Affective Social Buffering
Phase: NA
Conditions: Social Stress, Adolescent Behavior
Sponsor: University of Minnesota

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VERSION DATE: v.10 2024.12.18 
 
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Protocol Title 
The Role of Social Partners in Buffering Physiological and Brain 
Indicators of Threat-Related Stress in Children and 
Adolescents: An MRI Study of Learning and Social Support 
Principal 
Investigator/Faculty 
Advisor 
Kathleen M. Thomas, Ph.D. 
Institute of Child Development 
612-625-3389 
thoma114@umn.edu 
Student 
Investigator 
N/A 
 
  
 
 
 
Scientific 
Assessment 
Nationally-based, federal funding organizations 
IND/IDE # (if 
applicable) 
N/A 
IND/IDE Holder 
N/A 
Investigational Drug 
Services # (if 
applicable) 
N/A 
Version 
Number/Date: 
Version 10/ 2024.12.18 
 
PROTOCOL COVER PAGE 
 
 
 
 

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REVISION HISTORY 
 
Revision # 
Version Date 
Summary of Changes 
Consent Change? 
1 (v2) 
May 29, 2019 
Response to reviewer concerns 
Yes 
2 (v3) 
July 12, 2022 
The original protocol was written 
pre-grant award, with a planned 
mid-grant launch.  As a result of 
changes due to covid etc, the 
protocol has been updated. Re-
tooled for e-consent, remote 
buffers, and rather than 
counterbalancing two tasks, we 
will pilot and select one of the two 
tasks previously proposed; some 
measures were dropped for 
budgetary reasons. In response to 
the CR stipulations, we also 
updated consent/assent forms to 
conform to current language 
regarding: contact info, pregnancy 
info. We already noted that we will 
neither target nor exclude youth 
who have family members who are 
active members of military. 
Yes 
3 (v4) 
May 16, 2023 
Reduced manipulated conditions 
from 4 to 3 (alone, parent, friend). 
Removed unnecessary 
questionnaire. Updated fMRI task 
details. Updated biological 
samples. Increased participant 
payment. 
Yes 
4 (v4) 
Sept 21, 2023 
Added optional actigraphy 
measure. Added new recruitment 
methods. 
Yes 
6 (v6) 
Oct 27, 2023 
Added parent and child 
questionnaires.  
No 
7 (v7) 
Dec 7, 2023 
Added child questionnaire. 
No 
8 (v8) 
Jan 25, 2024 
Updated session length and hair 
assay language in consent form 
and protocol. Created consent 
addendum for participants who 
Yes 

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have already completed 
participation. 
9 (v9) 
March 7, 
2024 
Added recruitment method of 
mailing paper flyers. 
No 
10 (v10) 
December 
18th, 2024 
Increasing number of participants 
to enroll 
No 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Table of Contents 
 
1.0 
Objectives 
6 
2.0 
Background 
6 
3.0 
Study Endpoints/Events/Outcomes 
7 
4.0 
Study Intervention(s)/Investigational Agent(s) 
7 
5.0 
Procedures Involved 
8 
6.0 
Data and Specimen Banking 
14 
7.0 
Sharing of Results with Participants 
15 
8.0 
Study Population 
15 
10.0 
Local Number of Participants 
18 
11.0 
Local Recruitment Methods 
19 
12.0 
Withdrawal of Participants 
20 
13.0 
Risks to Participants 
21 
14.0 
Potential Benefits to Participants 
22 
15.0 
Statistical Considerations 
22 
16.0 
Confidentiality 
26 
17.0 
Provisions to Monitor the Data to Ensure the Safety of Participants 
27 
18.0 
Provisions to Protect the Privacy Interests of Participants 
28 
19.0 
Compensation for Research-Related Injury 
29 
20.0 
Consent Process 
29 
21.0 
Setting 
31 
22.0 
Multi-Site Research:    N/A 
31 
23.0 
Resources Available 
32 
24.0 
References 
32 
 

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ABBREVIATIONS/DEFINITIONS 
● fMRI: Functional Magnetic Resonance Imaging 
● CORT, AUCi: salivary cortisol, area under the curve from intercept 
● DHEA: dehydroepiandrosterone 
● EKG: Electrocardiogram 
● SCR:  Skin conductance response 
● vmPFC: ventromedial prefrontal cortex 
● ICD: Institute of Child Development 
 
 

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1.0 
Objectives 
1.1 Purpose:  
The goal of this project is to examine the neural systems underlying the 
physiological responses to threat in children and adolescents, and to 
examine the effectiveness of various social partners (parent, friend) in 
buffering youth from the physiological and brain effects of threat. 
2.0 
Background 
2.1 Significance of Research Question/Purpose & Existing Literature:  
Social buffering, a key concept in the psychobiology of stress, describes a 
phenomenon in which the presence and availability of social partners 
reduces activity of threat- and stress-mediating neurobiological and 
neuroendocrine systems (Gunnar, 2017). It is a key pathway through which 
social support reduces stress (Uchino et al., 2012) and enhances health 
(Kaplan et al., 1977). Impaired social buffering is a primary pathway 
through which adverse childhood experiences get under the skin to affect 
development (Hostinar et al, 2014).  Despite its centrality to stress or 
threat regulation, we know relatively little about the normative 
development of social buffering beyond the infant years. 
 
2.2 Preliminary Data:  
Prior behavioral data from our laboratories demonstrates that parents 
remain effective social buffers beyond infancy and throughout childhood. 
Among 9- and 10-year-olds, preparing for the Trier Social Stress Test (TSST) 
with a parent completely blocked elevations in cortisol to this social 
evaluative stressor (Hostinar et al., 2015). However, our results suggest 
that puberty results in a waning of parental HPA stress buffering 
effectiveness. Preparing for the TSST with the parent vs experimenter had 
no effect on 15- and 16-year-olds or on 11- to 14-year-olds who were at 
more advanced pubertal stages (Doom et al., 2016).  
 
Supportive friendships are associated with many positive outcomes for 
children, while their lack is associated with negative emotionality (Laursen 
& Hartl, 2013). However, counter to our initial expectations, our results 
showed that preparing for the TSST with a friend increased rather than 
reduced the cortisol response for adolescents (Doom et al., 2015). Having 
the friend “help” may actually have amplified effects of social evaluation. 
 
2.3 Study Aims:   
The current study examines the neural response, as well as the 
physiological responses to threat-related stress across the transition to 
puberty.  We use two standard threat learning paradigms that have been 

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used previously with adolescents. In this project, we will randomly assign 
youth to complete this testing under one of three social buffering 
conditions: presence of a parent, presence of a close friend, or no social 
support present (standard imaging condition). 
The long-term goal of our research is to enrich understanding of threat-
related stress regulation and its role in human development. The overall 
objective of this project is to understand normative processes in threat-
related stress regulation and their neurobiological correlates as children 
leave childhood and become more autonomous in adolescence. The 
central hypothesis of this project is that early adolescence is a period of 
time when parental buffering wanes in efficacy, leaving many youth 
experiencing a relative dearth of relationships that can provide powerful 
stress buffers.  
The aim of the current study is to delineate changes in brain activation in 
response to social buffering by parents and friends over the pubertal 
transition. 
Hypothesis #1: Parental social buffering will result in increased activation in 
brain regions associated with safety (e.g. vmPFC) and lower activation in 
brain regions involved in fear and pain (e.g., insula, amygdala, 
hypothalamus), as well as tighter coupling between the vmPFC and 
amygdala activity, but these relationships will wane with pubertal 
development.  
Hypothesis #2: Friend social buffering will result in less attenuation in the 
same brain regions, and less mPFC-Amygdala coupling than parental 
buffering, but will also become less effective with pubertal development. 
Hypothesis #3: Neural activity associated with social buffering will mediate 
the effectiveness of physiological buffering by social partners. 
 
3.0 
Study Endpoints/Events/Outcomes 
3.1 Primary Endpoint/Event/Outcome:  
Neural activity in vmPFC, insula, amygdala, and hypothalamus; salivary 
cortisol, heart rate, respiration, skin conductance  
3.2 Secondary Endpoint(s)/Event(s)/Outcome(s):  
Exploration of possible sex differences in across buffering conditions later 
in puberty but not earlier in puberty. 
4.0 
Study Intervention(s)/Investigational Agent(s) 
4.1 Description:  Note that there are no investigational agent(s) (e.g., drug, 
device) being evaluated. The study qualifies as an intervention due to 

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random assignment into social buffering condition, to determine the effect 
of social partner on threat-related stress responses. 
Child participants (10- to 15-year-olds) will be randomly assigned to one of 
three conditions during an fMRI version of a threat learning paradigm: (1) 
no social buffer, (2) presence of a parent, or (3) presence of a same-
sex/similar-age close friend. 
4.2 Drug/Device Handling:   N/A 
4.3 Biosafety:    N/A  
4.4 
Stem Cells:   N/A 
 
5.0 
Procedures Involved 
5.1 Study Design: The study design is cross-sectional.  150 target children (10- 
to 15-year-olds) will be randomly assigned to one of three conditions: (1) 
no social support (standard MRI scanning condition); (2) presence of a 
parent; or (3) presence of a close friend. Outcome measures will be 
compared across the three social buffering conditions. Pubertal stage will 
be included as a continuous variable in analyses of physiological and/or 
brain responses.   
Two tasks will be used during the MRI scanning. Both have been shown to 
activate stress-related regions of the brain in past studies and have been 
used by our labs. The order of these two tasks will be counterbalanced. 
Based on the results of piloting social buffering will EITHER be done with 
the presence of a buffer in person, or, displayed on screen via a live Zoom 
feed. All conditions will use the same modality. 
Study Overview:  
Visit 1: Parent and child will participate in an online video session (via 
Zoom) lasting approximately 2 hours.  Following a detailed overview of 
study and consent/assent procedures, participants will be asked to 
complete the following measures: 
Parent: 
● 
Questionnaire regarding family income, education level, and family 
composition 
● 
Parent-report questionnaire about child’s general health, behavior, 
and development (MacArthur Health Behavior Questionnaire; HBQ) 
● 
Parent-report MRI safety screening questionnaire about the child to 
rule out any contraindications for MRI scanning 
● 
[If the child is randomized to the parent-present buffering condition], 
self-report MRI safety screening questionnaire to rule out any 
contraindications for sitting in the MRI area 

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● 
Child life challenges question 
 
Children/adolescents:  
● 
Questionnaire about general health, behavior, and development 
(MacArthur Health Behavior Questionnaire; HBQ) 
● 
Questionnaire about their relationships with their parents and their 
best friend (Network of Relationships Questionnaire) 
● 
Questionnaire about anxiety in different contexts (State-Trait Anxiety 
Inventory for Children) 
● 
Pubertal development questionnaire (Morris and Udry, 1980) 
● 
Safety screening questionnaire to confirm safety in the MRI scanner 
● 
Perceived stress questionnaire 
● 
Child life challenges questionnaire 
 
Visit 2: Parent and child [and sometimes, child’s close friend] will come to 
the Center for Magnetic Resonance Research for a session lasting 
approximately 2 hours.  Note that if buffering occurs in person, we will 
invite the close friend to attend in person, and if buffering occurs on-line, 
we will invite them to attend via zoom. Following a session overview and 
consent/assent procedures, participants will be asked to complete the 
following measures: 
Parent: 
● 
Required safety screening questionnaire to confirm child’s safety in 
the MRI scanner 
● 
[If the child is randomized to the parent-present buffering condition], 
self-report MRI safety screening questionnaire to rule out any 
contraindications for sitting in the MRI area  
● 
[If the child is randomized to the parent-present buffering condition], 
sit next to scanner bed and listen to music, radio, audio book, podcast, 
etc. or read a book for duration of MRI scan (approx. 1 hour) OR sit in 
a separate room while connected via video conference call to research 
staff in the MRI console room for duration of MRI scan, chatting with 
the child periodically. 
 
Child: 
● 
Required safety screening questionnaire to confirm child’s safety in 
the MRI scanner 
● 
Saliva sampling at up to 10 time points throughout the testing session 
to assess levels of stress-sensitive compounds, including cortisol, alpha 
amylase. 
● 
Children chew on an absorbent sponge for 1-2 minutes to acquire 
each saliva sample. 

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● 
Practice in a pretend MRI machine to gain familiarity with the scanning 
environment and assess comfort in the scanner bore (i.e., rule out 
claustrophobia) 
● 
Standard non-invasive structural and functional MRI scanning for 1 
hour at 3-Tesla (clinical standard) field strength. 
● 
Collection of physiological data during the MRI scan (respiration belt, 
pulse oximeter and/or 3 EKG leads, MRI-safe skin conductance 
electrodes on the fingers or palm of one hand or foot). 
● 
Performance of two tasks during MRI scanning: one emotional faces 
matching task and one emotionally charged go-no go task. 
● 
Completion of a post-scan survey to assess perceived stress at various 
points throughout the session. 
● 
Optional: Wearing of an actigraphy device on the wrist for several days 
to record activity and sleep patterns. 
 
Child’s Friend: [Applicable only if the child is randomized to the friend-
present buffering condition] 
● 
MRI safety screening questionnaire to rule out any contraindications 
for sitting in the MRI area 
● 
Completion of the Network of Relationships questionnaire 
● 
Sit next to scanner bed and listen to music, radio, audio book, podcast, 
etc. or read a book for duration of MRI scan (approx. 1 hour) OR join 
the session online via video conference call to research staff in the 
MRI console room for duration of MRI scan , chatting with the first 
child periodically. 
● 
Note: Friend’s parent will complete a full consent procedure over the 
telephone or video conference call prior to the session and will return a 
signed consent and child safety-screening form via REDCap or by mail 
or with the participating family.  
 
5.2 Study Procedures: 
Social Buffering Manipulations: Will include 3 between-subjects conditions 
and both sexes. Participants will be randomly assigned to No Social Partner, 
Parent, or Close Friend conditions. Definition of Close Friend. We will ask 
the parent to have the child select the close same-sex friend they want to 
have accompany them. The parent then has to contact that child’s family 
and get their permission for us to contact them. We have successfully 
conducted studies in which the child brings their best friend.  
Affective Paradigms: Aversive Learning Paradigms:  Consent, assent, and 
questionnaires will occur during an online session (to reduce in-person 
requirements; minimize the impact of COVID on study feasibility.) At the 
session, after a reminder of procedures, youth will be separated from their 
parents. Youth will be taken to a private changing room to change into 

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hospital scrubs prior to moving to the MRI scanner. An MRI-safe respiration 
belt, pulse oximeter and EKG electrodes, and SCR electrodes will be 
positioned for physiological assessment and the child will be given ear 
plugs, a button response box for behavioral ratings, and an emergency call 
button. Youth will rest and/or watch a video during structural MRI 
sequences (6-10 minutes), then lie quietly with eyes open for a 5-10 minute 
scan with no video or audio material. The experimenter will then remind 
the child of the instructions for the affective task, and scanning will begin 
with the start of the task. (1) In the first task, one target face or shape is 
presented at the top of the screen and two option faces or shapes are 
presented at the bottom of the screen. One of the bottom faces shows the 
same emotion, or is the same shape, as the top face or shape and the 
participant is instructed to quickly identify which face or shape matches 
and select that option using the button box in their hand. The task includes 
five blocks of shape matching and four blocks of face matching, with each 
block containing six trials. Each trial presents the stimuli for 4.5 seconds 
and has an interstimulus interval of 0.5 seconds. At the end of the task, the 
child will be given a chance to move their bodies and rest their eyes before 
moving on to the second task. (2) In the second task, a target letter is 
presented in the center of the screen with a large image in the background. 
Participants are instructed to press a button on the button box if the letter 
is not an ‘X’ and refrain from pressing when they see the letter ‘X.’ The 
images presented behind the letters are sourced from the International 
Affective Picture System (IAPS) and are presented in blocks of positive, 
negative, neutral, negative, or scrambled (masked) images. The task 
includes two blocks of each affective category (positive, negative, neutral, 
and scrambled) in addition to two all-go blocks which present no ‘X’ and 
two rest blocks which present only a fixation cross. Each block contains six 
trials which present stimuli for 0.85 seconds with a variable interstimulus 
interval of 0.5-0.8 seconds. Following the second fMRI task, children will be 
removed from the scanner and will complete a stress questionnaire. Saliva 
samples will be collected every 10-20 minutes throughout the scanning 
session, including between scan segments, to assess dynamic changes in 
stress-related hormones. In the scanner, this involves moving the child 
partway out of the scanner bore and using a long collection sponge that 
hangs out of the mouth to avoid any possibility of choking since the child is 
lying supine.  We have used this method successfully in a previous study 
collecting saliva in the scanner environment.  Following the stress 
questionnaire, the child will change back into their own clothing and return 
to the lobby to complete additional questionnaires and wait for up to 30 
min while being sampled for stress hormones every 10 min.  
Pubertal Development: We will create a summary pubertal development 
index from parent & child report. A planned nurse exam was dropped due 

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to COVID and funding considerations. Parents & youth will complete the 
Morris and Udry (1980) pubertal development questionnaire, which allows 
placement in Tanner stages for hair, testicles, and breast development.  
 
Questionnaires.  
Demographic Questionnaire: Parents will complete information on pre-tax 
family income, education level of parent(s), composition of the household, 
and medications that the child regularly takes. 
Daily Diary: The parent and the youth both will report the following for the 
child that day: the time of wake up, medication usage, illness/fever, 
physical activity, and caffeine consumption.  
Relationship Quality:  Youth and their friend (when assigned to friend 
buffer condition) will complete the Network of Relationships Inventory 
(NRI, Furman & Buhrmester, 2009) for parents and their close friend. This 
questionnaire is based on an integration of attachment and Sullivanian 
theory and assesses three systems that are expected to be key in close 
relationships: attachment, caregiving and affiliation.  The scales are: 
companionship, seek safe haven, seek secure base, provide safe haven, 
provide secure base, conflict, antagonism and criticism. We will also 
include the full set of scales from the NRI,: Each scale consists of 3 items 
scored on a 5-point Likert scale. Psychometric qualities are high. Condition 
differences in quality of relationships will be examined and included as 
covariates if groups differ significantly. Otherwise, in follow-up analyses we 
will examine whether social buffering effects are stronger in relationships 
the child views as more supportive.  
MacArthur Health and Behavior Questionnaire (HBQ) for 9-18 Year-Olds 
(Parent and Child, 2.1):  This questionnaire was developed by Marilyn 
Essex based on the original HBQ for 4-8 year olds. We have used it many 
times. The scales we proposed to use here have high reliabilities (Cronbach 
alpha’s> .8). We will use the peer scales (acceptance/rejection, bullied, 
relational victimization, asocial, behavioral inhibition) and broad-band 
symptoms scales (Internalizing, Externalizing). These measures will be 
examined for balance across groups and if differences by condition, 
pubertal stage, or sex are found, will be entered as covariates.   
State-Trait Anxiety Index: A brief scale to assess current and general 
anxiety, which might be an important covariate. 
Self-Report of Stress: As a manipulation check, participants will rate their 
arousal and emotional state at various points during Visit 2. 
Morris and Udry Pubertal Development Questionnaire: The parent and 
youth will both report on hair, testes, and breast development. 

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Responses to Stress Questionnaire: Includes a checklist of stressors and 
how often they have occurred recently, then asks how respondents coped 
with each stressor listed and what coping methods were used more often. 
Child Life Events Inventory: Parent reports on past significant events in the 
child’s life that may have caused trauma, stress, or other relevant 
outcomes. 
Sleep Diary: Child reports on daily sleep habits over several days including 
timing of falling asleep and waking, perceived sleep quality, and sleep 
disruptions. This sleep diary would be completed once a day for up to 14 
days. 
Perceived Stress Scale: Child reports on perceptions of life stress over the 
past month.  
Child Life Challenges: Parent reports on their child’s global lifetime 
adversity exposure.  
Child Life Challenges: Child reports on their global lifetime adversity 
exposure. 
 
Physiological Measures. 
Stress Hormones: We will assay saliva samples for stress sensitive 
hormones: cortisol and alpha-amylase. Saliva will be collected using 
SalivaBio Oral Swabs (SOS) or passive drool and stored frozen at −20°C until 
shipped for assay. Medications will be recorded from parent report and 
classes of drug codes will be used to group meds by presumed mechanisms 
of action and will be entered as covariates (Granger et al., 2009). Systemic 
glucocorticoid medications are an exclusion criterion. For each analyte, we 
will calculate area under the curve from intercept (AUCi) to provide a single 
measure for the main analysis. 
Hair Hormones: Hair will be assayed for a full panel of stress, puberty, and 
sleep hormones. Hair cortisol provides a measure of the stress sensitive 
hormone cortisol indexed over approximately 3 months. We will take the ~ 
3 cm of hair next to the scalp on the posterior vertex. Based on an average 
hair growth rate of 1 cm/month, this represents hair grown over the 3-
month period prior to sampling. Wash and steroid extraction procedures 
will be conducted in Kirschbaum’s ISO lab, following their published 
protocol (Stalder & Kirschbaum, 2012, study II) with 7.5 mg of hair used for 
analysis. Participants will also complete a questionnaire about hair 
products and treatment that can be included as covariates. We anticipate 
that we will not be able to collect hair cortisol for some participants whose 
hair is too short or who refuse this sampling. In previous work, we have 

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had missing samples for ~ 10% of participants. We will deal with this using 
multiple imputation by chained equations (MICE) to impute missing data. 
Physiological Recording:  Participants will wear a combination of standard 
Siemens physiological recording equipment for the MRI environment 
(respiration belt, pulse oximeter, EKG leads). In addition, they will wear 
BioPac MRI-safe skin conductance electrodes. Samples will be collected 
throughout the entire MRI scan. Response measures will be computed by 
regressing aversive learning condition values on comparison values during 
the resting state scan and saving the residual as the response measure. If 
the parent and child opt-in, the child will wear an actigraphy device for 
several days. The actigraphy device uses accelerometers and gyroscopes to 
measure body movement. After completion of actigraphy data collection, 
the parent will return the device back to the researchers. 
All questionnaires and surveys are included in the ETHOS portal. Contact 
information and child age will be obtained from an existing registry of 
families who have expressed interest in child development research 
(specifically, parent name(s), address, phone number, email address, child 
name, child sex, and child birthdate). All other data will be acquired directly 
from the participants, and information obtained from the participant 
registry will be verified verbally with the parent. Birthdate will be collected 
from parent at the session so that a precise age of child can be obtained.  
 
5.3 Study Duration: Participants will participate in two sessions, one online, 
and the other in-person.  Each session will last approximately 2 hours. 
● 
All participants will be enrolled and assessed within approximately 
42-months. 
● 
Primary data analysis endpoints will be available within 6-12 months 
of the end of data collection (i.e., end of 5-year grant funding cycle). 
 
5.4 Individually Identifiable Health Information:  This study will collect health 
information directly from participants.  A HIPCO survey has been uploaded 
in the ETHOS portal. 
 
5.5   Use of radiation:    N/A 
5.6 Use of Center for Magnetic Resonance Research:    All MRI scanning will 
occur on a 3-Tesla MRI scanner managed by the CMRR. 
6.0 
Data and Specimen Banking 
6.1 Storage and Access: Saliva samples will be stored in a -20 degree freezer 
and hair samples will be stored in a dry dark secure location until they can 
be batched with other samples for hormonal assay.  Any material 

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remaining after the planned assays will be discarded.  No tissue or 
biological samples will be stored for future use or shared. 
 
Biospecimen samples will be identified only with a unique study ID, child 
sex, and sample number. No PHI will be associated with any biospecimen 
samples. Samples will not identify the study condition of interest (i.e., social 
buffering condition). 
6.2 Sharing: De-identified data will be shared with qualified researchers when 
requested for scientific purposes including replication of results, use in 
meta-analyses, or appropriate extension of analyses. No biospecimen 
samples will be saved or shared.   
7.0 
Sharing of Results with Participants 
7.1 Results will only be shared with participants in aggregate, with the 
following exceptions:   
 
Regarding hormonal or physiological measures in this study, abnormal 
values are commonly due to an interfering substance present in the sample 
(e.g., milk for salivary cortisol) or motion artifact in the case of EKG. 
Without a body of evidence, it seems unethical to alarm families given an 
out-of-range assay value and therefore we do not plan to share individual 
results with participant.  
In the event that the scanner operator or study personnel see something 
unusual in the MRI scan, the images will be submitted for review by a 
radiologist.  Images contain only the study ID number, participant’s age in 
years, participant sex, and test date.  In the event that the radiologist 
determines that the incidental finding should receive clinical follow-up, we 
will share this information with the parent(s) of the minor child participant 
with the recommendation to consult with the child’s primary care 
physician. Incidental findings from research scans are not clinically 
diagnostic and the images may not be added to the child’s medical record. 
8.0 
Study Population 
8.1 Inclusion Criteria: Healthy 10- to 15-year-olds. Additional inclusion criteria 
ensure that youth will be able to follow the study procedures (have 
sufficient vision, hearing, and language skills to provide verbal and written 
assent, see and read stimuli presented on the computer screen, and hear 
verbal instructions provided by the experimenter and/or judges).   
8.2 Exclusion Criteria for child participation: Premature birth (less than 37 
weeks), congenital and/or chromosomal disorders (e.g. cerebral palsy, FAS, 
mental retardation, Turner Syndrome, Down Syndrome, Fragile X), Autism 
Spectrum Disorders, history of serious medical illness (e.g., cancer, organ 

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transplant), youth using systemic glucocorticoids or beta-adrenergic 
medications, diagnosed psychiatric illness or psychotropic medication, 
seizure disorder or other neurological disorder, academic delay or 
individualized education plan (IEP), contraindications for MRI (implanted 
medical device; presence of non-removal metal in or on the body, including 
piercings, orthodontic braces or certain permanent retainers), known 
pregnancy, tattoo, or history of significant claustrophobia.  
8.3 Screening: Screening will occur by asking parents questions at the time of 
telephone recruitment. To reduce confounding of age and pubertal stage, 
we will use a stratified recruitment method. During recruitment, parents will 
be asked 3 questions about their child’s physical development, and children 
will be initially categorized into Pre/Early and Mid/Late Puberty. As cells fill, 
participants will no longer be enrolled into those cells. Also, to increase 
minority representation we will sample across zip codes with higher racial 
and ethnic minority representation.   
9.0 
Vulnerable Populations 
9.1 
Vulnerable Populations:  
 
Population / Group 
Identify whether any of the 
following populations will be 
targeted, included (not necessarily 
targeted) or excluded from 
participation in the study.  
Children 
Targeted Population 
Pregnant women/fetuses/neonates 
Excluded from Participation 
Prisoners 
Excluded from Participation 
Adults lacking capacity to consent 
and/or adults with diminished 
capacity to consent, including, but 
not limited to, those with acute 
medical conditions, psychiatric 
disorders, neurologic disorders, 
developmental disorders, and 
behavioral disorders 
Excluded from Participation 
Non-English speakers 
Excluded from Participation 
Those unable to read (illiterate) 
Excluded from Participation 

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Employees of the researcher 
Excluded from Participation 
Students of the researcher 
Excluded from Participation 
Undervalued or disenfranchised 
social group 
Included/Allowed to Participate 
Active members of the military 
(service members), DoD personnel 
(including civilian employees) 
Included/Allowed to Participate 
Individual or group that is 
approached for participation in 
research during a stressful situation 
such as emergency room setting, 
childbirth (labor), etc. 
Excluded from Participation 
Individual or group that is 
disadvantaged in the distribution of 
social goods and services such as 
income, housing, or healthcare. 
Included/Allowed to Participate 
Individual or group with a serious 
health condition for which there are 
no satisfactory standard treatments. 
Excluded from Participation 
Individual or group with a fear of 
negative consequences for not 
participating in the research (e.g. 
institutionalization, deportation, 
disclosure of stigmatizing behavior). 
Included/Allowed to Participate 
Any other circumstance/dynamic 
that could increase vulnerability to 
coercion or exploitation that might 
influence consent to research or 
decision to continue in research. 
Excluded from Participation 
 
9.2 
Additional Safeguards:  
Children are the targeted population for this study.  Others that have been 
checked as “allowed” above are not being intentionally included or 
excluded. For example, we will not inquire about someone’s membership 
in a disenfranchised group, nor whether a target child’s parent is on active 
duty in the military. Our careful consent/assent procedures, designed to 

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handle research with children, should be sufficient to cover other 
vulnerable participants who may inadvertently enter our study.  
All of the subjects will be children, ranging 10-15 years old. This study 
focuses on adolescents because our goal is an analysis of the impact of 
social buffering and puberty on aversive learning systems. Children have 
been the focus of our research for the past 30+ years. Our staff members 
are highly trained in working with children and families. The graduate 
students working on the project will be Child Psychology and/or Child 
Clinical Psychology Ph.D. students. The undergraduate research assistants 
will, for the most part, be majoring in child development.  
Much of the work will be performed at either the Institute of Child 
Development or the Center for Magnetic Resonance Research, centers 
specifically designed to be child-friendly. In addition to testing rooms, each 
site includes a playroom/waiting room.  While many of our participants will 
be “too old” for the playroom, they will likely find this a very welcoming 
environment.  
Our laboratory has been scanning both typically developing children and 
adolescents (ages 4-17) and those at risk due to early health or 
environmental risks for over two decades. We have conducted NIH funded 
research with child participants at the CMRR for the past 16 years. We are 
very experienced in putting children at ease in the MRI scanner and 
ensuring that children are aware that they can stop at any time.  We have 
completed a prior MRI study with 11- to 14-year-olds using a social stress 
paradigm; therefore, we are confident that we are well positioned to 
properly inform children about the procedures and to ensure that we are 
not stressing children beyond normal everyday levels. 
Parents provide consent, youth provide assent, and see below where that 
process is fully described to ensure that youth have full understanding and 
are free to decline participation. 
Individuals or groups with a fear of negative consequences for not 
participating in research are included/allowed to participate in this 
research study. These individuals are neither screened for nor excluded, 
but will be protected through our careful consent/assent procedures. 
During these procedures, as well as throughout the study sessions, 
participants are reminded that they are allowed to stop their participation 
at any time with no consequence. Our staff members are trained and 
experienced at noticing participants’ apprehension so that they may 
further discuss an individual’s participation in the study or stop the study 
procedures all together if necessary. 
 
10.0 Local Number of Participants 

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10.1 Local Number of Participants to be Consented:   We plan to analyze data 
from at least 150 target participants.  Of these, 50 will bring friends as 
social buffers. Thus, we plan to enroll at least 200 children. To account for 
attrition between sessions/replacement for incomplete data, up to 300 
children may be consented/enrolled. 
11.0 Local Recruitment Methods 
11.1 Recruitment Process: Families will be initially contacted by phone, email, or 
paper mail to invite them to participate. Paper mail will include the IRB-
approved recruitment flyer. Interested families will be provided a more 
detailed overview of the study procedures in a telephone call. If the family 
is interested in participating, the parent will verbally complete a screening 
interview to assess inclusion and exclusion criteria, and for those who are 
eligible, the first visit will be scheduled.  Copies of the consent and assent 
forms will be sent to families prior to the visit, along with directions to the 
University, and a reminder call will be a chance for families to confirm their 
interest in participation after having seen the consent forms. Souvenir 
brain images may be shared with families to increase interest. These 
images will only show a single cross-section of the brain.  
Regarding recruitment of a friend: The family of the participant will contact 
the family of the friend make sure that they have permission to give us their 
contact information. Parents of the friend will then be contacted by 
telephone and/or email to provide an overview of the study and the role of 
the friend. The friend’s parent will then be emailed a link to a consent & 
assent forms and MRI Safety Screening form (University of Minnesota 
REDcap) that they can complete and sign online before the session. If they 
do not respond to email in ~3 days, our recruiter will call them for a 
reminder. There will be phone and email contact information so that 
parents can ask any questions they have before their child comes to the lab. 
The assent form provided to the friend at the time of the study visit will also 
have a line that the parent of the main child participant can sign as a 
witness to the consent process.  We are currently using this process 
successfully in STUDY00006288. 
11.2 Identification of Potential Participants: Potential participants will be 
identified using the ICD registries of families interested in being contacted 
about research.  These families have agreed to be contacted, and once ICD 
Registry procedures have been followed, including proof of IRB approval, an 
encrypted list is provided to study staff. Paper and electronic flyers will be 
posted to supplement the list, and these materials may be physically posted 
as flyers, or electronically distributed via social media or list-servs such as 
CEHD. The Community Engagement and Education (CEEd) hub at the 
Masonic Institute for the Developing Brain (MIDB) will be used for 
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efforts. No protected records are involved. Study staff will make initial 
contact. 
The Institute of Child Development Participant Pool or IPP does not contain 
data that generates new knowledge. Indeed, maintaining the contact 
information of parents who have voluntarily expressed interest in child 
development research has been classified by the IRB as “Not Human 
Research” as defined by DHHS and FDA regulations and informed by HRP-
310. 
11.3 Recruitment Materials: We will recruit via telephone, email, and paper mail 
(as parents have given us these three means of contacting them).  We will 
also post flyers and put the flyer information on our website, other social 
media outlets, local organizations’ newsletters, and share via listserv so 
that families can contact us. Materials are uploaded in ETHOS. 
11.4 Payment: 
● 
Child Participant:  $40 debit or gift card for Visit 1; $60 in debit or gift 
card for Visit 2; $30 for optional actigraphy device 
● 
Parent of Child Participant: $25 in debit or gift card for Visit 1; $25 in 
debit or gift card for Visit 2 
● 
Friend of Child Participant: $40 in debit or gift card for Visit 2 
Cards will be given at the session to parent & child as specified. Where 
appropriate, due to remote buffering for example, we will mail cards. We intend 
to use Greenphire ClinCard prepaid debit cards for compensation and have 
included appropriate language in the consent document. We will provide a 
Target gift card instead of the ClinCard if parents object to the terms and 
conditions of the ClinCard. 
 
12.0 Withdrawal of Participants 
12.1 Withdrawal Circumstances: If participants are enrolled (consent signed) 
and subsequently determined to meet exclusion criteria, their data will be 
excluded from analyses. Children may be withdrawn from the study 
without their consent if we discover a safety concern for MRI scanning 
after consent/assent. Children may also be withdrawn if the experimenter 
believes that the child is excessively distressed by the affect paradigm or 
general MRI scanning procedures. Participants will receive full 
compensation for the session at which they are withdrawn. 
12.2 Withdrawal Procedures: If participants voluntarily withdraw, we will verify 
permission to retain and use any data collected to that point.  If 
participants wish to withdraw all permission, we will destroy any data 
collected from the participant but will keep their name in our study records 

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with a note that they withdrew. This procedure prevents us from 
inadvertently enrolling participants more than once. If the participant is 
withdrawn by the researcher, we will retain and use any data collected 
from the participant. 
12.3 Termination Procedures: Data will not be used after termination. 
13.0 Risks to Participants 
Foreseeable Risks:  
Questionnaires:  Youth or parents may experience concerns when 
completing questionnaires dealing with symptoms of behavior problems. 
Any time one works with youth there is the possibility that they will reveal 
information about abuse and/or indicate that they are thinking of hurting 
themselves or others. We are mandated reporters, a fact that is revealed in 
both consent and assent forms. However, in over a decade of working with 
children and youth drawn from our participant files, we have never 
encountered a reportable incident, so we consider this risk to be very low. 
Amygdala-activating Tasks. The tasks are designed to be uncomfortable, 
but not painful or distressing. Children encounter many types of affective 
stimuli in their everyday lives. Thus, we do not consider these stimuli to be 
out of the ordinary. To reduce risk, youth are told that they can always stop 
without prejudice if they choose. In addition, we carefully expose youth 
and their parents to example stimuli to ensure that all are comfortable 
with continuing.   
MRI Scanning: The primary risks associated with magnetic resonance 
scanning involve risk of injury or burn in the event that unsafe metallic 
objects are introduced into the scanner environment. Loose items have the 
potential to become projectiles while stationary metallic surfaces could 
build heat over time and cause a burn. In addition, implanted medical 
devices could become non-functional or experience torque that could move 
metal implants through body tissue. All participants are screened multiple 
times for the presence of metal in or on the body. Final screening occurs 
immediately prior to MRI scanning. 
 
There is no evidence for any long-term negative effect of MRI procedures on 
the body. The guidelines from the Bureau of Radiological Health (Food and 
Drug Administration) will be followed with regard to specific absorption rate 
(SAR) of energy (rf) and time-variant magnetic fields (dB/dt). All safety 
features of the clinically approved Siemens 3T scanner will be used to 
ensure that levels are well below the peripheral nerve stimulation threshold 
for both children and adults. 
The MRI procedure involves exposure to a high level of noise while in the 
scanner environment. Participants are required to wear sound attenuating 

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earplugs to protect their hearing. Beyond the risk to hearing, many 
participants find the scanner sounds annoying and monotonous.  We 
provide additional headphones and/or foam padding over the top of the 
earplugs to further attenuate the background noise.  This additional padding 
can become uncomfortable after long periods of time. 
To reduce anticipatory stress for the scanner experience and to ensure that 
children feel comfortable with all MRI procedures, youth will complete a 
mock MRI scan in an MRI simulator.  This pretend scanner has all of the 
primary features of the actual scanner, including an identically sized tunnel, 
the peripheral recording equipment, earplugs, headphones, video screen, 
audio system, button response box, and a speaker to play the scanner 
sounds around the tunnel, as if we were scanning, but with no magnetic 
field and no images produced. 
Saliva Sampling. The saliva is collected on an absorbent swab. Any time 
anything is put in the mouth there is the risk of choking. This is a greater 
concern when participants are lying supine in the MRI environment.  
However, we deliberately use long sponge rolls that that hang out of the 
mouth during saliva collection.  The child is instructed to hold on to the end 
sticking out of the mouth while chewing on the other end until it is soggy.  
The age of participants, 10-15 years, also reduces the risk of choking as 
these youth are better able to follow experimenter instructions and to 
realize when they are placing the sponge too far into their mouths to be 
comfortable. 
 
13.1 Reproduction Risks:  
The risks of strong magnetic fields for the developing human fetus are 
unknown.  To protect against these unknown but potential risks, we will 
screen all female youth (including youth serving as the friend social buffer) 
who have started menstruating for possible pregnancy. In addition, if 
buffers are in-person, they will be screened for possible pregnancy and 
exclude women who think they could be pregnant from serving as the 
social buffer (i.e., sitting in the MRI scanner room with the child). 
13.2 Risks to Others:   N/A 
14.0 Potential Benefits to Participants 
14.1 Potential Benefits: This study confers no direct benefits to participants. 
15.0 Statistical Considerations 
15.1 Data Analysis Plan:  
Preliminary Analyses to Determine Covariates Needed for Primary Analyses. 
The purpose of collecting data on potential covariates is to identify variables 

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that are correlated with both the independent variable (social buffering 
condition, sex, pubertal stage) and the primary outcome measures. Those 
covariates that are associated with the independent and outcome variables 
will be controlled for in the primary analyses. Those that are not will be used 
to describe the sample, but will not need to be included in the analyses to 
obtain unbiased estimates. For each potential covariate, we will conduct 
correlations or t-tests with pubertal stage, and t-tests, ANOVAs, or chi-
square tests (as appropriate) with sex and condition. If data do not confirm 
to modeling assumptions, we will use nonparametric analogs instead. 
 
Preliminary Analyses of Imaging Data to Yield Primary Outcome Measures: 
fMRI data will be analyzed in FSL software (version 6.0.1) to generate voxel-
wise contrast maps. Mean signal values will be extracted from these maps in 
a priori regions of interest for every participant using the methods described 
below. These preliminary analyses are used to generate the fMRI outcome 
variables of interest. The values of these outcome variables for each 
participant will be used in the general statistical analyses described in the 
next section. In total, these preliminary analyses will yield 6 primary task-
related outcome variables, and 6 primary connectivity-related outcome 
variables. 
 
Task-based analysis: In first level analyses (single subject), brain activity  will 
be modeled for each participant using a voxel-wise multiple regression 
analysis with a task predictor (aversive image vs. safe image), 24 regressors 
for head motion (Satterthwaite et al., 2013) and additional regressors to 
model censored data points (motion spikes). In subsequent analyses, we will 
explore the utility of including other time varying individual differences 
variables (e.g., heart rate, respiration) that are known to contribute to the 
BOLD signal. However, since these measures are expected to vary by stress 
condition, removing the effects of these physiological variables may 
unintentionally remove effects of interest. Particular care will be taken to 
reduce the confound of head motion by censoring (identifying as artifact) 
any data point with absolute motion greater than 2 mm (1 voxel) or a 
framewise displacement greater than 1 mm (1/2 voxel), as well as the 
immediately preceding and following data points. Data points with DVARS 
values exceeding the individual’s 75th percentile plus two times the 
interquartile range will also be censored. Participants with more than 25% 
of data points censored for artifact will be excluded from further analyses. 
This analysis will be used to identify brain regions differentially activated by 
aversive learning. Based on prior literature, we will examine activation in 5 a 
priori regions of interest, including “pain-related” brain regions (dorsal 
anterior cingulate [dACC], left and right anterior insula) and “safety-related” 
or regulatory brain regions (ventromedial prefrontal cortex [vmPFC], and 
left and right dorsolateral prefrontal cortex [dlPFC]). For each subject, the 

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mean percent change in MR signal (averaged across voxels) between 
aversive face/shape and the safe face/shape will be extracted from each of 
these 6 regions. This analysis will generate the 6 task-related brain 
activation outcome variables (listed in section 4.3) for each participant.  
 
Although these affect paradigms have been used previously, it is possible 
that the most prominent regions of activity for this task may not match the 
a priori regions identified from the prior literature. Therefore, we will also 
conduct an exploratory group-level analysis to identify primary regions of 
activation for the same aversive vs. safe stimuli (aversive image vs. safe 
image, cluster corrected p<.005). Because we expect that social buffering 
will alter the activity difference between aversive and safe periods, we will 
use only the control condition (no social partner) to identify regions 
activated by this task. In the resultant group-level map, we will identify up to 
4 primary regions of activation by selecting clusters that meet the cluster-
corrected p<.005 threshold, and then rank-ordering those regions by 
maximal z-statistic. Mean percent change in signal will be extracted for each 
participant (across all four buffering conditions) using anatomical masks 
derived from this analysis conducted in the control (no social partner) 
group. Therefore, exploratory analyses may generate up to 4 additional 
task-related MRI outcome variables. 
 
15.2 Power Analysis: For the power analysis for the primary analyses, we 
calculated the minimum detectable effect size (MDES) assuming n = 200, a 
= .05, and power = .80 with 12 covariates in our model. Power was 
estimated in R (R Core Team) using the pwr library (Champely, 2018).  A 
power analysis was performed to quantify the minimum detectable effect 
size (f2) for a three-way interaction between social buffering condition, sex, 
and pubertal stage within a multiple regression model controlling for the 
12 covariates, all two-way interactions, and the main effects (the most 
complex model considered in our primary analysis, see Equation 1 below). 
Assuming these conditions, we should be able to detect an f2 of 0.056. In 
other words, adding the three-way interaction to a model containing the 
covariates and all lower order interactions and main effects, the three-way 
interaction would need to explain an additional 5.6% of the variability to be 
detected. This falls between Cohen’s criteria for a small (f2 = .02) and 
medium effect (f2 = .15) (Cohen, 1988). With a sample size of 200, even if 
we lose 40 of the participants to movement artifact for the imaging 
analyses, assuming all the previous conditions but a sample size of 160, we 
will still have an f2 of .070, or still within a small-medium effect size. As the 
number of covariates decreases, the size of the MDES will decrease. 
Therefore, this represents a conservative estimate of the power for our 
primary analyses as we are likely to have fewer covariates in our models 

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and will use multiple imputation to correct for missing data. For the two-
way interaction and main effects models, described below, the power will 
be larger than these reported values as the models will be simpler. 
15.3 Statistical Analysis of MRI and non-MRI Outcome Variables:  
The following is a list of the analyses that we will perform on the outcome 
measures. They are classified as to whether they are the primary or 
mediational analyses. All statistical analyses will be performed in R. 
 
The primary analyses will consist of a series of multiple regression models. 
For each dependent variable, variation in neural activity, cortisol, and 
physiological responses, we will begin by testing the three-way interaction 
between social buffering condition, sex, and pubertal stage for all 
outcomes. If the three-way interaction is not significant, we will then 
examine all two-way interactions. If the two-way interactions are not 
significant, we will then examine main effects only models. If the three-way 
interaction is significant, we will not examine any other model. If condition 
is significant as an interaction or as a main effect, we will examine all 
pairwise comparisons using Tukey’s Honestly Significant Difference test. 
 
Multiplicity of testing. We will correct for multiple comparisons using the 
Benjamini-Hochberg (BH) correction. The BH method works by controlling 
the false discovery rate, and relative to the Bonferroni’s correction is more 
powerful, while still adequately protecting against Type I error (Williams, 
Jones, & Tukey, 1999). 
 
Mediational Analysis. When changes in activity of brain regions differ by 
condition or condition by pubertal status and we also see condition or 
condition by pubertal status differences for cortisol or physiological 
measures, then we will examine whether brain activity differences mediate 
the neuroendocrine or physiological differences. This will be examined 
using path analysis, while controlling for covariates identified in the 
preliminary covariate analysis, and will be estimated using the lavaan 
package in R (Rosseel, 2012). 
 
The significance of all indirect effects will be assessed using bias-corrected 
95% confidence intervals (Efron & Tibshirani, 1986), where confidence 
intervals that are non-overlapping with zero will be considered significant. 
 
Power Analysis for the Mediational Analysis: To assess power to detect 
mediation, we performed a Monte Carlo-based power analysis using 
simulated data. We varied the parameter values to roughly correspond to 
Cohen’s criteria for small, medium, and large effects. This resulted in a total 
of 27 conditions, which we examined at our target sample size of 200. Each 

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condition was replicated 2000 times and we calculated empirical power, 
which we defined as the proportion of times out of 2000 where we 
rejected the null hypothesis at α = .05. Given the exploratory nature of this 
analysis and to simplify the simulation for the power analysis, we omitted 
covariates and did not vary the effects for the other condition contrasts 
(i.e., they were always fixed to 0). Next, we fit a path model using the 
lavaan package in R.  
 
Our findings suggest that in order to detect mediation, the effect of 
condition on the mediator must be large and the effect of the mediator on 
the outcome must be medium or large. In addition, if the effect of 
condition on the outcome is not large, but small or medium sized, then we 
will be underpowered and would likely conclude complete mediation, 
when mediation may only be partial or non-existent. Based on this power 
analysis, it is likely that the MDES for the moderated mediation models will 
need to be at least medium to large as well. 
 
Missing data. Because we will replace participants who do not come in for 
session 2 after doing session 1 (which is rare in our experience), we should 
have 150 completed participants. Missing data, however, could arise from 
problems in obtaining good imaging data. Participants also often fail to fully 
complete questionnaires and/or refuse to provide information on 
household income. If there is missingness in any of the dependent or 
independent variables, multiple imputation will be used (Little & Rubin, 
2014; Peng, Harwell, Liou, & Ehman, 2006; Schafer & Graham, 2002). 
Variables related to the missingness and the variables missing data will be 
included in the missing data model. In the event that missingness is not at 
random (MNAR), we will investigate the use of models for non-ignorable 
missing data. 
15.4 Data Integrity: Biospecimen data will be assayed in labs that have standard 
quality control procedures.  MRI data reviewed for raw data quality and 
artifact detection.   
16.0 Confidentiality 
Data Security: Only authorized research staff will have access to the data, 
and all data will be stored in REDcap databases, secure servers (CMRR and 
Minnesota Supercomputing Institute) or in locked file cabinets. Names and 
contact information, and child birth date will be maintained in REDcap 
database to avoid enrolling participants more than once, to calculate age at 
test, and to allow us to contact families who have participated with overall 
study findings.  IRB and CMRR policies require that we maintain the MRI 
safety screening information for the same duration that we maintain 

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consent documents. These will be stored in locked file cabinets and/or 
uploaded to a secure REDcap database.   
A password-protected document linking name and ID number will be kept 
on REDCap until the conclusion of the study in case we must contact a 
family for any reason, including identification of an incidental MRI finding. 
This document will be destroyed at the conclusion of the study. 
All data files are de-identified, password protected, or maintained in secure 
storage locations (REDcap and/or University Box). All paper copies are 
stored in locked cabinets, and will be digitized/verified/destroyed. 
16.1 No documents will be placed in participants’ medical, employment, or 
educational records. 
17.0 Provisions to Monitor the Data to Ensure the Safety of Participants 
17.1 Data Integrity Monitoring: 
All of the planned procedures meet the NIH definition of minimal risk. The 
PI and Study Coordinator will meet regularly to ensure that all study 
protocols are followed and best practices are applied.  The Study 
Coordinator will bring any identified protocol violations to the PI in a timely 
fashion and team members will be retrained, if needed. 
17.2 Data Safety Monitoring: 
This project is considered a clinical trial because it involves a) random 
assignment to condition, and b) physiology or brain activity is being 
measured.  
There is no risk associated with the assignment to conditions (No Social 
Buffer, Close Friend, Parent). All conditions resemble naturally occurring 
social situations that children might experience, and there is no inherent 
danger to one condition over another as there would be for, say, a drug vs 
placebo trial.  
Further, in two of the conditions, the presence of another person is 
expected to provide a social buffer whose stress reduction potency by 
condition is what is being tested. All target youth experience the same 
stressor.  
The other potential risks of this study to which all of the participants are 
exposed regardless of condition are low, including, claustrophobia or fear of 
the MRI tunnel, loud noises in the scanner, and risk of choking during saliva 
collection.  
 
The steps taken to Reduce or prevent these risks include: use of proper 
saliva collection materials, and by completing thorough safety screening 
prior to MRI scanning, and providing youth with a simulated MRI experience 

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first. These risks are all managed by the routine processes monitored by our 
IRB and the CMRR Safety Committee. Adverse Events (Aes), including 
Serious Adverse Events (SAEs) such as deaths, hospitalizations, and life-
threatening events and Unanticipated Problems (Ups), will be managed and 
reported, as required, to the IRB and relevant monitoring authorities. All 
members of the project have received human subjects training and 
certification in FDA Good Clinical Practice through the Collaborative 
Institutional Training Initiative (CITI) curriculum. 
 
The following information will be collected to monitor risk: 
• The number of youth refusing the MRI scan. 
• Any instance of gagging during the saliva collection. 
 
How often will risks be assessed? Every three months we will examine the 
number of youth refusing the MRI scan and adjust procedures, if needed, to 
improve participant tolerance. In our experience, few youth of this age who 
have agreed to come in for testing end up refusing the MRI scan once they 
are here.  
 
The individuals responsible for trial monitoring and advising the appointing 
entity. Because of the low level of risk in this study, the PI, Dr. Kathleen 
Thomas will be responsible for monitoring and reporting events for this 
project. 
 
18.0 Provisions to Protect the Privacy Interests of Participants 
18.1 
Protecting Privacy: Once the participant has been tested, identifying 
information will be removed, and all materials will be identified with a 
participant number only. A master list with participant names, addresses 
and contact information will be maintained in secure REDcap database, 
separately from the data and study materials. A master list connecting links 
from study ID to participant name will be maintained in a password-
protected spreadsheet on RedCap and/or University Box Storage.  Only 
researchers involved in the study and having a legitimate reason for 
connecting data to an individual participant (e.g., in the event of a 
clinically-relevant incidental MRI finding) will have access to the file that 
merges participant number and personal information. Non-data study 
materials (e.g., consent forms, payment forms, MRI safety screening forms) 
will be maintained in a secure REDcap database separately from the de-
identified data. 
Considerable efforts are made to make participants comfortable, despite 
the nature of the stressor in the second visit.  Consent and all study 

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procedures will be conducted in private rooms.  During consent/assent 
multiple efforts are made to make it clear that participation is optional and 
that the child is in control of refusal; for example, payment is given at the 
start of the session rather than upon completion.  Collection of biological 
measures, placement of electrodes, and administration of sensitive 
questionnaires are all done with a “matter of fact” attitude to minimize any 
feeling of embarrasment.  Should a participant need to skip a portion or end 
a session early, experimenters are trained to accept this as a standard 
procedure when working with children rather than a problematic ending to 
the session. We have successfully guided hundreds of youth through these 
very procedures without undue problem, including longitudinal studies 
where the participants continue to return. 
Female participants who have begun menstruation will be verbally screened 
for possible pregnancy. If a child thinks she could be pregnant, we will not 
scan her. By Minnesota Statue, we are not allowed to break confidentiality 
to inform parents of a teen pregnancy. If a child tells us that she might be 
pregnant, we will offer her a urine pregnancy test to give her more 
information but will not require anyone to take a pregnancy test. If the test 
is positive, we will encourage her to tell her parents, and will offer to sit 
with the child while she informs a parent. If a child thinks she may be 
pregnant or has a positive pregnancy test, we will provide her with 
information on services that she can access to receive pre-natal care and 
counseling. 
18.2 
Access to Participants: This study does not involve access to participant 
medical records or other private records.  Any personal information 
collected in this study will be provided directly by the participants. 
19.0 Compensation for Research-Related Injury 
19.1 Compensation for Research-Related Injury: All of the proposed procedures 
meet the NIH definition of minimal risk.  In the event of an injury, costs will 
be charged to the participant or the participant’s medical insurance plan.  
19.2 Contract Language: N/A 
20.0 Consent Process 
20.1 Consent Process (when consent will be obtained):  
● 
All participants will be children, recruited by contacting parents.  Age 
will be verified with parent.  
● 
Parental permission will be obtained from: 
o 
One parent, even if the other parent is alive, known, competent, 
reasonably available, and shares legal responsibility for the care 
and custody of the child. 

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● 
The consent process will be conducted in an online secure Zoom 
session using RedCap econsent and assent forms. In the case of friends 
who act as buffers links to the forms will be sent to parents and 
verified prior to the session. 
● 
All youth will be part of the consent process, will be asked questions 
to verify understanding, including the question of “When are you 
allowed to skip a task or end the session?”.  All youth will provide 
written assent. 
● 
Only those individuals listed as personnel on the study will be allowed 
to obtain consent. All listed researchers have completed HIPAA and 
CITI training. Further, all consenters have experience 
consenting/assenting families and children in this age range for studies 
of similar procedures. Consenters are trained both to follow a bullet-
pointed list of key items as well as to sensitively discuss a range of 
concerns that parents or youth commonly raise. The consent process 
is a regular topic of weekly staff meetings, where researchers share 
experiences (without providing identifiable information) in order to 
help others learn novel approaches or gain tips for success.  
● 
In January of 2024 we created a consent addendum to expand on the 
allowed assays that could be performed on the child’s hair samples. 
This addendum would be sent to all participants that have completed 
participation in the study and provided a hair sample through the date 
of this modification’s approval (MODCR00004974). This expansion on 
allowed assays is also included in the consent form submitted for 
approval within this modification. 
20.2 Waiver or Alteration of Consent Process (when consent will not be 
obtained):     N/A 
20.3 Waiver of Written/Signed Documentation of Consent (when written/signed 
consent will not be obtained):    N/A 
20.4 Non-English Speaking Participants:    N/A 
20.5 Participants Who Are Not Yet Adults (infants, children, teenagers under 18 
years of age): 
● 
All participants will be children, recruited by contacting parents.  Age 
will be verified by the parent.  
● 
Parental permission will be obtained from: 
o 
One parent, even if the other parent is alive, known, competent, 
reasonably available, and shares legal responsibility for the care 
and custody of the child. 
● 
All participating youth will complete a written assent form to 
accompany the written parental consent described above, in 
accordance with Minnesota statutes. The investigator will review the 
consent form with both the youth and the parent, and will ask the 

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youth to tell them in their own words what will occur during the 
session (“In your own words, what is going to happen in today’s 
session?”). The investigator will then answer any questions the youth 
has and describe in more detail anything that she/he believes the 
adolescent does not fully understand. Each youth is reminded that 
they can stop at any time. We give them an “out” by saying, “Some 
people change their minds and decide this isn’t something they want 
to do right now, and that’s okay with us. Do you still want to go on?” If 
the youth does agree to participate in the study, s/he will sign an 
assent form. Similar procedures will be followed with the parents. 
Parents will be asked to describe in their own words what the child will 
be asked to do. If the parent seems unsure, the investigator will 
describe the procedures in another way and will provide corrective 
feedback. 
 
Recruitment procedures will make it clear that participation is 
voluntary and that families will still be able to participate in future 
research if they decline to participate in the current project. They may 
also remove their name from the participant database without 
prejudice if they wish. Parents will be mailed or emailed the consent 
forms and directions to the lab so that they have further time to make 
their decision before the session. Investigators will stress that parents 
and children are free to discontinue participation at any time without 
penalty. Further, compensation will be provided at the time of 
consent/assent, eliminating the chance that someone may feel 
compelled to continue for the compensation. 
20.6 Cognitively Impaired Adults, or adults with fluctuating or diminished 
capacity to consent:    N/A 
20.7 Adults Unable to Consent:    N/A  
21.0 Setting 
21.1 Research Sites:  
● 
Participants will be recruited from the ICD registry of potential 
participants, or through direct response to posted advertisements as 
described above.  
● 
Research will be conducted at the Institute of Child Development and 
the Center for Magnetic Resonance Research (CMRR). 
21.2 International Research:    N/A  
 
22.0 Multi-Site Research:    N/A 
 

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23.0 Resources Available 
23.1 Resources Available:  
● 
There are sufficient potential participants on the registry to meet the 
needs of the study design/cell count. In the age range we would need 
over the years of this proposal there are 14,529 girls and 15,284 boys 
from which to draw. 
● 
We anticipate that it will take approximately 2.5 years to complete 
recruitment and testing. We anticipate an additional 12 months 
needed to complete primary analyses of the MRI and physiological 
data. 
● 
The Institute of Child Development (ICD) and the Center for Magnetic 
Resonance Research (CMRR) are ideal facilities to collect these data, 
with appropriate rooms, parking, private consent areas, and 
sibling/family-friendly waiting areas. The ICD and CMRR have a 
realistic MRI simulator that will be critical for this study.  The CMRR 
houses 7 research-dedicated scanners, three of which are 3-Tesla 
scanners appropriate for the current study. Full-time staff maintain 
the32caners to ensure high-quality data acquisition, as well as linen 
services to provide necessary sheets, blankets, and hospital scrubs for 
participant safety and comfort.  
● 
We do not anticipate the need for either medical or psychological 
resources as risk is minimal in the current study. However, the ICD 
and CMRR all have First Aid and AED kits on-site, and the CMRR 
facilities are located on the medical campus of the University of 
Minnesota, providing close proximity to emergency medical services.   
● 
All staff & lab personnel, including undergraduate students working 
with de-identified data, undergo required and available ethics 
training.  The study coordinator has over 25 years of experience 
training staff and conducting behavioral and psychophysiological 
research with children and adolescents. Dr. Thomas has over 20 years 
of experience in designing and running MRI studies with child 
participants, and in mentoring junior scientists in MRI methods. The 
staff scientist has years of experience scanning children and 
supervising MRI data processing and analysis in Dr. Thomas’ lab. 
Weekly project meetings keep us adequately informed about any 
changes in protocol, updates to research procedures, or distribution 
of staff duties and functions. 
24.0 References 
Doom, J. R., Doyle, C., & Gunnar, M. R. (2016). Social stress buffering by friends in 
childhood and adolescence: Effects on HPA and oxytocin activity. Social Neuroscience, 
25, 1-14. 

PROTOCOL TITLE:  An MRI Study of Learning and Social Support 
VERSION DATE: v.10 2024.12.18 
 
Page 33 of 34 
Template Revised On: 02/28/2019 
Doom, J. R., Hostinar, C. E., VanZomeren-Dohm, & Gunnar, M. R. (2015). The roles of 
puberty and age in explaining the diminished effectiveness of parental buffering of 
HPA reactivity and recovery in adolescence. Psychoneuroendocrinology, 59, 102-11. 
Efron, B., & Tibshirani, R. (1986). Bootstrap methods for standard errors, confidence 
intervals, and other measures of statistical accuracy. Statistical Science, 1(1), 54–77. 
Gunnar, M. R. (2017). Social buffering of stress in development: A career perspective. 
Perspectives in Psychological Science, 12, 355-373. 
Hostinar, C. E., Johnson, A. E., & Gunnar, M. R. (2015). Parent support is less effective in 
buffering cortisol stress reactivity for adolescents compared to children. 
Developmental Science, 18, 281-297. 
Hostinar, C. E., Sullivan, R. M., & Gunnar, M. R. (2014). Psychobiological mechanisms 
underlying the social buffering of the hypothalamic-pituitary-adrenocortical axis: A 
review of animal models and human studies across development. Psychological 
Bulletin,140 (1), 256-282. 
Kaplan, B. H., Cassal, J. C., & Gore, S. (1977). Social support and health. Medical Care, 15, 
47-58. 
Laursen, B., & Hartl, A. C. (2013). Understanding loneliness during adolescence: 
developmental changes that increase the risk of perceived social isolation. Journal of 
Adolescence, 36, 1261-1268. 
Little, R. J., & Rubin, D. B. (2014). Statistical analysis with missing data. John Wiley & 
Sons. 
Neumann, D. L., Waters, A. M., Westbury, H. R., & Henry, J. (2008). The use of an 
unpleasant sound unconditional stimulus in an aversive conditioning procedure with 
8- to 11-year-old children. Biological Psychiatry, 79, 337-342. 
Peng, C.-Y. J., Harwell, M., Liou, S.-M., & Ehman, L. H. (2006). Advances in missing data 
methods and implications for educational research. Real Data Analysis, 31–78. 
Rosseel, Y. (2012). Lavaan: An R Package for Structural Equation Modeling. Journal of 
Statistical Software, 48(2). 
Schafer, J. L., & Graham, J. W. (2002). Missing data: Our view of the state of the art. 
Psychological Methods, 7(2), 147–177. 
Silvers, J. A., Lumian, D. S., Gabard-Durnam, L., Gee, D. G., Goff, B., Fareri, D. S., . . 
.Tottenham, N. (2016). Previous institutionalization is followed by broader 
amygdala-hippocampal-PFC network connectivity during aversive learning in human 
development. Journal of Neuroscience, 36, 6420-6430. 
Tzschoppe, J., Nees, F., Banaschewski, T., Barker, G. J., Büchel, C., Conrod, P. J., . . 
.IMAGEN Consortium (2014). Aversive learning in adolescents: modulation by 
amygdala- prefrontal and amygdala-hippocampal connectivity and neuroticism. 
Neuropsychopharmacology, 39, 875-884. 
Uchino, B. N., Bowen, K., Carlisle, M., & Birmingham, W. (2012). Psychological pathways 
linking social support to health outcomes: a visit with the “ghosts” of research past, 
present, and future. Social Science and Medicine, 74, 949-957. 
Williams, V. S., Jones, L. V., & Tukey, J. W. (1999). Controlling error in multiple 
comparisons, with examples from state-to-state differences in educational 
achievement. Journal of Educational and Behavioral Statistics, 24(1), 42–69. 

PROTOCOL TITLE:  An MRI Study of Learning and Social Support 
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Page 34 of 34 
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Spielberger, C. D. (1989). State-Trait Anxiety Inventory: Bibliography (2nd ed.). Palo Alto, 
CA: Consulting Psychologists Press. Spielberger, C. D., Gorsuch, R. L., Lushene, R., 
Vagg, P. R., & Jacobs, G. A. (1983). 
Stalder, T., & Kirschbaum, C. (2012). Analysis of cortisol in hair--state of the art and future 
directions. Brain and Behavioral Immunity, 26, 1019-1029.