to comply with the study protocol as judged by the investigators

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Cross over study design; Block randomized.
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Di-Leucine supplement<br>Participants will consume a amino acid supplement containing Di-Leucine following resistance exercise and their whole body anabolism will be determined over the subsequent 6 hours | Dietary Supplement: Di-Leucine Supplement<br>* Participants will consume a amino acid supplement containing Di-Leucine following resistance exercise and their whole body anabolism will be determined over the subsequent 6 hours<br>|
| Experimental: BCAA Supplement<br>Participants will consume a amino acid supplement containing branched chain amino acids (BCAA) following resistance exercise and their whole body anabolism will be determined over the subsequent hours | Dietary Supplement: BCAA Supplement<br>* Participants will consume a amino acid supplement containing branched chain amino acids (BCAA) following resistance exercise and their whole body anabolism will be determined over the subsequent hours<br>|
| Experimental: Collagen Supplement<br>Participants will consume a amino acid supplement containing collagen protein following resistance exercise and their whole body anabolism will be determined over the subsequent hours | Dietary Supplement: Collagen Supplement<br>* Participants will consume a amino acid supplement containing Collagen protein following resistance exercise and their whole body anabolism will be determined over the subsequent hours<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Whole-Body Protein Synthesis | Investigators will measure the enrichment of [13CO2] in the breath by isotope ratio mass spectrometry (IRMS) in atom percent excess (APE). The measurement of carbon dioxide production (VCO2) and stable isotope tracer enrichment in the breath allows for the assessment of the rate at which amino acids are used for energy (i.e., oxidized), rather than for protein synthesis (i.e., retained in the body) by calculating the fraction of expired CO2 that contains 13C. Leucine retention (umol/kg) will then be calculated from the difference between the known amount of leucine provided (ingested) and leucine oxidation (as determined from 13CO2 breath enrichment). | 6 hours | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Urinary Measures (Muscle Protein Breakdown) | Investigators will measure urinary 3-methylhistidine (3MH) as an indirect marker of muscle protein breakdown over the course of the trial (6 hours) through pooled urine collection vs baseline urine. | 6 hours | 
| Murine Cell-Based Experiments (ex-vivo experiments) -hypertrophy | Investigators will use human serum obtained from fasted and fed timepoints (15, 30, 45 and 60 minutes following beverage consumption) to condition cell culture media (20% volume). To determine the effects of using fasted and/or fed 'human-conditioned' culture media on cell growth, myotube diameter (hypertrophy) will be measured via microscopy following short term (4 hours) and long-term (24 hour) incubation in the human conditioned culture media. | 60 minutes | 
| Murine Cell-Based Experiments (ex-vivo experiments) - Protein Synthesis | Investigators will use human serum obtained from fasted and fed timepoints (15, 30, 45 and 60 minutes following beverage consumption) to condition cell culture media (20% volume). To determine the effects of using fasted and/or fed 'human-conditioned' culture media on cell protein synthesis, puromycin incorporation (measure of protein synthesis) will be measured via western blot. | 60 minutes | 
| Murine Cell-Based Experiments (ex-vivo experiments) - mTORC1 Signalling | Investigators will use human serum obtained from fasted and fed timepoints (15, 30, 45 and 60 minutes following beverage consumption) to condition cell culture media (20% volume). To determine the effects of using fasted and/or fed 'human-conditioned' culture media on cell anabolic signalling mTORC1 signalling will be measured via western blot. | 60 minutes | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Di-leucine, Branched chain amino acids, Collagen, Amino acids, anabolism, catabolism