Milk Protein Feeding After Aerobic Exercise in Older Adults With Pre-diabetes Taking the Biguanide Metformin
Phase: PHASE2
Conditions: Physical Activity
Sponsor: Colorado State University

 
PROTOCOL
Biomedical
Colorado State
University
Protocol # 15-5837H
 Date Printed: 11/05/2018
Amendment
1
Personnel Information
1
Subject Population
3
Study Location
3
General Checklist
3
Funding
5
Expedited Paragraphs
6
Purpose,Study Procedures,Background
7
Radioisotopes or Radiation Machines
11
Medical Equipment for Human Subjects and Laboratory Animals; Investigatiional Devices
12
Drugs, Reagents, or Chemicals
12
Subject Population (a-g)
13
Subject Population (h-m)
14
Risks
16
Benefits, Procedures to Maintain Confidentiality
20
Potential Conflict of Interest
20
Informed Consent
21
Assent Background
22
HIPAA
22
Attachments
23

 
PROTOCOL
Biomedical
Colorado State
University
Protocol # 15-5837H
 Date Printed: 11/05/2018
Obligations
29
Event History
29

Page 1 of 33
 
1
PROTOCOL
Biomedical
Colorado State
University
Protocol # 15-5837H
 Date Printed: 11/05/2018
Protocol Title:
Milk protein feeding after aerobic exercise in older adults with pre-diabetes taking the
biguanide metformin
Protocol Type:
Biomedical
Date Submitted:
10/04/2018
Approval Period:
10/04/2018-05/13/2019
Important Note:
This Print View may not reflect all comments and contingencies for approval. Please check the
comments section of the online protocol.
Questions that appear to not have been answered may not have been required for this submission.
Please see the system application for more details.
----------------------------------------------------------------------------------------------------
* * * Amendment * * *
 
 
Amendment (1. Complete this one-page form, 2.  Update the sections of your protocol that you are requesting to
amend, 3. Electronically "sign" your application by clicking the check box on the Obligations page, 4. Remember
to click "Submit Form" so that the IRB administrators receive your request.)
 
  
  
1. 
Summarize the proposed changes to the protocol in lay terms.
Change of principal investigator to Karyn Hamilton.
 
 
 
Proceed to the appropriate section(s) of the protocol and make your changes.
 
 
 
IMPORTANT NOTE ON AMENDING ATTACHED DOCUMENTS:  If you are requesting to amend a
file that has been previously attached and approved, you MUST:  1.  Delete the file that is currently
attached,  2. Browse on your computer,  and  3. Upload the revised file that you are requesting to use.
 
  
  
2. 
Indicate level of risk involved with the changes proposed.
(If level of risk has changed, please update the section 'Risks' in the protocol information.)
 
   
No Change
 
  
  
Approval Includes
 
  
  
3. 
List of sections (and questions) that have been changed/modified
---------------------------------------------------------------------------------------------
* * * Personnel Information * * *
IMPORTANT NOTE: Mandatory Personnel on a protocol are: Principal Investigator and Department Head. Only
the Principal Investigator can submit the protocol; although other personnel listed on the protocol can create the
protocol. Human Subjects Protection Training is mandatory for Principal Investigator, Co-Principal Investigator,
and Key Personnel (as defined by NIH). Training must be updated every three (3) years.
 
Principal Investigator  Mandatory
Name of Principal Investigator
Degree
Title

Page 2 of 33
 
(Faculty, Staff or Postdoc)
Hamilton, Karyn
Professor
Email
Phone
Fax
Karyn.Hamilton@ColoState.EDU
(970) 491-3961
Department Name
Campus Delivery Code
 
244
 
 Human Subjects Training Completed? PIs must complete training every three
(3) years.
Y
 
 
 
Department Head  Mandatory
Name of Department Head
Degree
Title
Braun, Barry
Ph.D.
Professor
Email
Phone
Fax
Barry.Braun@colostate.edu
(970)491-7875
Department Name
Campus Delivery Code
 
1582 Dept Hlth & Exer Sci
1582
 
 Human Subjects Training Completed?? Training is not required for
Department Head. Select "No" if you do not know if your Department Head has
completed training or not.
Y
 
Administrative Contact
Name of Administrative Contact,
Project Director, or Lab Coordinator
Degree:
Title
Biela, Laurie
BS
Research Associate III
Email
Phone
Fax
Laurie.Biela@colostate.edu
970-491-2242
970-491-0445
Department Name
Campus Delivery Code
 
1582 Dept Hlth & Exer Sci
1582
 
 Human Subjects Training Completed? Training is not required for
Administrative Contacts
Y
No training data is available.
 
Other Researcher or Key Personnel
Name of Other Researcher (NOTE:
Anyone listed in this role will have
View Mode access only)
Degree
Title
Leach, Heather
Assistant Professor
Email
Phone
Fax
Heather.Leach@colostate.edu
Department Name
Campus Delivery Code (CSU) or off-
campus mailing address
 
244
 
 Human Subjects Training Completed? Training is required for all Key
Personnel on NIH grants. Training must be updated every three (3) years.
Y

Page 3 of 33
 
No training data is available.
---------------------------------------------------------------------------------------------
* * * Subject Population * * *
 
 
Subject Population(s) Checklist
        Â Select All That Apply - Note that this is your Targeted Population :
X
 Adult Volunteers
 Decisionally Challenged
X
Elderly
 Employees
 Fetuses
 Long-Term Patients
 Mentally Disabled
 Minors (under 18)
 Pregnant Women
 Prisoners
 Soldiers
X
 Students
 Other (i.e., non-English Speaking or any population that is not specified above)
       
---------------------------------------------------------------------------------------------
* * * Study Location * * *
 
 
Study Location(s) Checklist
         Select All That Apply - NOTE: Check "Other" and input text: 1.) If your study location is not listed, or 2.) If
you would like to list details of your already-checked location (e.g., specific school within a school district)
Aims Community College
Colorado Department of Public Health & Environment
X
Colorado State University
Colorado State University - Pueblo Campus
Denver Public Schools
Greeley/Evans School District
Poudre School District
University of Colorado Health - North (Formerly -Poudre Valley Health System - PVHS)
Rocky Mountain National Park
Thompson School District
University of Colorado - Boulder
University of Colorado - Colorado Springs
University of Colorado - Denver
University of Colorado Health Sciences Center
University of Northern Colorado
Other (In the box below, list your study location if not checked above. You may also list details of your
already-checked location (e.g., specific school within a school district).
       
---------------------------------------------------------------------------------------------
* * * General Checklist * * *

Page 4 of 33
 
 
General Checklist
         Select All That Apply :
X
Proposed Start Date (cannot be before IRB approval):
July 1, 2015
X
Sponsored Project (Check if you will be funded OR if you have or plan to submit a grant application in
association with this protocol)
NSF Sponsored (Please upload mandatory Data Management Plan in the Attachment section)
X
FDA or EPA-regulated research.  Please contact the CSU Quality Assurance Manager, Cat Bens, at 970-
491-5445 to determine if your study is under Good Laboratory, Good Clinical, or Good Manufacturing
Practices (GLP, GCP, GMP).
 Training Grant
X
Clinical Trial.  To register your trial on Clinicaltrials.gov, please contact Cat Bens, CSU Quality Assurance
Manager and Clinical Trails Administrator at:  970-491-5445.
Project is associated with the Colorado School of Public Health - CSPH(faculty and/or student)
Cooperating/Collaborating Institution(s) Institution
where recruitment will occur OR Institution where
Collaborating PI will conduct associated research.
Interview
Questionnaire/Survey
X
 Subjects will be compensated for participation
Thesis or Dissertation Project
X
ÃÂ Radioisotopes/radiation-producing machines, even if standard of care.  Please contact Jim Abraham,
Radiation Safety Officer for questions related to use of all radiation-producing machine:  970-491-3736;
james.abraham@colostate.edu.  Upload your radiation-use approval  (if available) or your Radiation Safety
Training certificate in the attachment section.
X
Human blood, cells, tissues, or body fluids. You will need to obtain IBC approval if you check this box. For
information regarding IBC approval, contact Christine Johnson, IBC Coordinator:
christine.johnson@colostate.edu
 Tissues to be stored for future research projects
 Tissues to be sent out of this institution as part of a research agreement
Human Embryos. You will need to obtain IBC approval if you check this box. For information regarding IBC
approval, contact Christine Johnson, IBC Coordinator:  christine.johnson@colostate.edu
Human Embryonic Cells? Provide NIH Code Number(s)
or state that no federal funding will be used to support
this research. You may need to obtain IBC approval if
you check this box. For information regarding IBC
approval, contact Christine Johnson, IBC Coordinator:
christine.johnson@colostate.edu
 Use of Patient-related equipment? If Yes, specify what equipment is being used.
Medical equipment used for human patients/subjects also used on animals. For questions regarding animal
use approval, contact Elaine Kim, IACUC Senior Coordinator:  491-0236
Protocol involves studying potentially addicting drugs. For questions regarding approval for possession of
controlled substances, contact Chris Giglio, DRC Coordinator:  491-4830; Chris.Giglio@colostate.edu.
Investigational drugs, reagents, or chemicals (IND)
X
 Commercially available drugs, reagents, or other chemicals administered to subjects (even if they are not
being studied)
X
Investigational Device (IDE)
Cancer Subjects (e.g., clinical trials, behavior/prevention) or Cancer Tissues (e.g., blood, cells, body fluids).
You may need to obtain IBC approval if you check this box. For information regarding IBC approval, contact
Christine Johnson, IBC Coordinator:  christine.johnson@colostate.edu
Other (clarify in text box to the right)

Page 5 of 33
 
---------------------------------------------------------------------------------------------
* * * Funding * * *
 
Please complete this section if:  1. This protocol will be funded, 2. You have submitted or will submit a grant
application associated with this protocol.  Please be sure to input your PASS/SP1 number to assist Sponsored
Programs in setting up an account for your funds.
If this protocol is funded by the NIH or NSF, or will lead to the regulatory involvement of the FDA or EPA, please
be certain you are cognizant of any specific regulatory requirements for data acquisition, storage, retention and
sharing, as well as research expenditure allowability, with regard to this IRB protocol.
 
Funding Checklist
NONE
  
  
NOTE: Applicable Federal Grant Application, including competing renewals, must be attached in the
Attachment Section (#16). Applicable investigator's brochure and sponsor's protocol must also be
attached in section #16 for all industry-sponsored clinical trials.
 
Funding - Grants/Contracts
Funding Administered By
UNIVERSITY
CSU PASS #
121860
Sponsor's ID # (If known)
Funded By
Other
 
Dairy Management Inc.
Principal Investigator
Benjamin Miller
Grant/Contract Title if different from Protocol Title
Milk Protein Feeding After Aerobic Exercise in Older
Adults with Pre-Diabetes Taking the Biguanide
Metformin
For Federal projects, are contents of this protocol the
same as described in Federal proposal application?
N
Is this an Umbrella protocol?
N
Is this protocol under an Umbrella protocol?
Funding Administered By
UNIVERSITY
CSU PASS #
Sponsor's ID # (If known)
Funded By
Other
 
Dexcom Inc.
Principal Investigator
Grant/Contract Title if different from Protocol Title
For Federal projects, are contents of this protocol the
same as described in Federal proposal application?
Is this an Umbrella protocol?
Is this protocol under an Umbrella protocol?
Funding - Fellowships

Page 6 of 33
 
Funding - Other
 
Gift Funding
Dept. Funding
Other Funding
---------------------------------------------------------------------------------------------
* * * Expedited Paragraphs * * *
 
PLEASE READ:  This online application is for projects that will be reviewed by the IRB via the expedite or full-
board review process.  The criteria for expedited review are listed below. Review and check what expedite
criteria is/are appropriate for your project. NOTE:  If your research involves or may involve greater than minimal
risk, an element of deception, or is FDA-regulated research, do NOT check any of the expedited criteria listed
below.  Your protocol will then be reviewed by the full-board at their next regularly scheduled meeting.  If your
project meets the exempt criteria, please submit your exempt application via email to:
RICRO_IRB@mail.colostate.edu.  Information regarding exempt applications can be found here:
http://ricro.colostate.edu/IRB/ExemptReview.html
 
  Expedite Criteria:
 
1.
Clinical studies of drugs and medical devices only when condition (a) or (b) is met.
 
a) 
Research on drugs for which an investigational new drug application (21 CFR Part 31,32) is
not required. (Note: Research on marketed drugs that significantly increases the risks or
decreases the acceptability of the risks associated with the use of the product is not eligible
for expedited review.)
 
b) 
Research on medical devices for which                                                                                       
                             
 
      i) 
An investigational device exemption application (21 CFR Part 812) is not required; or
      ii) 
The medical device is cleared/approved for marketing and the medical device is being used in
accordance with its cleared/approved labeling.
 
2.
Collection of blood samples by finger stick, heel stick, ear stick, or venipuncture as follows:
 
a) 
From healthy, nonpregnant adults who weigh at least 110 pounds. For these subjects, the
amounts drawn may not exceed 550 ml in an 8- week period and collection may not occur
more frequently than 2 times per week; or
 
b) 
From other adults and children, considering the age, weight, and health of the subjects, the
collection procedure, the amount of blood to be collected, and the frequency with which it will
be collected. For these subjects, the amount drawn may not exceed the lesser of 50 ml or 3
ml per kg in an 8-week period and collection may not occur more frequently than 2 times per
week.
 
3.
Prospective collection of biological specimens for research purposes by non-invasive means.
 
4.
Collection of data through non-invasive procedures (not involving general anesthesia or sedation)
routinely employed in clinical practice, excluding procedures involving x-rays or microwaves. Where

Page 7 of 33
 
medical devices are employed, they must be cleared/approved for marketing. (Studies intended to
evaluate the safety and effectiveness of the medical device are not generally eligible for expedited
review, including studies of cleared medical devices for new indications.)
 
Examples:
a) 
Physical sensors that are applied either to the surface of the body or at a distance and do not
involve input of significant amounts of energy into the subject or an invasion of the subject's
privacy;
b) 
Weighing or testing sensory acuity;                                                                                              
           
c) 
Magnetic resonance imaging;                                                                                                      
     
d) 
Electrocardiography, electroencephalography, thermography, detection of naturally occurring
radioactivity, electroretinography, ultrasound, diagnostic infrared imaging, doppler blood flow,
and echocardiography;
e) 
Moderate exercise, muscular strength testing, body composition assessment, and flexibility
testing where appropriate given the age, weight, and health of the individual.
 
5.
Research involving materials (data, documents, records, or specimens) that have been collected, or
will be collected solely for nonresearch purposes (such as medical treatment or diagnosis). (NOTE:
Some research in this paragraph may be exempt from the HHS regulations for the protection of
human subjects. 45 CFR 46.101(b)(4). This listing refers only to research that is not exempt.)
 
6.
Collection of data from voice, video, digital, or image recordings made for research purposes.
 
7.
Research on individual or group characteristics or behavior(including, but not limited to, research on
perception, cognition, motivation, identity, language, communication, cultural beliefs or practices, and
social behavior) or research employing survey, interview, oral history, focus group, program
evaluation, human factors evaluation, or quality assurance methodologies. (NOTE: Some research in
this category may be exempt from the HHS regulations for the protection of human subjects. 45 CFR
46.101(b)(2) and (b)(3). This listing refers only to research that is not exempt.)
 
 
---------------------------------------------------------------------------------------------
* * * Purpose,Study Procedures,Background * * *
 
Original Protocol Number (e.g., 07-226H)
   
Title (Please indicate if the protocol title is different from the proposal title)
Milk protein feeding after aerobic exercise in older adults with pre-diabetes taking the biguanide metformin
 
      Complete Sections 1 - 16. Specify N/A as appropriate. Do not leave any required sections blank.
1. Purpose of the study
a)
Provide a brief lay summary of the project in <200 words.  The lay summary should be readily
understandable to the general public, and is, for example, what would be released to a newspaper if
requested.
We propose that post-exercise milk protein feeding will enhance the mitochondrial protein synthesis
(biogenesis) response to an exercise-training program. In addition, we propose that this stimulatory effect
of protein feeding will overcome the potential blunting effect of metformin on exercise responses. We will
investigate these outcomes over a 12-week exercise-training program in older adults with pre-diabetes
with or without metformin treatment.
Sub-study:
1. CGM in Individuals with prediabetes undergoing an exercise-training program with or without Metformin.
2. CGM in healthy control individuals to compare to individuals with prediabetes.

Page 8 of 33
 
 
b)
What does the Investigator(s) hope to learn from the study?
We hope to:
a) provide the first evidence that post-exercise milk protein supplementation is effective in diseased
population (i.e., older adults with pre-diabetes) and those on medications (i.e., metformin).
b) demonstrate that the potential inhibitory effects of metformin on aerobic exercise adaptations can be
opposed by milk protein feeding.
Sub-study
1. Glucose Profiles via CGM in pre diabetic individuals before, during and after the intervention (i.e.,
aerobic exercise training with or without metformin with post-exercise carbohydrate or
carbohydrate+protein feeding as listed below in 2a).
2: Glucose profiles via CGM in healthy individuals. Healthy individuals will complete baseline procedures
but will not complete an intervention.
 
c)
Proposed Start Date (may not precede IRB approval date):
August 1, 2015
 
2. 
Study Procedures (If this is a student project, the methods section of the thesis or dissertation proposal
must be attached in section #16 - Attachment section.)
 
a)
In lay language, describe all the procedures, from screening through end-of-study, that the human subject
must undergo in the research project, including study visits, drug treatments, randomization and the
procedures that are part of standard of care.  Please note: Do NOT respond "See Attachment Section." If
you would like to add tables, charts, etc., attach those files in the Attachment section (#16).
Study Overview:
Pre-diabetic and healthy participants will first undergo a medical questionnaire, resting ECG, graded
exercise test with ECG and assessment of VO2max, anthropometric measurement (height, weight and
body composition by DEXA scanning), an oral glucose tolerance test (OGTT), and a muscle biopsy.
Prediabetic participants will be screened to eliminate those lactose intolerant, medications known to affect
the primary outcomes, medications or recent procedures contraindicated with metformin  or not pre-
diabetic. Subjects who meet these criteria will be randomized so that subject characteristics are matched
between groups, while the study team and participants will remain blinded to treatment (i.e., randomized,
double-blind study design). Subjects will be assigned to exercise + carbohydrate + placebo, exercise +
protein + placebo, exercise + carbohydrate + metformin, or exercise + protein + metformin.
A minimum of two days apart from the VO2max test, skeletal muscle will be sampled from pre-diabetic and
healthy participants. During these visits, pre-diabetic subjects will have the option to participate in a study
to test the impact of one counseling session on physical activity after completion of the exercise training
and to identify individual, socio-cultural and environmental determinants of physical activity 12 weeks after
the end of the exercise intervention. If the subjects wish to participate they will complete questionnaires to
measure sociodemographic information, physical activity, self-efficacy, planning, self-regulation and
outcome expectancy of exercise, and perceptions of neighborhood support at baseline, completion of
exercise training and 12 weeks after the exercise training.
Pre diabetic and healthy participants will also have the option to participate in a study to wear a Dexcom
generation 6 (G6) continuous glucose monitor (CGM) to measure glucose values every 5 minutes. Healthy
participants will wear the device once for up to 10 consecutive days. Pre-diabetic participants will wear the
device for up to 10-consecutive days at three different time points: before, during and after the 12-week
exercise intervention for a total of 30 days wearing the CGM. The Dexcom G6 CGM is for investigational
use only and is not FDA approved. The G6 is modeled after commercially available, FDA approved CGM
devices. The differences between the G6 and commercially available devices are a reduced calibration
scheme, meaning less finger stick blood glucose measurements for calibration;  a longer wear for up to 10
days for the G6 vs. 7 days for commercially available product. Last, commercially available devices could
give false high glucose readings if individuals took acetaminophen. The G6 sensor contains updated
sensor technology aimed to block the interference of acetaminophen.
After randomization into placebo or metformin groups, the pre-diabetic subjects will begin a 12-wk exercise

Page 9 of 33
 
intervention consisting of 3 days per week of aerobic exercise training with post-exercise consumption of a
20 gram protein beverage or isocaloric carbohydrate beverage. We have completed this supplementation
protocol in previous studies and a currently ongoing investigation.
At the 8-wk time point, pre-diabetic subjects will begin the daily consumption of heavy water (D2O) for the
assessment of mitochondrial and individual protein synthesis. At the completion of the training period, the
pre-diabetic subjects will repeat all assessments performed prior to training including the DEXA, VO2max,
OGTT and muscle sample. Muscle samples obtained pre- and post-training will be used to evaluate
mitochondrial function using high-resolution respirometry, western blot analysis, and stable isotope
incorporation (post only).
Healthy participants will complete the medical history questionnaire, resting ECG, graded exercise testing
(ECG and VO2max), DEXA, OGTT, CGM, and muscle biopsy.
Detailed Procedures:
Oral glucose tolerance test (OGTT): An OGTT is the most common clinical procedure to determine
impaired glucose tolerance and indices of insulin sensitivity. First, an intravenous catheter will placed in an
arm or hand vein to obtain blood samples for analysis of plasma glucose and glucoregulatory hormones.
Blood will be sampled before, 5, 10, 20, 30, 45, 60, 90, and 120 min after completing the consumption of
75 gram glucose beverage. The fasted blood sample will also be used for a blood chemistry panel that
includes electrolytes, glucose, in addition to kidney and liver related variables (including creatinine, eGFRif
AFR AMER, eGFR, Albumin, Bilirubin, Alk Phos, AST, Uric Acid, ALT, A/G ratio, etc.).Subjects will not be
allowed to participate until a medical doctor has signed-off on the blood screening. A 6-week blood draw
will be taken to confirm that kidney and liver variables have not changed. Additional blood draws may be
obtained per physician discretion.
Body Composition: Height, weight will be recorded. Percent body-fat, fat-free mass, total and regional
adipose tissue mass, and total bone mineral density will be determined using whole-body dual energy x-
ray absorptiometry (DEXA) as we have previously used.
VO2Max: Subjects will undergo a 12-lead electrocardiogram (ECG) examination at rest and during a
graded exercise test on a stationary bicycle until volitional fatigue, in order to determine maximal oxygen
uptake (VO2max). ECG measurements will be made continuously, and blood pressure and ratings of
perceived exertion will be recorded. These tests will be performed in the Human Performance/Clinical
Research Laboratory (HPCRL) following standard procedures.
Skeletal Muscle Biopsy: Skeletal muscle biopsies will be performed using well-established IRB approved
SOPs. The belly of the vastus lateralis will be anesthetized locally with 1% lidocaine without epinephrine. A
small incision (~4-mm in length) will be made and a Bergstrom needle inserted. Negative pressure will be
applied to the inner channel of the needle using a sterile 30-ml plastic syringe. Benjamin Miller, Ph.D.,
Adam Konopka, Ph.D., or Matthew Hickey Ph.D. will perform the skeletal muscle biopsies. Collectively, the
investigative team has performed this procedure on over 1,000 subjects. A muscle sample will be obtained
once before and after the 12-week exercise training (a total of 2 muscle biopsies).
Exercise Training: The exercise-training program will be performed within the facilities for the Noon Hour
Fitness Program at CSU according to our previously published procedures in older individuals. Exercise
will be performed on a treadmill, stationary bicycle, or elliptical and include warm-up (15 min at a heart rate
corresponding to 60% of max ) and workout stages (30 min, progression from 65 to 85% of max heart rate
by wk 5). The participants will wear heart rate monitors to ensure proper heart rate ranges and the speed,
grade or resistance of equipment will be changed to maintain heart rate at the specified percentage of max
HR. A CPR trained technician will supervise each exercise session to monitor adherence to the exercise
prescription and ensure subject safety. The drinks will be consumed immediately after exercise under
supervision.
Nutritional Intervention: Subjects will consume either a protein-carbohydrate or carbohydrate drink with a
consistency and temperature similar to a smoothie or milk shake immediately after each exercise bout.
The protein-carbohydrate drink will be greek yogurt+fruit (~300 kcal, 63g carbohydrate, 20g protein, 1g fat)
and the carbohydrate will be fruit (~300 kcal, 82g carbohydrate, 2g protein and 1g fat).
Metformin and Placebo: We use a titration dosing protocol in which the amount of metformin starts at 500
mg/day in week 1 and increases by 500 mg/day/week until reaching 2000 mg/day by week 4. 2000 mg/day
is the standard clinical dose and used previously by the investigative team. The titration dosing scheme
and taking metformin/placebo with meals will help reduce the most common side effects (i.e.,
gastrointestinal discomfort). If participants experience gastrointestinal discomfort, the dose will be lowered
to 1500 mg/day. If participants have a body weight of 75 kg or less the maximal dose will be 1500 mg/day
to decrease the risk of gastrointestinal discomfort.
Stable Isotope Labeling: The subjects will consume D2O (70%; Cambridge Isotope Laboratories, Andover

Page 10 of 33
 
MA, USA) during the last four-week period of the 12 week intervention. D2O is a stable, non-radioactive
isotope that is indistinguishable from normal water,  A target of 1–2% enrichment is achieved during a one-
week priming stage (50 ml of D2O 3x/d = 150 ml/d) and maintained for three weeks (50 ml of D2O 2x/d =
100 ml/d) as we have previously performed.
Optional Continous Glucose Monitoring (CGM): CGM is a commonly used device to continuously measure
glucose values in healthy control and diabetic populations in both research and clinical applications. Only
trained members of the research team will place and remove the CGM devices following a standard
operating procedure and user guide.  The sensor pod will be secured to the skin of the abdomen using
medical grade adhesive. Once secure, a plunger will insert the sensor underneath the skin to record
interstitial glucose values every 5 minutes. After careful removal of the applicator, a transmitter will be
attached to the secured sensor pod. The transmitter sends data to a receiver. The receiver is smaller than
the size of a cell phone to easily fit in a pocket or bag. The receiver needs to be within 20 feet of the device
to continuously transmit data. The receiver will be set to blinded mode so the participant will not know their
glucose values. The device can be worn up to 10 days. After each 10-day wear a trained member of the
research team will remove the sensor. During the 10-day wear the device will  prompt the participant to
calibrate. After verbal and written instruction, participants will be shown how to perform a finger stick,
measure blood glucose using the glucose meter provided and enter the glucose value into the receiver to
calibrate the device. During the 10-day wear, participants will be provided a log to record the timing and
amount of medications, physical activity, food and drink.
Optional Physical Activity Behavior Counseling: After completion of the 12-week exercise intervention,
participants will be randomized to 1) receive one session of physical activity behavior counseling (PABC),
or 2) no counselling control (CON).The counseling session will consist of strategies based on social-
cognitive theory hypothesized to increase physical activity adherence. Session activities will include 1)
discussion of the benefits of physical activity, 2) discussion of evidence based recommendations for
frequency, intensity, time and type of physical activity for reducing the risk of type 2 diabetes and
associated co-morbidities, 3) setting individual physical activity goals, 4) identifying and discussing barriers
and facilitators for physical activity, and 5) identifying strategies to overcome barriers. The counseling
session will be completed in person while the follow-up questionnaires 12 weeks after completing the
exercise training can be done via phone or email.
Healthy participants will complete the medical history questionnaire, resting ECG, graded exercise testing
(ECG and VO2max), DEXA, OGTT, CGM, and muscle biopsy.
 
b)
Explain why human subjects must be used for this project.
We are interested in improving human health using recommendations by American Diabetes Association
with dairy protein supplementation and therefore need to use human subjects.
 
c)
Alternative Procedures. If the proposed study is a clinical trial of a drug, vaccine, device or treatment,
describe alternative procedures, if any, that might be advantageous to the subject. Describe the important
potential risks and benefits associated with the alternative procedure(s) or course(s) of treatment. Any
standard treatment that is being withheld must be disclosed. This information must be included in the
consent form.
No alternative procedures
 
d)
If the proposed study is a clinical trial of a drug, vaccine, device or treatment, will it be possible to continue
the more (most) appropriate therapy for the subject(s) after the conclusion of the study?
Clinical parameters obtained during the study will be available to the participants to share with their
primary physician. Then the subject and primary physician can decide on the most appropriate therapy for
the subject after the conclusion of the study.
 
e)
Study Endpoint. If the proposed study is a clinical trial of a drug, vaccine, device or treatment, what are the
guidelines or end points by which you can evaluate the alternative treatments during the study? If one
treatment proves to be clearly more effective than another (or others) will the study be terminated before
the projected total subject population has been enrolled? When will the study end if no important
differences are detected?
No alternative treatments
 
f)
State if deception will be used. If so, provide a rationale and describe debriefing procedures. Submit a

Page 11 of 33
 
debriefing script in the Attachment Section (#16).
No deception will be used.
 
3. Background
a)
Describe past experimental and/or clinical findings leading to the formulation of the study, if applicable.
Physical activity and the biguanide drug Metformin have been shown independently to reduce the
incidence of T2D. Interestingly, these data also suggest in adults over the age of 60 metformin had
marginal efficacy while physical activity lowered incidence of T2D by 70%. Currently, governing agencies
recommend physical activity and metformin together for individuals with pre-diabetes of all ages to prevent
the progression to overt T2D. Until recently, the interaction between exercise and metformin has been
largely understudied. Members of our investigative team have previously showed that metformin does not
enhance but may blunt the glucoregulatory and cardiovascular response to acute and chronic exercise,
which is especially relevant to older adults. A purported mechanism of action is that metformin inhibits
mitochondrial respiration, which likely limits VO2max and mitochondrial protein synthesis. We have
previously demonstrated that post-aerobic exercise dairy feeding can improve VO2max to a greater extent
than post-exercise carbohydrate beverage. Therefore, we feel dairy supplementation may override the
potential inhibition of metformin on exercise adaptations and maximize the benefits for older adults. To
date, real-time continuous glucose monitoring is limited in individuals at risk of T2D. Moreover, glucose
profiles via CGM during or in response to exercise, metformin and/or post-exercise nutrition have yet to be
fully characterized. Much clinical research is limited to measurements in the clinical research setting while
real-time glucose monitoring performed in both the standardized clinical research setting and in free-living
scenarios is novelty of the current study design that will close a gap in knowledge regarding
glucoregluatory effects of exercise, metformin and/or standardized post-exercise nutrition.
 
b)
Describe any animal experimentation and findings leading to the formulation of the study, if applicable.
---------------------------------------------------------------------------------------------
* * * Radioisotopes or Radiation Machines * * *
 
 
You selected NO for Radioisotopes in the General Checklist. If you would like to add Radioisotopes, change the
selection to YES in general Checklist.
 
4. 
Radioisotopes or Radiation Machines Please note: For projects requiring radiation procedures, please
contact the CSU Radiation Control Office (RCO). For more information see:
http://www.ehs.colostate.edu/WRad/Home.aspx :
 
 
a) 
If applicable, summarize in lay language the radiographic diagnostic and therapeutic procedures
associated with this protocol.
Body composition will be measured by dual energy x-ray absorptiometry (DEXA) before and after
the 12 weeks of exercise in pre diabetic participants. In healthy individuals, one DEXA scan will be
performed.
  b) 
Are the radiation procedures being performed a normal part of the clinical management for the medical
condition that is under study (Standard of Care) or are the procedures being performed because the
research subject is participating in this project (extra CT scans, more fluoroscopy time, additional Nuclear
Medicine Studies, etc.,) (Not Standard of Care)? If some procedures are Standard of Care and some are
Not Standard of Care, check both boxes.
 
  
NOT STANDARD OF CARE
X
STANDARD OF CARE
  
If it is not standard of care, complete the rest of
this section. Provide the CSU RCO approval
information
If it is only standard of care, skip the rest of this
section.
 

Page 12 of 33
 
 
CSU Radiation Control Office approved protocol number:
 
 
CSU Radiation Control Office protocol approval date:
 
For more information, see the RCO website at:http://www.ehs.colostate.edu/Wrad/home.aspx or Contact:
James Abraham, Radiation Safety Officer, at 970-491-3736.
---------------------------------------------------------------------------------------------
* * * Medical Equipment for Human Subjects and Laboratory Animals; Investigatiional Devices * * *
 
 
5. Medical Equipment for Human Subjects
 
  
If medical equipment is being used for human subjects/patients, describe this equipment and indicate if the
use is normal practice for the population under study. You may have already described this equipment in
the Study Procedures section. If you have already listed this information in the Study Procedures Section,
please do not duplicate this information here. In the space below, input N/A if not applicable, indicate if this
is already listed in the Study Procedures Section, or describe the equipment.
Continous Glucose Monitor (CGM) described in study procedures and in the attached investigator
brochure and user guide. The Dexcom G6 CGM is for investigational use and is not FDA approved.
 
 
6. Investigational Devices
         Please list in the space below all Investigational Devices to be used on Subjects.
Investigational Devices
Investigational Devices
Describe the device(s) to be used
DEXCOM G6 Continuous Glucose Monitoring (CGM) System. CGM can supplement, but does not replace,
blood glucose monitoring. Previous versions of the device (Dexcom SEVEN PLUS, G4 PLATINUM, and G5
Mobile CGM) have been classified as a Class III device according to 21 CFR Part 814. The device’s
product code is MDS. Class III devices generally are those for which insufficient information exists to
determine that general or special controls are sufficient to provide a reasonable assurance of safety and
effectiveness.
Device Name
DEXCOM G6 Continuous Glucose Monitoring (CGM) System
Manufacturer
Dexcom, Inc.
Significant risk
Y
Non-Significant risk
IDE #
Rationale for Non-Significant Risk Device
Device is not exempt from requirements in 21 CFR 812. Device is intended as an implant? Device is
purported or represented to be for use supporting or sustaining human life? Device used of substantial
importance in diagnosing, curing, mitigating, or treating disease, or otherwise preventing impairment of
human health? Device presents a potential for serious risks to the health, safety, or welfare of a subject? If
yes to any, then considered SR device.
If a non-significant risk device study is indicated, provide rationale for the device being non-significant risk.
Please state if you need IRB guidance on whether this is a significant or non-significant risk device.
---------------------------------------------------------------------------------------------
* * * Drugs, Reagents, or Chemicals * * *
 

Page 13 of 33
 
 
7. Drugs, Reagents, or Chemicals
 a)
Please list in the space below all investigational drugs, reagents or chemicals to be administered to
subjects during this study.
 
 b)
Please list in the space below all commercial drugs, reagents or chemicals to be administered to subjects
during this study.
 
Drug Name
Metformin
Source (e.g., Pharmacy, Sponsor, etc.,)
Pharmacy
If not premixed, where will the material be mixed and
by whom
Manufacturer
IND # (if available)
Dosage
Maximum dose of 1500 mg/day
Administration Route
Oral
N
 Are these new or different uses of these
commercially available drugs, reagents, or
chemicals?
N
IND Regulations
 
Please read the IND Statements
---------------------------------------------------------------------------------------------
* * * Subject Population (a-g) * * *
 
 
8.  
Subject Population - In the space below, please detail the participants that you are requesting to recruit
(include requested participant number and description of each group requested).  (Input N/A if not
applicable)
a)
Requested Participant Description (Include number of participants that you plan to study and description of
each group requested, if applicable).
We plan to recruit 120 men and women with an estimation that 15-25% of participants will not qualify (i.e.,
screen out) and 15-25% of participants will not complete the study (i.e. drop out). We plan to complete the
study in 60 men and women aged >55 years who are prediabetic defined as impaired fasting glucose (100
to 126 mg/dl), HbA1c (5.7-6.4%), family history of type 2 diabetes, and/or impaired glucose tolerance
defined as 2 hour postprandial blood glucose of 140 to 200 mg/dl. Participants will be eligible if they meet
any 1 of these criteria.
10 healthy individuals, 18-45 years old who have normal glucose values (fasting glucose (<100mg/dl),
HbA1c (<5.7%), 2 hour postprandial blood glucose (<140mg/dl)) or no family history of type 2 diabetes will
be recruited.
Sub-study: 20 pre diabetic participants and 10 healthy individuals will complete the study.
Subpopulation of [xx] participants will be recruited for Physical Activity Behavior Counseling (previously
approved in Amendment 3.
 
b)
What is the rationale for studying the requested group(s) of participants?

Page 14 of 33
 
Individuals at risk for T2D are the intended population for metformin; CGM; and Counseling studies. We
will use a healthy control group to compare to individuals at risk for T2D.
 
c)
If applicable, state the rationale for involvement of potentially vulnerable subjects to be entered into the
study, including minors, pregnant women, economically and educationally disadvantaged, or decisionally
impaired subjects. Specify the measures being taken to minimize the risks and the chance of harm to the
potentially vulnerable subjects.
We will not enroll minors, pregnant women, economically and educationally disadvantaged, or decisionally
impaired subjects.
 
d)
If women, minorities, or minors are not included, a clear compelling rationale must be provided. Examples
for not including minors: disease does not occur in children; drug or device would interfere with normal
growth and development; etc.
Minors are not included because the cost-benefit ratio is not clear and therefore inappropriate for this
study.
 
e)
State if any of the subjects are students, employees, or laboratory personnel. They should be presented
with the same written informed consent. If compensation is allowed, they should also receive it.
If any of the subjects are students, employees, or laboratory personnel they will be presented with the
same written informed consent and compensation.
 
f) 
Describe how potential subjects will be identified for recruitment (e.g., chart review, referral from individual's
treating physician, those individuals answering an ad). How will potential participants learn about the
research and how will they be recruited (e.g., flyer, email, web posting, telephone, etc.)? Attach recruitment
materials in the Attachment Section (#16). Important to remember: potential subjects may not be contacted
before IRB approval.
Potential subjects will be recruited initially from the Colorado State and Fort Collins community via email,
electronic, newspaper, radio, or bulletin advertisements. Colorado State employees, local physicians and
interested participants may be emailed for recruitment and communication purposes. Physicians will be
contacted so if they have prediabetic patients they can be informed of the research study which includes
standard care treatment options (i.e., physical activity and metformin). Text for advertisements will be
obtained from the attached flyer. Information for local physicians is also attached.
 
g) 
If applicable, provide rationale for the inclusion of healthy volunteers in this study. Specify any risks to
which these healthy volunteers may possibly be exposed. Specify the measures being taken to minimize
the risks and the chance of harm to these volunteers.
We will include healthy volunteers to compare to individuals at risk for T2D. No additional risks are known
for healthy individuals compared to those at risk for T2D. Healthy individuals will not perform the
intervention.
---------------------------------------------------------------------------------------------
* * * Subject Population (h-m) * * *
 
 
8. Subject Population (Input N/A if not applicable)
h)
Inclusion and Exclusion Criteria (e.g., Participants must have 20/20 vision, Participants must be 30-45
years of age, etc.)
 
 
  
   
Identify inclusion criteria.
We plan to recruit 120 men and women with an estimation that 15-25% of participants will not
qualify (i.e., screen out) and 15-25% of participants will not complete the study (i.e. drop out).
We plan to complete the study in 60 men and women aged >55 years who are prediabetic
defined as impaired fasting glucose (100 to 126 mg/dl), HbA1c (5.7-6.4%), family history of type
2 diabetes, and/or impaired glucose tolerance defined as 2 hour postprandial blood glucose of
140 to 200 mg/dl. Participants will be eligible if they meet any 1 of these criteria.

Page 15 of 33
 
10 healthy participants will be recruited who are 18-45 years old who have normal glucose
values (fasting glucose <100mg/dl, HbA1c <5.7%, 2 hour postprandial blood glucose <140
mg/dl) and no family history of T2D.
 
 
  
   
Identify exclusion criteria.
Medications contraindicated with Metformin (Dofetilide, Lamotrigine, Pegvisomant, Somatropin,
Trimethoprim, Trospium, Gatifloxacin, Cephalexin, Cimetidine, Dalfampridine)
Recent (less than 6 weeks) or planned imaging that requires IV contrast
Renal failure, creatinine ≥ 1.3 mg/dL in men or ≥ 1.2 mg/dL in women
ALT levels exceed 52 IU/L
Heart, Kidney or Liver Disease
Type I or Type II Diabetes
Anti-coagulant therapy (warfarin/heparin)
Lung/respiratory dysfunction
Medications affecting primary outcomes
Lactose Intolerant
Tobacco Use
Heavy Alcohol Use
Cancer
Lidocaine Allergy
EKG that is nondiagnostic for ischemia, such as: Complete Left bundle branch block or
nonspecific Intraventricular conduction delay with ST/T changes; LVH with significant ST
changes; Wolff-Parkison-White syndrome;  ventricular pacemaker; digoxin therapy with greater
than 1 mm ST depression or
Arrythmias, such as: Uncontrolled rate in atrial fibrillation; nonsustained ventricular tachycardia.
 
i)
Describe your screening procedures. Attach your screening document(s) (e.g., health history
questionnaire) in the Attachment Section (#16).
Blood draw for chemistry panel, health questionnaire, medical history questionnaire, oral glucose tolerance
test, resting and exercise EC.
 
j)
Describe how you will be cognizant of other protocols in which subjects might be participating. Please
explain if subjects will be participating in more than one study.
Subjects will be instructed to communicate with investigators should they consider or begin to participate in
other studies. Subjects participating in studies that may confound the results of the present investigation
will be asked to withdraw from the the present investigation.
 
k)
Compensation. Explain the amount and schedule of compensation, if any, that will be paid for participation
in the study. Compensation includes food, gift cards, money, tokens, etc. Include provisions for prorating
payment, if applicable.  Compensation should be prorated if several activities are involved for different time
periods (e.g., $10 for session #1, and $10 for session #2).
Pre diabetic ubjects will be paid $500 at the completion of the study. Proration will occur at $20 per oral
glucose tolerance test and $50 per muscle biopsy. Participants that complete the CGM portion of this study
will receive additional compensation of $100 because of the additional time required for this study
component.
Healthy participants will be compensated $50 for the time spent completing the oral glucose tolerance test
(OGTT) and up to 10 days of CGM.
 
l)
Costs. Please explain any costs that will be charged to the subject.
Subjects will not incur any costs.
 
m)
Estimate the probable duration of the entire study. This estimate should include the total time each subject
is to be involved and the duration the data about the subject is to be collected (e.g., This is a 2-year study.
Participants will be interviewed 3 times per year; each interview will last approximately 2 hours.  Total
approximate time commitment for participants is 12 hours).  These times should be consistent with the time
commitment listed on the consent document.
After initial screening Pre-diabetic subjects will undergo a 12 week intervention followed by post-
intervention assessments. Total time demand for each subject is estimated at 15 weeks with a total

Page 16 of 33
 
commitment of approximately 46 hours over that 15 weeks. If participants chose to participate in the
optional physical activity behavior counseling there will be additional time spent completing questionnaires
(less than 1 hr) at baseline, immediately after completion of exercise training and 12 weeks after
completion of exercise training but these can be completed via email or phone. Additionally, participants
will be randomized to complete 1 session of in person physical activity behavior counseling (approximately
1 hour).
Pre-diabetic subjects who choose to participant in wearing the Dexcom G6 CGM will wear it for up to 10
days at a time on three separate occasions: before, during and after the 12-week intervention.
Healthy subjects who participant in wearing the Dexcom G6 CGM will wear it for up to 10 days on one
occasion.
---------------------------------------------------------------------------------------------
* * * Risks * * *
 
 
9. Risks (Input N/A if not applicable)
     
US Department of Health & Human Services (HHS) Regulations define a subject at risk as follows: "...any
individual who may be exposed to the possibility of injury, including physical, psychological, or social injury,
as a consequence of participation as a subject in any research, development, or related activity which
departs from the application of those accepted methods necessary to meet his needs, or which increases
the ordinary risks of daily life, including the recognized risks inherent in a chosen occupation or field of
service."
 
 a) 
PI's evaluation of the overall level of Risk. (Please check one: minimal or > minimal.)
Minimal (everyday living)
Y
> Minimal (greater than everyday living)
 
 b)   
For the following categories include a scientific estimate of the frequency, severity, and reversibility of
potential risks. Wherever possible, include statistical incidence of complications and the mortality rate of
proposed procedures. Where there has been insufficient time to accumulate significant data ON risk, a
statement to this effect should be included. (In describing these risks in the consent form to the subject, it is
helpful to use comparisons which are meaningful to persons unfamiliar with medical terminology.) Address
any risks related to:
 
 
   
   
1.   
Use of investigational devices. Please include the clinical adverse events (AEs) associated with
each of the devices with an indication of frequency, severity and reversibility. This information
can often be found in the Investigator(s) brochure.
Dexcom CGM G6 is not an FDA approved device. This is currently available for research and
investigational purposes. Please see IB in attachments and notes in protocol notes. Wearing of
device should not be placed through baggage x-ray machine or body scanner. Rarely, the
sensor wire may break or detach from the sensor pod. These do not pose a significant medical
risk, but participants should contact the Study Coordinator within 24 hours if experiencing a
broken wire. Risks include infection, bleeding, pain or skin irritations (e.g., redness, swelling,
bruising, itching, scarring or skin discoloration) from the sensor and wearing the adhesive
patch.
 
 
   
   
2   
Use of investigational drugs. Please include the clinical AEs associated with each of the drugs
with an indication of frequency, severity and reversibility. This information can often be found in
the Investigator(s) brochure.
n/a
 

Page 17 of 33
 
 
   
   
3   
Use of commercially available drugs, reagents or chemicals. Please include the clinical AEs
associated with each of the drugs with an indication of frequency, severity and reversibility. This
information can often be found in the package insert provided by the manufacturer.
Metformin has been commonly used as a diabetes medication for many years, both within and
outside of the US. It is safely used by nearly 50 Million people on a daily basis in the US.
Like most medication there are possible side effects: From the MAYO CLINIC:
Risks:
More common
Abdominal or stomach discomfort
Cough or hoarseness
Decreased appetite
Diarrhea
Fast or shallow breathing
Fever or chills
General feeling of discomfort
Lower back or side pain
Muscle pain or cramping
Painful or difficult urination
Sleepiness
Less common
Anxiety
Blurred vision
Chest discomfort
Cold sweats
Coma
Confusion
Cool, pale skin
Depression
Difficult or labored breathing
Dizziness
Fast, irregular, pounding, or racing heartbeat or pulse
Feeling of warmth
Headache
Increased hunger
Increased sweating
Nausea
Nervousness
Nightmares
Redness of the face, neck, arms, and occasionally, upper chest
Seizures
Shakiness
Shortness of breath
Slurred speech
Tightness in the chest
Unusual tiredness or weakness
Wheezing
Rare
Behavior change similar to being drunk
Difficulty with concentrating
Drowsiness
Lack or loss of strength
Restless sleep
Unusual sleepiness
 
 
   
   
4   
When performing procedures, please include all investigational, non-investigational and non-
invasive procedures (e.g., surgery, blood draws, treadmill tests).
Catheter Placement for OGTT (beginning and end of study): Minor discomfort of having the

Page 18 of 33
 
needle inserted and taped to an arm. In about 1 in 10 cases, a small amount of bleeding will
occur under the skin that will cause a bruise. The risk of forming a blood clot in the vein is about
1 in 100, and the risk of significant blood loss is 1 in 1,000. This procedure is currently CSU-IRB
approved and active.
Muscle biopsy (beginning and end of study): The risk of allergic reaction to lidocaine and
lidocaine toxicity is extremely low (0 in the ~1500 cases witnessed by PI). There is a small risk
of infection at the site of incision. There is a small risk of the incision reopening/bleeding after
the subject leaves the lab. All subjects will have a small scar at the incision site. The rate and
degree of healing varies considerably, but it is expected that scars will be difficult to see within
6-12 months after the procedure.
Consumption of Heavy Water: Brief periods (< 30 minutes) of dizziness have been reported in a
small number of people (< 5%) after drinking special water. This is due to the special water
being heavier than normal water. Heavier water can temporarily disrupt your inner ear ability to
control balance, producing a sensation of dizziness.
Exercise and Exercise Testing (VO2max): There is a very small chance of an irregular
heartbeat during exercise (< 1% of all subjects).  Other rare risks of a stress test are heart
attack (< 5 in 10,000) and death (<2 in 10,000).  Wearing a mouthpiece and nose-clip during
the graded exercise test can sometimes cause dryness in the mouth and mild discomfort.
CGM: The insertion of the CGM is a very quick procedure that causes no more discomfort than
the blood sampling or finger prick. The sensor sits beneath the skin and the participant should
not feel the senor once it is inserted. There is a small risk of infection, bleeding, pain or skin
irritations (e.g., redness, swelling, bruising, itching, scarring or skin discoloration) after inserting
the CGM sensor and wearing the adhesive patch. To minimize the risk of discomfort,
researchers will clean the area of skin with sterile alcohol swabs. The sensors and applicator
are shipped in sterile packaging and will be inserted within 5 minutes of opening package. Over
325 G6 CMG devices have been studied in 241 subjects across 9 independent clinical study
sessions. There were 5 total adverse events reported in these studies that included reddening,
irritation, infection and shooting pain at the site of the insertion or adhesive site that resolved
within 24 hours. No serious adverse device effects or unanticipated adverse device effects
were reported. A detailed report can be found in the investigational brochure in Section 4
Existing Clinical Data.
Fingersticks: Participants will likely experience brief (<30s) physical discomfort during the finger
stick procedure when the lancet goes into your finger. In about 10% of the cases a small
amount of bleeding under the skin will produce a bruise. A small scare may persist for several
weeks. The risk of local infection is less than 1 in 1,000 and is minimized by using the alcohol
(70% ethanol) swabs provided.
 
 
   
   
5  
Radioisotopes/radiation-producing machines(e.g., X-rays, CT scans, fluoroscopy).
Body Composition by DEXA scanning -
A small amount of radiation exposure (0.05 mRem) associated with the DEXA that is less than
1/20 of a typical chest x-ray. No immediate short-term or long-term risk to adults from this
radiation dose. This procedure is currently CSU-IRB approved and active.
 
 
c)      
For the following categories, include an estimate of the potential risk, if applicable.
 
 
   
   
1.   
Physical well-being.
None anticipated
 
 
   
   
2.   
Psychological well-being.
None anticipated

Page 19 of 33
 
 
 
   
   
3.   
Economic well-being.
None anticipated
 
 
   
   
4.   
Social well-being.
None anticipated
 
 d) 
In case of overseas research, or working with a specific race/ethnicity in the United States, provide
background on what experience the Investigator(s) have with the proposed population. Describe
qualifications/preparations that enable the Investigator(s) to evaluate cultural appropriateness and
estimate/minimize risks to subjects.
N/A
 
 e) 
Special Precautions. Describe the planned procedures for protecting against or minimizing potential risks. If
appropriate, include the standards for termination of the participation of the individual subject. Discuss
plans for ensuring necessary medical or professional intervention in the event of adverse effects to the
subjects.
The potential risks of the proposed studies will be minimized by: a) using only safe, well-established
procedures; b) constant, personal monitoring of each experimental session by the investigators and their
respective staffs; and c) the availability of emergency equipment and medicine. Students and other
subordinate personnel on the research team will undergo the required human subjects training and will be
very carefully monitored in their technique and interactions with subjects.
Venous catheter risks
Venous catheters will only be placed by experienced phlebotomists.
Muscle Biopsy
The skin will be disinfected with Betadine and all instruments will be sterilized to reduce the risk of
infection.
Exercise
Qualified personel will supervise your initial resting and exercise ECG to determine if you are healthy
enough to perform regular exercise. During the exercise intervention a CPR trained exercise physiologist
will supervise every exercise session.
CGM
CGM will only be placed and removed by trained personnel following standard operating procedures. The
skin will be disinfected using sterile alcohol swabs.
 
 f) 
Data Safety Monitoring                                                                                          
    
N
Is there a Data Safety Monitoring Board (DSMB)?
 
 
   
   
   
If yes, describe its role and indicate who set up the Data Safety Monitoring Board (e.g., sponsor
or Protocol Director).
 
 
   
   
   
Describe the data and safety monitoring plan developed to ensure the safety of participants and
the validity and integrity of research data. Monitoring should be commensurate with risks and
with the size and complexity of the trials.
---------------------------------------------------------------------------------------------

Page 20 of 33
 
* * * Benefits, Procedures to Maintain Confidentiality * * *
 
 
10. Benefits (Input N/A if not applicable)
a)
Describe the potential benefit(s) to be gained by the subjects. If there is no direct benefit to the subjects
describe how the results of the study may benefit society or a particular group.
The pre diabetic and healthy individuals will learn about their physiology, body composition and general
cardiovascular risk factors determined from blood samples and ECG. Although there are no guaranteed
benefits from participating in this study, we are providing the first line of care for preventing the progression
of prediabetes to overt Type 2 Diabetes.  Therefore, the anticipated benefits of the research are a positive
effect on glucose metabolism, aerobic fitness, quality of life, and performance of daily activities.
 
11. Procedures to Maintain Confidentiality
a)
Describe the procedures that protect the privacy of the subjects and maintain the confidentiality of the data.
If a linked list is used, explain when the linked list will be destroyed.  Provide a sample of the code that will
be used, if applicable.
Research records and data will be stored on file in locked cabinets in the investigators' office/laboratory, or
digitally on a password protected central server. Only members of the research team will have access to
these records. The materials will be archived for a minimum of three years after completion of the project.
All data will be coded; e.g. XYZ123456
 
b)
If information derived from the study will be provided to the subject's personal physician, a government
agency, or any other person or group (other than the research team), describe to whom the information will
be given and the nature of the information, if applicable.
Information derived from this study will be available for the subjects to provide to their personal physician.
 
c)
Specify where and under what conditions study data will be kept, how samples will be labeled, who has
access to the data, and what will be available and to whom.  Federal regulations require that study data
and consent documents be kept for a minimum of three (3) years after the completion of the study by the
PI.  For longitudinal projects and federally regulated studies, the PI may be required to keep the data and
documents for a longer time period.
Research records and data will be stored on file in locked cabinets in the investigators' office/laboratory, or
digitally on a password protected central server. Only members of the research team will have access to
these records. The materials will be archived for a minimum of three years after completion of the project.
All data will be coded; e.g. XYZ123456. De-identfied data from subjects who participate in the CGM
substudy will be shared with Dexcom, Inc.
---------------------------------------------------------------------------------------------
* * * Potential Conflict of Interest * * *
 
 
12. Potential Conflict of Interest
Although you have already submitted CSU's official Conflict of Interest form (FCOI/COI/COC) to the
University, it is the IRB's responsibility to ensure that conflicting interests related to submitted protocols do
not adversely affect the protection of participants or the credibility of the human research protection program
at CSU. Please answer questions a-d below.  Please note that if you indicate that you have a potential
financial or professional conflict of interest in relation to this protocol, your CSU FCOI/COI/COC Reporting
Form must reflect this potential conflict.  Link to CSU's Conflict of Interest policy:
:http://www.facultycouncil.colostate.edu/files/manual/sectiond.htm#D.7.7
a)
N
In connection with this protocol, do you or any of the protocol investigators or their
immediate family members (i.e., spouse and legal dependents, as determined by the IRS)
have a potential financial or professional conflict of interest?
b)
N/A
If you do have a potential conflict of interest, is this reported in your current
FCOI/COI/COC?

Page 21 of 33
 
c)
N/A
If you do have a potential conflict of interest, is there a management plan in place to
manage this potential conflict?
d)
N/A
If you do have a potential conflict of interest, is this potential conflict of interest included in
your consent document (as required in the Management Plan)?
 
If you have reported a possible conflict of interest, the IRB will forward the title of this protocol to your Research
Associate Dean to complete your COI file. 
 
For more information on CSU's policy on Conflict of Interest, please see the Colorado State University
Academic Faculty and Administrative Professional Manual Sections D.7.6 & D.7.7.
:http://www.facultycouncil.colostate.edu/files/manual/sectiond.htm#D.7.7
 
Link to CSU's Conflict of Interest Policy: http://www.provost.colostate.edu/index.asp?url=faculty_affairs.
---------------------------------------------------------------------------------------------
* * * Informed Consent * * *
 
 
13. Informed Consent
NOTE: In order to complete this protocol, you must upload either a Consent Form or an Alteration of
Consent Form (i.e., Cover Letter or Verbal Script) OR (if neither of those apply to your project) you must
complete the Waiver of Consent information.
In the space below, please provide consent process background information for each Consent Form(s),
Alteration of Consent Form(s), or Waiver(s).
Informed Consent
  Title
15-5837H CGM Consent 02242017
  Consent Information Type
Consent
  Sponsor's Consent Version Number: (if any)
Consent Form Template
X Attachment
 
15-5837 CGM Informed Consent
24Feb2017 (1)
<a href='http://ricro.colostate.edu/IRB/ConsentAssentTemplates.html' target=_blank Consent Form
Samples
Who is obtaining consent? The person obtaining consent must be knowledgeable about the study and
authorized by the PI to consent human subjects.
How is consent being obtained?
What steps are you taking to determine that potential subjects are competent to participate in the decision-
making process?
  Title
15-5837H Main Study 24April2017 Stamped
  Consent Information Type
Consent
  Sponsor's Consent Version Number: (if any)
Consent Form Template
X Attachment
 
15-5837H Informed Consent
24April2017_Main Study RICRO
STAMPED (1)

Page 22 of 33
 
<a href='http://ricro.colostate.edu/IRB/ConsentAssentTemplates.html' target=_blank Consent Form
Samples
Who is obtaining consent? The person obtaining consent must be knowledgeable about the study and
authorized by the PI to consent human subjects.
How is consent being obtained?
What steps are you taking to determine that potential subjects are competent to participate in the decision-
making process?
  Title
Healthy Individual Consent 11July2017
  Consent Information Type
Consent
  Sponsor's Consent Version Number: (if any)
Consent Form Template
X Attachment
 
15-5837 Informed Consent
Healthy Individuals (1)
<a href='http://ricro.colostate.edu/IRB/ConsentAssentTemplates.html' target=_blank Consent Form
Samples
Who is obtaining consent? The person obtaining consent must be knowledgeable about the study and
authorized by the PI to consent human subjects.
How is consent being obtained?
What steps are you taking to determine that potential subjects are competent to participate in the decision-
making process?
---------------------------------------------------------------------------------------------
* * * Assent Background * * *
 
 
14. Assent Background  (Complete if applicable)
All minors must provide an affirmative consent to participate by signing a simplified assent form, unless the
Investigator(s) provides evidence to the IRB that the minor subjects are not capable of assenting because of
age, maturity, psychological state, or other factors.
See sample consent/assent forms at  http://ricro.colostate.edu/IRB/ConsentAssentTemplates.html
Provide assent process background information, in the space below, for each Assent Form, Alteration Form
(i.e., Cover Letter or Verbal Script), and Waiver.
Assent  Background
---------------------------------------------------------------------------------------------

Page 23 of 33
 
* * * HIPAA * * *
 
 
15. HIPAA
Are you using PHI*? (See definition below)
N
Colorado State University is a hybrid entity and does not have a research-related HIPAA policy. If you will be
working with a HIPAA covered entity (e.g., Poudre Valley Health System), you will need to follow their HIPAA
guidelines. If your project will involve a HIPAA-regulated entity, in the Attachment section (#16) please attach that
entity's required HIPAA consent and/or each waiver of authorization or alteration of authorization requested (e.g.,
waiver of authorization for access to medical records). Include HIPAA authorization language in the consent
document(s) as appropriate (e.g., when enrolling subjects).  
*Protected Health Information (PHI) is health information with one or more of the following identifiers. For
more information see: http://www.hhs.gov/ocr/hipaa/
  1.
Names
  2.
Social Security numbers
  3.
Telephone numbers
  4.
All geographic subdivisions smaller than a State, including street address, city, county, precinct,
zip code, and their equivalent geocodes, except for the initial three digits of a zip code, if,
according to the current publicly available data from the Bureau of the Census: (1) The
geographic unit formed by combining all zip codes with the same three initial digits contains
more than 20,000 people; and (2) The initial three digits of a zip code for all such geographic
units containing 20,000 or fewer people is changed to 000
  5.
All elements of dates (except year) for dates directly related to an individual, including birth date,
admission date, discharge date, date of death; and all ages over 89 and all elements of dates
(including year) indicative of such age, except that such ages and elements may be aggregated
into a single category of age 90 or older
  6.
Fax numbers
  7.
Electronic mail addresses
  8.
Medical record numbers
  9.
Health plan beneficiary numbers
10.
Account numbers
11.
Certificate/license numbers
12.
Vehicle identifiers and serial numbers, including license plate numbers
13.
Device identifiers and serial numbers
14.
Web Universal Resource Locations (URLs)
15.
Internet Protocol (IP) address numbers
16.
Biometric identifiers, including finger and voice prints
17.
Full face photographic images and any comparable images; and
18.
Any other unique identifying number, character, or code (note this does not mean the unique
code assigned by the Investigator(s) to code the research data)
---------------------------------------------------------------------------------------------
* * * Attachments * * *
 
16. Attachments
Attach relevant documents here. These could include: Collaborating Investigator's IRB approval and
approved documents; Conflict of Interest information; Debriefing Script; Grant/Sub-contract; HIPAA
Authorization Form from HIPAA-covered entity; Interview/Focus Group Questions; Investigator's Brochure;
Letters of Agreement/Cooperation from organizations who will help with recruitment; Methodology section of
associated Thesis or Dissertation project; Questionnaires; Radiation Control Office approval material;
Recruitment Material (e.g., flyers, email text, verbal scripts); Sponsor's Protocol; Surveys; Other files
associated with the protocol (you can upload most standard file formats: xls, pdf, jpg, tif, etc.)  Please be
sure to attach all documents associated with your protocol. Failure to attach the files associated with the

Page 24 of 33
 
protocol may result in this protocol being returned to you for completion prior to being reviewed. Students:
Be sure to attach the Methods section of your thesis or dissertation proposal. If this protocol is associated
with a grant proposal, please remember to attach your grant.
To update or revise any attachments, please delete the existing attachment and upload the revised document to
replace it. 
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
1 page summary for clinicians v. 14May2015
Document Name
1 page summary for clinicians v. 14May2015
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
Verbal Recruitment 14May2015
Document Name
Verbal Recruitment 14May2015
Document Type
Questionnaire/Survey
Attachment
medical questionaire
Document Name
medical questionaire
Document Type
SOP
Attachment
B-01-R1 Biopsy SOP
Document Name
B-01-R1 Biopsy SOP
Document Type
SOP
Attachment
B-02-R1 Biopsy Needle Cleaning
Document Name
B-02-R1 Biopsy Needle Cleaning
Document Type
SOP
Attachment
HP-05-R1 Reporting AEs SOP
Document Name
HP-05-R1 Reporting AEs SOP
Document Type
SOP
Attachment
D2O-01-R1 Prep and Dispensing SOP
Document Name
D2O-01-R1 Prep and Dispensing SOP
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
Dairy Metformin Ad
Document Name
Dairy Metformin Ad
Document Type
Letters of Agreement.Cooperation
Attachment
Physician Support Letter
Document Name
Physician Support Letter

Page 25 of 33
 
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
CTS_Materials_Ad Examples
Document Name
CTS_Materials_Ad Examples
Document Type
Questionnaire/Survey
Attachment
BARSE
Document Name
BARSE
Document Type
Questionnaire/Survey
Attachment
EXSE_Baseline
Document Name
EXSE_Baseline
Document Type
Questionnaire/Survey
Attachment
EXSE
Document Name
EXSE
Document Type
Questionnaire/Survey
Attachment
IPAQ_English_self-admin_short
Document Name
IPAQ_English_self-admin_short
Document Type
Questionnaire/Survey
Attachment
NEWS
Document Name
NEWS
Document Type
Questionnaire/Survey
Attachment
Socioeconomic questionnaire
Document Name
Socioeconomic questionnaire
Document Type
Questionnaire/Survey
Attachment
Planning, Self-Regulation and Expectancy
Document Name
Planning, Self-Regulation and Expectancy
Document Type
Other Protocol Material
Attachment
Report - Draft 4
Document Name
Report - Draft 4
Document Type
Questionnaire/Survey
Attachment
15-5837H  Medical Screening form 19Feb2016
Document Name
15-5837H  Medical Screening form 19Feb2016

Page 26 of 33
 
Document Type
Other Protocol Material
Attachment
AE DC464 GI
Document Name
AE DC464 GI
Document Type
Other Protocol Material
Attachment
AE SR312 GI
Document Name
AE SR312 GI
Document Type
Other Protocol Material
Attachment
AE TA334 GI
Document Name
AE TA334 GI
Document Type
Other Protocol Material
Attachment
AE TA334 pink colored urine
Document Name
AE TA334 pink colored urine
Document Type
IBC Approval Letter
Attachment
IBC approval
Document Name
IBC approval
Document Type
Other Protocol Material
Attachment
AE AC693 GI present prior to start of intervention
Document Name
AE AC693 GI present prior to start of intervention
Document Type
Investigator's Brochure
Attachment
G6 Investigator Brochure
Document Name
G6 Investigator Brochure
Document Type
Investigator's Brochure
Attachment
CONTOUREZ-User-Guide-ENGLISH
Document Name
CONTOUREZ-User-Guide-ENGLISH
Document Type
SOP
Attachment
CGM SOP 9.1.16
Document Name
CGM SOP 9.1.16
Document Type
Investigator's Brochure
Attachment
Microlet2
Document Name
Microlet2
Document Type
Investigator's Brochure

Page 27 of 33
 
Attachment
G6 Blinded US_Study Instructions_User Guide
Document Name
G6 Blinded US_Study Instructions_User Guide
Document Type
Questionnaire/Survey
Attachment
Participant_Pre Screening 01Sept2016 v.2
Document Name
Participant_Pre Screening 01Sept2016 v.2
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
Dairy-Metformin Advertisement Example
Document Name
Dairy-Metformin Advertisement Example
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
Participant Testimonial
Document Name
Participant Testimonial
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
new recruiment flyer 1
Document Name
new recruiment flyer 1
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
new recruiment flyer 2
Document Name
new recruiment flyer 2
Document Type
Recruitment Material (e.g., flyers, email text, verbal
scripts)
Attachment
VIDEO PRODUCTION OUTLINE
Document Name
VIDEO PRODUCTION OUTLINE
Document Type
Other Protocol Material
Attachment
GCP protocol v 1.0 7 Decmber2016 v1.0
Document Name
GCP protocol v 1.0 7 Decmber2016 v1.0
Document Type
Other Protocol Material
Attachment
DOA CGM 03Feb2017
Document Name
DOA CGM 03Feb2017
Document Type
Grant/Sub-Contract
Attachment
Colorado State University - CTSA IIIS-2016-039 (11-
01-2016) (003)

Page 28 of 33
 
Document Name
Colorado State University - CTSA IIIS-2016-039 (11-
01-2016) (003)
Document Type
Grant/Sub-Contract
Attachment
CSU Dexcom Proposal 8.4.16
Document Name
CSU Dexcom Proposal 8.4.16
Document Type
Other Protocol Material
Attachment
15.5837H NSR DETERMINATION 02242017
Document Name
15.5837H NSR DETERMINATION 02242017
Document Type
Other Protocol Material
Attachment
15.5837 SR-NSR determination
Document Name
15.5837 SR-NSR determination
Document Type
Other Protocol Material
Attachment
Adverse Event Log 21April2017
Document Name
Adverse Event Log 21April2017
Document Type
Other Protocol Material
Attachment
CGM Adverse Event Log 03Feb2017
Document Name
CGM Adverse Event Log 03Feb2017
Document Type
Other Protocol Material
Attachment
Protocol Deviation CGM applied twice for pre
measurements 04April2017 signed
Document Name
Protocol Deviation CGM applied twice for pre
measurements 04April2017 signed
Document Type
Other Protocol Material
Attachment
Protocol Deviation consent signed
04March2017signed
Document Name
Protocol Deviation consent signed
04March2017signed
Document Type
Other Protocol Material
Attachment
Adverse Event Log 01June2017
Document Name
Adverse Event Log 01June2017
Document Type
Other Protocol Material
Attachment
CGM Adverse Event Log 01June2017
Document Name
CGM Adverse Event Log 01June2017

Page 29 of 33
 
---------------------------------------------------------------------------------------------
* * * Obligations * * *
 
 
Obligations (Researcher's Responsibilities)
 
The Principal Investigator is ultimately responsible for the conduct of the project. Obligations of the Principal
Investigator are:
 
Conduct the research involving human subjects as presented in the protocol, including modifications, as
approved by the Department and Institutional Review Board. Changes in any aspect of the study (for example
project design, procedures, consent forms, advertising materials, additional key personnel or subject populations)
will be submitted to the IRB for approval before instituting the changes (PI will submit the "Amendment/Revision"
form);
 
Provide all subjects a copy of the signed consent form, if applicable. Investigators will be required to retain signed
consent documents for three (3) years after close of the study;
 
Maintain an approved status for Human Subjects Protection training. Training must be updated every three (3)
years (Contact RICRO to check your current approval/renewal dates). For more information:  Human Subjects
Training Completed?
 
Submit either the "Protocol Deviation Form" or the "Report Form" to report protocol Deviations/Violations,
Unanticipated Problems (UPs) and/or Adverse Events (AEs) that occur in the course of the protocol. Any of these
events must be reported to the IRB as soon as possible, but not later than five (5) working days.  Note that if an
event resulted in life threatening injury or death OR an event resulted in substantive harm to the safety, rights or
welfare to human subjects, this must be reported to the IRB within 24 hours;
 
Submit the "Continuing Review" Form in order to maintain active status of the approved protocol. This form must
be submitted annually at least four (4) weeks prior to expiration, five (5) weeks for protocols that require full
review. If the protocol is not renewed before expiration, all activities must cease until the protocol has been re-
reviewed;
 
Notify the IRB that the study is complete by submitting the "Final Report" form.
 
  
  
Dr. Karyn Hamilton
X
The Principal Investigator has read and agrees to abide by the above obligations.
---------------------------------------------------------------------------------------------
* * * Event History * * *
Event History
Date
Status
View Attachments
Letters
04/29/2015
NEW FORM CREATED
05/15/2015
NEW FORM SUBMITTED
Y
05/15/2015
NEW FORM PANEL
ASSIGNED

Page 30 of 33
 
05/15/2015
NEW FORM REVIEWER(S)
ASSIGNED
05/18/2015
NEW FORM REVIEWER(S)
ASSIGNED
07/01/2015
NEW FORM REVIEWER(S)
ASSIGNED
07/27/2015
NEW FORM REVIEWER(S)
ASSIGNED
08/13/2015
NEW FORM MOVED
08/13/2015
NEW FORM APPROVED
Y
Y
08/24/2015
AMENDMENT 1 FORM
CREATED
08/24/2015
AMENDMENT 1 FORM
SUBMITTED
Y
08/24/2015
AMENDMENT 1 FORM
SUBMITTED
Y
08/25/2015
AMENDMENT 1 FORM
REVIEWER(S) ASSIGNED
10/07/2015
AMENDMENT 1 FORM
MOVED
10/07/2015
AMENDMENT 1 FORM
APPROVED
Y
Y
10/27/2015
AMENDMENT 2 FORM
CREATED
10/27/2015
AMENDMENT 2 FORM
SUBMITTED
Y
11/04/2015
AMENDMENT 2 FORM
REVIEWER(S) ASSIGNED
12/03/2015
AMENDMENT 2 FORM
APPROVED
Y
Y
12/03/2015
AMENDMENT 2 FORM UNDO
APPROVED
12/03/2015
AMENDMENT 2 FORM
MOVED
12/03/2015
AMENDMENT 2 FORM
APPROVED
Y
Y
12/14/2015
AMENDMENT 3 FORM
CREATED
12/14/2015
AMENDMENT 3 FORM
SUBMITTED
Y
12/15/2015
AMENDMENT 3 FORM
REVIEWER(S) ASSIGNED
01/06/2016
AMENDMENT 3 FORM
REVIEWER(S) ASSIGNED
01/21/2016
AMENDMENT 3 FORM
MOVED
01/21/2016
AMENDMENT 3 FORM
APPROVED
Y
Y
02/19/2016
AMENDMENT 4 FORM
CREATED
02/19/2016
AMENDMENT 4 FORM
SUBMITTED
Y
02/22/2016
AMENDMENT 4 FORM
REVIEWER(S) ASSIGNED
03/09/2016
AMENDMENT 4 FORM
REVIEWER(S) ASSIGNED

Page 31 of 33
 
03/16/2016
DEVIATION 1 FORM
CREATED
03/16/2016
DEVIATION 1 FORM
SUBMITTED
Y
03/17/2016
DEVIATION 2 FORM
CREATED
03/17/2016
DEVIATION 2 FORM
SUBMITTED
Y
03/18/2016
DEVIATION 2 FORM
REVIEWER(S) ASSIGNED
03/18/2016
DEVIATION 2 FORM
REVIEWER(S) ASSIGNED
03/18/2016
AMENDMENT 4 FORM
REVIEWER(S) ASSIGNED
03/21/2016
AMENDMENT 4 FORM
MOVED
03/21/2016
AMENDMENT 4 FORM
APPROVED
Y
Y
04/15/2016
DEVIATION 2 FORM
REVIEWER(S) ASSIGNED
04/15/2016
DEVIATION 1 FORM
REVIEWER(S) ASSIGNED
04/20/2016
CONTINUING REVIEW 1
FORM CREATED
04/27/2016
CONTINUING REVIEW 1
FORM SUBMITTED
Y
05/13/2016
CONTINUING REVIEW 1
FORM REVIEWER(S)
ASSIGNED
05/17/2016
DEVIATION 1 FORM
APPROVED
Y
N
05/17/2016
DEVIATION 2 FORM
APPROVED
Y
N
06/03/2016
CONTINUING REVIEW 1
FORM APPROVED
Y
Y
07/08/2016
REPORT 1 FORM CREATED
07/11/2016
REPORT 1 FORM
SUBMITTED
Y
07/18/2016
REPORT 1 FORM
REVIEWER(S) ASSIGNED
07/18/2016
REPORT 1 FORM
REVIEWER(S) ASSIGNED
08/10/2016
REPORT 1 FORM
APPROVED
Y
N
09/01/2016
AMENDMENT 5 FORM
CREATED
09/02/2016
AMENDMENT 5 FORM
SUBMITTED
Y
09/09/2016
AMENDMENT 5 FORM
REVIEWER(S) ASSIGNED
09/12/2016
AMENDMENT 5 FORM
REVIEWER(S) ASSIGNED
10/10/2016
AMENDMENT 5 FORM
REVIEWER(S) ASSIGNED
10/13/2016
REPORT 2 FORM CREATED

Page 32 of 33
 
10/14/2016
REPORT 2 FORM
SUBMITTED
Y
10/14/2016
REPORT 2 FORM
REVIEWER(S) ASSIGNED
11/02/2016
AMENDMENT 5 FORM
MOVED
11/02/2016
AMENDMENT 5 FORM
APPROVED
Y
Y
11/02/2016
AMENDMENT 5 FORM UNDO
APPROVED
11/02/2016
AMENDMENT 5 FORM
APPROVED
Y
Y
11/30/2016
AMENDMENT 6 FORM
CREATED
11/30/2016
AMENDMENT 6 FORM
SUBMITTED
Y
12/07/2016
AMENDMENT 6 FORM
REVIEWER(S) ASSIGNED
12/14/2016
AMENDMENT 6 FORM
MOVED
12/14/2016
AMENDMENT 6 FORM
APPROVED
Y
Y
01/05/2017
DEVIATION 3 FORM
CREATED
01/05/2017
DEVIATION 3 FORM
SUBMITTED
Y
01/06/2017
DEVIATION 3 FORM
REVIEWER(S) ASSIGNED
01/07/2017
REPORT 2 FORM
APPROVED
Y
N
01/11/2017
DEVIATION 3 FORM
REVIEWER(S) ASSIGNED
02/02/2017
AMENDMENT 7 FORM
CREATED
02/03/2017
AMENDMENT 7 FORM
SUBMITTED
Y
02/10/2017
AMENDMENT 7 FORM
REVIEWER(S) ASSIGNED
02/13/2017
DEVIATION 4 FORM
CREATED
02/13/2017
DEVIATION 4 FORM
SUBMITTED
Y
02/22/2017
AMENDMENT 7 FORM
REVIEWER(S) ASSIGNED
02/22/2017
AMENDMENT 7 FORM
REVIEWER(S) ASSIGNED
02/24/2017
AMENDMENT 7 FORM
REVIEWER(S) ASSIGNED
02/27/2017
AMENDMENT 7 FORM
MOVED
02/27/2017
AMENDMENT 7 FORM
APPROVED
Y
Y
03/23/2017
AMENDMENT 8 FORM
CREATED
04/24/2017
AMENDMENT 8 FORM
SUBMITTED
Y

Page 33 of 33
 
04/25/2017
AMENDMENT 8 FORM
REVIEWER(S) ASSIGNED
04/26/2017
AMENDMENT 8 FORM
APPROVED
Y
Y
05/04/2017
CONTINUING REVIEW 2
FORM CREATED
05/04/2017
CONTINUING REVIEW 2
FORM SUBMITTED
Y
05/12/2017
CONTINUING REVIEW 2
FORM REVIEWER(S)
ASSIGNED
05/19/2017
CONTINUING REVIEW 2
FORM APPROVED
Y
Y
05/25/2017
DEVIATION 3 FORM
APPROVED
Y
N
07/05/2017
AMENDMENT 9 FORM
CREATED
07/11/2017
AMENDMENT 9 FORM
SUBMITTED
Y
07/14/2017
AMENDMENT 9 FORM
REVIEWER(S) ASSIGNED
07/18/2017
DEVIATION 4 FORM MOVED
07/18/2017
DEVIATION 4 FORM
APPROVED
Y
N
07/24/2017
AMENDMENT 9 FORM
REVIEWER(S) ASSIGNED
07/27/2017
DEVIATION 5 FORM
CREATED
07/27/2017
AMENDMENT 9 FORM
APPROVED
Y
Y
07/27/2017
AMENDMENT 9 FORM UNDO
APPROVED
07/27/2017
AMENDMENT 9 FORM
APPROVED
Y
Y
07/27/2017
DEVIATION 5 FORM
SUBMITTED
Y
08/09/2017
DEVIATION 5 FORM
REVIEWER(S) ASSIGNED
08/28/2017
DEVIATION 5 FORM
APPROVED
Y
N
04/04/2018
CONTINUING REVIEW 3
FORM CREATED
05/09/2018
CONTINUING REVIEW 3
FORM SUBMITTED
Y
05/15/2018
CONTINUING REVIEW 3
FORM MOVED
05/15/2018
CONTINUING REVIEW 3
FORM APPROVED
Y
Y
10/03/2018
AMENDMENT 10 FORM
CREATED
10/04/2018
AMENDMENT 10 FORM
SUBMITTED
Y
10/04/2018
AMENDMENT 10 FORM
APPROVED
Y
Y

Statistical Plan  
Data are presented as mean ± standard error of the mean (SEM) and a significance 
was set a priori at P<0.05. Normality was confirmed by the D’Agostino and Pearson 
test. A two-way ANOVA (Treatment Group x Time) with repeated measures for time 
was performed. Upon a significant effect, a Holm-Sidak’s post-hoc test was used for 
multiple comparisons. When comparing the change (∆) from pre to post-intervention 
between groups, data were analyzed using an unpaired t-test. Pearson’s correlation 
coefficient (r) was used to determine associations between dependent variables. 
GraphPad Prism 7.0 was used to perform statistical analysis