Berries, Inflammation, and Gut Microbiome

Study Overview
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Brief Summary
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The primary objectives are to characterize changes in indices of systemic and gut inflammation, assess host- and microbial-derived metabolite pools, and describe and link functional metagenomics and metatranscriptomic alterations in the gut microbiome with metabolite and inflammatory outcomes after acute (24hr) and chronic (4 week) intake of anthocyanins and ellagitannins from strawberry and red raspberries compared to a control diet (negative control), FOS (positive control, non-polyphenol, carbohydrate-based fermentable fiber/pre-biotic), or combination diet (berry composite + FOS) in human participants with low-grade inflammation.

Detailed Description
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The proposed study will be conducted in humans according to Good Clinical Practice (GCP) guidelines. All subjects will review and sign an Informed Consent Form approved by the Illinois Institute of Technology's Institutional Review Boards (IRB) prior to screening. This is a single-center, randomized, single-blinded, 4-arm, placebo-controlled clinical trial. It features an acute single exposure 24-h protocol and a chronic 4-week daily intake protocol to evaluate the effects of anthocyanins and ellagitannins from strawberry and red raspberry intake compared to a control diet, FOS, or FOS and berries combined on gut inflammation, in adults with chronic low-grade inflammation. A planned sample size of 88 will be enrolled in the study. This study will require one initial screening visit, a pre-study visit, and 5 study visits. This study will take 5 weeks per subject to complete. The initial screening visit will provide subject with their site-specific, IRB-approved informed consent document prior to the start of any study-related procedures. Subject eligibility will be determined through a high sensitivity C-Reactive Protein (hs-CRP) marker value, anthropometric measurements, vital signs, fasting blood glucose test (finger prick), and completion of a survey related to general eating, health, mood and exercise habits. Eligible subjects will follow a limited polyphenolic diet throughout the duration of their participation, although stricter guidelines will be imposed during the 3 days prior to a study visit that involved blood/urine/fecal collection. Shopping lists and meal plans will be provided to subjects, along with counseling by the investigator's study investigators, to help subjects adhere to the limited polyphenolic diet. The trial will initiate with a 3-day food diary to assess background (pre-study) dietary intake followed by counseling to follow a diet relatively low in (poly)phenolic rich-beverages/foods, which will be maintained for the duration of the experiment. After an initial 7-day wash-in period on the limited polyphenolic diet, subjects will be randomized to 1 of 4 treatment sequences in the form of a drink: a mixed berries diet (raspberries and strawberries), a FOS diet (non-polyphenol, carbohydrate-based fermentable fiber/pre-biotic), a combination diet (mixed berries + FOS), or a control diet (negative control). Subjects will receive the same treatment for the duration of the study. Treatment codes will be maintained by the principal investigator. Each visit will last  2 hours and subjects will be required to remain at the Clinical Nutrition Research Center for the duration of the visit. Fasting blood samples will be collected at the 0 and 24h time points via a butterfly needle placed by a certified phlebotomist.

Official Title
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Polyphenols and Human Gut Microbiome Interactions: Role of in Ameliorating Inflammation

Conditions
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Healthy, Overweight or Obesity

Intervention / Treatment
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* Dietary Supplement: Mixed berries
* Dietary Supplement: FOS
* Dietary Supplement: Control
* Dietary Supplement: Combination


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria C-Reactive Protein (hs-CRP) > 1.0 and ≤ 10 ng/L BMI ≥ 25 kg/m2 Aged 20-60 years Able to provide informed consent and comply with study procedures Willing to maintain stable body weight and follow his/her habitual diet and physical activity patterns throughout the trial. Judged by the Investigator to be in general good health on the basis of medical history and screening laboratory tests. People with no documented disease condition that would interfere with the study endpoints (ie., CVD, diabetes, hypertension, major organ diseases) or taking medication or dietary supplements that may interfere with study endpoints Exclusion Criteria Current smoker or marijuana user No history or presence of atherosclerosis/cardiovascular disease, inflammatory disease, diabetes mellitus, or other systemic diseases, psychological or psychiatric disorders that may interfere with study outcomes. Taking any medications and/or supplements that would interfere with outcomes of the study (i.e., lipid-lowering medications, anti-inflammatory drugs, etc) Take part in prebiotics, probiotics or drugs active on gastrointestinal motility or a laxative of any class within 1 month Unstable use of any medication/supplement, this could include marijuana used as -needed for medical reasons Have a history of cancer, except for non-melanoma skin cancer in past 5 years Addicted to drugs and/or alcohol (>4 drinks/day) Have been exposed to any non-registered drug product within last 30 days. Working overnight (e.g. 3rd shift of overnight workers) Excessive exercisers or trained athletes Have allergies/intolerances to any of the study products. Extreme dietary habits (ie. vegetarian/vegan) Excessive coffee/tea drinker (>4 cups/day) Actively losing weight/ trying to lose weight (unstable body weight fluctuations of > 5 kg in 3 months) Donated blood within last 3 months Participated in a clinical trial within 1 month Female who is pregnant, planning to be pregnant, breastfeeding Any condition the Investigator believes would interfere with his or her ability to provide informed consent or comply with the study protocol, or which might confound

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 60 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: Other
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Mixed Berries<br>Strawberry and red raspberry composite served as a frozen drink | Dietary Supplement: Mixed berries<br>* Strawberry and red raspberry<br>|
| Active Comparator: FOS<br>Non-polyphenol, carbohydrate-based fermentable fiber/pre-biotic served as a frozen drink | Dietary Supplement: FOS<br>* Fructo-oligosaccharide<br>|
| Active Comparator: Combination<br>Mixed berry composite + FOS served as a frozen drink | Dietary Supplement: Combination<br>* Mixed berry composite + FOS<br>|
| Placebo Comparator: Control<br>Placebo similar in color to mixed berry supplement without any polyphenols served as a frozen drink | Dietary Supplement: Control<br>* Placebo similar in color to mixed berry supplement without any polyphenols<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in plasma biomarkers and measures of inflammation concentration: Nrf2/ NF-κB -response between 4 treatments | Plasma biomarkers and measures of inflammation: Nrf2/ NF-κB activation in monocytes will be analyzed. | Baseline to 4 weeks | 
| Changes in plasma biomarkers and measures of inflammation concentration-response between 4 treatments | Inflammatory cytokines (Interleukin-6 (IL-6), High Sensitivity C-Reactive Protein (hs-CRP), Tumor Necrosis Factor-α (TNF-α)) will be measured. | Baseline to 4 weeks | 
| Changes in plasma biomarkers and measures of inflammation concentration (GLP-2)-response between 4 treatments | Analysis of GLP-2 will be done in plasma samples before and after chronic exposure to the study foods. | Baseline to 4 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in gut inflammatory markers between 4 treatments. | The concentration of calprotectin, zonulin, and IgA in fecal samples will be determined by enzyme-linked immunosorbent assay (ELISA) before and after chronic exposure to study foods. | Baseline to 4 weeks | 
| Changes in Bile acids between 4 treatments. | Bile acids in plasma and fecal samples will be determined using UHPLC-QQQ-MS. | Baseline to 4 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Strawberry, Raspberry, Metabolic responses, Gut microbiome, Inflammatory markers