Fruit and Vegetable Biomarker Discovery

Study Overview
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Brief Summary
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The purpose of this research is to find a set of markers in the blood and/or urine that can be linked to consumption of specific fruits and vegetables. This will allow for better understanding of the link between diet and health-related outcomes. Furthermore, the results of this study will lead to the development of new methods to evaluate the nutritional status of individuals in both community and clinical settings. Food frequency questionnaires and diaries/recalls can be affected by intentional or unintentional misreporting, and thus can create errors in determining nutritional status. This study will lead to the development of an objective way to assess the consumption of specific fruits and vegetables by the general population.

Detailed Description
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The purpose of this research is to find a set of markers in the blood and/or urine that can be linked to consumption of specific fruits and vegetables. This will allow for better understanding of the link between diet and health-related outcomes. Furthermore, the results of this study will lead to the development of new methods to evaluate the nutritional status of individuals in both community and clinical settings. Food frequency questionnaires and diaries/recalls can be affected by intentional or unintentional misreporting, and thus can create errors in determining nutritional status. This study will lead to the development of an objective way to assess the consumption of specific fruits and vegetables by the general population. This study includes 3 aims to identify and then validate urine and plasma biomarkers associated with fruit and vegetable intake. Aim 1 will use a randomized crossover dietary intervention to determine the dose and time response of metabolites in plasma and urine associated with exposure to a mixture of MyPlate fruits and vegetables. Aim 2 will use a parallel design, controlled diet intervention to determine whether the biomarkers of fruit and vegetable intake determined in Aim 1 are predictors of consumption in the context of a defined dietary pattern. Aim 3 will be a cross-sectional validation arm that determines if the biomarkers of food intake developed in Aims 1 and 2 have predictive value reflecting recent and habitual consumption of these foods in a heterogeneous and diverse population.

Official Title
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University of California (UC), Davis Dietary Biomarkers Development Center

Conditions
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Biomarkers

Intervention / Treatment
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* Other: Aim 1 Meal Challenge 1
* Other: Aim 1 Meal Challenge 2
* Other: Aim 1 Meal Challenge 3
* Other: Aim 2 Typical American Diet
* Other: Aim 2 Typical American Diet Plus Test Foods
* Other: Aim 2 Dietary Guidelines for Americans Diet Plus Test Foods


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Body Mass Index (BMI) 18.5-39.9 kg/m^2 Willingness to provide urine and have blood drawn Exclusion Criteria: Pregnancy or lactation Allergy or aversion to foods provided in test diet History of gastrointestinal disorders including the following: Ulcerative colitis or Crohn's disease Celiac sprue Hereditary non-polyposis colorectal cancer (HNPCC) Familial adenomatous polyposis Pancreatic disease Liver disease Previous gastrointestinal resection or bariatric surgery Bleeding disorders that preclude blood draws Recent hospital admissions (past 6 months) for myocardial infarction (MI), cerebrovascular accident (CVA), or congestive heart failure (CHF) Cardiovascular disease (CVD) under physician guided therapy that is not medically stable Cancer under radiation or chemotherapy treatment that is active or within 6 months of treatment Weight change (± 5% in the last 3 months) Regular alcohol intake of > 2 drinks/day (equivalent to 720 mL of beer, 240 mL of wine, or 90 mL of spirits), and unwilling to abstain during feeding periods Use of tobacco and/or marijuana, hookahs, e-cigarettes, or vapes and not willing to abstain during feeding periods Use of illicit drugs and not willing to abstain during feeding periods BMI >40 kg/m^2 Regular (daily to weekly) use of over-the-counter (OTC) weight-loss aids and unwilling or unable to stop taking these during feeding periods Regular (daily to weekly) use of OTC anti-inflammatories and unwilling or unable to stop taking these during feeding periods Unwilling to stop taking OTC dietary supplements that interfere with the test foods being studied including pills, chewables, liquids or powders for the following (may continue if vitamin supplement is prescribed by a medical doctor): Protein supplements Soy Fiber Flaxseed Fish oil (including cod liver oil) Probiotics Carotenoids Selenium Other antioxidants Other phytochemicals Glucosamine Chondroitin Oral or IV antibiotic use in the past 6 months (could defer participation until 6 months post-completion of course of antibiotics) Seated blood pressure >140/90 mmHg Fasting clinical lab values outside of the following ranges: Fasting Glucose: 54-125 mg/dL Urea: 6-50 mg/dL Creatinine: 0.4-1.3 mg/dL Estimated Glomerular Filtration Rate (eGFR): >60 mL/min Sodium: 133-146 mmol/L Alanine Transaminase (ALT): 5-60 U/L Aspartate Aminotransferase (AST): 5-40 U/L Alkaline Phosphatase (ALP): 20-135 U/L Total Bilirubin: 0.0-1.9 mg/dL Total Protein: 5-9.0 g/dL Albumin: 3.5-5.9 d/L Low-density Lipoprotein (LDL) Cholesterol: <160 mg/dL Triglycerides: <500 White Blood Cells: 3-10.5 Kl/µL Hematocrit: 35-48 g/dL (women), 37.5-49 g/dL (men) Current use of the following prescription medications: Diuretics Steroids (oral): daily oral any dose within 1 month of study (except for oral contraceptive pills) Opiates: any use within 1 month of study Hypolipidemic agents that affect GI or renal function (ie. fibrates) Hypoglycemic medications other than metformin (ie. insulin, SGLT2 inhibitor, α-glucosidase inhibitor) Psychiatric medications that affect metabolism/renal function (anti-psychotics, lithium) Biologics/immune modulators (ie. rheumatoid arthritis, psoriasis, other rheumatologic/hematologic active disease) Anti-coagulants (coumadin, heparin, Eliquis, etc.) HIV/HAART medications (dyslipidemia inducing) Inability to freely give informed consent

Ages Eligible for Study
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Minimum Age: 18 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: Basic Science
Allocation: Non-Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Aim 1: crossover, Aim 2: parallel, Aim 3: observational
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Aim 1 Group 1<br>Order of Treatments: Meal Challenge 1, Meal Challenge 2, Meal Challenge 3 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 1 Group 2<br>Order of Treatments: Meal Challenge 1, Meal Challenge 3, Meal Challenge 2 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 1 Group 3<br>Order of Treatments: Meal Challenge 2, Meal Challenge 3, Meal Challenge 1 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 1 Group 4<br>Order of Treatments: Meal Challenge 2, Meal Challenge 1, Meal Challenge 3 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 1 Group 5<br>Order of Treatments: Meal Challenge 3, Meal Challenge 1, Meal Challenge 2 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 1 Group 6<br>Order of Treatments: Meal Challenge 3, Meal Challenge 2, Meal Challenge 1 | Other: Aim 1 Meal Challenge 1<br>* A meal challenge will be provided that contains 1 serving of peaches and 2 servings of strawberries as a smoothie and 2 servings of tomatoes and 1 serving of carrot as a vegetable dish.<br>Other: Aim 1 Meal Challenge 2<br>* A meal challenge will be provided that contains 1 serving of banana and 2 servings of peaches as a smoothie and 1 serving of tomatoes and 2 servings of green beans as a vegetable dish.<br>Other: Aim 1 Meal Challenge 3<br>* A meal challenge will be provided that contains 2 serving of bananas and 1 serving of strawberries as a smoothie and 1 serving of green beans and 2 servings of carrots as a vegetable dish.<br>|
| Experimental: Aim 2 Typical American Diet<br> | Other: Aim 2 Typical American Diet<br>* The diet will provide sufficient calories for weight maintenance and will contain refined grains and low amounts of fruits and vegetables, representative of the typical American diet. No bananas, strawberries, peaches, green beans, carrots, or tomatoes will be provided.<br>|
| Experimental: Aim 2 Typical American Diet Plus Test Foods<br> | Other: Aim 2 Typical American Diet Plus Test Foods<br>* This diet will provide sufficient calories for weight maintenance and will contain refined grains and low amounts of fruits and vegetables, representative of the typical American diet. Bananas, strawberries, peaches, green beans, carrots, and tomatoes will be provided in amounts less than or equal to 1 serving per day each.<br>|
| Experimental: Aim 2 Dietary Guidelines for Americans Diet Plus Test Foods<br> | Other: Aim 2 Dietary Guidelines for Americans Diet Plus Test Foods<br>* This diet will provide sufficient calories for weight maintenance and will contain whole grains and high amounts of fruits and vegetables, representative of the Dietary Guidelines for Americans. Bananas, strawberries, peaches, green beans, carrots, and tomatoes will be provided in amounts greater than or equal to 1 serving per day each.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in urine metabolomics profile | Urine metabolites will be measured by ultra-performance liquid chromatography mass spectrometry (UPLC-MS) before and after consumption of no, low, or high doses of study fruits and vegetables. Change will be assessed by the appearance of and change in levels of dietary-derived metabolites, which are candidate dietary biomarkers. | Aim 1: day 3, 8, 13; overnight fasted and post prandial 0-2 hours, 2-4 hours, 4-6 hours, 6-8 hours, 8-24 hours. Aim 2: day 3, 9, 10; Aim 3: day 4. | 
| Change in plasma metabolomics profile | Plasma metabolites will be measured by ultra-performance liquid chromatography mass spectrometry (UPLC-MS) before and after consumption of no, low, or high doses of study fruits and vegetables. Change will be assessed by the appearance of and change in levels of dietary-derived metabolites, which are candidate dietary biomarkers. | Aim 1: day 3, 8, 13; overnight fasted and post prandial 1 hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 7 hour, 8 hour, 24 hour. Aim 2: days 3, 9, 10. Aim 3: day 4. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Diet adherence | Intake of test and non-test meals/foods will be monitored through weigh-backs by study personnel in Aim 1 and participant self report in Aim 2 to identify deviations in intake of fruits/vegetables of interest. | Aim 1: day 3, 8, 13. Aim 2: day 3, 6, and 9. | 
| Habitual consumption of fruits and vegetables | 3-month General Nutrition Assessment Food Frequency Questionnaires (GNA FFQ) will be administered at site visit 1 for each aim. | Day 1 | 
| Acute consumption of fruits and vegetables | An Automated Self-Administered 24-Hour (ASA24) 3-day food record will be obtained over 2 weekdays and 1 weekend day during Aim 3. | Days 1, 2, 3. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Metabolomics, Diet, Fruit, Vegetables, Biomarker, Urine, Blood