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This study will be a randomised, double-blind trial where participants will be randomly assigned to consume either a i) vegan whey protein; ii) a whey protein; or iii) a non-protein carbohydrate control supplement for 3 days following strenuous eccentric knee extension exercise. For the duration of the study period, participants will orally consume deuterated water stable isotope tracer (D2O). This, together with repeated blood sampling and muscle biopsies will allow for the calculation of free-living myofibrillar protein synthesis rates under conditions of rest and over 24-, 48- and 72-hours post-exercise recovery.
Participants will be asked to attend 7 experimental visits at the School of Sport, Exercise and Rehabilitation Sciences at the University of Birmingham. The first of the 7 visits will be to assess participant eligibility and to gain written informed consent, where the remaining 6 visits will involve experimental procedures. Briefly, participants will be randomly assigned to ingest one of three nutritional supplements (i: vegan whey protein isolate; ii: whey protein isolate; iii: a non-protein carbohydrate control), 3 times a day for a total of 3 days following an acute bout of unilateral lower-limb eccentric exercise. The study will be run in a double-blinded fashion, where the researchers and participants will both be unaware of nutritional treatment allocation. The amount of protein in each supplemental beverage is prescribed based on total body mass (0.3 g of protein per kg of body mass). As well, for 3 days before and 3 days after the exercise bout, participants will be provided with a standardised diet to consume. For the 3 days before the exercise bout, all participants will consume an identical diet, with a protein intake of 1 g per kg body mass and diets will be energy-matched between treatment groups. For the subsequent 3 days, participants assigned to each of the protein supplementation groups (whey protein or vegan whey protein) will have a daily protein intake of 1.9 g per kg body mass, whereas participants assigned to the carbohydrate control group will maintain protein intake at 1 g per kg body mass. Measures of muscle function, soreness and thickness will be taken before and at various time points after the eccentric exercise, details of which can be found below. To assess the muscle protein synthetic response to each nutritional treatment, participants will orally consume a stable isotope tracer ("heavy water") for a period of 9 days, and muscle biopsies and saliva samples will be taken periodically. An outline of each experimental visit is outlined below.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Vegan whey protein | Experimental | 3 x daily supplementation of 0.3 g per kg body mass of vegan whey protein isolate for 3 days following an acute bout of lower-body unilateral resistance exercise |
|
| Dairy whey protein | Active Comparator | 3 x daily supplementation of 0.3 g per kg body mass of dairy whey protein isolate for 3 days following an acute bout of lower-body unilateral resistance exercise |
|
| Carbohydrate placebo | Placebo Comparator | 3 x daily supplementation of isocaloric carbohydrate placebo for 3 days following an acute bout of lower-body unilateral resistance exercise |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Stable isotope tracer consumption | Other | Daily consumption of oral deuterium oxide (0.3 ml per kg of lean body mass) to equilibrate in body water and become slowly incorporated into muscle protein, enabling measurement of muscle protein synthesis rates |
| Measure | Description | Time Frame |
|---|---|---|
| Myofibrillar protein synthesis rates (iMyoPS) | 1. Changes in myofibrillar protein synthesis rates of the exercised leg, both over time (1, 24, 48 and 72 hours) during post-exercise recovery and between supplemental groups (vegan whey, whey protein, and non-protein carbohydrate control). Determined by assessing the incorporation of deuterium in muscle, relative to saliva and blood deuterium enrichment. | 3 days pre-exercise and 3 days post-exercise |
| Muscle function | 4. Changes in leg muscle architecture followed by leg strength and neural activation will be assessed pre- and at 1, 8-, 24-, 48- and 72-hours post-exercise. Muscle architecture will be assessed using ultrasound and specifically, shear-wave elastography. Change in isometric strength will be assessed on a Biodex dynamometer at a set knee angle while simultaneously assessing neural activation. Participants will be asked to perform a maximal voluntary contraction (MVC) for 5 seconds while neural activation is measured using the interpolated twitch method. There will be 3 measurements for MVC and neural activation with 2 minutes rest between each attempt on visit | pre- and 1, 4, 24, 48 and 72 hr post-exercise |
| Intramuscular anabolic signalling | 2. Changes in the total protein content and phosphorylation status of key anabolic signalling proteins, both over time post-exercise and between the supplemental groups. Assessed by Western Blotting of muscle biopsy samples | pre- and 1-hr and 24-hr post-exercise |
| Muscle morphology | 3. Changes in muscle structure and morphology at pre- and at 1, 24-, 48- and 72-hours post-exercise will be assessed via immunofluorescence microscopy of muscle biopsy samples. | pre- and 1, 24, 48 and 72 hr post-exercise |
| Measure | Description | Time Frame |
|---|---|---|
| Muscle soreness | Subjective ratings of muscle soreness will be measured via completion of validated visual analogue scales (no muscle soreness 0 - worst imaginable muscle soreness 100 mm) | pre- and 1, 4, 24, 48 and 72 hr post-exercise |
| Blood markers of muscle damage |
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Inclusion Criteria:
Exclusion Criteria:
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| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| School of Sport, Exercise and Rehabilitation Sciences, University of Birmingham | Birmingham | West Midlands | B15 2TT | United Kingdom |
| PubMed Identifier | Type | Citation | Retractions |
|---|---|---|---|
| 41500363 | Derived | Rogers LM, Korzepa M, Belfield AE, Quinlan JI, Wallis GA, Breen L. Muscle Recovery and Myofibrillar Protein Synthesis after Damaging Exercise with Recombinant Bovine beta-Lactoglobulin, Dairy-Derived Whey or Carbohydrate Supplementation in Young Healthy Adults. J Nutr. 2026 May;156(5):101321. doi: 10.1016/j.tjnut.2025.101321. Epub 2026 Jan 5. |
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| ID | Term |
|---|---|
| D063806 | Myalgia |
| ID | Term |
|---|---|
| D009135 | Muscular Diseases |
| D009140 | Musculoskeletal Diseases |
| D009468 | Neuromuscular Diseases |
| D009422 | Nervous System Diseases |
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| ID | Term |
|---|---|
| D015502 | Absorptiometry, Photon |
| D001800 | Blood Specimen Collection |
| D055070 | Resistance Training |
| D019587 | Dietary Supplements |
| ID | Term |
|---|---|
| D011859 | Radiography |
| D003952 | Diagnostic Imaging |
| D019937 | Diagnostic Techniques and Procedures |
| D003933 | Diagnosis |
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| DXA scan | Radiation | Whole body DXA scan to assess lean body mass for prescription of deuterium oxide tracer |
|
| Controlled diet | Behavioral | Participant will be asked to consume a provided 6-day diet package that will be prepared by the research team. This package will include 3 meals per day plus snacks |
|
| Saliva sample | Other | Obtained from participants by asking them to spit into a tube each day during the study, for 11 days in total. |
|
| Blood sample | Other | Resting venous blood sample from a forearm vein on 6 separate occasions |
|
| Muscle biopsy | Procedure | A highly experienced researcher will sample a small section of muscle (~200 mg) from the vastus lateralis muscle under local anaesthetic (5 ml, 1% Lidocaine) |
|
| Resistance exercise | Behavioral | participants will complete 8 sets of 10 repetitions of unilateral eccentric knee extensions on an isokinetic dynamometer. |
|
| Isometric strength test | Other | Bilateral isometric strength will be measured using a well-established protocol on an isokinetic dynamometer. |
|
| Supplement | Dietary Supplement | Consumption of 3 supplements a day for 3 days following resistance exercise. All supplements are commercially available. |
|
Plasma concentrations of systemic markers of muscle damage (creatine kinase, lactate dehydrogenase etc) will be measured from blood samples |
| pre- and 1, 4, 24, 48 and 72 hr post-exercise |
| D059352 | Musculoskeletal Pain |
| D010146 | Pain |
| D009461 | Neurologic Manifestations |
| D012816 | Signs and Symptoms |
| D013568 | Pathological Conditions, Signs and Symptoms |
| D003720 |
| Densitometry |
| D010783 | Photometry |
| D002623 | Chemistry Techniques, Analytical |
| D008919 | Investigative Techniques |
| D013048 | Specimen Handling |
| D019411 | Clinical Laboratory Techniques |
| D011677 | Punctures |
| D013514 | Surgical Procedures, Operative |
| D005081 | Exercise Therapy |
| D012046 | Rehabilitation |
| D000359 | Aftercare |
| D003266 | Continuity of Patient Care |
| D005791 | Patient Care |
| D013812 | Therapeutics |
| D026741 | Physical Therapy Modalities |
| D064797 | Physical Conditioning, Human |
| D015444 | Exercise |
| D009043 | Motor Activity |
| D009068 | Movement |
| D009142 | Musculoskeletal Physiological Phenomena |
| D055687 | Musculoskeletal and Neural Physiological Phenomena |
| D005502 | Food |
| D000066888 | Diet, Food, and Nutrition |
| D010829 | Physiological Phenomena |
| D019602 | Food and Beverages |