The Role of Altered Nutrient Partitioning in Food Reward
Phase: NA
Conditions: Overweight and Obesity
Sponsor: Virginia Polytechnic Institute and State University

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 1 of 49 
Revised: March 5, 2019 
INSTRUCTIONS: 
●
Use this “TEMPLATE PROTOCOL (HRP-503)” to prepare a study protocol
outlining your research plan.
●
Depending on the nature of your study, some major sections might not be
applicable to your research. If so, simply mark as “N/A.” For example, a simple
survey might have many sections with “N/A.”  For subsections (e.g., 1.x or 8.x)
you can mark as “N/A”if you are certain that the subsection is not applicable.
●
Once the IRB/HRPP approves your submission, your latest approved version of
the protocol will be stored in the IRB Protocol Management online system.
●
If your research plan changes and you need to modify the protocol, please submit
an amendment to Protocol Management with the requested modifications.
Download your current protocol from Protocol Management and indicate the
changes/revisions using the track changes feature in order to make review of the
modifications easier to follow. If you are unable to use track changes, please
create a new paragraph wherever you need to make a change, and indicate
“Amendment: Date” before making a change to any section. Protocol
management will store the older versions of your protocol if the IRB or HRPP
staff need to compare them during the review.
PROTOCOL TITLE: 
Include the full protocol title.  
The role of altered nutrient partitioning in food reward 
PROTOCOL NUMBER: 
Include the number assigned in Protocol Management (verify this has been added 
before submitting protocol to HRPP).  
#21-964 
PRINCIPAL INVESTIGATOR: 
Full Name and Degrees: Alexandra Gold DiFeliceantonio, PhD 
Department: HNFE 
Telephone Number: 5405262285 
Email Address: dife@vt.edu 
FUNDING: 
Sponsor(s): NIH UL1TR003015 
Funded already or in the proposal phase?: Funded already as a sub-award to 
Virginia Tech, pending receipt of funding from NIH NCATS CTSA UL1TR003015 
Is Virginia Tech the primary awardee or the coordinating center of this grant or 
contract? If not, list the primary institution: University of Virginia  
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
1 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 2 of 49 
Revised: March 5, 2019 
 
 
VERSION NUMBER/DATE:  
Include the version number and date of this protocol. Versions should start at 1.0. 
 
2.0 
 
REVISION HISTORY:  
Use this table to keep track of changes. Add more rows as needed. 
 
Revision 
# 
Version Date 
Brief Summary of Changes  
(i.e., the different sections) 
Consent 
Change? 
1 
10/12/22 
Section 6.0: updated information about where 
and how recruitment materials will be used 
Section 11.0: Updated details about blood draw 
protocol to be consistent with the types of 
blood tubes we are able to procure 
Section 11.0: updated compensation amounts 
for each visit to equal ~$20/hr 
 
2 
4/26/23 
Section 12.0: Inclusion Criteria: changed 
inclusion criterion for fMRI scan from BMI 
25-35 kg/m2 to BMI >25 m/kg2 
no 
 
 
 
 
 
 
 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
2 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 3 of 49 
Revised: March 5, 2019 
Table of Contents 
1.0 
Study Summary 
4 
2.0 
Objectives 
4 
3.0 
Background 
5 
4.0 
Study Endpoints 
7 
5.0 
Study Design and Statistical Analysis Plan 
8 
6.0 
Setting 
8 
7.0 
Study Intervention(s)/Investigational Agent(s) 
9 
8.0 
Procedures Involved 
12 
9.0 
Data and Specimen Long Term Storage and Use 
21 
10.0 Sharing of Results with Subjects 
23 
11.0 Study Timelines 
23 
12.0 Inclusion and Exclusion Criteria 
24 
13.0 Vulnerable Populations 
26 
14.0 Number of Subjects 
27 
15.0 Recruitment Methods 
28 
16.0 Withdrawal of Subjects 
30 
17.0 Risks to Subjects 
32 
18.0 Potential Benefits to Subjects 
36 
19.0 Data Management and Confidentiality 
36 
20.0 Provisions to Protect the Privacy Interests of Subjects 
39 
21.0 Provisions to Monitor the Data to Ensure the Safety of Subjects 
40 
22.0 Compensation for Research Related Injury 
41 
23.0 Economic Burden to Subjects 
41 
24.0 Consent Process 
42 
25.0 Process to Document Consent in Writing 
45 
26.0 Resources Available 
46 
27.0 Multi-Site Research 
49 
 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
3 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 4 of 49 
Revised: March 5, 2019 
1.0 
Study Summary 
 
Study Title 
The role of altered nutrient partitioning in food reward 
 
Study Design 
This is a within-subjects crossover study design that involves 
a conditioning paradigm in which participants are exposed to 
novel flavored beverages paired with sugars and non-
nutritional sweeteners, and assessment of energy expenditure 
and blood metabolites will be performed during the 
conditioning paradigm. A subset of participants will undergo 
fMRI measurements and metabolic chamber measurements. 
Primary Objective 
To assess the feasibility of measuring reward response 
following a flavor-nutrient conditioning paradigm across the 
normal to obese body mass index range and in states of 
altered metabolic health 
Secondary 
Objective(s) 
To develop an appropriate protocol for measuring metabolic 
flexibility in a "flex" metabolic chamber 
Study Population 
The general US population 
Sample Size 
15 
Research 
Intervention(s)/ 
Investigational 
Agent(s)  
Blood measurements, resting and postprandial energy 
expenditure and substrate oxidation, questionnaires, 
magnetic resonance imaging 
Study Duration for 
Individual 
Participants 
6-7 weeks in total: 
Week 1: Training Session (1 hour) 
Weeks 2-4: Behavior Sessions (4 sessions x 2.5 hours each) 
Week 5: Post-testing Session with fMRI (only a subset of 
participants; 3 hours) 
Week 5: Post-testing Session without fMRI (a majority of 
participants; 1 hour) 
Weeks 6-7: Metabolic chamber sessions (2 sessions x 6 
hours each, with 5-day washout between sessions) 
4 days total of controlled feeding with food provided by the 
metabolic kitchen 
 
Acronyms and 
Definitions  
REE: resting energy expenditure; MRI: magnetic resonance 
imaging; FBRI: Fralin Biomedical Research Institute; 
CHBR: Center for Health Behavior Research 
 
 
2.0 
Objectives 
2.1 Describe the purpose, specific aims, or objectives of this study: 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
4 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 5 of 49 
Revised: March 5, 2019 
The first aim of this study is to assess the feasibility of measuring reward response 
following a flavor-nutrient conditioning paradigm across the normal to obese body mass 
index range and in states of altered metabolic health. The second aim is to develop a 
protocol for measuring metabolic flexibility in a "flex" metabolic chamber over a 6-hour 
measurement.  
 
 
 
2.2 State the hypotheses to be tested:  
 
We hypothesize that reinforcement learning will be impaired with increasing obesity and 
that metabolic flexibility can be determined utilizing data produced from a 6-hour 
measurement inside a "flex" metabolic chamber.  
 
 
3.0 
Background 
3.1 Summarize the relevant prior research on this topic and gaps in current 
knowledge within the field of study: 
 
There is conflicting evidence as to whether people with obesity find foods more 
reinforcing. People with obesity exhibit a greater motivation to consume palatable foods 
that does not habituate over time compared to healthy weight individuals (Saelens and 
Epstein 1996; Epstein et al. 2007; Temple et al. 2009); however, they do not report liking 
food more (Wall et al. 2020). Obesity is also associated with decreased sensitivity to 
post-ingestive signals (Beutler et al. 2020), possibly due to dietary history and exposure 
(Small and DiFeliceantonio 2019). Further, there is evidence for impaired reinforcement 
learning and habituation in studies from both humans with obesity and animals (Kroemer 
and Small 2016; Coppin et al. 2014; Reichelt, Morris, and Westbrook 2014; Johnson and 
Kenny 2010). Additionally, many hormones that are altered in states of obesity and 
metabolic dysfunction, such as insulin, influence food reinforcement. For example, a 
recent study in humans revealed central insulin signaling influences striatal dopamine 
tone (Kullmann et al. 2021). However, there is no experimental evidence in humans that 
post-ingestive learning is altered in states of obesity. The experiments in this proposal 
aim to fill that gap. 
The normal process of energy metabolism involves a dynamic and coordinated switch 
between primarily fat and carbohydrate as oxidative fuel substrates, depending upon 
nutritional or physiological condition. This process of adjusting substrate oxidation to 
match substrate availability and energy demand is termed “metabolic flexibility” (Kelley 
and Mandarino 1990). Metabolic inflexibility, on the other hand, is caused by nutrient 
overload in the mitochondria and results in altered nutrient partitioning in which a 
consistent mix of oxidative substrates, regardless of nutritional or physiological 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
5 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 6 of 49 
Revised: March 5, 2019 
circumstances, is utilized (Muoio 2014). Metabolic inflexibility has been identified in 
obesity (GM et al. 2012; Hulver et al. 2003), type 2 diabetes(Kelley and Mandarino 
1990), insulin resistance (Kelley et al. 1999; van der Kolk et al. 2016), and  other 
conditions characterized by dysregulated energy homeostasis. Metabolic inflexibility is 
putatively a key influence in the pathogenesis of peripheral insulin resistance, as 
biomarkers of altered fat oxidation have been observed prior to the development of 
insulin resistance (Jans et al. 2011; Heilbronn et al. 2007). If flavor nutrient conditioning 
is dependent upon some mechanism involved in the pathway of nutrient 
sensing, transport, and utilization, altered nutrient partitioning due to metabolic 
inflexibility may provide insight into the relationship between post-ingestive metabolic 
signals and food reward in obesity and metabolic disease.  
 
Indirect calorimetry is a powerful tool for determining energy expenditure and 
macronutrient oxidation in response to a mixed-macronutrient diet or meal. In other 
words, it allows for a versatility in measurement of metabolic flexibility to both fat and 
carbohydrate in a physiologically relevant manner. Indeed, a new method for assessing 
metabolic flexibility to a high-carbohydrate or high-fat meal using indirect calorimetry in 
a large, whole-room metabolic chamber has recently been published (McDougal et al. 
2020); however, the reproducibility of this method across metabolic chambers and 
clinical populations has not yet been determined. Furthermore, this methodology is 
designed around use of a 27 m3 chamber, which requires a relatively long duration of 
measurement for an appropriate temporal resolution of measured O2 and CO2 
concentrations to determine energy expenditure and macronutrient oxidation (Chen et al. 
2018). The Fralin Biomedical Research Institute houses two state-of-the-art metabolic 
chambers. The smaller of the two chambers, approximately 4.6 cubic meters, allows for 
more rapid and accurate measurement of O2 and CO2 compared to large chambers and 
is specifically designed for conducting dynamic metabolic studies over shorter 
measurement durations. This smaller “flex” chamber is one of only a few in the world, 
and as such, no methodologies to assess metabolic flexibility specifically in the “flex” 
chamber have been developed. Thus, we are poised to take advantage of novel 
technology to lead the development of a new method to assess a key component of 
metabolic health.  
 
 
 
3.2 Describe any relevant preliminary data: 
 
This is a new project without preliminary data collected. 
 
 
3.3 Based on the existing literature, provide the scientific or scholarly 
rationale for and significance of your research and how will it add to 
existing knowledge: 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
6 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 7 of 49 
Revised: March 5, 2019 
 
Although substantial advances in obesity treatment have been made in recent decades, 
obesity remains a public health epidemic. This may in part be due to the fact that 
although current treatments are effective in improving metabolic health and reducing 
body weight in the short term, strategies that promote long-term maintenance 
of metabolic health and reduced body-weight remain elusive. Obesity is therefore a 
chronic disease that requires lifelong attention. From a public health perspective, 
developing long-term effective treatments is an essential medical research goal. The gut-
brain axis has long been recognized as an important regulator of metabolism and energy 
homeostasis; however, only recently have advances in research tools allowed for a 
better understanding of the complex mechanistic integration of gut and brain signals. The 
fidelity of the gut-brain axis presents a novel and potentially powerful therapeutic target 
for obesity and metabolic diseases. In this proof-of concept pilot, we propose to probe the 
gut-brain axis as a target for obesity intervention by testing the fidelity of the gut-brain 
axis 1) across a range of body mass index (BMI) and 2) in states of altered metabolic 
health (independent of BMI). The data from this proposal will provide evidence for the 
gut-brain axis as a therapeutic target in obesity treatment and allow the development of 
state-of-the-art tools to measure metabolic health at Virginia Tech, essential steps 
towards applying for federal funding. 
 
 
4.0 
Study Endpoints 
4.1 Describe the primary and secondary study endpoints. See links below for 
discussion of study endpoints and how they may differ from study 
objectives. These are most common in clinical trials but are sometimes 
applicable to other types of biomedical research, as well as social, 
behavioral, or educational research. See link below for a discussion. 
https://docs.google.com/document/d/1Wocz7K7a0hCQJPPO_khh5l1
SQQjhGDDGHzcOPRHR5Tw/edit?usp=sharing  
 
Primary study endpoints are blood metabolite measurements, indirect calorimetry 
measurements, drink preference and ratings, and brain activation measured by fMRI.  
 
 
4.2 Describe any primary or secondary safety endpoints. These should be 
included for all studies that are greater than minimal risk. (Minimal risk: 
The probability and magnitude of harm or discomfort anticipated in the 
research that are not greater in and of themselves than those ordinarily 
encountered in daily life or during the performance of routine physical or 
psychological examinations or tests.): 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
7 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 8 of 49 
Revised: March 5, 2019 
There are no safety endpoints. 
 
 
5.0 
Study Design and Statistical Analysis Plan 
5.1 Describe the basic study design/approach (e.g., qualitative study using five 
focus groups of first year students to describe assimilation into the 
university community; randomized controlled trial of a behavioral change 
intervention to increase dietary intake of whole grains; pre- post-test 
evaluation of new pedagogical techniques to improve adult literacy): 
 
This study consists of two main aims. In the first aim, a within-subjects design will be 
used to compare the rewarding properties of two beverages, one containing 75kcal of 
sugar and one containing 0 kcal sucralose in individuals with normal weight and with 
excess adiposity or high body mass index. In a subset of participants, neural activation to 
each of the beverage tastes measured by fMRI will be compared. In the second aim, a 
within-subjects design will be used to compare metabolic response to a high-
carbohydrate and high-fat test meal using a 6-hour metabolic chamber stay. 
 
 
 
 
5.2 Describe corresponding data analysis plan/approach (e.g., content analysis 
of focus group transcripts; descriptive analysis followed by linear 
regression modeling; nonparametric analysis of pre- and post-test 
measures): 
 
The first aim will use linear mixed effects models to compare ratings of the beverages 
and control for confounding variables. Linear regression will be used to relate blood and 
energy expenditure measures to liking and other subjective ratings. Standard 
preprocessing and statistical techniques for fMRI data will be used to compare beverages 
and look for relationships between neural activity and physiological measures. The 
second aim will use linear mixed effects models to compare energy expenditure and 
substrate oxidation to the test high-carbohydrate and high-fat meals measured in the 
metabolic chamber. Measures of metabolic flexibility will include changes in respiratory 
exchange ratio in response to each meal, the time to reach peak respiratory exchange ratio 
after meal consumption, slope from baseline to peak respiratory exchange ratio, and rate 
and slope of the return to baseline respiratory exchange ratio.  
 
 
6.0 
Setting 
6.1 Describe the sites or locations where your research team will conduct the 
research.  Consider each of the items listed below: 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
8 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 9 of 49 
Revised: March 5, 2019 
● 
Identify where your research team will identify and recruit potential 
subjects. 
● 
Identify where the team will perform the research procedures. 
● 
Describe the composition and involvement of any community 
advisory board(s). 
● 
For research conducted in other locations, describe: 
o 
Site-specific regulations or customs affecting the research at 
those locations. 
o 
Local scientific and ethical review structure at those locations.  
Examples include work in other cultures or ethnic groups 
(within or outside of the U.S.) and work with churches. The 
HRPP will provide additional guidance for international 
research. 
 
 
Approved study recruitment materials will be distributed in the follow ways. Flyers will 
be posted on bulletin boards and approved posting sites throughout Roanoke, Blacksburg, 
and surrounding areas. Digital advertisements will be displayed on DiFeliceantonio lab 
and CHBR social media, VT News, and in list serves with permission.  
 
All advertisements will include a link to the lab website where there will be approved 
language describing the research study. Interested participants will fill out a general 
screening survey. All screening surveys will be delivered through Ripple. This screening 
survey will be used to determine study eligibility. Once the general screening survey has 
been completed, participants who may be eligible for this study will be contacted via 
email (see attachment 'Screening email'). If participants choose to complete the study-
specific screening form and are eligible for this study, they will be contacted to schedule 
a consent session. 
 
The study procedures for the study will all take place at the Fralin Biomedical Research 
Institute (FBRI) at VTC campus in Roanoke, VA. The consent session and metabolic cart 
measurement sessions will occur in the Center for Health Behavior Research (CHBR) at 
the FBRI. Flex calorimeter sessions and controlled feeding will occur in the metabolic 
chamber and metabolic kitchen, respectively, at the FBRI at VTC. 
 
 
 
7.0 
Study Intervention(s)/Investigational Agent(s) 
7.1 Describe the study interventions (including behavioral interventions) and/or 
investigational agents (e.g., drugs or devices) to be used in this study.  Consider 
each of the items listed below:  
● 
Drug/Device Handling: If the research involves drugs or devices, 
describe your plans to store, handle, and administer the drugs or 
devices so that they will be used only on subjects, and only by 
authorized investigators. 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
9 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 10 of 49 
Revised: March 5, 2019 
● 
Describe whether any of the following will be used: microwaves, X-
rays, DEXA scans, general anesthesia, or sedation 
● 
If control of the drugs or devices used in this protocol will be 
accomplished by following an established, approved organizational 
SOP (e.g., Research Pharmacy SOP for the Control of 
Investigational Drugs, etc.), please reference the SOP in this section. 
 
 
Beverage Solutions:  
This study will involve the use of 2 beverage solutions, which will be given to each 
participant. Beverages include citric acid, water, flavor, and either: 
1. 0 kcal sucralose (to match the sweetness of the calorie-containing sugar beverage) 
2. 75kcals sugars 
 
Sucralose, sugars, and citric acid are common food and beverage additives that nearly 
every participant would have consumed previously in their normal diet. 
 
The flavors used will be 0.002% acerola, 0.5% bilberry, 0.1% horchata, 0.1% lulo, 0.2% 
yuzu. 0.1% papaya, 0.1% chamomile, 0.1% aloe vera, 0.1% mamey, and 0.2% maqui 
berry (Bell Labs Flavors and Fragrances,IL, USA, product numbers: 33.81940, 15.80182, 
132.81478, 141.14606. 101.29478, 102.82506, 141.31243, 141.31480, 46.29969 and 
13.32059). 
 
Foods: 
This study will involve consumption of high carbohydrate (60% carbohydrate, 20% fat) 
and high fat (60% fat, 20% carbohydrate) test meals. In addition, a standardized diet 
(50% carbohydrate, 30% fat) will be provided for 2 days preceding each test meal 
measurement day. All of the foods included are typical foods nearly every participant 
would have consumed in their normal diet. Participants will be screened for food 
allergies/intolerances and excluded if any of these foods are included in the study diet or 
test meals.  
 
Device Handling: 
This research involves use of a metabolic chamber and metabolic cart. Both are located in 
spaces with restricted, proxy access. This research also involves the use of an MRI 
scanner. The MRI scanner is located in a space in the Human Neuroimaging Lab with 
restricted, proxy access, behind biometrically secure doors.   
 
 
 
 
7.2 List the name of all drugs (including any vitamins, supplements, herbs, or 
nicotine) to be used in the study.  Indicate whether they have FDA 
approval, and list any limitations for their use:  
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
10 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 11 of 49 
Revised: March 5, 2019 
No drugs or supplements will be used in this study. 
 
 
7.3 List all devices, how they will be used, their purpose in the study, and if 
they will be used in a manner consistent with their approved uses. If they 
will be used in ways that are not yet FDA approved, indicate whether they 
need an IDE or a determination that they are exempt from the IDE 
Determination. If a determination of significant risk or non-significant risk 
is needed for any of the devices, include the researcher’s recommendation 
for each of those devices:  
 
Metabolic Chamber and Cart: Participants will undergo metabolic measurements in a 
metabolic chamber and metabolic cart in a manner consistent with approved uses. 
 
Magnetic Resonance Imaging: A subset of participants will receive an MRI scan. The 
Siemens 3T scanner has been approved by the FDA and will be used in a manner 
consistent with approval. 
 
 
7.4 If the drug is investigational (has an IND) or the device has an IDE or a 
claim of abbreviated IDE (non-significant risk device), include the 
following information: 
● 
Identify the holder of the IND/IDE/abbreviated IDE. 
● 
Explain procedures followed to comply with sponsor requirements 
for FDA regulated research for the following: 
 
Applicable to: 
FDA Regulation 
IND Studies 
IDE studies 
Abbreviated 
IDE studies 
21 CFR 11 
X 
X 
 
21 CFR 54 
X 
X 
 
21 CFR 210 
X 
 
 
21 CFR 211 
X 
 
 
21 CFR 312 
X 
 
 
21 CFR 812 
 
X 
X 
21 CFR 820 
 
X 
 
 
 
N/A 
 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
11 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 12 of 49 
Revised: March 5, 2019 
 
 
 
 
8.0 
Procedures Involved 
8.1 Describe and explain the study design: 
 
This is a crossover design and within-subjects analysis plan where participants receive all 
conditions during the flavor-nutrient conditioning paradigm of Aim 1. A subset of 
participants will also undergo fMRI scanning for Aim 1. All participants will undergo the 
controlled feeding diet and metabolic chamber measurements for Aim 2.  
 
 
8.2 Provide a description of: 
● 
All research procedures being performed  
● 
If the study has more than one procedure, session, and/or subject 
population, describe each procedure, session, and/or study 
population separately. For complex studies, you are encouraged to 
include a figure or chart. 
 
 
Study Overview:  
Approximately 15 people from the general population will be recruited for Aim 1 
(without fMRI) and Aim 2 of this randomized crossover design study conducted at FBRI. 
Of these 15 people, 5 will also undergo fMRI scanning as a part of Aim 1. This will be an 
approximately 6- to 7-week study. Participants will report to the research laboratory in 
Week 1 for the consent and training session. They will return to the research laboratory to 
complete 2 behavior sessions per beverage and a final post-testing session in Aim 1. 
Participants who are asked to complete the fMRI scan will undergo this measurement 
during the final post-testing session. For Aim 2, participants will report to the metabolic 
kitchen at FBRI for 2 days preceding each chamber measurement day to collect food to 
be consumed for each of those days. On each metabolic chamber measurement day, they 
will report to the metabolic chambers located in 4 Riverside. There will be a 5-day 
washout period between the first metabolic chamber session and controlled feeding for 
the second metabolic chamber session.  
 
Recruitment and Screening: 
Approved study recruitment materials will be distributed in the follow ways. Flyers will 
be posted on bulletin boards and approved posting sites throughout Roanoke, Blacksburg, 
and surrounding areas. Digital advertisements will be displayed on DiFeliceantonio lab 
and CHBR social media, VT News, and in list serves with permission.  
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
12 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 13 of 49 
Revised: March 5, 2019 
All advertisements will include a link to the lab website where there will be approved 
language describing the research study. Interested participants will fill out a general 
screening survey. All screening surveys will be delivered through Ripple. This screening 
survey will be used to determine study eligibility. Once the general screening survey has 
been completed, participants who may be eligible for this study will be contacted via 
email (see attachment 'Screening email'). If participants choose to complete the study-
specific screening form and are eligible for this study, they will be contacted to schedule 
a consent session. 
 
AIM 1:  
 
Consent and Training Session 
Informed Consent 
Participants will be asked to sign a consent form upon arrival to the CHBR. They will be 
given ample time to review and ask questions prior to signing the document. After 
research personnel have answered the participant's questions and the consent form has 
been signed, participants will begin the training session. 
 
Anthropometrics and Clinical Measures 
Participants' height and weight and hip and waist circumferences will be measured. A 
fingerstick blood sample will be collected for point-of-care hemoglobin A1C assessment 
to ensure eligibility. Blood pressure will be collecting using an automated 
sphygmomanometer. Body composition will be assessed using bioelectrical impedance 
analysis.  
 
Flavor Ratings 
The participants will first be trained by the researcher on how to use the standard 
perceptual rating scales (Bartoshuk et al., 2004). Then participants will taste and make 
ratings of beverages using the standard rating scales. After the participant has completed 
the tasting protocol, the researcher will determine two flavors that were equally liked and 
equally unfamiliar to the participant. These flavors will be used in the beverages the 
participants will consume during the behavioral conditioning sessions (one beverage per 
2 sessions). In addition, participants will complete a triangle test to determine whether 
they can detect a difference between sugar- and sucralose-sweetened beverages.  
 
Mock MRI Training 
Participants will also complete a "mock" MRI training, in which they learn to swallow 
small amounts of liquids while lying supine. (Note: most people have no difficulty 
learning to swallow small quantities of liquid while lying down). This training will be 
performed in an MRI simulator that is the same bore size and similar environment to the 
real scanner, but does not contain a magnetic field. Participants who have difficulty with 
supine swallowing or who express discomfort in the simulated fMRI environment may be 
excluded from the study or only undergo portions of the study that do not involve fMRI 
scanning (i.e., Aim 1 without the fMRI scan and Aim 2). 
 
Dietary, Physical Activity, and Health Behavior Questionnaires 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
13 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 14 of 49 
Revised: March 5, 2019 
After the training session, participants will receive an email containing links to dietary- 
physical activity- and behavior-related questionnaires in REDCap to complete prior to the 
first behavior session. It is estimated that it will take the participant approximately 30 
minutes to complete all of the questionnaires. These questionnaires will be sent using the 
automated invitation function in REDCap, which will generate an email containing 
unique links for each survey for each participant. Only participant email addresses will be 
stored as participant identifiers to track survey responses for data collection. The 
automated invitations will also contain information about how to contact appropriate lab 
personnel to opt out of further participation if a participant wishes to do so.  
 
Behavior Sessions 1 and 3 (interchangeable with Behavior Sessions 2 and 4, 
respectively): 
Participants will come to the CHBR after a 4 hour fast. Participants will also be asked to 
abstain from moderate-to-vigorous physical activity for at least 24h prior to their session. 
They will first rate their internal states on standard scales.  
 
An IV catheter will then be placed in participants' arms for blood draws at seven time 
points during the 1-hour session: at baseline and 10-, 15-, 20-, 30-, 40-, and 60-minutes of 
the session. For the IV catheter placement, a small plastic tube will be placed in one of 
the veins in the medial cubital fossa area of the arm; this tube will stay in place for the 
duration of the 1-hour testing session. No more than three attempts will be made to insert 
the IV catheter during each blood draw testing session. Once the catheter is inserted, a 
baseline sample of 2 tubes of blood will be drawn: a 4 ml tube for EDTA plasma, and a 5 
ml tube for serum.  In the case of difficulty inserting the IV catheter and obtaining blood 
at the baseline time point, we will evaluate whether to continue the protocol or not. If the 
participant is willing, venous blood draw samples may be collected instead at baseline, 
30, and 60 minutes. 
 
Participants will then consume a single 355ml conditioning beverage (with the 
appropriate conditioning stimuli and flavor as determined during the consent and training 
session) within 5 minutes and rate internal states immediately after consumption. Two 
tubes of blood (one 4 ml EDTA plasma, one 5 ml serum) will be collected at 10-, 15-, 20-
, 30-, 40-, and 60-minutes following beverage consumption. In total, 14 tubes of blood 
will be drawn during the session (approximately 63 mls of blood). The IV catheter line 
will be flushed with normal saline between each blood draw to prevent coagulation in the 
catheter tubing. Once the 60-minute sample has been drawn, the catheter will be removed 
and the venipuncture site secured with sterile tape or a bandage. 
 
At the end of this session, participants will be given 2 additional bottles containing 355 
ml of the beverage consumed during this session. Participants will be instructed to 
consume one beverage 1 hour before dinner that evening and the other beverage 1 hour 
before lunch the following day. Participants will also be asked to complete a short survey 
regarding consumption of each of these beverages. An automated email containing links 
to a survey about the dinner and lunch beverage consumption will be sent using the same 
REDCap automated email function described above.  
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
14 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 15 of 49 
Revised: March 5, 2019 
Behavior Sessions 2 and 4 (interchangeable with Behavior Sessions 1 and 3, 
respectively): 
Participants will come to the CHBR after a 4 hour fast. Participants will also be asked to 
abstain from moderate-to-vigorous physical activity for at least 24h prior to their session. 
This session will occur within the same week as Behavioral session 1 (or 3, as 
appropriate) with at least one day in between the sessions. They will first rate their 
internal states on standard scales.  
 
Participants will then rest for approximately 30 minutes in preparation for the fasting 
indirect calorimetry measurement, which requires participants to be in a calm, restful 
state for accurate measurement. Participants will then undergo an indirect calorimetry 
measurement using a metabolic cart for approximately 30 minutes. 
 
Indirect calorimetry determines gas exchange (i.e., oxygen consumption and carbon 
dioxide expiration) at the level of the lungs, and these measures are used to calculate 
energy expenditure and fat and carbohydrate utilization. During the test, participants lie 
supine while breathing normally into a transparent plastic hood. There are two holes on 
the hood, one that allows the inflow of room air into the hood, and another that allows for 
the extraction of gases into the gas analyzer via a vacuum tube (see picture below). The 
hood is then sealed around the participant's shoulders to prevent any gas from escaping 
during the test. Indirect calorimetry using a metabolic cart is a common, noninvasive 
assessment conducted routinely in both research and clinical nutrition practice. There is 
no more than minimal risk associated with this method of indirect calorimetry, and 
participants may wear normal clothes during the procedure. Since the hood is clear 
plastic, participants will be able to see and communicate with research personnel 
throughout the test. Should a participant become uncomfortable with the test, they may 
choose to stop the procedure at any time.  
 
 
 
Following completion of the approximately 30-minute fasting indirect calorimetry 
measurement, participants will be given a 355 ml conditioning beverage (with the 
appropriate conditioning stimuli and flavor as determined during the consent and training 
session) to consume within 5 minutes. Participants will then rate their internal states on 
standard scales, and indirect calorimetry will be performed again for approximately 1 
hour.  
 
At the end of this session, participants will be given 2 additional bottles containing 355 
ml of the beverage consumed during this session. Participants will be instructed to 
consume one beverage 1 hour before dinner that evening and the other beverage 1 hour 
before lunch the following day. Participants will also be asked to complete a short survey 
regarding consumption of each of these beverages. An automated email containing links 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
15 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 16 of 49 
Revised: March 5, 2019 
to a survey about the dinner and lunch beverage consumption will be sent using the same 
REDCap automated email function described above.  
 
Post-Testing Session (for participants NOT undergoing fMRI portion of the protocol):  
Participants will come to the CHBR after a 4 hour fast. Participants will also be asked to 
abstain from moderate-to-vigorous physical activity for at least 24h prior to their session. 
They will first rate their internal states on standard scales.  
 
They will then taste and rate on standard scales the 2 flavors they consumed as part of the 
conditioning beverages throughout the protocol. After the rating, they will be given 
access to bottles containing the two they consumed during the protocol sessions and 
allowed to drink as much or as little as they want from the 2 bottles for 30 minutes. 
Consumed amounts of each beverage will be measured and recorded. Lastly, they will be 
offered 2 additional bottles containing the same conditioning beverages and allowed to 
choose one to take home with them in a forced-choice test.  
 
Post-Testing Session (for participants who ARE undergoing fMRI portion of the 
protocol):  
Participants completing the fMRI portion of the protocol will first report to the CHBR 
after a 4 hour fast. Participants will also be asked to abstain from moderate-to-vigorous 
physical activity for at least 24h prior to their session. They will first rate their internal 
states on standard scales. Then, they will be escorted to the Human Neuroimaging 
Laboratory in 2 Riverside for the fMRI scan.   
 
During this scan, participants will lie on a table that slides into the bore of the scanner. 
Using a head coil, the functional images will be acquired using an EPI bold sequence. 
Because the process is noisy (due to the sounds generated as the magnetic field gradients 
are changed), participants will wear a pair of headphones and disposable earplugs. During 
the entirety of the scan, subjects will be able to communicate with the experimenter via a 
speaker and microphone system. If subjects become uncomfortable or claustrophobic 
while in the scanner, they will be able to squeeze an emergency ball to get the attention of 
the experimenters who will then immediately remove them from the scanner. They can 
then withdraw from the study if they are unable to complete the MRI.  
 
While in the magnet, subjects will taste small amounts of liquid stimuli that will be 
presented according to a standard paradigm we have successfully used before in taste 
MRI studies (Small et al., 2003; Small et al., 2004). In brief, hydraulic pumps will be 
used to pump solutions through plastic beverage tubing attached to a mouthpiece to the 
subject lying in the scanner. The mouthpiece will be sterilized and new beverage tubing 
will be used for each subject. The mouthpiece will be anchored to the head coil so that 
the end comfortably rests on the anterior tongue, just inside the subjects’ mouth. This will 
not interfere with breathing through the mouth and subjects will always be able to breathe 
through their noses. To ensure participants can breathe through their nose, they will be 
asked to close their mouth and take one full breath (inhale and exhale) through their nose. 
If they are unable to breathe normally through their nose on the day of the scan, the scan 
will be rescheduled.  The solutions will be dispensed through one tube at a time. The 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
16 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 17 of 49 
Revised: March 5, 2019 
overall flow rate of solution into the mouth will be very slow (0.5ml of a liquid over 3-5 
seconds). Each taste will be followed by a rinse of a tasteless solution.  
 
After the fMRI is completed, they will be given access to bottles containing the two they 
consumed during the protocol sessions and allowed to drink as much or as little as they 
want from the 2 bottles for 30 minutes. Consumed amounts of each beverage will be 
measured and recorded. Lastly, they will be offered 2 additional bottles containing the 
same conditioning beverages and allowed to choose one to take home with them in a 
forced-choice test.  
 
AIM 2:  
 
Controlled Feeding: 
Participants will consume a standardized diet (50% carbohydrate, 30% fat) for 2 days 
prior to each metabolic chamber session. Total daily energy needs for each participant 
will be estimated using average resting energy expenditure determined during completion 
of Behavior Sessions 2 and 4 in Aim 1 and an activity factor derived from the physical 
activity questionnaire completed as part of the Consent and Training Session in Aim 1. 
The 2-day diet will be developed by a registered dietitian, and all foods will be prepared 
and assembled in the metabolic kitchen located in 4 Riverside.  
 
Participants will report to the metabolic kitchen to pick up coolers of food. Participants 
will be provided with all foods and drinks, except water, for the 2 days preceding each 
metabolic chamber measurement and instructed to consume all foods and beverages 
provided in the coolers.  
 
Metabolic Chamber Sessions: 
Participants will report to the metabolic chamber lab in 4 Riverside after a >8 hour 
(overnight) fast. Participants will also be asked to abstain from moderate-to-vigorous 
physical activity for at least 24h prior to their session.  
 
Participants will then enter the flex metabolic chamber, which is outfitted with a bed, 
pillows, and a TV, to rest quietly during the fasting measurement. The fasting 
measurement will take approximately 50 minutes. After a fasting measurement, 
participants will be instructed to consume a high-carbohydrate (60% carbohydrate, 20% 
fat) or high-fat (60% fat, 20% carbohydrate) test meal within 5 minutes. Participants will 
then rest inside the chamber, without sleeping, for 5 hours during the post-consumption 
measurement. 
 
The metabolic chamber operates under the same indirect calorimetry assumptions and 
principles as the metabolic cart, but the size of the chamber (4.6 cubic meters) allows for 
longer measurements with better comfort for participants compared with the metabolic 
cart. There are no known risks associated with this method of indirect calorimetry, and 
participants may wear normal clothes during the procedure. The chamber is outfitted with 
a 2-way intercom system, and one wall is clear plastic; participants will be able to see and 
communicate with research personnel throughout the test. Should a participant become 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
17 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 18 of 49 
Revised: March 5, 2019 
uncomfortable with the test, they may choose to stop the procedure at any time. (See 
picture below.) 
 
 
 
 
 
 
 
 
8.3 Describe: 
● 
Procedures or safeguards intended to reduce the probability and 
magnitude of risks. (For example: Reducing the risk of injury in a 
virtual reality study either by having the subjects sit during the study 
or by providing an obstacle-free space for walking.) 
● 
Be sure to describe all drugs and devices used in the research, when 
they will be administered or used, and their purpose.  
● 
Methods used to collect data about subjects. Please upload all data 
collection forms to Protocol Management. Some common examples 
are: 
● Screening questionnaires  
● Survey(s), including online surveys 
● Demographic questionnaire(s) 
● Interview guide(s), e.g., questions or pool of questions for semi-
structured interviews  
● Focus group guide(s) 
● Other documents used to collect data  
 
 
Procedures or safeguards intended to reduce the probability and magnitude of risks:  
 
Indirect calorimetry 
There are no known risks associated with either method of indirect calorimetry. However, 
some people may feel uncomfortable or anxious with the enclosed clear plastic hood 
placed over their head and shoulders during the metabolic cart measurement. Subjects 
will be able to breathe normally during the entirety of the testing procedure. They will 
also be able to see and communicate with the experimenter and will be able to tell the 
experimenter if they would like to terminate the measurement. Some people may feel 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
18 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 19 of 49 
Revised: March 5, 2019 
uncomfortable enclosed in a sealed room during the metabolic chamber measurement. 
Subjects will be able to breathe normally during the entirety of the test procedure, and a 
large window comprises most of one wall of the chamber. Participants will be able to see 
and communicate with the experimenter during the entirety of the measurement and will 
be able to tell the experimenter if they would like to terminate the measurement. 
 
MRI 
Every effort will be made to ensure the subject's comfort and to reduce any minimal risk. 
First and foremost, subjects will be thoroughly screened to make sure that MRI 
contraindications are not present. If, at any time, subjects experience discomfort, 
dizziness, or claustrophobia during the scan, they will have access to an emergency 
squeeze bulb which they can squeeze to get the attention of the experimenters. Subjects 
who squeeze this bulb will be immediately removed from the MRI machine. 
Additionally, the staff operating the MRI will have completed Advanced MRI/Operator 
Training which is required for those persons wishing to conduct research on the Human 
Neuroimaging Lab (HNL) MRIs. Training consists of a safety training, full-day advanced 
user training, scanning observation, and hands-on practice, supervised scanning. Topics 
include safety and emergency procedures, subject preparation and screening, scanner set-
up and operation, and troubleshooting.  Each person must also complete at least 4 hours 
of observation (to include 2 different studies) and at least 16 hours of supervised scanning 
(to include 4 different studies). Certification is granted upon completion of the training 
and passing of a competency exam.  
 
An MRI Safety Screening Form will be completed by each participant and reviewed by 
trained staff in order to ensure there are no contraindications to MRI scanning. 
 
Catheter Blood Draw 
Some pain or discomfort may be experienced when the catheter is inserted in the vein, 
but this should persist for only a short time. During the blood draws, there may be pain 
and/or bruising at the site where the IV catheter is inserted. In about 1 in 10 or 10% of the 
cases, a small amount of bleeding under the skin will cause bruising. The risk of a blood 
clot forming in the vein is about 1 in 200, while the risk of infection or significant blood 
loss is 1 in 1000. There is a small risk of the vein becoming inflamed and/or painful in 
the hours or days after the needle is removed. To reduce this risk, all catheter placements 
and blood draws will be performed by a trained and skilled medical technologist, 
phlebotomist, or nurse. In addition, a maximum of three attempts will be made to insert 
the IV catheter at different sites. If the participant feels faint during or after a blood draw, 
research personnel will immediately have the participant lie back in the phlebotomy 
chair. 
 
Fingerstick Blood Sample 
The risks associated with performing a fingerstick to collect a capillary blood sample are 
minimal. The likely risks involve pain and bleeding in the fingertip at the collection site. 
Rarely, an infection may occur at the puncture site. To minimize these risks, research 
team members will be trained on the proper technique for obtaining a capillary blood 
sample and care for the site after the sample is obtained.  
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
19 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 20 of 49 
Revised: March 5, 2019 
 
 
 
 
8.4 What data will you collect during the study and how you will obtain them? 
Please include descriptions of electronic data collection, database 
matching, and app-based data collection: 
 
Ripple will be used to collect screening information. Information about the beverages 
participants will be consuming at home as well as dietary, physical activity, and behavior 
information will be collected through REDCap. MRI safety information will be collected 
via paper form (attached). Perceptual ratings will be collected using a laptop or tablet and 
custom software written in Matlab. Blood glucose will be measured using a Hemocue 
point-of-care system, and insulin will be measured using the Immunolite 1000 
Immunoassay System or ELISA. Brain images will be acquired during fMRI scans. Gas 
exchange values will be collected using a standard metabolic cart from ParvoMedics and 
a metabolic chamber from MEI Research, Ltd. 
 
 
 
8.5 Who will transcribe or code audio and/or video recordings?: 
 
There will be no audio or video recordings. 
 
 
8.6 Include a description of any deception to be used in the study. Include 
justification for the use of deception (why the deception is necessary), 
describe the debriefing process, and describe how the study meets all the 
following criteria  for alteration of consent (deception is considered an 
alteration of informed consent): 
• The research involves no more than minimal risk to the subjects 
• The alteration will not adversely affect the rights and welfare of the 
subjects 
• The research could not practicably be carried out without the 
alteration/deception 
• (Optional but encouraged in most cases) Subjects will be provided with 
additional pertinent information after participation (i.e., debriefing for 
studies involving deception)  
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
20 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 21 of 49 
Revised: March 5, 2019 
There will be no deception. 
 
 
 
8.7 If the study involves long-term follow-up (once all research related 
procedures are complete), describe what data will be collected during the 
follow up period and when it will occur:  
 
There are no long-term follow ups. 
 
 
 
 
 
 
9.0 
Data and Specimen Long Term Storage and Use 
9.1 If you will store data or specimens for future use, describe where you will 
store the data or specimens, how long they will be stored, and how and by 
whom the data or specimens will be accessed: 
 
Indirect calorimetry data and 24-hour food recall data will be stored on a secure network 
drive provided by FBRI. Behavioral (e.g., last time food eaten before measurement, last 
bout of exercise before measurement, etc.), anthropometric, responses to questionnaires, 
and pertinent energy expenditure variables collected during indirect calorimetry 
measurement will be stored in FBRI's instance of REDCap.Data will be stored 
indefinitely for future analysis and use in developing future protocols using indirect 
calorimetry methods. Only individuals with FBRI access to secure network drives and 
FBRI's instance of REDCap will have access to stored data. Plasma and/or serum aliquot 
samples will be frozen and stored for future analysis of insulin. As it is more cost 
effective to run an ELISA or Immulite 1000 with a full number of wells, these samples 
will be stored until they can be analyzed in batch. Additional aliquots of plasma and/or 
serum will be stored indefinitely after the completion of data analysis to allow for 
replicate testing to ensure data quality and for additional analyses as the field moves 
forward.  
 
 
9.2 For specimens, list the data to be stored or associated with each 
specimen: 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
21 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 22 of 49 
Revised: March 5, 2019 
Only a 4 digit number will be used to label data and the date collected. 
 
 
9.3 Describe the procedures to release data or specimens outside of the 
research team, including the process to request a release, approvals 
required for release, who can obtain data or specimens, and what data 
will be provided with specimens: 
 
No specimens will be released.  
 
 
9.4 Describe the identifiers to be included with stored data or specimens, as 
well as any key or code that could be used to make them identifiable. 
Describe where the code will be stored, who will have access to it, and 
when it will be destroyed: 
 
Data will be coded using a 4 digit number. Participant IDs will be created sequentially 
starting at 9999. The key linking the participant name or identifiable information and 
the code will be stored on an encrypted server managed by the FBRI IT department. 
This server can only be accessed by those on the research team. All study data will be 
kept on a separate server from identifiable information, also encrypted. Any paper 
documents generated that contain other identifiers, such as name, phone number, or 
email will be stored in a locked file cabinet accessible only by authorized staff.   
Identifiable participant information will be destroyed 5 years after publication or at the 
written request of the participant.  
 
Per Virginia Tech policy, in order to compensate participants, social security number (or 
Taxpayer ID) and street address will be collected via a W-9 form or W-8BEN. The 
participant address and SSN will be entered into the ClinCard systems, which is a HIPAA 
compliant payment system approved by Virginia Tech. After entry, this form will be 
given to and stored by the central administration at the FBRI to be used if it is required 
that a tax statement be sent to the participant, based on Federal Income Tax regulations. 
Study staff will not store any documents with SSN or address used for payment purposes. 
 
 
9.5 Please select the identifiers you will obtain (whether directly from 
participants or from another source), including but not limited to: 
 
☒ 
Name 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
22 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 23 of 49 
Revised: March 5, 2019 
☒ 
Geographical subdivisions smaller than a state, including street address, 
city, county, precinct, zip code, and equivalent geocodes (note, the initial 
three digits of a zip code are not considered identifiable) 
☒ 
Elements of dates (except year) for dates directly related to an individual, 
including birth date, admission date, discharge date, date of death, and 
single year of age over 89 and all elements of dates (including year) 
indicative of such age (note, such ages and elements may be aggregated 
into a single category of age 90+) 
☒ 
Phone numbers 
☐ 
Fax numbers 
☒ 
Electronic mail addresses (e-mail) 
☒ 
Social Security numbers 
☐ 
Medical record numbers 
☐ 
Health plan beneficiary numbers 
☐ 
Account numbers 
☐ 
Certificate/license numbers 
☐ 
Vehicle identifiers and serial numbers, including license plate numbers 
☐ 
Device identifiers and serial numbers 
☐ 
Web Universal Resource Locators (URLs) 
☐ 
Internet protocol (IP) address numbers 
☐ 
Biometric identifiers, including finger and voice prints (audio recording) 
☐ 
Full face photographic images and any comparable images (including 
video recording) 
☐ 
Student record number or identification number   
☐ 
User name for online or computer accounts 
☐ 
Any other unique identifying number, characteristic, or code (note this 
does not mean the unique code assigned by the investigator to code the 
data): Click here to explain.   
 
10.0 Sharing of Results with Subjects 
10.1 Describe whether you will share results (study results or individual subject 
results, such as results of investigational diagnostic tests, genetic tests, or 
incidental findings) with subjects or others (e.g., the subject’s primary care 
physician). If so, describe how you will share the results and include this 
information as part of the consent document. Upload materials you will use 
to explain the results to subjects: 
 
Results of the study will not be shared with the participants as they are de-identified prior 
to analysis and/or publication 
 
 
 
11.0 Study Timelines 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
23 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 24 of 49 
Revised: March 5, 2019 
11.1 Describe: 
● 
The duration of an individual subject’s participation in the study (for 
example, 1 hour, 2-4 weeks, 3-5 years). 
● 
The amount of time expected to enroll all study subjects (weeks, 
months, years, etc.) 
● 
The amount of time expected for the investigators to complete this 
study including primary data analyses. 
 
Consent and Training Session: 2 hours 
Behavior Sessions 1-4: 2 hours each (8 hours total) 
Post-Testing Session without fMRI: 1 hour 
Post-Testing Session with fMRI: 3 hours 
Metabolic Chamber Sessions 1 and 2: 6 hours each (12 hours total) 
 
Each participant is expected to be enrolled for approximately 8-10 weeks.  
The study, including recruitment, enrollment, and primary data analysis, is expected to 
last 1 year. 
 
 
 
12.0 Inclusion and Exclusion Criteria 
12.1 Describe how you will screen individuals for eligibility. When will 
screening occur and what procedures will you use? Upload any screening 
scripts or surveys to Protocol Management:  
 
Participants will complete an online screening survey after they have indicated interest in 
participating in the study. A link to the Ripple survey will be sent to participants via 
email, or they can access the link through a written web address or QR code on our flyer. 
Additionally, social media posts will contain the web address. Participants undergoing 
fMRI will also complete an MRI screening form. 
 
 
 
12.2 Describe the eligibility criteria that define who will be included and who 
will be excluded from enrollment for each procedure of your study. 
Include any geographic criteria (e.g., Virginia Tech undergraduate 
students, a national sample of adults with engineering degrees, minors 
aged 8-12 in the New River Valley, university faculty in Virginia and 
Paris, France): 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
24 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 25 of 49 
Revised: March 5, 2019 
 
Inclusion Criteria: 
1. Age 18-45 years 
2. BMI between 18.5-40 kg/m2  
3. Not pregnant or planning to become pregnant during study participation 
4. Residing in the Roanoke area and/or willing/able to attend sessions at FBRI 
5. Able to speak and write in English 
6. Specific to fMRI scan only: BMI >25 kg/m2 
 
Exclusion Criteria: 
1. Current inhaled nicotine use 
2. History of alcohol dependence. 
3. Current or past diagnosis of diabetes, or thyroid problems. 
4. Hemoglobin A1C >5.7% 
5. Taking medications known to influence study measures (including antiglycemic 
agents, thyroid medications, sleep medications) 
6. Active medical or neurologic disorder. 
7. Recent change in body weight (gain or loss of > 5 lbs within the past 3 months) 
8. Current shift work (typical pattern of work/activity overnight) 
9. Previous weight loss surgery 
10. Adherence to a special diet within the past 3 months (e.g., low-carb or ketogenic diet, 
exclusion of food groups/specific macronutrients, intermittent fasting, etc.)  
11. Allergy to any food or ingredient included in the study diets, meals, or beverages 
12. Currently pregnant or planning to become pregnant during study participation 
13. Claustrophobia 
14. Contraindications for MRI, including pacemaker, aneurysm clips, neurostimulators, 
cochlear or other implants, metal in eyes, regular work with steel, etc. (Note: This is an 
fMRI-specific exclusion criterion. Participants may be allowed to participate in all other 
study sessions and measures that do not involve fMRI.)  
15. Contraindications for bioelectrical impedance analysis, specifically implanted devices 
 
 
12.3 Indicate specifically whether you will include or exclude each of the 
following special populations: (You may not include members of these 
populations as subjects in your research unless you indicate them in the 
description of your subject population.) 
● 
Minors, as defined by state law where the study is performed 
(infants, children, teenagers) 
● 
Pregnant women (can be included in minimal risk studies by 
mentioning in section 13.1) 
● 
Prisoners (including all incarcerated individuals) 
● 
Adults not capable to consent on their own behalf 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
25 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 26 of 49 
Revised: March 5, 2019 
We will exclude minors, prisoners, pregnant women and adults not able to consent on 
their own behalf.  
 
 
 
 
 
 
 
13.0 Vulnerable Populations 
13.1 If the research involves individuals who are vulnerable to coercion or 
undue influence, please describe additional safeguards you will include to 
protect their rights and welfare. Consider the applicable items listed 
below: 
● 
If the research involves Virginia Tech students, indicate whether 
these are students of any of the investigators. If so, describe whether 
the activities will take place during class time as part of the 
curriculum and the steps you will take to reduce the possibility that 
students feel obliged to participate in order to improve their course 
grade. The HRPP can provide further guidance as needed. Describe 
whether you will request access to student records (e.g., SAT, GPA, 
GRE scores). 
● 
If the research involves employees of Virginia Tech or the research 
sponsor, describe steps you will take to ensure that the employees 
are freely participating and describe how their data will be 
protected from inspection by their supervisors.  
● 
If the research involves Virginia Tech NCAA athletes, you must 
obtain approval from the athletic department.  
● 
For research involving Montgomery County Public Schools, you 
must obtain county approval (after obtaining contingent Virginia 
Tech approval). Other locales have different requirements; please 
check on these and describe here. Approval is typically granted by 
the superintendent, principal, and classroom teacher (in that order). 
Approval by an individual teacher is insufficient. School approval, in 
the form of a letter or a memorandum should be uploaded as a 
supporting document. 
● 
If the research involves pregnant women, review “CHECKLIST: 
Pregnant Women (HRP-412)” to ensure that you have provided 
sufficient information in this protocol. 
● 
If the research involves prisoners, review “CHECKLIST: Prisoners 
(HRP-415)” to ensure that you have provided sufficient information 
in this protocol. 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
26 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 27 of 49 
Revised: March 5, 2019 
● 
If the research involves persons who have not attained the legal age 
for consent to treatments or procedures involved in the research 
(minors), review the “CHECKLIST: Minors (HRP-416)” to ensure 
that you have provided sufficient information in this protocol. 
● 
If the research involves cognitively impaired adults, review 
“CHECKLIST: Cognitively Impaired Adults (HRP-417)” to ensure 
that you have provided sufficient information in this protocol. 
 
 
All subjects will be informed of the purpose of the study, the potential value of the study 
to society, the lack of value to the subject personally, and all potential risks to the subject. 
Study staff will explain to the subject that he or she is under no obligation whatsoever to 
participate, and that if he or she wishes to discontinue their involvement at any time 
during the study, he or she is free to do so without penalty. Employees will be told that 
their employment status and evaluation will in no way be affected by their decision to 
participate or not participate in the study. Students will be advised that their decision to 
participate will not affect their student status, grades, coursework, or extra-curricular 
activities. 
 
 
 
 
 
14.0 Number of Subjects 
14.1 Indicate the total number of subjects to be enrolled and how this number 
was determined (e.g., sample size calculation [show], number of available 
subjects in a finite pool, number of tests funding award would allow):  
 
We plan to consent approximately 25 participants with the intent of having 15 
participants complete all sessions of Aims 1 and 2, and we anticipate having 5 of those 
participants also complete an fMRI scan. These numbers were determined based on our 
pilot grant award amount, the sample size we anticipate needing to generate 
pilot/feasibility data, and the anticipation of ~40% either dropping out or being excluded 
after enrollment (e.g., unable to determine flavors equally liked and unfamiliar for the 
conditioning paradigm, hemoglobin A1C outside of inclusion range, scheduling 
challenges, etc.).  
 
 
 
14.2 If this is a multi-site study, indicate the number of subjects to be enrolled 
at this site and the total to be enrolled from all sites: 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
27 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 28 of 49 
Revised: March 5, 2019 
Single site. 
 
 
14.3 If applicable, indicate the number of potential subjects you expect to 
screen for enrollment, and the number of subjects you will need to 
complete the research procedures: 
 
We anticipate screening via the online screening survey approximately 150 potential 
participants. We anticipate enrolling and screening approximately 25 participants during 
our Consent and Training Session. We intend to have 15 participants complete all 
procedures except the fMRI scan. Of those 15 participants, we intend to have 5 
participants complete the fMRI scan. 
 
 
 
14.4 If the study has more than one procedure, indicate the total number of 
subjects to undergo each procedure separately: 
 
Consent and Training Session (flavor tasting/rating, anthropometrics, fingerstick sample 
for HbA1C): 25 participants 
Behavior Sessions 1 and 3 (IV catheter blood draw): 15 participants 
Behavior Sessions 3 and 4 (metabolic cart): 15 participants 
Post-Test Session without fMRI (flavor and beverage tasting/rating): 15 participants 
Post-Test Session with fMRI (flavor and beverage tasting/rating and fMRI scan): 5 
participants 
Metabolic Chamber Sessions 1 and 2: 15 participants 
 
 
15.0 Recruitment Methods 
15.1 Describe when, where, and how you will recruit potential subjects: 
 
Approved study recruitment materials will be distributed in the follow ways. Flyers will 
be posted on bulletin boards and approved posting sites throughout Roanoke, Blacksburg, 
and surrounding areas. Digital advertisements will be displayed on DiFeliceantonio lab 
and CHBR social media, VT News, and in list serves with permission.  
 
All advertisements will include a link to the lab website where there will be approved 
language describing the research study. Interested participants will fill out a general 
screening survey. All screening surveys will be delivered through Ripple. This screening 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
28 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 29 of 49 
Revised: March 5, 2019 
survey will be used to determine study eligibility. Once the general screening survey has 
been completed, participants who may be eligible for this study will be contacted via 
email (see attachment 'Screening email'). If participants choose to complete the study-
specific screening form and are eligible for this study, they will be contacted to schedule 
a consent session. 
 
 
Opt-In Database 
Individuals who complete the screening survey in response to other studies in the lab will 
have the opportunity to opt-in to being notified of future studies our lab is conducting. 
One question on the screening survey asks if individuals would like to be contacted if 
they potentially qualify for any future studies. Individuals who have opted-in to being 
notified of future studies receive an email containing information about the study and an 
email address for contacting the research study team. The email will be sent through 
Ripple, the secure web application we are using for storage and management of 
personally identifying information of research participants. Individuals who are interested 
in scheduling a consent session will undergo a brief screening to ensure they have had no 
changes to their health history or other screening survey questions since they first 
completed the survey that would preclude them from participating. 
 
Carilion Clinic Recruitment 
Flyers will be posted throughout Carilion Clinic waiting rooms and offices. Physicians 
and other providers will also be notified of this study and asked to encourage individuals 
who may qualify to complete the screening survey. 
 
 
 
15.2 Describe the source of subjects (for example, clinic patients with specific 
conditions, students in the library, community members at a gathering, or 
members of a local gym): 
 
Participants will be recruited from the general public. 
 
 
15.3 Describe the methods that you will use to identify potential subjects: 
 
Responses on the online screening form will be used to identify potential subjects.  
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
29 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 30 of 49 
Revised: March 5, 2019 
15.4 Describe materials that you will be use to recruit subjects. Attach copies 
of these documents with this protocol in Protocol Management and be 
sure to include the IRB protocol number on each document.  
● For flyers, attach the final copy of printed flyers.  
● For Virginia Tech News, Facebook postings and ads, newspaper ads, 
websites, MTurk/SONA/online survey systems, etc., attach the final 
wording and graphics to be used.  
● For email recruitments, please include the subject line.  
● For advertisements meant for audio broadcast, please submit the 
wording of the advertisement prior to taping (to avoid having to re-
record with approved language) and submit the final recorded version 
for IRB review before use.  
● Describe any compensation to subjects. Separate compensation into 
appropriate categories, such as: reimbursement for expenses, time and 
effort, and additional incentives for study participation.  For each 
category, specify the amount (including any pro-rated amount), schedule, 
and method of payment. 
 
See above in 15.3 for a list of recruitment materials and methods that will be used. 
 
Consent and Training Session (2 hrs) - $40 
Behavior Sessions 1-4 (2 ½ hours each) - $50each session, up to $200 total 
Post-Test Session (1 hour) - $20  
fMRI Scan (2 hours) - $40 
Metabolic Chamber Sessions 1-2 (6 hours each)- $120 each session, up to $240 total 
 
Total (with fMRI scan): up to $540 
Total (without fMRI scan): up to $500 
 
Participants will be paid via ClinCard, a prepaid Mastercard that we will provide them at 
the end of the first session. Participants will be compensated at the end of each visit using 
the ClinCard system.  
 
 
 
 
16.0 Withdrawal of Subjects 
16.1 Describe circumstances under which you anticipate subjects could be 
withdrawn from the research without their consent: 
 
As is standard with MRI research, if subjects experience unusual sensations, they will be 
withdrawn. In some cases, due to the magnetic field (as described in risks below) subjects 
may experience peripheral nerve stimulation, e.g. tingling or twitching. They will be 
withdrawn from the study if this occurs.  
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
30 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 31 of 49 
Revised: March 5, 2019 
 
If participants are able to detect the maltodextrin in the triangle test, they will not 
continue in the study and will be compensated for the training session. 
 
Other potential reasons for withdrawal without participant consent include information on 
the screening questionnaire that would make participating unsafe for the participant, 
failure to follow the study protocol as instructed (e.g., not fasting for measurements, 
failure to consume beverages at home at the appropriate times, consuming foods or 
beverages outside of those provided by the study on controlled feeding days, etc).  
 
 
 
16.2 If applicable, describe any procedures for orderly termination (e.g., 
discontinuation of a study drug or debriefing after a behavioral 
intervention): 
 
There are no study drugs or behavioral interventions. 
 
 
16.3 Describe procedures that you will follow when subjects withdraw from the 
research, including partial withdrawal from procedures with continued 
data collection (e.g., participant declines to continue with regular blood 
draws, but continues with periodic behavioral questionnaires): 
 
The following steps will be taken in the event a participant withdraws from the study 
while undergoing the study protocol and informs us they want their information 
destroyed: 
1. The participant database will be updated that the participant withdrew (for all 
withdrawals). 
2. Participant will be compensated for their time and sessions completed up to time of 
withdrawal. There will be no partial withdrawal. 
3. All participant identifying information and data collected up to time of withdrawal will 
be removed from study database(s).  
4. All stored blood samples collected on the participant will be removed from the storage 
freezer and discarded.  
5. All paperwork associated with the participant will be shredded.  
 
 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
31 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 32 of 49 
Revised: March 5, 2019 
17.0 Risks to Subjects 
17.1 List the reasonably foreseeable risks, discomforts, hazards, or 
inconveniences to the subjects related the subjects’ participation in the 
research. Include for the IRB’s consideration a description of the 
probability, magnitude, duration, and reversibility of the risks. Consider 
physical, psychological, social, legal, privacy, and economic risks. Do not 
indicate “No risk” or “N/A.” Instead, for studies with very low risk (e.g., 
anonymous online questionnaire on a mundane topic) indicate “The 
investigators are not aware of any risks from participation in this study.” 
or “No more than risks than are found in everyday life.” The example 
consent form presents a tabular method for risk information, which you 
can also use here. Common risk types include: 
● Physical (e.g., potential for pain, discomfort, infection) 
● Psychological (e.g., potential for stress, discomfort, and/or 
embarrassment) 
● Social (e.g., potential for discrimination or stigmatization and 
disruption of personal and family relationships) 
● Legal (e.g., potential for disclosure of illegal activity, negligence) 
● Privacy (e.g., potential for personal information being accessed, used, 
or disclosed without the subjects’ knowledge or consent, breach of 
confidentiality/security) 
● Economic (e.g., potential for individuals to lose access to economic 
services, employment, insurability) 
 
Food and beverages 
All components of the beverage and food stimuli are readily available outside the 
laboratory (grocery stores, etc.) and are likely already consumed by the participants. 
There are no known risks associated with these stimuli. 
 
Indirect calorimetry 
There are no known risks associated with the noninvasive indirect calorimetry methods 
we will be using to measure gas exchange. These non-invasive procedures are simple and 
painless, and normal clothes can be worn during the procedures. For the metabolic cart 
procedure, subjects will be lying back in a chair and a transparent "canopy hood" will be 
placed over their face and neck. The subjects will then be asked to relax and breathe 
normally during the entirety of the testing. During the testing, the study staff will be able 
to see and hear participants so should they, at any time during the testing, become 
uncomfortable with the hood or chair, they may choose to stop the procedure. For the 
metabolic chamber procedure, subjects will rest quietly on a hospital bed inside an 
enclosed chamber for the duration of the test. They will be asked to relax and breathe 
normally during the entirety of testing. During the testing, study staff will be able to see 
participants, so should they, at any time during the testing, become uncomfortable, they 
may choose to stop the procedure. 
 
MRI 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
32 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 33 of 49 
Revised: March 5, 2019 
The risks associated with fMRI are the same as those with conventional MRI. Movement 
or heating of metallic implants is a potential risk, therefore subjects will be screened to 
exclude people with metallic implants, fragments, or pacemakers. Some individuals may 
experience claustrophobic reactions in the scanner. Any subject experiencing 
claustrophobia will be removed from the scanner immediately. 
The fMRI scanner produces loud noises which could be harmful to the subject. 
Participants will be required to wear headphones and earplugs to minimize the noise 
generated by the scanner. 
 
The Siemens 3 T scanner has been approved by the FDA. However, there may be 
additional risks associated with scanning at 3.0 T compared to the conventional clinical 
scanners in the 1.5-2.0 T range. These include: 
1. Effect of the static field. There is no conclusive evidence for irreversible or hazardous 
bioeffects to acute, short-term exposures of humans up to 2.0 T (Shellock and Kanal, 
1996). Studies have indicated some side-effects at 4.0 T, namely unusual sensations 
including nausea, vertigo, and metallic taste (Schenck, 1991). However, there is no 
evidence that this is either irreversible or harmful. If subjects experience unusual 
sensations, they will be withdrawn. 
 
2. Effect of the gradient field. MRI operates by rapidly changing small additional fields, 
called gradients. This will induce small electrical currents in any conductor, and thus 
could theoretically induce mild peripheral nerve stimulation. However, this is not 
substantially different at higher magnetic fields since the gradients are separate from the 
main magnet. There is no evidence that the effect of the gradients is any different at 3 T 
than at 1.5 T. However, if subjects experience peripheral nerve stimulation, e.g. tingling 
or twitching, they will be withdrawn. 
 
3. Effect of the RF electromagnetic field. The higher magnetic field strength requires that 
higher RF frequency pulses are used to excite the protons in the subject's brain. The 
limits of RF energy that can be safely given to humans has been clearly defined by the 
FDA: a). The exposure to RF energy below the level of concern is an SAR of 0.4 W/kg or 
less averaged over the body, and 8.0 W/kg or less spatial peak in any 1 g of tissue, and 
3.2 W/kg or less average over the head; or b). The exposure to RF energy that is 
sufficient to produce a core temperature increase of 1 degree C and localized heating to 
no greater extent than 38 degrees C in the head, 39 degrees C in the trunk, and 40 degrees 
C in the extremities, except for patients with impaired systemic blood flow and/or 
perspiration. We will adhere to the recommendations for the head, which is also 
monitored by a Siemens built-in monitor. 
 
4. Acoustic Noise Levels. Rapid changes in the currents in the gradient coils of the MRI 
scanner produce significant levels of acoustic noise. The levels of noise range 
approximately between 65 and 95 dB, but could have higher peaks. Communications 
with subjects takes place with the standard pneumatic headphones provided by the 
scanner manufacturer. These headphones provide some degree of noise reduction. Foam 
ear plugs with a Noise Reduction Rating of 31 dB will be provided.  
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
33 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 34 of 49 
Revised: March 5, 2019 
Subjects may experience discomfort, such as anxiety during the MRI scanning. Subject 
comfort will be reassessed and subjects will be allowed to discontinue any session at any 
time. 
 
The subject may communicate with the experimenter at any time: 1) before, in between, 
or after a scan via an intercom device between the scanner and control room, and 2) 
during a scan via a pneumatic squeeze bulb located in the scanner that triggers an alarm 
in the control room. Subjects who report discomfort and wish to discontinue their 
participation will be immediately withdrawn from the scanner. Subjects who are 
withdrawn from the study will be compensated for their participation to that point. 
 
IV Catheter Blood Draws 
All catheter placements and draws will be performed by a trained and skilled medical 
technologist, phlebotomist, or nurse in the appropriate laboratory setting with proper 
antiseptic technique. However, there are still some risks of this procedure. Some pain or 
discomfort may be experienced when the catheter is inserted in the vein, but this should 
persist for only a short time. During the blood draws, there may be pain and/or bruising at 
the site where the IV catheter is inserted. In rare cases infection or blood clot may occur 
at the IV catheter site. These are the same risks associated with IV catheter placement in 
any medical setting.  
 
Fingerstick Blood Sample 
The risks associated with performing a fingerstick to collect a capillary blood sample are 
minimal. The likely risks involve pain and bleeding in the fingertip at the collection site. 
Rarely, an infection may occur at the puncture site. To minimize these risks, research 
team members will be trained on the proper technique for obtaining a capillary blood 
sample and care for the site after the sample is obtained.  
 
Bioelectrical Impedance Analysis 
The risks associated with having body composition measured with bioelectrical 
impedance analysis are minimal. The likely risks include potential interference with 
electronic medical device implants, including pacemakers and defibrillators. To minimize 
these risks, medical device implants are an exclusion criterion for study participation.  
 
 
 
17.2  Indicate the measures you will use to minimize risks and monitor subjects 
for safety. (e.g., asking a subject at regular intervals to rate how they are 
feeling from 1 to 10, or to slowly crouch in order to check their balance.) 
 
Indirect calorimetry 
Indirect calorimetry sessions (Behaviors Sessions 2 and 4, and Metabolic Chamber 
Sessions) will be performed by trained personnel. Subjects who become uncomfortable 
during the measurements can withdraw at any time during the procedure. 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
34 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 35 of 49 
Revised: March 5, 2019 
MRI 
In order to minimize noise from the MRI scanner that can be potentially harmful for the 
subject, all subjects will be required to wear ear protection while in the scanner. The 
subject may communicate with the experimenter at any time: 1) before, in between, or 
after a scan via an intercom device between the scanner and control room, and 2) during a 
scan via a pneumatic squeeze bulb located in the scanner that triggers an alarm in the 
control room. Subjects who report discomfort and wish to discontinue their participation 
will be immediately withdrawn from the scanner. Subjects who are withdrawn from the 
study will be compensated for their participation to that point. 
 
Advanced MR/Operator Training:  Required for those persons wishing to conduct 
research on the HNL MRIs. Training will consist of presentations, observation, and 
hands-on practice. Topics will include safety and emergency procedures, subject 
screening and preparation, scanner set-up and operation, and troubleshooting.  Each 
person must also complete at least 4 hours of observation (to include 2 different studies) 
and at least 16 hours of supervised scanning (to include 4 different studies). Certification 
will be granted upon completion of the training and passing of a competency exam 
 
Fingerstick Blood Draw 
To reduce the risks associated with performing a fingerstick, research personnel will train 
participants on the proper technique for performing a fingerstick blood draw and care for 
the site after the sample is obtained. Participants will also be given separate, single-use 
blood collecting devices to be used for each blood draw.  
 
IV Catheter Blood Draws 
To reduce the risks associated with inserting an IV catheter and drawing blood from the 
catheter, only skilled medical technologists, nurses and/or other medical professionals 
will insert the IV catheters and draw the blood samples. In addition, a maximum of three 
attempts will be made to insert the catheter in the participants' arm.  
 
 
 
17.3 If applicable, indicate which procedures might have risks to the subjects 
that are currently unforeseeable. This will be rare, and usually applicable 
when testing a new drug or device or a new use of an existing drug or 
device: 
 
There are no new devices or drugs used in this study. 
 
 
17.4 If applicable, indicate which procedures might have risks to an embryo or 
fetus should the subject be or become pregnant: 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
35 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 36 of 49 
Revised: March 5, 2019 
No procedures have a known risk to a fetus or embryo. 
 
 
17.5 If applicable, describe risks to others who are not subjects (e.g., collection 
of sensitive health data that might affect sexual partners if disclosed, 
mandatory reporting of abuse, DNA testing that might affect family 
members or relationships): 
 
N/A 
 
 
18.0 Potential Benefits to Subjects 
18.1 Describe the potential benefits that individual subjects might experience 
from participating in the research. Include the probability, magnitude, and 
duration of the potential benefits, as this will be useful to the IRB’s 
risk:benefit analysis. Do not include benefits to society or others. Do not 
list monetary or non-monetary compensation for participation, as this is 
not a benefit These should be included in section 2 or 3 of this document: 
 
There is no direct benefit to the individual subject for participation. 
 
 
18.2 If applicable, specify that there are no anticipated direct benefits for 
participants:  
 
There is no direct benefit to the individual subject for participation. 
 
 
 
 
 
 
19.0 Data Management and Confidentiality 
19.1 Describe procedures that you will use for quality control to ensure validity 
of collected data: 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
36 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 37 of 49 
Revised: March 5, 2019 
 
All data will be collected and analyzed by trained research personnel. They will be 
trained in MRI safety, Human Subjects Protections, and standard laboratory procedures 
(perceptual ratings, etc.). 
 
 
19.2 Describe any existing data or biospecimens you will obtain as part of this 
study. Include:  
● 
Variables or samples to be obtained  
● 
Source of the data or specimens 
● 
Your authorization to access or receive the data or biospecimens 
● 
Whether the data or biospecimens are publicly available 
● 
Whether the data or specimens you receive will contain identifiers  
 
This study collects new data. 
 
 
19.3 Describe the steps that you will take to handle and secure study data 
during data collection, storage, use, and transmission.  Include 
information about training of study staff, authorization of access, 
password protection, encryption, physical controls, certificates of 
confidentiality, separation of identifiers and data, etc.: 
 
Each subject will be assigned a coded designation. All study data to be used in analysis 
will be de-identified using this coded designation which will contain no biographical data 
that can be linked to individuals.  
 
Participant personal information and the coded designation will be kept in Ripple™, a 
secure web application designed for the storing and management of personally 
identifying information of research participants. Ripple was initially developed at the 
University of Michigan to provide a user-friendly, web-based secure interface where 
research teams can centralize the storage and management of research participants’ 
personal information, including name, participant ID, demographics, and study workflow 
(e.g., appointments). Participant information managed with Ripple is private and secure. 
This information is kept in fully encrypted format inside dedicated databases that are 
segregated from other Ripple accounts and thus only authorized study staff will have 
access to the study data. Likewise, Ripple infrastructure complies with the privacy and 
security guidelines of the Health Insurance Portability and Accountability Act (HIPAA), 
including 2048-bit data encryption in transit and at rest, automatic logoff, audit trail, daily 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
37 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 38 of 49 
Revised: March 5, 2019 
backups in triplicate dedicated servers, firewall, custom access permission for lab 
members, zxcvbn password strength estimation, and enterprise administrative safeguards 
to prevent unauthorized staff from accessing participant information. Furthermore, Ripple 
is used only for storing personally identifiable information of participants and is not used 
to capture other research data (e.g., questionnaires, health records, etc.). This ensures that 
the personally identifiable information and research data are segregated. Only personnel 
who are direct members of the research team will have access to personally identifiable 
information stored in Ripple. The link between the coded designation and personal 
information will only exist on this secure platform. 
 
MRI images will be stored on secured FBRI and VT ARC servers.  
The behavioral stream of data acquired during experiments will be collected and recorded 
using an encrypted FBRI laptop, desktop computer, or tablet and then transferred to an 
FBRI server. 
 
 
19.4 For multi-site studies, describe how data or specimens will be handled 
and secured for each site (e.g., central or disseminated data storage, data 
coordinating center): 
 
This is a single site study. 
 
 
19.5 Describe the plan for data disposition following the conclusion of the 
study (e.g., long term maintenance of data, data destruction methods).  
● 
What information will be included in the long term storage of data 
or specimens? 
● 
How long will the data or specimens be stored? 
● 
Where and how data or specimens will be stored? 
● 
Who will have access to the data or specimens during long term 
storage? 
● 
Who is responsible for receipt or transmission of the data or 
specimens? 
● 
How will data or specimens be shared or transported? 
● 
When and how will personal identifiers be destroyed? 
 
 
Data and specimens may be archived indefinitely for future analysis. All data and 
specimens will be de-identified, coded and contain no specific biographical information 
which can be related back to an individual. The 'key' will be stored in a database which 
cannot be accessed unless specifically authorized. Deidentified data may be made 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
38 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 39 of 49 
Revised: March 5, 2019 
available publicly upon publication as required by funding agencies and journals. All 
staff are informed of HIPAA Security and Privacy rules. 
 
 
 
 
20.0 Provisions to Protect the Privacy Interests of Subjects 
20.1 Describe the steps that you will take to protect subjects’ privacy interests. 
“Privacy interest” refers to a person’s desire to place limits on with whom 
they interact or to whom they provide personal information (e.g., 
collecting the minimal amount of private information required to complete 
the study, protecting the data once it is obtained):  
 
Only the information required to complete the study, document participation or 
compensate the participant will be collected. Once obtained, these data will be stored 
securely, accessible only by authorized personnel. For instance, documents with a 
participant name (such as the Consent Form) will be stored in a separate, locked file 
cabinet from files containing participant coded identifiers. Study keys will only be 
accessible to authorized staff requiring access, such as trained research coordinators 
interacting with the participant for the study session. 
 
 
20.2 Describe steps that you will take to make subjects feel at ease with the 
research situation in terms of the questions being asked and the 
procedures being performed. “At ease” does not refer to physical 
discomfort, but the sense of intrusiveness a subject might experience in 
response to questions, examinations, and procedures (e.g., use of a same 
gender investigator to place sensors on the torso, a private changing area 
if clothing must be changed, sensitivity when discussing  pregnancy testing 
with subjects, making it clear on surveys that participants can discontinue 
at any time, not asking questions about private or sensitive issues unless 
necessary for the research): 
 
Research staff will be trained to check with participants throughout all procedures to 
ensure they are comfortable with all proceedings. Before all procedures, participants will 
be reminded they can stop at any time. After obtaining written consent for the study, 
research staff will always ask for verbal consent before each procedure.  
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
39 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 40 of 49 
Revised: March 5, 2019 
20.3 Describe how you plan to access existing sources of information about the 
subjects (e.g., medical records, grades) and how you will protect 
participant privacy through the data security plan: 
 
We will not access existing sources of information. 
 
 
20.4 Describe any required reporting that might occur as a result of your 
research questions, study populations, and data collection methods. 
Examples for Virginia and Virginia Tech include: 
• Any suspicions (e.g., circumstantial, disclosed) of child abuse (physical, 
emotional, sexual) and neglect  
• Sexual discrimination and/or sexual violence that involves a student 
• Disclosure or signs of intention to harm oneself (i.e., suicidal ideation 
and/or plan) 
• Disclosure or signs of desire to harm others (i.e., homicidal ideation 
and/or plan) 
• Suspected abuse, neglect or exploitation of vulnerable adults (e.g., 
individuals with a disability, elderly persons) 
 
 
We do not foresee any circumstance where this type of required reporting would come up 
during our study. 
 
 
 
 
21.0 Provisions to Monitor the Data to Ensure the Safety of Subjects 
Safety monitoring is required when research involves greater than minimal risk 
and is sometimes appropriate for other studies. 
21.1 Describe: 
● 
The plan to periodically evaluate the data collected regarding both 
harms and benefits to determine whether subjects remain safe  (e.g., 
periodic reporting to the IRB, establishing a data monitoring 
committee, reporting data monitoring committee findings to the IRB 
and the sponsor). 
● 
What data you will review, including safety data, unexpected events, 
and data that show the ability to produce the intended results. 
● 
How the safety information will be collected (e.g., with case report 
forms, at study visits, by telephone calls with subjects). 
● 
The frequency of data collection, including when safety data 
collection starts. 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
40 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 41 of 49 
Revised: March 5, 2019 
● 
Who will review the safety data and with what frequency. 
● 
The statistical tests for analyzing the safety data to determine 
whether harm is occurring. 
● 
Any conditions that will trigger an immediate suspension of the 
research (e.g., a serious adverse event). 
 
 
The Principal Investigator (PI) or study staff will be responsible for ensuring participants’ 
safety on a daily basis and for reporting Serious Adverse Events and Unanticipated 
Problems to the Institutional Review Board. The PI will be informed of serious adverse 
events as soon as they occur by the study staff and will report any serious adverse events 
and unanticipated problems to the IRB. Safety information will be collected at study 
visits by research personnel. This safety information will be reviewed weekly by the PI 
during a weekly lab meeting. A serious adverse event would trigger immediate 
suspension of the research.  
 
 
 
 
 
22.0 Compensation for Research Related Injury 
22.1 If the research involves more than minimal risk to subjects, describe the 
available compensation in the event of research-related injury, if any: 
 
This study does not present more than minimal risk. 
While the risk involved in this study are minimal, in the event that a research-related 
event or injury does occur, participants will be treated appropriately according to the 
nature of the event or injury. If participants are no longer able to continue in the study, 
they will receive compensation in accordance with the type and number of sessions 
completed.  
 
 
 
22.2 Provide a copy of contract language, if any, relevant to compensation for 
research-related injury. At Virginia Tech, this is most common for 
sponsored research: 
 
This study does not present more than minimal risk. 
 
23.0 Economic Burden to Subjects 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
41 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 42 of 49 
Revised: March 5, 2019 
23.1 Describe any costs that subjects might be responsible for because of 
participation in the research, including any uncompensated costs for items 
such as transportation, missed work, and childcare: 
 
Participants are compensated for their time. They can make arrangements to participate 
outside of work and child-care needed hours.  
 
 
24.0 Consent Process 
24.1 Indicate the process by which you will obtain consent for study 
participation. Please upload all consent, parental permission, and assent 
forms, documents, and scripts referenced in this section to Protocol 
Management. 
Describe the following: 
● 
Where the consent process will take place (e.g., clinic waiting area, 
classroom, online) 
● 
The time interval between sharing the consent information with the 
prospective subject and obtaining consent. For lab, interview, and 
focus group studies, the Virginia Tech IRB prefers that subjects have 
at least 24 hours to review the consent form and study information 
before the appointment where consent will be obtained. For simple 
online survey studies, you can typically present the consent 
information immediately before subjects begin participation. 
● 
If applicable, processes to ensure ongoing consent or assent (e.g., 
for multiple sessions; for research in which a minor will turn 18 
during the study; for longitudinal research with minors who will 
later be asked to provide or affirm their assent). 
● 
Please review “SOP: Informed Consent Process for Research (HRP-
090)” for recommended procedure. Describe your process, being 
sure to include: 
o 
The name and role of all study personnel who will be trained 
and certified by the PI to conduct the consent process 
o 
The time that will be devoted to the consent discussion 
o 
Steps that you will take to minimize the possibility of coercion 
or undue influence 
o 
Steps that you will take to gauge or ensure the subjects’ 
understanding 
 
Consent for the screening questionnaire will be documented by completion of the survey 
by the participant. Information about the study, instructions for completing the survey, 
next steps for participation, and what will happen with their information if they are/are 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
42 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 43 of 49 
Revised: March 5, 2019 
not (i.e., screen fail) determined to be eligible to participate will be provided in the 
"Introduction" section of the survey. 
 
During the first session (Consent and Training Session), consent for participating in the 
study will be obtained on-site by research personnel in a quiet, private room or waiting 
area. Consent materials will be emailed to the participants for their review at least 24hrs 
in advance of their first appointment. 
 
Participants will be given as much time as they request to review the consent documents.  
 
This study involves multiple sessions and a single consent form will be signed that details 
all sessions. Participants will provide their consent for all sessions at the first session, but 
will be informed they may withdraw from the study at any time. Verbal consent will be 
obtained before each subsequent session.  
 
Staff will devote as much time as requested to the consent process. Typically, review of 
the consent form takes approximately 10 minutes. Participants will then be given time to 
review the form independently and ask any questions they may have. Staff will confirm 
with the participant that they have had all questions answered prior to the participant 
signing the consent form. 
 
In order to determine understanding, staff will discuss each section with the participant, 
verifying their understanding by verbal confirmation throughout.   
 
 
 
Non-English Speaking Subjects 
● 
Indicate what language(s) other than English are understood by 
prospective subjects or representatives. 
● 
If non-English speakers will be recruited, describe the process you 
will use to ensure that the oral and/or written consent information 
provided will be in a language that they understand.  
● 
If you translate consent forms and study materials, please provide a 
certified translation of the form as well as the certification 
document. 
● 
Indicate the spoken language that study personnel obtaining consent 
will use. Describe how you will assess fluency of personnel obtaining 
consent to ensure that the translation is accurate. 
 
 
We will recruit only English-speaking participants. 
 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
43 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 44 of 49 
Revised: March 5, 2019 
Waiver or Alteration of Consent Process (consent will not be obtained, 
required information will not be disclosed, or the research involves 
deception) 
● 
Review the “CHECKLIST: Waiver or Alteration of Consent Process 
(HRP-410)” to ensure you have provided sufficient information for 
the IRB to make these determinations (i.e., that it meets the criteria 
for a waiver or alteration of the consent process). 
 
We do not waive consent. 
 
 
 
Subjects who are not yet adults (minors: infants, children, teenagers) 
● 
Describe the criteria that you will use to determine  legal age for 
consent to treatments or procedures involved in the research under 
the applicable law of the jurisdiction in which the research will be 
conducted (e.g., in Virginia, individuals under the age of 18 years). 
o 
For research conducted in Virginia, review “SOP: Legally 
Authorized Representatives, Minors, and Guardians (HRP-
013)” to determine which individuals in the state meet the 
definition of “minor.” 
o 
For research conducted outside of the state, please describe 
the legal requirements for the definition of “minor.”  
● 
Describe the process for obtaining parental permission.  
o 
Permission from one parent is acceptable for studies that 
involve no greater than minimal risk OR involve greater than 
minimal risk but present the prospect of direct benefit to the 
minor subject. 
o 
Permission from both parents is required in all other 
cases (unless one parent is deceased, unknown, 
incompetent, or not reasonably available, or when only 
one parent has legal responsibility for the care and 
custody of the minor). 
● 
Describe whether you will obtain permission from individuals other 
than parents or Legally Authorized Representatives, and if so, who 
will be allowed to provide permission. Describe the process you will 
use to determine these individuals’ authority to consent to the 
minor’s general medical care. 
● 
Indicate whether you will obtain assent from all, some, or none of 
the minors. If you will obtain assent from some minors, indicate 
which minors will be required to assent. Consider chronological age 
and intellectual capacity when determining who will be required to 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
44 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 45 of 49 
Revised: March 5, 2019 
provide assent (e.g., infants are unable to assent. However, 
teenagers are likely able to read and sign an assent form). 
● 
When assent of minors is obtained, describe whether and how you 
will document it.  Will minors sign an assent form or give verbal 
assent? 
● 
Attach parental permission and minor assent forms or scripts in 
Protocol Management.  
 
 
Only adults 18 years or older will be eligible to participate in the study. 
 
 
 
Adults Unable to Consent  
● 
Describe the process you will use to determine whether an individual 
adult is capable of consent. 
● 
List the individuals from whom you will obtain permission in order 
of priority (e.g., durable power of attorney for health care, court 
appointed guardian for health care decisions, spouse, and non-
minor child). 
o 
For research conducted in the Virginia, review “SOP: Legally 
Authorized Representatives, Minors, and Guardians (HRP-
013)” to determine which individuals in the state meet the 
definition of “legally authorized representative.” 
o 
For research conducted outside of Virginia, please describe 
the legal requirements for obtaining permission from a legally 
authorized representative in the state where the research will 
occur. 
● 
Describe the process for assent of the subjects. 
o 
Indicate whether you will require assent from all, some, or 
none of the subjects. If some, indicate which subjects will be 
required to assent and which will not. 
o 
If you will not obtain assent from some or all subjects, please 
provide justification for not obtaining assent. 
o 
Describe whether and how you will document assent.  
 
Adults must be able to consent themselves. 
 
 
25.0 Process to Document Consent in Writing 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
45 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 46 of 49 
Revised: March 5, 2019 
25.1 Consult “SOP: Written Documentation of Consent (HRP-091)” for 
recommended procedures, and describe whether and how consent of the 
subject will be documented in writing: 
 
We do not request to waive written consent. See details in 8.0 Procedures Involved for 
details about consent and training session. In brief, participants will sign the informed 
consent document in the presence of an investigator in the first study session. They will 
be emailed a copy of the consent document at least 24 horus before the first session, and 
the investigator will review the protocol with the participant and answer any questions 
prior to signing the consent document. Only authorized research personnel will print the 
consent form from the IRB Protocol Management site to ensure the most recent 
document is being used. Copies of the signed consent document will be provided to 
participants.  
 
 
25.2 If the research presents no more than minimal risk of harm to subjects and 
involves no procedures for which written documentation of consent is 
normally required outside of the research context, you can request that the 
IRB waive the requirement to obtain written documentation of consent 
(e.g., consent to participate is indicated by pressing a button for an online 
questionnaire – after the consent information is presented and before the 
questionnaire begins): 
 
We do not request to waive written consent.  
 
 
25.3 If you will document consent in writing, attach a consent document with 
places for signatures. If you will obtain consent, but not document consent 
in writing, please attach the consent script or text. Review “CHECKLIST: 
Waiver of Written Documentation of Consent (HRP-411)” to ensure that 
you have provided sufficient information. You should use “TEMPLATE 
CONSENT DOCUMENT (HRP-502)”to create the consent document or 
script: 
 
Consent for this project is attached to this application. 
 
 
 
 
26.0 Resources Available 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
46 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 47 of 49 
Revised: March 5, 2019 
26.1 Describe the resources available to conduct the research. For example, as 
appropriate: 
● 
Describe the PI’s availability to supervise the research. 
● 
Justify the feasibility of recruiting the required number of suitable 
subjects within the agreed recruitment period. For example, how 
many potential subjects do you have access to? What percentage of 
those potential subjects do you need to recruit? 
● 
Describe the time that you will devote to conducting and completing 
the research. 
● 
Describe your facilities. 
● 
Describe the availability of medical or psychological resources that 
subjects might need as a result of an anticipated or unanticipated 
consequence of participation in the research. 
● 
Describe your process to ensure that all persons assisting with the 
research are adequately informed about the protocol, the research 
procedures, and their duties and functions (e.g., training plans, 
detailed study notebooks). 
 
The PI is a research-dedicated (100%) Faculty member at Virginia Tech and the Fralin 
Biomedical Research Institute. The majority of the PI's time is spent supervising and 
analyzing ongoing research.  
 
Fralin Biomedical Research Institute 
Dr. DiFeliceantonio’s lab is part of the Fralin Biomedical Research Institute at Virginia 
Tech Carilion (FBRI at VTC), located in Roanoke, Virginia. The Research Institute was 
founded approximately eight years ago as part of a commitment to biomedical sciences 
by Virginia Tech, and was originally called the Virginia Tech Carilion Research Institute. 
The name changed to Fralin Biomedical Research Institute at VTC in late 2018 following 
a transformative gift of fifty million dollars by the Fralin Family. Primary institute faculty 
have a primary faculty appointment within a traditional department at Virginia Tech, but 
are funded through and housed at the FBRI. Primary faculty comprise a broad 
interdisciplinary group working in a variety of areas such as neural computation, 
psychiatric and developmental disorders, genetics, substance abuse and molecular 
virology and biology. The FBRI also provides to the faculty an administrative staff of 
individuals with expertise in grants management, personnel management, supply 
ordering/tracking, web systems management, desktop support, laboratory animal care, 
IRB support, software development and facilities and operations support and planning as 
well as general administrative support.  
 
DiFeliceantonio Lab Resources 
Dr. DiFeliceantonio’s lab is equipped to draw blood using an IV catheter system, analyze 
blood samples for glucose, perform indirect calorimetry, train participants in the MRI 
scanner environment, and measure participant’s height and weight. 
Equipment includes: 
Stadiometer - Measurement of height 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
47 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 48 of 49 
Revised: March 5, 2019 
Bioelectrical Impedance Analysis equipment- Noninvasive measurement of body 
composition (% body fat) 
Hemoglobin A1C analyzer 
Scale - Measurement of weight 
Parvo-Medics Metabolic Cart – Measurement of gas exchange (O2 consumption, CO2 
expiration) 
Hemocue Analyzer – Measurement of whole blood glucose (venous) 
Centrifuge 
MR Simulator (i.e., "mock" MRI scanner; Psychology Science Tools) 
Laptop Computer for data collection 
Two Tablets for data collection (Lenovo) 
Custom gustometer for delivery of liquids in simulated MR environment 
3 workstations for research personnel 
Laptop with Nutrition Data System for Research (NDSR) software and license 
 
 
Computing Resources 
The Fralin Biomedical Research Institute (FBRI) at Virginia Tech Carilion (VTC) houses 
the following shared computing resources available: 
Dell / Intel HPC Cluster (20 nodes, 960 Core, 3,840 GB RAM, 40Gb Interconnect) – 
SLURM; IBM iDataPlexLinux Cluster (60 node, 720 core Intel Xeon based, 1,440GB 
RAM, GigE Interconnect) - PBS; 10+ dedicated Linux servers (48 core AMD Opteron-
based, 192GB RAM) are available for image and data analysis; Virtualized infrastructure 
using VMware vSphere, virtual machine environment available for general compute and 
image and data analysis; 40Gbit/s storage connectivity for research data; 10 Gbit/s 
Internal Local Area Network between file servers and cluster; 8Gbit/s Fibre Channel 
Storage Area Network; 10 Gbit/s Wide Area Connection for access to Virginia Tech 
main campus / Internet; 1.4 Petabytes of NAS centralized disk storage; 250 Terabytes of 
SAN-attached centralized disk storage; 400 Terabyte library-based tape backup; Nightly 
backups and snapshots; 2.4Ghz / 5Ghz secure wireless network; 1 Gbit/s commodity 
Ethernet network; Data Analysis Tools: MATLAB, SPM12, AFNI, FSL, MRIcro, 
xjView, R, SAS, Prism Graphpad, SPSS; Productivity Tools: Adobe Suite, Microsoft 
Office Suite, vi, vim, emacs 
 
MRI Scanning Resources (FBRI) 
Virginia Tech has three research-dedicated Siemens 3T MR scanners (2 Siemens 
Magnetom TIM Trios, and 1 PRISMA-FIT) available. Each scanner bay is equipped with 
the following stimulation and response interfaces: 
behavioral response: two-hand, eight-button optical response pads with USB, serial, and 
TTL output (Current Designs, Inc.) 
video stimulation: rear-projection video display (Hitachi CP-SX635) 
corrective lenses for use with video stimulation: MR-compatible frames with insertable 
polycarbonate lenses (prescriptions range from -8.00 to +8.00) (Solo Bambini) 
stimulus delivery: dedicated computers for experiment presentation (Dell Optiplex 980) 
audio delivery: MRI compatible headphones. 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
48 of 49

PROTOCOL (STUDY) TITLE AND NUMBER 
VT Research Protocol version 1.0.1 
Page 49 of 49 
Revised: March 5, 2019 
Metabolic Chamber Resources (FBRI) 
Virginia Tech has 2 research-dedicated chambers of whole-room indirect calorimetry 
available (MEI Research, Ltd). The larger room is equipped for 24-hour measurements, 
and the smaller room is equipped for shorter-term measurements.  
 
 
 
27.0 Multi-Site Research 
Contact the HRPP for multi-site research (involving multiple institutions) and 
the details required for this section will be provided. Otherwise, indicate N/A. 
 
This is a single site study. 
 
 
Virginia Tech Institutional Review Board Protocol No. 21-964 
Approved April 27, 2023 
49 of 49