Healthy
Conditions
Brief summary
This is a research study about the effects of blueberries on the health of children who do not eat much fruits and vegetables. By doing this study, we hope to find out what nutrients from blueberries appear in the blood and urine of the child
Detailed description
This is a double blinded randomized control trial aiming to characterize the profile of polyphenol-derived metabolites in plasma and urine for children ages 11-12 years supplemented short term with wild blueberry powder. The study will assess if a short-term supplementation (5 days) with wild blueberry powder impacts immune responses as well as markers for cardiovascular and bone health. Up to 44 children will be recruited. The participants will attend three visits at Arkansas Children's Nutrition Center.
Interventions
DIETARY_SUPPLEMENTWild blueberry containing foods
Foods that contain wild blueberry supplement to deliver up to 30 grams of blueberry powder
DIETARY_SUPPLEMENTPlacebo containing foods
Foods that do not contain the blueberry powder but a placebo
Sponsors
United States Department of Agriculture (USDA)
National Institute of General Medical Sciences (NIGMS)
Arkansas Children's Hospital Research Institute
Study design
Allocation
RANDOMIZED
Intervention model
PARALLEL
Primary purpose
PREVENTION
Masking
TRIPLE (Subject, Caregiver, Investigator)
Masking description
Double blinded randomized control
Eligibility
Sex/Gender
ALL
Age
11 Years to 12 Years
Healthy volunteers
Yes
Inclusion criteria
* Eats \<1.5 cup of fruit and \<2.0 cup of vegetables per day * Boys or girls * All ethnicities * All BMIs
Exclusion criteria
* Known allergy to blueberries * Epilepsy * Asthma * Chronic kidney disease * Hormonal disease * Autoimmune disease * Bleeding disorders * Chronic infections * Type 2 and Type 1 diabetes mellitus * Attention deficit hyperactivity disorder * Opposition defiant disorder * Autistic spectrum disorder * Pre-existing medical conditions or medications as determined by the investigators to affect the outcomes of interest * If the participants/parents perceive barriers with adhering to low-polyphenol meals * Parent or child refusal to stop nutritional supplements * Parent or child refusal to have blood drawn
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention | Measurements were conducted hourly for 5 hours; the cumulative number of metabolites characterized over the 5 hours is reported | Fasting Blood and urine were collected at visit two and three following an overnight fasting. At visit three, children consumed a single dose of the study product and plasma and urine were collected |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Number and Percentage of Lymphocytes Cluster of Differentiation 4 (CD4), Lymphocytes Cluster of Differentiation 8 (CD8) and Natural Killer (NK) Cells | up to 2 weeks | Mitogen stimulated blood will be stained with combinations of fluorochrome-labeled antibodies and analyzed by flow cytometry. Absolute and relative number of CD4, lymphocytes CD8 and natural killer (NK) will be compared between groups. |
| T-lymphocyte Responsiveness to Immune Stressors | up to 2 weeks | Blood will be collected following an overnight fast after 5 days on the intervention. Whole blood will be cultured and cells stressed with E. coli lipopolysaccharide (LPS) to induce cytokine release from immune cells. Cytokines will be assayed using electrochemiluminescence based detection platform with multiplexed immunoassays, and their levels compared between study groups. |
| Endothelial Function by Brachial Artery Flow Mediated Dilation | up to 2 weeks | Endothelial-dependent flow-mediated vasodilation of the brachial artery in the fasting state was measured using high resolution ultrasound at enrollment and after 5 days on the intervention. This technology uses sound waves (ultrasound) to visualize the diameter of the brachial artery. A baseline image of the brachial artery will be recorded at rest. A blood pressure cuff will be placed around the lower arm and the cuff will be inflated to 50 mmHg above systolic pressure for 5 minutes, followed by rapid deflation of the cuff. The diameter of the brachial artery will be measured from ultrasound images captured throughout the 8 minute recording protocol. The percent change in the brachial artery will be compared between and within groups. |
Countries
United States
Participant flow
Recruitment details
Forty-four children were enrolled at the Arkansas Children's Nutrition Center, with the first participant enrolled on April 23, 2021, and the last participant on August 4, 2022
Pre-assignment details
After enrollment, the children were placed on a low-polyphenol diet for 48 hours. Following this period, they were assigned to consume either wild-blueberry products (n = 22) or placebo-containing products (n = 22).
Participants by arm
| Arm | Count |
|---|---|
| Wild Blueberry Supplement 30 grams of wild blueberry powder per day in foods items provided to study participants
Wild blueberry containing foods: Foods that contain wild blueberry supplement to deliver up to 30 grams of blueberry powder | 22 |
| Placebo Supplement Food items without wild blueberry power were provided to study participants.
Placebo containing foods: Foods that do not contain the blueberry powder but a placebo | 22 |
| Total | 44 |
Withdrawals & dropouts
| Period | Reason | FG000 | FG001 |
|---|---|---|---|
| Overall Study | Dropped or lost to follow up | 4 | 2 |
Baseline characteristics
| Characteristic | Placebo Supplement | Total | Wild Blueberry Supplement |
|---|---|---|---|
| Age, Continuous | 11.94 years STANDARD_DEVIATION 0.6 | 11.94 years STANDARD_DEVIATION 0.6 | 11.94 years STANDARD_DEVIATION 0.6 |
| BMI percentile | 50.7 percentile STANDARD_DEVIATION 30.6 | 63.5 percentile STANDARD_DEVIATION 32.9 | 72.2 percentile STANDARD_DEVIATION 30.6 |
| Diastolic blood pressure percentile | 51 percentile STANDARD_DEVIATION 20 | 52 percentile STANDARD_DEVIATION 21 | 53 percentile STANDARD_DEVIATION 22 |
| Ethnicity (NIH/OMB) Hispanic or Latino | 1 Participants | 1 Participants | 0 Participants |
| Ethnicity (NIH/OMB) Not Hispanic or Latino | 21 Participants | 42 Participants | 21 Participants |
| Ethnicity (NIH/OMB) Unknown or Not Reported | 0 Participants | 1 Participants | 1 Participants |
| Flow mediated dilation of the brachial artery | 11.6 Percentage of artery diameter STANDARD_DEVIATION 4.8 | 11.6 Percentage of artery diameter STANDARD_DEVIATION 4.5 | 11.8 Percentage of artery diameter STANDARD_DEVIATION 4.2 |
| Peak VO2 | 35.9 ml/kg/min STANDARD_DEVIATION 10.9 | 34.5 ml/kg/min STANDARD_DEVIATION 10.2 | 33.1 ml/kg/min STANDARD_DEVIATION 9.4 |
| Race (NIH/OMB) American Indian or Alaska Native | 0 Participants | 0 Participants | 0 Participants |
| Race (NIH/OMB) Asian | 0 Participants | 0 Participants | 0 Participants |
| Race (NIH/OMB) Black or African American | 4 Participants | 6 Participants | 2 Participants |
| Race (NIH/OMB) More than one race | 1 Participants | 1 Participants | 0 Participants |
| Race (NIH/OMB) Native Hawaiian or Other Pacific Islander | 0 Participants | 0 Participants | 0 Participants |
| Race (NIH/OMB) Unknown or Not Reported | 1 Participants | 3 Participants | 2 Participants |
| Race (NIH/OMB) White | 16 Participants | 34 Participants | 18 Participants |
| Region of Enrollment United States | 22 Participants | 44 Participants | 22 Participants |
| Sex: Female, Male Female | 11 Participants | 22 Participants | 11 Participants |
| Sex: Female, Male Male | 11 Participants | 22 Participants | 11 Participants |
| Systolic blood pressure percentile | 48 percentile STANDARD_DEVIATION 29 | 47 percentile STANDARD_DEVIATION 28 | 45 percentile STANDARD_DEVIATION 27 |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | 0 / 22 | 0 / 22 |
| other Total, other adverse events | 2 / 22 | 2 / 22 |
| serious Total, serious adverse events | 0 / 22 | 0 / 22 |
Outcome results
Primary
Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention
Fasting Blood and urine were collected at visit two and three following an overnight fasting. At visit three, children consumed a single dose of the study product and plasma and urine were collected
Time frame: Measurements were conducted hourly for 5 hours; the cumulative number of metabolites characterized over the 5 hours is reported
Population: Phenolic-derived metabolites were characterized in the samples using an ultra-high performance liquid chromatography method coupled with mass-spectrometry/mass spectrometry multiple reactions monitoring
| Arm | Measure | Group | Value (NUMBER) |
|---|---|---|---|
| Wild Blueberry Supplement | Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention | Plasma | 13 Counts of metabolites |
| Wild Blueberry Supplement | Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention | Urine | 25 Counts of metabolites |
| Placebo Supplement | Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention | Plasma | 0 Counts of metabolites |
| Placebo Supplement | Phenolic Acid Metabolites Measured in Plasma and Urine After Dietary Intervention | Urine | 3 Counts of metabolites |
Secondary
Endothelial Function by Brachial Artery Flow Mediated Dilation
Endothelial-dependent flow-mediated vasodilation of the brachial artery in the fasting state was measured using high resolution ultrasound at enrollment and after 5 days on the intervention. This technology uses sound waves (ultrasound) to visualize the diameter of the brachial artery. A baseline image of the brachial artery will be recorded at rest. A blood pressure cuff will be placed around the lower arm and the cuff will be inflated to 50 mmHg above systolic pressure for 5 minutes, followed by rapid deflation of the cuff. The diameter of the brachial artery will be measured from ultrasound images captured throughout the 8 minute recording protocol. The percent change in the brachial artery will be compared between and within groups.
Time frame: up to 2 weeks
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Wild Blueberry Supplement | Endothelial Function by Brachial Artery Flow Mediated Dilation | 9.4 percentage change in arterial diameter | Standard Deviation 3.4 |
| Placebo Supplement | Endothelial Function by Brachial Artery Flow Mediated Dilation | 9.4 percentage change in arterial diameter | Standard Deviation 3 |
p-value: <0.05Wilcoxon (Mann-Whitney)
Secondary
Number and Percentage of Lymphocytes Cluster of Differentiation 4 (CD4), Lymphocytes Cluster of Differentiation 8 (CD8) and Natural Killer (NK) Cells
Mitogen stimulated blood will be stained with combinations of fluorochrome-labeled antibodies and analyzed by flow cytometry. Absolute and relative number of CD4, lymphocytes CD8 and natural killer (NK) will be compared between groups.
Time frame: up to 2 weeks
Secondary
T-lymphocyte Responsiveness to Immune Stressors
Blood will be collected following an overnight fast after 5 days on the intervention. Whole blood will be cultured and cells stressed with E. coli lipopolysaccharide (LPS) to induce cytokine release from immune cells. Cytokines will be assayed using electrochemiluminescence based detection platform with multiplexed immunoassays, and their levels compared between study groups.
Time frame: up to 2 weeks