Eat 2 ounces of walnuts provided by the study. Eating any other nuts is to be avoided. Background diet will be low in polyphenols.

Eating any other nuts is to be avoided. Background diet will be low in polyphenols.

Healthy

Normal Weight

Overweight

Obese

Study participants will eat 2 ounces of walnuts each day.

Walnut

Nut-Free

Ragle Human Nutrition Research Center

Response of Gut Microbiota and Cardiometabolic Biomarkers to Consumption of Walnuts

Changes in fecal metagenomics will be assessed via Illumina sequencing

Fecal secondary bile acids will be measured by LC-QTOF/MS

Short-chain fatty acids in fecal matter will be measured by LC-MS/MS

Measure for changes in the levels of urolithins in separated plasma; measured in units of ug/uL. Blood will be assessed at each of the 4 study visits.

Measure for changes in the levels of calprotectin in collected fecal matter; measured in units of ug/g. Feces will be assessed at each of the 4 study visits.

Measure for changes in the levels of HDL and LDL in separated plasma; measured in units of mg/dL. Blood will be assessed at each of the 4 study visits

Measure for changes in PON-1 activity in separated plasma at each study of the 4 study visits. Activity will be calculated based on linear rates of hydrolysis of phenyl acetate to phenol

Changes in CETP activity will be measured at each of the 4 study visits. The isolated plasma will be co-incubated with donor and acceptor molecules using a commercially available assay kit.

hsCRP will be measured in blood at each study visit

SAA will be measured in blood at each study visit

ICAM and VCAM will be measured in blood at each study visit using a commercially available ELISA kit.

In-vitro: media from LPS stimulated monocytes will be used to measure gene expression and cytokines Measurement of changes in PBMC derived cytokines as a marker of inflammation at each of the 4 study visits. Assessment will be based on an increase or decrease in concentration in pg/ml.

Use a computer program to check for study dietary requirements. Subjects are to avoid olives and olive containing products during study protocol periods. This will be done using a subject self-recorded 3-day food record submitted to study personnel.

Measure waist (cm) and hip circumference (cm) before the start of drawing blood on study day protocols.

Measure weight (kg) before the start of drawing blood on study day protocols

Measure blood pressure (mmHg) before the start of drawing blood on study day protocols

University of California, Davis

After screening, participants will avoid foods high in ellagic acid. These foods include pomegranates, hazelnuts, pistachios, strawberries, raspberries, blackberries, oak-aged wines, spirits, and walnuts (besides the ones given by researchers); a complete list will be provided to the subjects. Participants will then return to research facility and provide urine and stool samples, as well as a set of 3-day dietary records. Then, they will start to consume 2 ounces of walnuts per day for 21 days with their usual diet. At the end, they will collect another urine and stool sample as well as another set of dietary records, and then come in for the scheduled colonoscopy where they will be asked to provide biopsy specimens. That completes the intervention and participation in the study.

Colo-rectal Cancer

Colon Cancer

Diet Habit

Participants will consume 2 ounces of walnuts for 21 days

Walnuts

University of Connecticut Health Center

Ellagic Acid, Urolithin and Colonic Microbial Communities Affected by Walnut Consumption

Urolithin will be measure in urine using a mass spectrometer to characterize this population into three know urolithin phenotypes, Uro-A, Uro-B and Uro-0, after walnut consumption.

Assess biomarkers in colonic mucosa from biopsy samples collected at colonoscopy that are associated with the three different urolithin metabotype following walnut consumption. This biomarkers is a total of 287 genes involved in cellular apoptosis and proliferation, inflammation and senescence, including TIMP1 (used for power calculations), cytokine and T cell and B cell signaling genes, as well as markers of lymphocyte subsets and immune checkpoint pathways and targets, providing a wide range of functional annotation groups.

Microbiome will be analyzed before and after walnut consumption with the goal to identify microbes responsible for urolithins production. DNA will be extracted from fecal samples and the V1-3 hyper-variable region of bacterial 16S rRNA genes will be sequenced to analyze bacterial community structure. Although 16S rRNA gene sequencing allows inexpensive bacterial identification to the genus/species level, mWGS will be employed in order to achieve finer classification (strain level), and identify other microbes, including viruses, fungi and small eukaryotes.

Daniel Rosenberg

Professor

UConn Health

American Institute for Cancer Research

California Walnut Commission

The Jackson Laboratory

This will be the comparative arm, of 6 weeks before and after the study participant is on their habitual diet

Experimental Arm of 12 weeks of Walnut Intake, with study visits at baseline (prior to walnut intake) and after 6 and 12 weeks of 40g of Walnut Intake.

Overweight and Obesity

Endothelial Dysfunction

Cardiovascular Risk Factor

Walnut Intake

Department of Nutrition

Influence of Walnut Intake on Vascular Function and Metabolism

The Influence of Daily Walnut Intake on Vascular Function and Associated Changes in Lipid Mediators and Primary Metabolites.

Digital microvascular function as measured by the EndoPAT2000

Circulating levels of non-esterified fatty acids

Circulating levels of non-esterified oxylipins

Conjugated and unconjugated Ellagitanin-derived metabolites

Nitric oxide metabolites produced from the intervention

w Will include total platelet number and mean platelet volume

Will assess fasting cholesterol and triglyceride levels

Will assess fine facial wrinkles and redness

Open-arm study, comparing the effects of 12 weeks of 40g of walnut intake to 6 weeks of the study participant's habitual diet.

The excretor status will be determined by analysis for urolithin A glucuronide in 24 hour urine after one dose of POMx. A blood sample and stool sample will be obtained before administering the extract.

The excretor status was determined by analysis of urolithin A glucuronide in 24 hour urine after one dose of POMx. A blood sample and stool sample were obtained before administering the extract.
PomX: 1000 mg POMx daily for 4 weeks

Urolithin Excretors

The excretor status was determined by the analysis of urolithin A glucuronide in 24 hour urine after one dose of POMx. A blood sample and stool sample were obtained before administering the extract.
PomX: 1000 mg POMx daily for 4 weeks

Non Excretors

Total

Sex: Female, Male

Race/Ethnicity, Customized

Body Mass Index (BMI)

Difference of percentage of total microflora between urolithin excretors and non-excretors after consumption of pomegranate extract for 4 weeks

Microbial phylum abundance (% of total) in urolithin excretors after consumption of 1000 mg pomegranate extract daily for 4 weeks

Microbial Phylum Abundance in Urolithin Excretors

Microbial phylum abundance (% of total) in urolithin excretors after consumption of 1000 mg pomegranate extract daily for 4 weeks.

Microbial Phylum Abundance in Non Excretors

Change in Percentage of Total Microflora Between Urolithin Excretors and Non-excretors

Overall Study

Zhaoping Li, Professor

microbial phylum abundance in urolithin excretors after consumption of 1000 mg pomegranate extract daily for 4 weeks

Microbial Phylum Abundance in Urolithin Non Excretors

Urolith

PomX

Effects of Pomegranate Extract on Intestinal Flora

Variations in Urolithin Production by the Microbiome After Pomegranate Extract Administration: Understanding Metabolism and Responsiveness to Pomegranate Ellagitannins

urolithin/orah

Response of Gut Microbiota and Cardiometabolic Biomarkers to Consumption of Walnuts

Ellagic Acid, Urolithin and Colonic Microbial Communities Affected by Walnut Consumption

Influence of Walnut Intake on Vascular Function and Metabolism

Effects of Pomegranate Extract on Intestinal Flora

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