Carbohydrate Availability and microRNA Expression

• Sponsor: United States Army Research Institute of Environmental Medicine (U.S. Fed)
• Overall Status: Completed
• CT.gov ID: NCT03250234
• Collaborator: (none)
• Enrollment: 12
• Location: 1
• Arms: 2
• Actual Duration (Months): 9
• Patients Per Site Per Month: 1.3

Study Details

Study Description

Brief Summary

This investigation will examine the impact of skeletal muscle glycogen stores on skeletal muscle and circulating microRNA expression and exogenous carbohydrate oxidation.

Primary Objective Determine the influence of carbohydrate availability (e.g., glycogen depletion and repletion) on skeletal muscle microRNA expression, and if changes in circulating microRNA are reflective of changes in skeletal muscle microRNA.

Secondary Objective Determine how initiation of exercise with adequate or low glycogen stores effects exogenous carbohydrate efficiency.

Condition or Disease	Intervention/Treatment	Phase
Glycogen Depletion	Other: Adequate Carbohydrate; Other: Low Carbohydrate	N/A

Detailed Description

Twelve, normal weight, healthy, physically active men and women will complete a glycogen depletion protocol, cycling at various intensities until failure. Participants will then consume a carbohydrate (CHO: 1 g/kg/hr) or an energy free control (CON) beverage matched for taste and color during the first 3-hr post glycogen depletion. For the remainder of the day participants will consume a diet designed to adequately (6.0 g CHO/kg/d) or inadequately (1.2 g CHO/kg/d) replenish glycogen stores. The following day participants will to complete 80-min of steady-state (~60% VO2peak) cycle ergometry consuming carbohydrate at a rate of 1.8 g/min enriched with a stable isotope of carbon to determine exogenous carbohydrate efficiency when glycogen stores are low or adequate at the onset of exercise.

Following a minimum 7-d washout period volunteers will return to the laboratory to complete the second arm of the investigation. To ensure glycogen stores are similar between volunteers on testing day, 48-hrs prior to testing all volunteers will complete a glycogen depletion protocol and then consume a diet providing 6.0 g/kg/d carbohydrate to replenish glycogen stores.

Study Design

• Study Type: Interventional
• Actual Enrollment: 12 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Masking: None (Open Label)
• Primary Purpose: Basic Science
• Official Title: Influence of Carbohydrate Availability on Skeletal Muscle and Circulating microRNA Expression
• Actual Study Start Date: Aug 1, 2017
• Actual Primary Completion Date: May 3, 2018
• Actual Study Completion Date: May 3, 2018

Arms and Interventions

Arm	Intervention/Treatment
Other: Adequate Carbohydrate Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d	Other: Adequate Carbohydrate; During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion
Other: Low Carbohydrate Non-nutritive control beverage. Low carbohydrate diet 1.2 g/kg/d	Other: Low Carbohydrate; During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free beverage. For the day participants will consume a diet of 1.2 g/kg/d carbohydrate following glycogen depletion

Outcome Measures

Primary Outcome Measures

1. Fold Change in microRNA Expression From Baseline [24 hours]

   Determine the influence of low or adequate glycogen on expression of skeletal muscle and circulating microRNA. Expression of microRNA was determined using real time polymerase chain reaction analysis. microRNA expressions were calculated as a fold change relative to baseline.

Secondary Outcome Measures

1. Exogenous Carbohydrate Oxidation [80 minutes]

   Determine how initiation of exercise with adequate or low glycogen stores effects exogenous carbohydrate efficiency. Exogenous carbohydrate oxidation was measured using an oral 13C-glucose isotope that was consumed in the study carbohydrate drink during steady-state cycling. Breath samples were collected during exercise and analyzed using isotope ratio mass spectrometry to calculate exogenous carbohydrate oxidation rates.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years to 39 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Men and women aged 18 - 39 years

• Weight stable (±5 lbs) for at least 2 months prior to the start of the study

• Body mass index (BMI) between 18.5-30 kg/m2

• Recreationally active based on assessment of physical activity history 2-4 days per week aerobic and/or resistance exercise

• Refrain from taking any NSAIDS (i.e., aspirin, Advil®, Aleve®, Naprosyn®, or any aspirin-containing product for 10 days before and at least 5 days AFTER each muscle biopsy. (*Tylenol® or acetaminophen is ok to use if needed for discomfort)

• Refrain from the use of alcohol and nicotine for the duration of the study

• Females must be on contraception (e.g., oral birth control, NuvaRing®, Depo Provera®, etc.)

• Supervisor approved leave status for federal civilian employees working within the US Army Natick Soldier Systems Center

Exclusion Criteria:

• Metabolic or cardiovascular abnormalities, gastrointestinal disorders (i.e., kidney disease, diabetes, cardiovascular disease, etc.)

• Disease or medication (i.e., diabetes medications, statins, corticosteroids, etc) that affects macronutrient utilization and/or the ability to participate in strenuous exercise

• Allergies or intolerance to foods (including but not limited to lactose intolerance/milk allergy), vegetarian practices, or medications (including, but not limited to, lidocaine or phenylalanine) to be utilized in the study

• Anemia (HCT < 38) and Sickle Cell Anemia/Trait

• Abnormal prothrombin time (PT)/ partial thromboplastin time (PTT) test or problems with blood clotting

• Present condition of alcoholism, use of nutritional/sports supplements, anabolic steroids, or other substance abuse issues

• Musculoskeletal injuries that compromise the ability to exercise

• Blood donation within 8 weeks of beginning the study

• Pregnancy

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	USARIEM	Natick	Massachusetts	United States	01760

Sponsors and Collaborators

• United States Army Research Institute of Environmental Medicine

Investigators

None specified.

Study Documents (Full-Text)

• Study Protocol and Statistical Analysis Plan - May 23, 2019

More Information

Publications

None provided.

• Responsible Party: United States Army Research Institute of Environmental Medicine
• ClinicalTrials.gov Identifier: NCT03250234
• Other Study ID Numbers: 17-17H
• First Posted: Aug 15, 2017
• Last Update Posted: Sep 2, 2021
• Last Verified: Aug 1, 2021

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No
• Keywords provided by United States Army Research Institute of Environmental Medicine: microRNA, carbohydrate oxidation

Study Results

Participant Flow

Pre-assignment Detail	This was a cross over design. 12 participants enrolled and completed both study arms.

Arm/Group Title	Adequate Carbohydrate First, Then Low Carbohydrate	Low Carbohydrate First, Then Adequate Carbohydrate
Arm/Group Description	Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d first, then Low carbohydrate diet 1.5 g/kg/d Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion 7 day washout Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free (0 kcal) beverage For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion	Non-nutritive control beverage. Low carbohydrate diet 1.5 g/kg/d first, then adequate carbohydrate diet 6.0 g/kg/d Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free (0 kcal) beverage. For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion 7 day washout Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion
Period Title: Overall Study
STARTED	6	6
COMPLETED	6	6
NOT COMPLETED	0	0

Baseline Characteristics

Arm/Group Title	Adequate Carbohydrate First, Then Low Carbohydrate	Low Carbohydrate First, Then Adequate Carbohydrate	Total
Arm/Group Description	Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion	Non-nutritive control beverage. Low carbohydrate diet 1.5 g/kg/d Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free beverage. For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion	Total of all reporting groups
Overall Participants	6	6	12
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	6 100%	6 100%	12 100%
>=65 years	0 0%	0 0%	0 0%
Sex: Female, Male (Count of Participants)
Female	0 0%	0 0%	0 0%
Male	6 100%	6 100%	12 100%
Race and Ethnicity Not Collected (Count of Participants)
Count of Participants [Participants]	0 0%
Region of Enrollment (Count of Participants)
United States	6 100%	6 100%	12 100%

Outcome Measures

1. Primary Outcome

Title	Fold Change in microRNA Expression From Baseline
Description	Determine the influence of low or adequate glycogen on expression of skeletal muscle and circulating microRNA. Expression of microRNA was determined using real time polymerase chain reaction analysis. microRNA expressions were calculated as a fold change relative to baseline.
Time Frame	24 hours

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Adequate Carbohydrate	Low Carbohydrate
Arm/Group Description	Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion	Non-nutritive control beverage. Low carbohydrate diet 1.5 g/kg/d Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free (0 kcal) beverage. For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion
Measure Participants	12	12
Mean (Standard Deviation) [fold change]	0.73 (.2)	1.36 (1.35)

2. Secondary Outcome

Title	Exogenous Carbohydrate Oxidation
Description	Determine how initiation of exercise with adequate or low glycogen stores effects exogenous carbohydrate efficiency. Exogenous carbohydrate oxidation was measured using an oral 13C-glucose isotope that was consumed in the study carbohydrate drink during steady-state cycling. Breath samples were collected during exercise and analyzed using isotope ratio mass spectrometry to calculate exogenous carbohydrate oxidation rates.
Time Frame	80 minutes

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Adequate Carbohydrate	Low Carbohydrate
Arm/Group Description	Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion	Non-nutritive control beverage. Low carbohydrate diet 1.5 g/kg/d Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free (0 kcal) beverage. For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion
Measure Participants	12	12
Mean (Standard Deviation) [g/min]	.87 (.16)	.84 (.14)

Adverse Events

Time Frame	3 weeks
Adverse Event Reporting Description	Hamstring cramping
Arm/Group Title	Adequate Carbohydrate		Low Carbohydrate
Arm/Group Description	Carbohydrate beverage (1 g/kg/hr) Adequate carbohydrate diet 6.0 g/kg/d Adequate Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a carbohydrate (1 g/kg/hr) beverage For the day participants will consume a diet of 6.0 g/kg/d carbohydrate following glycogen depletion		Non-nutritive control beverage. Low carbohydrate diet 1.5 g/kg/d Low Carbohydrate: During the 3-hr recovery period after glycogen depletion participants will consume a nutrient free (0 kcal) beverage. For the day participants will consume a diet of 1.5 g/kg/d carbohydrate following glycogen depletion
All Cause Mortality
	Adequate Carbohydrate		Low Carbohydrate
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/12 (0%)		0/12 (0%)
Serious Adverse Events
	Adequate Carbohydrate		Low Carbohydrate
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/12 (0%)		0/12 (0%)
Other (Not Including Serious) Adverse Events
	Adequate Carbohydrate		Low Carbohydrate
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	1/12 (8.3%)		1/12 (8.3%)
Musculoskeletal and connective tissue disorders
Hamstring Cramping	1/12 (8.3%)	1	1/12 (8.3%)	1

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.