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Development of Oral Amino Acid Tracers to Study Protein Turnover in Humans

Not Applicable
Completed
Conditions
Healthy Adult Males
Interventions
Dietary Supplement: Crystalline amino acids to assess myofibrillar protein synthesis rates
Dietary Supplement: Crystalline amino acids to assess whole-body protein turnover, amino acid oxidation, and net protein balance
Registration Number
NCT04887727
Lead Sponsor
University of Toronto
Brief Summary

Previous studies have used a combination of oral L-\[1-13C\]leucine and intravenous labeled L-\[5,5,5-2H3\]leucine to assess the acute postprandial changes in whole-body protein turnover (12, 19). Intravenous and dietary-labeled amino acid tracers have also been used in tandem to assess rates of myofibrillar protein synthesis in response to bolus protein ingestion and resistance exercise (46). By validating whole-body net balance to myofibrillar protein synthesis, our proposed multi-tracer approach will develop minimally invasive models to study protein turnover in a variety of populations in which traditional infusions and/or repeated blood samples are not possible (i.e. pediatric, free-living populations).

Detailed Description

The primary objective of the proposed study is to validate the use of a novel oral tracer model to accurately and reliably measure myofibrillar protein synthesis. It is hypothesized that oral L-\[1-13C\]leucine and L-\[ring-2H5\]phenylalanine, ingested as a bolus to mimic an intravenous 'pulse dose' administration (51, 65), will reveal similar rates of myofibrillar protein synthesis when compared to traditional intravenous L-\[5,5,5-2H3\]leucine infusion. Moreover, it is hypothesized that both methods will reveal the expected graded changes in myofibrillar protein synthesis in response to feeding and resistance exercise (i.e. fasted\<feeding\<exercise \& feeding).

The secondary objective of the proposed study is to develop and validate non-invasive models to measure whole-body amino acid oxidation and net balance in response to feeding and resistance exercise. It is hypothesized that whole-body net balance, as determined by a novel oral tracer model (i.e. L-\[1-13C\]leucine) , will align with traditional intravenous tracer methodology (i.e. L-\[5,5,5-2H3\]leucine) and reveal the expected physiological changes in whole-body protein turnover in response to feeding and resistance exercise (i.e. fasted\<feeding\<exercise \& feeding).

Recruitment & Eligibility

Status
COMPLETED
Sex
Male
Target Recruitment
22
Inclusion Criteria
  • Male
  • Currently performing structured physical activity 2-5 days per week
Exclusion Criteria
  • Unable to safely perform exercise as per PARQ+ guidelines
  • Currently using tobacco products
  • Currently using or have history of anabolic steroid use
  • Diagnosed with medical condition including type 2 diabetes, cancer, heart disease
  • Unable to abstain from supplement use (HMB, branched chain amino acids, phosphatidic acid) for at least three weeks prior to trial
  • currently using medications known to affect protein metabolism e.g. corticosteroids, NSAID, prescription-strength acne medication
  • allergic to local anesthetics
  • female: Hormonal fluctuations associated with the menstrual cycle have been reported to alter protein metabolism during exercise and may influence indices of the post-exercise myofibrillar protein synthetic response. Accordingly, the study will include males to ensure a stable hormonal environment and to increase the homogeneity of the physiological response.

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
Healthy, recreationally-active adult males - Muscle TrialCrystalline amino acids to assess myofibrillar protein synthesis ratesSubjects receive 0.25g/kg crystalline amino acids modelled after egg protein, enriched with L-\[1-13C\]leucine and L-ring-\[2H5\]phenylalanine with a primed-constant infusion of L-\[555-2H3\]leucine to assess myofibrillar protein synthesis rates
Healthy, recreationally-active adult males - Whole Body TrialCrystalline amino acids to assess whole-body protein turnover, amino acid oxidation, and net protein balanceSubjects receive 0.25g/kg crystalline amino acids modelled after egg protein, enriched with L-\[1-13C\]leucine with a primed-constant infusion of L-\[555-2H3\]leucine to whole-body protein turnover, amino acid oxidation, and net protein balance
Primary Outcome Measures
NameTimeMethod
Myofibrillar protein synthesis rates5 hours

Myofibrillar protein synthesis rates assessed by oral and intravenous tracers during Fast, Fed, and Ex-Fed

Secondary Outcome Measures
NameTimeMethod
Muscle anabolic signalling2 and 5 hours

Immunoblotting for mTORC1/key downstream targets of mTORC1

Whole-body protein turnover5 hours

Whole-body protein turnover assessed by oral and intravenous tracers during Fast, Fed, and Ex-Fed

Amino acid oxidation and net protein balance5 hours

Amino acid oxidation and net protein balance assessed by oral tracers during Fast, Fed, and Ex-Fed. Net protein balance is derived from the difference between amino acid intake (known) and total amino acid oxidation over the 5h measurement period.

Amino acid transporter expression2 and 5 hours

Immunoblotting for amino acid transporters LAT1/SNAT2

Trial Locations

Locations (1)

Goldring Centre for High Performance Sport at the University of Toronto

🇨🇦

Toronto, Ontario, Canada

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