Cardiometabolic Benefits of Potatoes Mediated Along the Gut-Vessel Axis in Adults With Metabolic Syndrome
Phase: NA
Conditions: Cardiovascular Diseases, Endotoxemia, Endothelial Dysfunction, Postprandial Hyperglycemia, Gut Health
Sponsor: Ohio State University

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 1
Cardiometabolic Benefits of Potatoes Mediated Along the Gut-Vessel Axis in Adults with Metabolic 
Syndrome 
 
I. 
Objectives 
 
The objective of this application is to establish evidence that incorporation of potatoes into a diet 
pattern based on the Dietary Guidelines for Americans (DGA) enhances cardiometabolic health in 
persons with metabolic syndrome (MetS) by improving microbiota- and host-mediated metabolic 
responses along the gut-vessel axis. Our central hypothesis is that the inclusion of potatoes, as part 
of a DGA-based dietary pattern, will potentiate gut barrier function and attenuate postprandial 
hyperglycemia (PPH)-mediated impairments in vascular endothelial function (VEF). To test this, we 
will complete the following objectives: 1) define changes in gut barrier function in association with 
improved gut microbiota composition, increased fecal short chain fatty acid (SCFA) production, and 
decreased serum endotoxin, 2) define changes in postprandial glycemic responses and endotoxemia, 
and 3) define changes in gut hormones that promote glycemic control and changes in markers of 
oxidative stress in relation to improvements in endothelial vascular function, all following 2-week 
potato consumption. Upon completing this study, we anticipate novel evidence demonstrating gut-
level benefits of a potato-containing diet to lower cardiometabolic risk in MetS adults.      
 
 
II. 
Procedures 
 
A. Study Design 
We will enroll equal numbers of male and female MetS adults (n = 30; 18-50 y) from the Columbus, 
OH area as we detailed1 to complete a 2-arm, randomized controlled, cross-over trial. Participants 
will be randomized in 3-unit blocks to receive a DGA-based diet containing Russet Burbank potatoes 
(350 g/d; 260 kcal; 12.4 g resistant starch) or an energy-matched bagel that is nearly devoid of 
resistant starch. Our approach closely follows recommendations of the DGA2 and entails high 
scientific rigor by providing all foods during each 14-d intervention. On day 0, 7, and 14 of each 
period, coinciding with when participants obtain prescribed foods, we will assess anthropometrics 
(weight, height, waist circumference) and blood pressure. At baseline (day 0) of each trial, fasting 
glucose, insulin, and endotoxin will be measured to verify stable health status. Following each 14-d 
intervention, participants will provide a fecal sample and undergo testing in the fasted state for 
metabolic endpoints, gut barrier function, and VEF. Then, they will receive a glucose challenge (75 
g) to examine VEF in relation to PPH at 30 min intervals for 2 h. Gut permeability probes (sucralose, 
erythritol, lactulose, mannitol) will also be co-administered on day 14, and urine collected for 24-h. 
After completing a 2-wk washout period (i.e. to restore microbiota composition3), participants will 
receive the alternate treatment (i.e. DGA diet + potato or bagel) and repeat all procedures. 
 
B. Study Subjects 
 
Enrollment Criteria. MetS adults will be studied because of their impaired VEF in association with 
glucose intolerance.4 We will enroll those who are unmanaged (i.e. no drug therapy) and they will be 
required to meet these established MetS criteria5 to improve homogeneity of study outcomes: i) 
fasting glucose 100-126 mg/dL, ii) waist circumference >102 cm for men or >88 cm for women, and 
iii) fasting triglyceride >150 mg/dL. We anticipate that they may also fulfill other MetS criteria, 
specifically HDL-C (<40 mg/dL for men, <50 mg/dL for women) and elevated blood pressure6 
(>120/80 mmHg); these will be considered for co-variate analysis. Other inclusion criteria are: i) 
non-smoker, ii) non-dietary supplement user (>1-mo), iii) free of gastrointestinal disorders, CVD, 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 2
cancer, and iv) no recent use of antibiotics, or any medications affecting glycemia, lipidemia, or blood 
pressure. Participants having any of these exclusion criteria will not be enrolled: i) use of anti-
inflammatory agents or probiotics, ii) vegetarian, gluten intolerant, carbohydrate-restricted diet, iii) 
alcohol intake >2 drinks/d, or iv) ≥7 h/wk of aerobic activity. 
 
Power Calculation. Plasma glucose and FMD during the 2-h postprandial period (AUC0-120 min) and 
fasting endotoxin following 14-d controlled feeding have been selected as the primary outcome 
variables. These endpoints had no gender differences (P>0.05) in our prior studies. PPH was also 
inversely related (P<0.001) to FMD.4,7 MetS adults had improved FMD following low-fat milk 
ingestion compared with rice milk (AUC0-3 h: 1150±145 vs 908±118, CV=56%) and attenuated PPH.4 
For this study, we predict that FMDAUC will be 25% greater in DGA+Potato during the oral glucose 
challenge on day 14. Thus, 26 MetS subjects would be needed to detect a treatment effect with >80% 
power (P<0.05). We will therefore enroll 30 subjects to account for unexpected attrition and increase 
power. A similar power calculation was performed using endotoxin measured from MetS vs. healthy 
adults.1 To detect a 25% predicted lowering of endotoxin, 22 MetS subjects would be needed. Based 
on our preliminary data, DGA+Potato could lower endotoxin by increasing vitamin C status in 
addition to alleviating gut dysbiosis in a resistant starch-dependent manner. This secondary 
hypothesis will be tested by measuring vitamin C on day 14 using our routine HPLC-ECD method.1  
Statistical Analysis Plan 
Most data in will be analyzed by linear mixed effects models (LMMs) with random effect for subjects 
(to account for repeated measures) and fixed effects for gender, time period, and their interactions. 
Multivariate regression analysis will define pairwise correlations between study variables in the 
presence or absence of potential study covariates. Statistical significance for all analyses will be set 
at P<0.05. 
C. Detailed Study Procedures 
 
Overview of Study Procedures. Potential participants who call the study center in an anonymous 
manner for more information as well as those identified through ResearchMatch will be given a brief 
description about the study and asked a few questions to determine their eligibility (see Phone Script 
attachment). If they meet the eligible criteria, they will be invited to the study center for a screening 
meeting. During the meeting, the Informed Consent (see Informed Consent attachment) will be 
explained and provided for them to review. The participant will then be given the opportunity to 
review the Informed Consent form. If he/she chooses to participate in the study, they will then be 
asked to provide written consent. Women choosing to participate will also be asked to complete the 
Menstrual History QuestionnaireAfter receiving informed consent, the participants’ height, weight, 
waist circumference, and blood pressure will be measured. Additionally, a small fasting blood sample 
will be collected for blood chemistry analysis. If they are not fasted at least 10 hours, they will be 
asked to come back in the fasted state at time of mutual convenience. These blood results in 
combination with anthropometric parameters will determine the participant’s eligibility. Eligible 
participants who agree to proceed with the study will then complete two 2-week intervention periods 
in a randomized order, each followed by a 2-h postprandial trial. We estimate that completion of all 
study procedures will take ~6-12 weeks per participant. Each step of the study procedure will be 
discussed in detail below: 
 
Screening Meeting. Potential participants who have met the initial criteria of the study (based on the 
telephone interview) will be invited to the study center at a mutually convenient time. During this 
time, the participant and a member of the research team will meet in a private, quiet conference room 
or office. The individual will be provided the informed consent form, and its contents will be 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 3
described to the potential participant. The participant will then can review it, and if they choose to 
participate in the study, they will be asked to provide written consent. Although the participant will 
be asked to sign the informed consent, the participant will be told that they will not be asked to 
participate if their body measurements, blood pressure, or plasma chemistries (see Enrollment 
Criteria) do not meet the study criteria. If the participant has provided consent, we will then measure 
the participant’s height, weight, waist circumference, and blood pressure. Next, if the participant is 
fasted for at least 10 hours, we will ask if a trained individual can draw a small blood sample (10 mL; 
1 tube) so that we may measure blood chemistries (glucose, HDL-cholesterol, triglyceride). All 
samples will be coded to maintain participant anonymity. If the participant’s anthropometrics, blood 
pressure, or plasma chemistries do not meet the study criteria, they will be told that they do not meet 
the study criteria.  
 
Potential participants who meet the criteria will be contacted within a few days after their screening 
meeting to provide them with their blood and body measurement results and inform them of their 
eligibility to participate in the study. Consistent with our CLIA exemption, blood results will be 
provided in a categorical manner (i.e. “normal”, “marginally high”, “high”) rather than providing 
actual blood concentrations of lab values (see Subjects Results Sheet). Potential participants having 
any blood values outside of the “normal range” will be directed/encouraged to follow-up with their 
own physician. Those having body measurements, blood pressure, and blood values within 
acceptable limits (see Inclusion/Exclusion criteria), will be invited to participate in the study. Subjects 
will be included or excluded based on a best fit of the inclusion and exclusion criteria (an example of 
best fit would be if a potential subject says he/she exercises 8 h/wk, which is close to our exclusion 
criteria of 7 h/wk, they might still be included in the study if they meet all other inclusion and 
exclusion criteria more closely than other potential subjects). Participants will be read one phone 
script if they qualify and another phone script if they do not qualify (see Participant Eligibility Phone 
Script attachment). Potential participants who qualify for the study will be communicated a message 
as follows: “Congratulations! You have been selected for participation in our study based on your 
blood testing results. A study investigator will be telephoning you to invite you to the testing session. 
Would you like to know your blood chemistry results?” Subjects not selected for study will be told 
the following message: “You have not been chosen for our study, but thank you for your interest.”  
This message will be followed by an explanation why they were not chosen, such as lab values outside 
of the range we are looking for: “Your blood testing data is…..we were looking for participants who 
had levels less than …..”.  “We can provide you a copy of your results if you would like…..how 
would you like us to provide them to you?”. “You should also consider sharing these results with 
your physician.”  
 
If a participant is telephoned and is unavailable, a message will be left requesting a callback at a 
convenient time or that at a member of the study team will try calling again at a later time. No 
confidential or sensitive information will be shared with third parties or left on answering machines. 
 
Intervention timeline. Each participant will complete a 2-arm (DGA + Potato; DGA + Bagel), 
randomized, cross-over study. For this study, each participant will visit the study center a total of 12 
times (6 times per study arm, including 1) baseline measurements of anthropometrics, serum glucose, 
insulin, endotoxin, and pick-up of meals for days 1-3, 2) pick-up of meals for days 4-7 2) pick-up of 
meals for days 8-11 and measurement of anthropometrics, 3) pick-up of meals for days 11-14 and 
stool collection kit, 4) 2 h postprandial testing, and 5) return samples from 24-h urine collection. 
Interventions will be separated by two weeks (for women, interventions will be separated by 2, 6, 10 
etc. weeks to ensure vascular testing will be performed at the same time of the menstrual cycle). 
 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 4
Baseline visit. On day 0 of each trial arm, participants will report in the fasted state (10-12 hours) to 
the study center for assessment of height, weight, waist circumference, blood pressure, serum 
glucose, serum insulin, and serum endotoxin. They will receive 3-4 days of DGA meals plus potatoes 
(350g/day) or bagels (1/day = 95g). 
 
Dietary intervention. Participants will be provided all foods for the 14-d intervention period. After 
the baseline visit, participants will report back to the study center on days ~3, 7, and 10 to pick up 
their meals for the proceeding 3-4 days. On day 7, anthropometrics (height, weight, and waist 
circumference) will also be measured to ensure weight maintenance. To standardize resistant starch 
(~12 g/d/350 g potato) and promote dietary compliance, potatoes will be prepared in several ways 
that minimize resistant starch degradation8 (e.g. Day 1 = whole baked, Day 2 = mashed, Day 3 = 
seasoned wedges, Day 4 = sliced, with soy milk and vegan cheese) without creating a food product 
that is contraindicative to a DGA diet (e.g. potato chips, deep-fat fried). The prepared potatoes will 
be refrigerated by the participants until ready for consumption, at which point they may be 
microwaved if desired. Participants will consume the potato daily, with a meal of their choice. 
Participants will be provided non-sodium seasoning (e.g. pepper blend) or may consume the potato 
with other foodstuffs (e.g. eggs at breakfast). Similarly, participants may choose to co-ingest the bagel 
with other provided foods. 
 
Test trials.  At the end of the intervention period (day 14), participants will report back to the 
laboratory in the fasted state (10-12 hours) for assessment of height, weight, waist circumference, 
blood pressure, serum glucose, serum insulin, and serum endotoxin. Participants will return a stool 
sample collected on day 13 of the intervention period at home. Participants will ingest a sugar solution 
containing 75 g glucose, which we and others have routinely shown to induce PPH, oxidative stress, 
and VED. This will be co-ingested with 1 g sucralose,1 g erythritol, 1 g mannitol, 5 g lactulose for 
assessment of gut permeability. Then, at 30 min intervals during the 2 h postprandial period, we will 
assess blood markers and brachial artery flow-mediated dilation. During each study visit, participants 
will have access to drinking water and use of the restroom as needed. Urine will be collected during 
the 2 h postprandial period in the laboratory, and participants will be instructed on urine collection 
for when they return home following their laboratory visit. Participants will return to the study center 
the following day to drop off their 24-h urine collection containers. To ensure that the order by which 
each participant completes each trial is randomized, a random sequence generator 
(http://www.random.org/sequences/) will be used to determine the order by which each participant 
will undergo the two trials. 
 
Sugar Probe Test. On the morning of testing at day 14, participants will arrive at the study center 
after abstaining from food and only consuming water for 10-12 hours. A sugar probe test will be 
conducted as described.9 Upon arrival, participants will be asked to empty their urinary bladder. 
Subsequently, they will then be asked to ingest a drink containing 1 g sucralose, 1 g erythritol, 1 g 
mannitol, and 5 g lactulose dissolved in a 75 g glucose solution (240 mL). Within 2-hours after 
ingesting the sugar test beverage, participants will be asked to consume an additional 500 mL of 
water. Additionally, participants will be provided with standardized sucrose-, erythritol-, lactulose-, 
and mannitol-free meals (e.g. English muffin, butter, scrambled eggs) during the 24-h period. All 
foods consumed during the 24-hour period will be free of artificial sweeteners. Urine will be collected 
from 0-5 and 6-24 hours in sterile containers. For each urinary collection period, participants will be 
provided 2-L urine collection containers and an insulated bag with ice packs to store collected urine 
until they return their samples the next day. 
 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 5
Sampling Handling. At each time point of the postprandial trial and once at baseline, a blood sample 
(20 mL; 1.4 tablespoons x 5 time points = 100 mL or 0.42 cups) will be collected into evacuated 
blood collection tubes. Collecting a total of 100 mL or 0.42 cups during the postprandial trial and 
once at baseline is necessary to ensure adequate amount of plasma for each time point to accurately 
analyze levels of glucose, insulin, endotoxin and gut hormones. During blood collection, participants 
may feel an initial pain when inserting the needle, bruising around the insertion area, lightheadedness, 
or fainting, which are common when donating blood. However, we do not foresee any additional 
significant risks for collecting this amount of blood over 2 hours, other than the risks stated 
previously.  
 
Throughout the span of the study (~8-12 weeks in duration depending on participant/investigator 
availability) which consists of four weeks total of controlled feeding, a blood collection at wk 0 and 
2 and then repeated following a ≥2 week washout period, and 1 screening day, we will be collecting 
a total of ~210 mL or ~0.88 cups) of blood. Urine will be collected in provided containers (VWR) 
containing 10% thymol to inhibit bacterial growth. We will then have the participants return their 
urine samples to the study center or coordinate with study personnel to meet at a public and mutual 
location to return their samples within 24-h after collection. Feces will be collected using a 
commercial commode specimen collection system (Fisher Scientific). Briefly, the collection kit 
consists of the necessary materials (e.g. gloves, waste bag) for participants to easily and hygienically 
collect their stool without contaminating the sample or themselves. Volumes of urine will be recorded 
and fecal mass and observations will be recorded based on the Bristol Stool Chart.10  
 
During each blood collection, plasma will be obtained by centrifugation and then transferred to 
cryogenic storage tubes. Serum samples will be obtained by allowing the blood to clot, followed by 
centrifugation and transfer to cryogenic storage tubes. Tubes will be stored at -80° C until analysis 
can be completed. Analyses will include plasma glucose, insulin, triglyceride, total cholesterol, 
nitrite, nitrate, arginine, ADMA, SDMA, MDA, CCK, serum endotoxin. Urine will be stored at -
80ºC until analysis can be completed. Analysis from urine will include sucralose and erythritol. 
Fecal samples will be stored at -80ºC until analysis can be completed. Feces analysis will include 
microbiota composition and fecal short chain fatty acids (butyrate, acetate, propionate). Remaining 
plasma, serum, urine, and fecal samples not used for these analyses will be archived for 5 years at 
-80 C in the event we decide to measure additional inflammatory, antioxidant, or microbiota related 
markers. Appropriate notation has been made in the informed consent to alert participants that we 
will be archiving specimens and that they have the right to refuse our use of these specimens for 
future analyses. Lastly, approval from OSU IRB will be sought via a protocol amendment prior to 
the analysis of any additional biomarkers not specified herein. 
 
Privacy/Confidentiality. For all data and records that are a part of this study, a number (i.e. code) 
will be assigned to each participant and will only be available to research personnel. Any records 
containing the names of participants will be stored in a locked filing cabinet or on a password 
protected computer in the PI’s laboratory or office. Research personnel under the supervision of the 
PI and the PI himself will be the only individuals that have access to this information. The names of 
participants will not be used for publication in any form. The records will be maintained until the 
data are published, up to a maximum of five years. All archived samples will be coded, but the key 
linking the code to each participant’s identifiable information will have been destroyed. In addition, 
participants will be instructed that their participation in this study is voluntary and that they may 
withdraw at any time without prejudice. Data (biochemical values) obtained from this study will 
stored on a computer in the PI’s laboratory. In addition, a backup of digital data will be stored on the 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 6
PI’s computer in his office. Both computers are password protected and both doors are locked when 
work areas are not in use. 
 
D. Internal Validity 
 
For all data and records that are a part of this study, a number (i.e. code) will be assigned to each 
participant. This will minimize measurement bias when performing analysis on dietary records, and 
biochemical markers because all samples/records will be coded. The codes will only be broken once 
data analysis has been completed and verified by the PI. 
 
F.   Medical Safety Plan 
All aspects of the clinical study will be conducted in Dr. Bruno’s clinical lab located in in 262D 
Campbell Hall. Participants will be fasted for 10-12 hours prior to each study visit. We recognize that 
certain risks associated with fasting include: hypoglycemia, weakness, and fainting. This duration of 
fasting is consistent with guidelines set forth by the American Diabetes Association to minimize risk 
to the individual62 when determining fasting blood glucose concentrations.  
 
Risks related to hypoglycemia are anticipated to be low due to participants’ already elevated fasting 
blood glucose (study entry criteria = 100-126 mg/dL). They will also be ingesting a glucose beverage 
containing 75 g of glucose immediately following the fasting period. Throughout each visit to the 
study center, participants will be closely monitored by Emily Shaw and Geoffrey Sasaki (Doctoral 
Students), both of whom are members of the research team. They will monitor the safety and well-
being of participants for any signs and symptoms of hypoglycemia including: confusion, dizziness, 
irritability, weakness, headaches, and fainting. Emily Shaw is certified by the Red Cross in CPR and 
basic first aid. Additionally, Dr. Bruno (PI; 325 Campbell Hall) has an academic office in close 
proximity to the study center and has significant experience coordinating clinical research studies 
involving overnight fasting, and other dietary- and carbohydrate-challenges, thereby supporting the 
competency of our research team in managing potential adverse events relating to fasting glucose and 
glycemic responses.  
 
Consistent with our prior studies of similar design, and to ensure participant safety, all procedures 
throughout each study visit will occur while positioned on a hospital bed in the prone position. In the 
event that a participant was to become weak, dizzy, or faint, they would already be ideally positioned 
to minimize risks associated with these symptoms. In the event that hypoglycemia-related symptoms 
occur, the study would be terminated to allow the participant to recover.  We are prepared to provide 
pre-packaged beverages and snacks containing simple carbohydrates (e.g. Gatorade, fruit juice, apple 
sauce, crackers) that will allow for rapid restoration of blood glucose. These food items will be stored 
in a refrigerator, located in close proximity (262 Campbell Hall), that is dedicated for foods used in 
research studies. Participant status (e.g. attentiveness, skin color) will be monitored in our clinical 
laboratory to ensure recovery. The clinical area where participants undergo vascular assessment is 
also equipped with first-aid measures (e.g. smelling salt) and all study team members are trained to 
assist with basic first-aid application if needed. 
 
Any adverse hypoglycemic response that might occur during this study will receive care 
commensurate to the symptoms.  For example, if a participant were to faint, then smelling salts would 
be administered along with a carbohydrate-containing food upon regaining consciousness. The study 
PI would also be contacted.   
 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 7
Alternatively, should a more severe hypoglycemic response occur (e.g. contusion or laceration 
relating to fainting), the research team would immediately contact medical services (i.e. 911). For 
non-life threatening emergencies, any OSU students participating in these studies would be directed 
to the Wilce Student Health Center.  For other emergencies or those participants who are not OSU 
students, individuals would be directed to the Wexner Medical Center either by transporting them 
directly or requesting ambulance service. Regardless of the complexity of the adverse events, the 
research team would monitor the participant in the interim, provide palliative care as appropriate, and 
follow-up after any medical care has been provided. 
 
 
 
 
 
 
 
Meal Pattern A: 
 
Meal Pattern B: 
 
Meal Pattern C: 
 
Meal Pattern D: 
Breakfast 
Boiled eggs 
 
Grapenuts cereal  
Whole wheat 
waffles 
 
Scrambled eggs 
Blueberries 
 
Soy milk 
 
Peanut butter 
 
Diced green bell 
pepper 
Oat milk 
 
Mandarin 
oranges 
 
Tropical fruit cup  
Diced onion 
English muffin  
 
English muffin 
 
Fruit & nut bar 
 
Whole wheat 
tortilla 
Butter spread 
 
Honey 
 
 
 
Fruit & nut bar 
Raisins 
 
Butter spread 
 
 
 
 
Lunch: 
Turkey breast 
 
Spinach 
 
Spaghetti 
 
Rotini pasta 
Wheat bread 
 
Sunflower oil 
 
Chicken 
meatballs 
 
Ground beef 
Baby carrots 
 
Whole wheat 
bread 
 
Marinara sauce 
 
Marinara sauce 
Fruit & nut bar 
 
Tuna 
 
 
 
 
Cheese slice 
 
Mayonnaise 
 
 
 
 
 
 
Diced pears 
 
 
 
 
Dinner: 
Grilled chicken 
 
Taco shells 
 
Turkey burger 
 
Tilapia filets 
Wheat bun 
 
Shredded 
chicken 
 
Wheat bun 
 
Brown rice 
Tomato sauce 
 
Grapes 
 
BBQ crisps 
 
Spinach 
Riced cauliflower 
 
 
 
Au gratin 
potatoes or 
bagel* 
 
Sunflower oil 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
 8
Broccoli 
 
Grape tomatoes 
 
 
 
Diced pineapple 
Red apple 
 
Potato wedges 
or bagel* 
 
 
 
Mashed potatoes 
or bagel* 
Sun Chips 
 
 
 
 
 
 
Baked potato or 
bagel* 
 
 
 
 
 
 
Snacks 
Rice cakes 
 
Celery 
 
Craisins 
 
Granola 
Peanut butter 
 
Peanut butter 
 
Cashews 
 
Clementines 
 
 
Granola 
 
 
 
Soy milk 
 
 
 
III. 
Literature Cited 
 
1. 
Mah E, Sapper TN, Chitchumroonchokchai C, Failla ML, Schill KE, Clinton SK, Bobe G, Traber MG, Bruno 
RS. α-Tocopherol bioavailability is lower in adults with metabolic syndrome regardless of dairy fat co-
ingestion: a randomized, double-blind, crossover trial. Am J Clin Nutr. 2015;102(5):1070-80. doi: 
10.3945/ajcn.115.118570 
2. 
USDA. Dietary Guidelines for Americans 2015-2020. 2015. 
3. 
David LA, Maurice CF, Carmody RN, Gootenberg DB, Button JE, Wolfe BE, Ling AV, Devlin AS, Varma 
Y, Fischbach MA, Biddinger SB, Dutton RJ, Turnbaugh PJ. Diet rapidly and reproducibly alters the human 
gut microbiome. Nature. 2014;505(7484):559-63. doi: 10.1038/nature12820 
4. 
Ballard KD, Mah E, Guo Y, Pei R, Volek JS, Bruno RS. Low-fat milk ingestion prevents postprandial 
hyperglycemia-mediated impairments in vascular endothelial function in obese individuals with metabolic 
syndrome. J Nutr. 2013;143(10):1602-10. doi: 10.3945/jn.113.179465 
5. 
Alberti KG, Eckel RH, Grundy SM, Zimmet PZ, Cleeman JI, Donato KA, Fruchart JC, James WP, Loria 
CM, Smith SC, Jr., International Diabetes Federation Task Force on E, Prevention, Hational Heart L, Blood 
I, American Heart A, World Heart F, International Atherosclerosis S, International Association for the Study 
of O. Harmonizing the metabolic syndrome: a joint interim statement of the International Diabetes Federation 
Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart 
Association; World Heart Federation; International Atherosclerosis Society; and International Association 
for the Study of Obesity. Circulation. 2009;120(16):1640-5. doi: 10.1161/CIRCULATIONAHA.109.192644 
6. 
Whelton PK, Carey RM, Aronow WS, Casey DE, Jr., Collins KJ, Dennison Himmelfarb C, DePalma SM, 
Gidding S, Jamerson KA, Jones DW, MacLaughlin EJ, Muntner P, Ovbiagele B, Smith SC, Jr., Spencer CC, 
Stafford RS, Taler SJ, Thomas RJ, Williams KA, Sr., Williamson JD, Wright JT, Jr. 2017 
ACC/AHA/AAPA/ABC/ACPM/AGS/APhA/ASH/ASPC/NMA/PCNA Guideline for the Prevention, 
Detection, Evaluation, and Management of High Blood Pressure in Adults: Executive Summary: A Report 
of the American College of Cardiology/American Heart Association Task Force on Clinical Practice 
Guidelines. J Am Coll Cardiol. 2018;71(19):2199-269. doi: 10.1016/j.jacc.2017.11.005 
7. 
Mah E, Noh SK, Ballard KD, Matos ME, Volek JS, Bruno RS. Postprandial hyperglycemia impairs vascular 
endothelial function in healthy men by inducing lipid peroxidation and increasing asymmetric 
dimethylarginine:arginine. J Nutr. 2011;141(11):1961-8. doi: 10.3945/jn.111.144592 

PI: Richard Bruno 
The Ohio State University 
IRB Protocol #2018H0265 
Last Approval Date: March 2, 2021 
 
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8. 
Gormley R WT. Effects of boiling, warm-holding, mashing and cooling on the levels of enzyme-resistant 
potato starch. Int J Food Sci Technol. 1999;34:15. doi:  
9. 
Mitchell CM, Davy BM, Halliday TM, Hulver MW, Neilson AP, Ponder MA, Davy KP. The effect of 
prebiotic supplementation with inulin on cardiometabolic health: Rationale, design, and methods of a 
controlled feeding efficacy trial in adults at risk of type 2 diabetes. Contemp Clin Trials. 2015;45(Pt B):328-
37. doi: 10.1016/j.cct.2015.10.012 
10. 
Lewis SJ, Heaton KW. Stool form scale as a useful guide to intestinal transit time. Scand J Gastroenterol. 
1997;32(9):920-4. doi: 10.3109/00365529709011203