Education, Faculty of
Kinesiology, School of
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UBCV
Hicks, Hana
Master of Science - MSc
University of British Columbia
Long-term physical activity and exercise have been shown to mitigate neurobiological responses to acute psychosocial stressors. There is also evidence that engaging in a single bout of exercise has immediate effects on reducing neurobiological reactivity to acute psychosocial stressors. However, much of this previous work applied definitions of intensity zones on American College of Sports Medicine’s global prescription recommendations, not based on individualized, physiologically meaningful cut-off points for demarcating the intensities at which participants exercised. This project expands on prior evidence by investigating the extent to which a single bout of exercise, performed at light, moderate, or vigorous intensities – established with meaningful indicators of changes in metabolic needs – alters the neurobiological response to a subsequently induced psychosocial stressor. It is hypothesized that vigorous intensity, defined as the midway point between the first and second ventilatory thresholds, will dampen cortisol output following a subsequently induced psychological stressor, compared to moderate (defined as being 10% below the VT1) and light (set at 1.7km/hour) intensities. Moderate intensity is expected to dampen the cortisol trajectory as compared to the light intensity, but to a lesser extent than vigorous. To achieve this, participants complete a modified maximal cardiopulmonary exercise test (CPET) to determine their individual ventilatory thresholds. Participants then return to the lab a second time to complete the experiment, whereby they are randomized to complete a 30-minute exercise at their randomized intensity zone, followed by the Trier Social Stress Test 45 minutes later. Multilevel Growth Curve Analysis determined that participants in the vigorous group had significantly slower rates of increased cortisol compared to both the light and moderate groups. Results also indicated no significant differences in cortisol trajectories between the light and moderate groups, apart from the fact that those in the moderate group had lower estimated starting points prior to starting the TSST. Results suggest that the current findings may be a result of the negative feedback loop, if they hold when the complete sample is analyzed. Examination of other psychological and physiological responses to psychosocial stressors will help elucidate the extent to which exercise mitigates stress reactivity in its more comprehensive conceptualization.
Effects of Exercise Intensities on Cortisol Release Following the Induction of a Psychological Stressor by Hana Hicks B.A., Saint Mary’s University, 2018 B.A.H., Saint Mary’s University, 2021 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Kinesiology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) December, 2024 © Hana Hicks, 2024 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: Effects of Exercise Intensities on Cortisol Release Following the Induction of a Psychological Stressor submitted by Hana Hicks in partial fulfilment of the requirements for the degree of Master of Science in Kinesiology Examining Committee: Dr. Eli Puterman, Professor, Kinesiology, UBC Supervisor Dr. Mark Beauchamp, Professor, Kinesiology, UBC Supervisory Committee Member Dr. Frances Chen, Professor, Psychology, UBC Supervisory Committee Member iii Abstract Long-term physical activity and exercise have been shown to mitigate neurobiological responses to acute psychosocial stressors. There is also evidence that engaging in a single bout of exercise has immediate effects on reducing neurobiological reactivity to acute psychosocial stressors. However, much of this previous work applied definitions of intensity zones on American College of Sports Medicine’s global prescription recommendations, not based on individualized, physiologically meaningful cut-off points for demarcating the intensities at which participants exercised. This project expands on prior evidence by investigating the extent to which a single bout of exercise, performed at light, moderate, or vigorous intensities – established with meaningful indicators of changes in metabolic needs – alters the neurobiological response to a subsequently induced psychosocial stressor. It is hypothesized that vigorous intensity, defined as the midway point between the first and second ventilatory thresholds, will dampen cortisol output following a subsequently induced psychological stressor, compared to moderate (defined as being 10% below the VT1) and light (set at 1.7km/hour) intensities. Moderate intensity is expected to dampen the cortisol trajectory as compared to the light intensity, but to a lesser extent than vigorous. To achieve this, participants complete a modified maximal cardiopulmonary exercise test (CPET) to determine their individual ventilatory thresholds. Participants then return to the lab a second time to complete the experiment, whereby they are randomized to complete a 30-minute exercise at their randomized intensity zone, followed by the Trier Social Stress Test 45 minutes later. iv Multilevel Growth Curve Analysis determined that participants in the vigorous group had significantly slower rates of increased cortisol compared to both the light and moderate groups. Results also indicated no significant differences in cortisol trajectories between the light and moderate groups, apart from the fact that those in the moderate group had lower estimated starting points prior to starting the TSST. Results suggest that the current findings may be a result of the negative feedback loop, if they hold when the complete sample is analyzed. Examination of other psychological and physiological responses to psychosocial stressors will help elucidate the extent to which exercise mitigates stress reactivity in its more comprehensive conceptualization. v Lay Summary Stress is a part of daily life, the effects of which can have far reaching complications on one’s health, both physically and psychologically. Due to the unavoidable nature of stress, ways to combat its harmful effects are crucial. This study will expand on previous evidence that suggests that exercise and physical activity can help reduce the reactivity of the stress response. Specifically, this study will examine how different exercise intensity levels (light, moderate, and vigorous), determined by meaningful metabolic points, impact the cortisol stress response to a subsequent psychological stressor. vi Preface This thesis was conducted in the Fitness, Aging, and Stress Laboratory at the University of British Columbia. The aim of this thesis is to analyze the cortisol reactivity and recovery from a psychological stressor following a randomized 30-minute bout of exercise from 42 participants of the Exercise and Social Interaction Trail (EAST) which is still on-going. The EAST study is approved under the University of British Columbia’s Clinical Research Ethics Board (certificate #: H20-02065) and is registered on the Open Science Frame (OSF: https://doi.org/10.17605/OSF.IO/PJZ6C) under the preregistration registry after data collection had started but prior to analysis. I am the lead investigator on the EAST project with Dr. Eli Puterman as the Principal Investigator and am responsible for the launch of the study (i.e., design, pilots, ethics, etc.), research assistant recruiting, hiring, scheduling, and training, participant recruitment, overseeing and leading all data collection, and analysis. My committee members Dr. Frances Chen, and Dr. Mark Beauchamp not only helped guide this process but have both contributed to previous work that the parent EAST study is expanding upon. Luke Peddie also played a role in the creation of the study, Dr. Gabriel Zieff acted as our physiologist, and Brook Haight, Ria Gill, and Isabella Ravagli all acted as the study coordinator at different timepoints of the study. Ben Hives also played a major role in the data analysis. Although the data for the thesis has been completed the EAST study data collection is still in progress which I intent to continue to lead. In no particular order, Max Abercrombie, Ria Gill, Annie Li, Rebecca Dutchak, Zachary Kwan, Adam Chan, Julia Dancey, Samantha Nix, Eva D’Odorico, Priya Dhaliwal, Chloe Kim, Jordan McLeod, Chris Lee, Tanvi Pandhi, Grace Poblete, Dennis Tanujava, Steven Kim, Jesper de Baat, Joshua Quon, Kodie Harkins, Megan vii Liu, Ashvin Dhaliwal, Grace Wang, Charlotte Drennan, Sarah Barghouthi, Reatu Gelaye, Ruth Liu, Rachel Lam, Rachelle Tweed, Jasmeen Tiwana, Luai El-Khatib, Mylla Hosseinzadeh, Jayda Anderson, Agatha Leung, Sunny Kwok, and Jordana Hadfield all participated in recruitment and data collection. No publication has been submitted from this project to date. viii Table of Contents Abstract ......................................................................................................................................... iii Lay Summary .................................................................................................................................v Preface ........................................................................................................................................... vi Table of Contents ....................................................................................................................... viii List of Tables ..................................................................................................................................x List of Figures ............................................................................................................................... xi List of Abbreviations .................................................................................................................. xii Acknowledgements .................................................................................................................... xiv Dedication .....................................................................................................................................xv Chapter 1: Introduction and Purpose .......................................................................................... 1 1.1 Defining an Acute Stressor ............................................................................................... 2 1.2 Neurobiological Pathways ................................................................................................ 4 1.3 Response Outcomes from Single Bout of Exercise .......................................................... 6 1.4 Cross-stressor Adaptation Hypothesis .............................................................................. 7 1.5 Limitations with the Literature ....................................................................................... 13 1.6 The Current Study ........................................................................................................... 13 1.7 Hypothesis....................................................................................................................... 14 Chapter 2: Methodology, Study Protocol, and Planned Analysis ............................................. 15 2.1 Participants ...................................................................................................................... 15 2.2 Study Design ................................................................................................................... 19 2.3 Measures ......................................................................................................................... 20 2.4 Procedures ....................................................................................................................... 23 Chapter 3: Data Analytic Approach .......................................................................................... 34 3.1 Power Calculations ......................................................................................................... 34 3.2 Statistical Analysis .......................................................................................................... 34 3.3 Hypotheses ...................................................................................................................... 36 Chapter 4: Results ..................................................................................................................... 37 4.1 Participants ...................................................................................................................... 37 4.2 Protocol Adherence ......................................................................................................... 39 4.3 Salivary Cortisol Assays ................................................................................................. 39 4.5 Unconditional Means Model ........................................................................................... 39 4.6 Level-1 Growth Model; cortisol response as a function of time .................................... 40 4.7 Level-2 Growth Model; Cortisol response between groups ........................................... 40 4.8 Missing Data ................................................................................................................... 43 Chapter 5: Discussion ............................................................................................................... 44 5.1 Glucocorticoid Negative Feedback Loop ....................................................................... 44 5.2 Demarcating exercise intensity levels ............................................................................. 45 5.3 Understanding the stress response .................................................................................. 48 5.4 Future Directions ............................................................................................................ 53 5.5 Limitations ...................................................................................................................... 55 5.6 Conclusion ...................................................................................................................... 56 References .....................................................................................................................................57 Appendices ....................................................................................................................................69 Appendix A Recruitment Advertisements ......................................................................... 69 A.1.1 Advertisement Poster ........................................................................................................................... 69 ix A.1.2 Advertisement Slide ............................................................................................................................... 70 A.1.3 In-Class Advertisement Script ........................................................................................................... 71 A.1.4 HSP Advertisement ................................................................................................................................ 72 A.1.5 Initial Contact Email ............................................................................................................................... 73 A.1.6 Phone Screener ........................................................................................................................................ 75 A.1.7 Reminder Emails ..................................................................................................................................... 80 Appendix B Consent Forms .............................................................................................. 83 B.1 Cash Honorarium Consent Form .......................................................................................................... 83 B.2 HSP Consent Form ...................................................................................................................................... 94 Appendix C Deception Form ........................................................................................... 105 Appendix D Debriefing Protocol ..................................................................................... 109 D.1 Cash Debriefing Protocol ...................................................................................................................... 109 D.2 HSP Debrief Protocol .............................................................................................................................. 113 D.3 HSP Take-home Debrief Form ........................................................................................................... 117 Appendix E Receipts ....................................................................................................... 118 E.1 Cash Receipt ............................................................................................................................................... 118 E.2 HSP Receipt ................................................................................................................................................. 119 Appendix F Data Collection Forms ................................................................................. 120 F.1 Data Collection Forms Day 1 ............................................................................................................... 120 F.2 Data Collection Forms Day 2 ............................................................................................................... 125 Appendix G Trier Social Stress Test Script .................................................................... 133 Appendix H Questionnaires ............................................................................................ 136 H.1 Demographic Questionnaires ............................................................................................................. 136 H.2 Current Health Status ............................................................................................................................ 137 H.3 Pre-Exercise Responsibility Checklist ............................................................................................ 138 H.4 Physical Activity Readiness Questionnaire for Everyone (PAR-Q+) ................................. 139 H.5 L-CAT (Leisure Time Categorical Activity Scale) ....................................................................... 143 H.6 Centre for Epidemiology Studies – Depression Scale – Revised ......................................... 144 H.7 Perceived Stress Scale (PSS) ............................................................................................................... 145 H.8 Rating of Perceived Exertion (RPE-Scale) .................................................................................... 146 H.9 Felt Arousal Scale (FAS) ........................................................................................................................ 147 H.10 Feeling Scale ............................................................................................................................................ 148 H.11 Negative & Positive Affect Questionnaire (PANAS-X) ........................................................... 149 H.12 Task Appraisal Questionnaires ....................................................................................................... 150 H.13 Thoughts Questionnaire .................................................................................................................... 152 Appendix I Pre-exercise Explanation .............................................................................. 153 I.1 Full Pre-Exercise Explanation Day 1 ................................................................................................. 153 I.2 Full Pre-Exercise Explanation Day 2 ............................................................................................... 155 Appendix J Lab Manual Forms ....................................................................................... 158 J.1 COSMED Cheat-Sheet ............................................................................................................................... 158 J.2 BIOPAC Set-up ............................................................................................................................................ 160 x List of Tables Table 1: Demographic Statistics ................................................................................................... 38 Table 2: Cortisol Demographic Statistics .................................................................................... 40 Table 3: Unconditional Means Model .......................................................................................... 40 Table 4: Coefficients with Alternate Group Coding .................................................................... 43 xi List of Figures Figure 1: Salivette® Instructions for use (Sarstedt, Rommeldsorf, Germany) ............................ 22 Figure 2: First Assessment Day Timeline ..................................................................................... 25 Figure 3: Second Assessment Day Timeline ................................................................................ 32 Figure 4: Group and Individual Trajectories of Cortisol .............................................................. 42 xii List of Abbreviations AED Automated External Defibrillator ANOVA Analysis of Variance CPET Cardiopulmonary Exercise Test CPR Cardiopulmonary Resuscitation CO2 Carbon Dioxide ECG Electrocardiogram FAST Fitness Aging and Stress fMRI Functional Magnetic Resonance Imaging Scan HSP Human Study Pool (UBC Psychology) HPA Hypothalamic-Pituitary-Adrenal HRR Heart Rate Reserve L-CAT Stanford Leisure-Time Activity Categorical Item LICT Light Intensity Continuous Training MICT Moderate Intensity Continuous Training MIST Montreal Imaging Stress Test PAR-Q+ 2019 Physical Activity Readiness Questionnaire for Everyone PSS Perceived Stress Scale RPE Rate of Perceived Exertion SAM-p Sympathomedullary Pathway TSST Trier Social Stress Test TSST-G Trier Social Stress Test-Group UBC University of British Columbia VICT Vigorous Intensity Continuous Training VO2 Volume of Oxygen Consumption xiii V̇T Ventilatory Threshold V̇T1 First Ventilatory Threshold VT2 Second Ventilatory Threshold VO2max Maximal Oxygen Consumption xiv Acknowledgements I’d like to take a moment to extend my deepest thanks and admiration to Dr. Eli Puterman who took a chance on me over three years ago, providing me with an opportunity of a lifetime to be able to study and practice research here at the University of British Columbia. I would also like to thank my committee members, Drs. Mark Beauchamp and Frances Chen, for their patience and flexibility around my unorthodox timeline. This project would not be possible without my team, of whom there are many: Julia Dancey, Adam Chen, Zach Kwan, Annie Li, Eva D’Odorico, Ria Gill, Jordan Mcleod, Priya Dhaliwal, Grace Poblete, Rebecca Dutchak, Samantha Nix, Jesper de Baat, Joshua Quon, Steven Kim, Dennis Tanujaya, Kodie Harkins, Megan Liu, Ashvin Dhaliwal, Grace Wang, Charlotte Drennan, Sarah Barghouthi, Reatu Gelaye, Ruth Liu, Rachel Lam, Chris Lee, Rachelle Tweed, Jasmeen Tiwana, Luai El-Khatib, Mylla Hosseinzadeh, Jayda Anderson, Agatha Leung, Sunny Kwok, and Jordana Hadfield thank you for your time and effort. A special shout out to two individuals who were always willing to help lighten any load I had. Max Abercrombie and Brook Hadwen: thank you for taking on all you did and for being such amazing supports. To Ben Hives, without whom I’d still be staring a pile of numbers unable to comprehend their meaning. To Brook Haight, whose unwavering professionalism and organization is to be admired: thank you for always weathering the storm with me and for sharing laughs when it seemed impossible. Working with your best friend is something I hope everyone gets to experience at least once. To my friends and family coast to coast, thank you for being in my corner, always. With a specific shoutout to my mom, Kimberley Hicks. And to Jackson, who never failed to show up with a smile and a sandwich: I wouldn’t be where I am today without you. xv Dedication Dedicated to everyone I burst into tears in front of when they asked me if I was “okay”. 1 Chapter 1: Introduction and Purpose “Stress” is a ubiquitous experience of human existence. According to Epel and colleagues (2018), stress is a multidimensional concept dependent on dynamic interactions between individuals and their environments and is both physiological and psychological in nature. Stress can be experienced when exposed to both acute and chronic stressors. Acute stressors are intense but brief exposures that elicit an unconscious or conscious cognitive appraisal ultimately evoking a physiological response. These can include daily experiences like rush-hour traffic or arguments with a family member or close friend. Chronic stressors, on the other hand, are prolonged exposure to a stress-inducing circumstance, such as providing care to a family member with a chronic health condition, or the repeated, frequent exposure to acute stressors on a daily basis (Epel et al., 2018; Hobfoll, 2001). Exposure to stressors, regardless of their duration or intensity, can elicit neurobiological responses, including activation of the sympathetic-adrenal medullary pathway (SAM-p) and the hypothalamus-pituitary-adrenal axis (HPA-axis). When activated acutely, activation of these neurobiological systems is not inherently harmful; rather, these responses are evolutionary mechanisms that support the body in its attempt to manage its resources to effectively meet the demands of the stressor. It is evidenced, though, that prolonged activation of these neurobiological mechanisms, as seen with chronically present stressors, yields both psychological and/or physical damage resulting in a wide range of health complications. These health problems include increases in systemic inflammation (Kivimaki & Steptoe, 2018), promotion or progression of non-communicable diseases (Miller et al., 2007), and early mortality (Kopp & Réthelyi, 2004). Chronic exposure to physiological stress has also been associated with psychological disorders, like major depressive disorder and anxiety disorders (Miller et al., 2007) 2 and increased engagement in maladaptive health behaviours (e.g., smoking, poor diet, decreased sleep, lower physical activity levels, etc.), ultimately perpetuating a self-destructive cycle impacting overall health (Cohen et al., 2007; O’Connor et al., 2021). Therefore, finding approaches to mitigate the negative consequences of high stress is important. Physical activity and exercise have been shown to mitigate neurobiological responses to acute stressors (Wunsch et al., 2019; Luger et al., 1987 Puterman et al., 2011; Rimmele et al, 2009, 2007; Traustadotóttir et al., 2005). There is also evidence that engaging in a single bout of exercise has immediate effects on reducing neurobiological reactivity to an acute psychosocial stressor (Zschucke et al., 2015; Wood et al., 2018; Caplin et al., 2021). However, much of this previous work did not investigate physiologically meaningful cut-points for demarcating the intensities at which their participants engaged, and thus the physiological underpinning of the effects of acute exercise at different intensities remains elusive. 1.1 Defining an Acute Stressor According to Epel and colleagues (2018), stressors are characterized by a set of characteristics, with the most prominent being its timeframe or duration. An acute stressor is specifically defined as an eliciting event that is processed cognitively and affectively, either consciously or unconsciously, that is time-bound and brief, and that exerts a physiological effect. Examples of such daily events are job interviews, public talks, or arguments with close friends or family members. The idea of acute stressors was first discussed in 1936, when pioneer stress researcher Hans Selye identified multifaceted physiological responses to “acute non-specific nocuous agents,” such as surgery, exercise, and element and cold/heat exposures (Selye, 1936). He labelled these nocuous physical agents as “stressors” and the body’s responses as “stress” (Selye, 3 1936). Selye determined these responses to be adaptive in nature, both effective and crucial for survival. In his work, Selye also highlighted that the physiological response to the stressors, while crucial, could also be detrimental during prolonged activation. In his later writings and research, Selye also suggested that psychological stressors can also activate similar physiological responses (Selye, 2013). In 1984, Lazarus and Folkman differentiated cognitive processes through which an acute psychological event is experienced as either a threat or challenge. When the individual appraises that they have the necessary social, behavioural, or psychological resources to manage the demands of the event, the event is appraised as a challenge. On the other hand, when the individual appraises that they do not have the necessary resources to manage the stressor, the event is appraised as a threat. According to Lazarus and Folkman (1984), depending on the appraisal as challenge or threat, the individual then consciously or unconsciously determines the coping strategy to employ to manage the stressor, as well as experiences an emotional reaction to the stressor that aligns with either threat or challenge. In 2000, Blascovich and Mendes (2000) introduced the Biopsychosocial Model of Threat and Challenge, integrating Selye’s (1936) physiological responses and Lazarus and Folkman’s (1984) psychological responses to an acute stressor into a unified theory. The Model posits that individuals undergo an immediate appraisal process when exposed to a goal-relevant physical or psychological acute stressor, which determines how they will respond to the stressor (Blascovich & Mendes, 2000). One of the Biopsychosocial Model principles argues that the evaluation of a stressor as either a challenge or threat triggers distinct physiological changes required to manage the stressor (Blascovich et al., 2004, Blascovich & Tomaka, 1996). A challenge appraisal triggers an increase in heart rate and ventricle contractility through the release of norepinephrine 4 and epinephrine. A threatened appraisal also activates the Hypothalamic-Pituitary-Adrenal (HPA) axis resulting in the release of cortisol, which in turn suppresses cardiac output and increases total peripheral resistance. These different, yet specific, physiological reactions highlight the important implications of the appraisal processes that follow a stressor and their impact on physiological responses to acute stressors. 1.2 Neurobiological Pathways The wide physiological changes that occur across the body when exposed to an acute stressor occur are the result of the activation of two neurobiological pathways (Godoy et al., 2018; McEwen, 2008), namely the sympathetic-adrenal-medullary pathway (SAM-p) and the hypothalamus-pituitary-adrenal axis (HPA-a). Activation of these pathways releases a cascade of hormones in support of effective management of the stressor. The SAM-p is the initial fast-acting physiological response, known for fight-or-flight, with the adrenal medulla releasing epinephrine (i.e., adrenaline) and norepinephrine (i.e., noradrenaline) into the bloodstream. Release of these catecholamines leads to increased heart rate, glycolysis (break down of glucose in the liver, and conversion to energy), decreased digestion, and elevated awareness (Cannon, 1929; McEwen, 2008). When a psychosocial stressor is experienced as threat, the HPA-axis is activated shortly after the activation of the SAM-p, releasing a cascade of hormones that result in amplified and longer-lasting physiological and behavioural responses (Godoy et al., 2018; McEwen, 2008; Sothmann et al., 1996). This response is initiated by the secretion of corticotropin-releasing hormone (Gunnar & Quevedo, 2006) which activates release of adrenocorticotropin from the anterior pituitary (Gunnar & Quevedo, 2006). Adrenocorticotropic hormone is released into the bloodstream and stimulates the release of the glucocorticoid cortisol from the adrenal cortex 5 (Gunnar & Quevedo, 2006; Sothmann et al., 1996). Cortisol plays an important role in the body as it is a primary regulator for a wide range of different physiological processes, including anti-inflammatory responses and metabolism of nutrients (Sothmann et al., 1996). One critical function of cortisol is to initiate gluconeogenesis, the conversion of proteins and lipids to glucose for energy (Del Corral et al., 1998). This incremental spike in cortisol can be captured through salivary cortisol measures which provide an accurate representation of unbound, active cortisol (Do et al., 2021; O’Connor & Corrigan, 1987). While the activation of the HPA-axis is necessary for energy release, it is equally as necessary to ensure that energy stores are not cleared too quickly and maintained upon continued presence or termination of the stressor. According to rodent studies (Gjerstad et al., 2018), there are two temporal negative feedback responses. One is a fast-acting response that happens within seconds or minutes that inhibits glucocorticoid release through suppressing CRH and adrenocorticotropin secretion from the paraventricular nuclei and anterior pituitary (Gjerstad et al., 2018). The other one is a delayed response, taking hours which also inhibits corticotropin-releasing hormone secretion in the paraventricular nuclei and suppresses proopiomelanocortin within the pituitary corticotrope cells (de Kloet et al., 2005; Gjerstad et al., 2018; McEwen, 2008). However, this built-in system designed to limit exposure to catabolic and immunosuppressive activity is not immune to dysregulation. Frequent exposure to acute stressors (i.e., daily exposures to interpersonal conflict, financial problems) or a severe stressor (e.g., natural disaster), or extended exposure to a continued stressor (e.g., caregiving) can lead to periods of elevated glucocorticoids that overwhelm the feedback loop (hypercortisolism). In addition to the potential inability to inhibit cortisol levels, frequent exposure can also have a 6 blunting effect limiting the production of cortisol altogether (hypocortisolism) resulting in a host of other health complications (Guilliams & Edwards, 2010). 1.3 Response Outcomes from Single Bout of Exercise Given the impact of a single psychosocial stressor, it is important to understand ways to effectively mitigate the psychological and physiological responses to the stressor. It has been well established that regular participation in exercise has a wide range of benefits on physical health (e.g., reduction in noncommunicable diseases like cardiovascular disease, diabetes, and all-cause mortality), as well as mental and cognitive health, and overall well-being (Pedersen & Saltin, 2015). The literature has also demonstrated that individuals who engage in habitual exercise or have higher fitness levels, when compared to less active and lower fit individuals, have mitigated stress responses, including physiological responses to acute psychological stressors (Puterman et al., 2011; Rimmele et al, 2009, 2007; Traustóttir et al., 2005). Research on the immediate effects of a single bout of exercise on a subsequent acute stress exposure is more limited. Based on the few pre-existing studies (see section 1.1.4), there is evidence to suggest that a single bout of exercise alters the HPA-axis stress response (Zschucke et al., 2015; Wood et al., 2018; Caplin et al., 2021). However, gaps still exist within the literature regarding the impacts of specific exercise intensities on the HPA-axis. To address this, there needs to be an understanding of HPA-axis reactivity to a single bout of exercise. 1.3.1 Neuroendocrine Response to Single Bout of Exercise A single bout of exercise can elicit a cortisol response, the degree to which is determined by intensity and duration of the bout (Duclos & Tabarin, 2010; Viru & Viru, 2004; Do et al., 2021; Athanasiou et al, 2023). Specifically, research indicates that a single bout of exercise activates release of cortisol when individuals surpass the first ventilatory threshold (VT1). Two 7 ventilatory thresholds exist. The first, VT1, corresponds to the first point when respiration rate increases in order to clear extra CO2 produced by the rise in lactate in the blood. VT1 is the transition point between moderate and vigorous intensity. The second ventilatory threshold (VT2), also known as the respiratory compensation point, is the second inflection point when minute ventilation increases at a greater rate for volume of exhaled CO2 and lactate level is at a maximum. VT2 is the transition from vigorous to very vigorous or severe intensity (Keir et al., 2018). There is evidence that changes in SAM-p and HPA-a responses to a single bout of exercise correspond to lactate levels and VTs (Schwarz & Kindermann, 1990; Urhausen et al., 1994; Chicharro et al., 1998). 1.4 Cross-stressor Adaptation Hypothesis As indicated earlier, Selye (1936) suggested that the body’s response to a demand (stressor) is specific in the sense that a global physiological response occurs as a reaction to the stressor; however, the stressor itself can be nonspecific. This idea of the specificity of a response and the non-specificity of an influencing agent is the foundation for the cross-stressor adaptation hypothesis (Sothmann et al., 1996). The cross-stressor adaptation hypothesis suggests that the neurobiological response to a homotypic stressor (the same type of stressor) is habituated to repeated exposure of the stressor, whereas heterotypic stressors (dissimilar in nature) can produce either habituated or more extreme responses after repeated exposure (Sothmann, 2006; Sothmann et al., 1996). The notion that neurobiological responses to homotypic stressors habituate with repeated exposure is attributed to the associative learning process which argues that the adaptation process contains a temporal component and occurs by associating two stimuli (classical conditioning) or associating a stimulus and response (operant conditioning) (McCarty et al., 1992). Heterotypic stressors, on 8 the other hand, are linked to non-associative learning: There is a neuroadaptation that occurs when exposed to repeated stimuli that are dissimilar in nature and therefore cannot be explained by classical or operant conditioning (Sothmann et al., 1996). 1.4.1 Cross-stressor adaptation through exercise training: Quasi-experiments Several studies have explored the effect of chronic exposure to exercise on neurobiological responses to acute psychosocial stressors in individuals using quasi-experimental designs (Puterman et al., 2011; Rimmele et al., 2007, 2009; Traustadóttir et al., 2005). One study compared trained elite sportsmen (athletes on national teams) to untrained men who completed less than 60 minutes of weekly activity and found that the trained elite athletes had lower neurobiological reactivity to a subsequent psychological stressor (lower cortisol and lower heart rate) compared to their untrained counterparts (Rimmele et al., 2007). In a follow-up study, Rimmele and colleagues (2009) compared elite athletes, amateur athletes, and untrained men and demonstrated that the elite sportsmen had the lowest cortisol and heart rate reactivities in response to the psychological stressor compared to the other two groups of men. Additionally, the amateur athletes had lower heart rate responses compared to the untrained men, but similar cortisol reactivity. Traustadóttir and colleagues (2005) compared the HPA-axis response to a series of cognitive challenges in twelve older (mean age = 66.6) fit women (those scoring above average on a VO2max test), fourteen older (mean age 66.3) unfit women (those scoring average or below average on a VO2max test) women, and 10 younger unfit women (mean age = 27.9). The older unfit group had significantly higher adrenocorticotropic hormone levels at baseline and throughout the trial compared to the other two groups (P < 0.01). The older unfit group also demonstrated significantly higher cortisol output during the cognitive tasks compared to the unfit 9 younger and fit older groups (P < 0.05). The results also indicated that older fit women had similar cortisol output to the younger unfit women suggesting that greater HPA-axis reactivity to psychological stressors may be associated with aging, but moderated by aerobic fitness levels (Traustadóttir et al., 2005). In general, these studies expose a trend suggesting that fitness and physical activity levels can play an important role in the reactivity to and recovery from psychological stressors. 1.4.2 Cross-stressor adaptation through exercise training: Randomized-controlled trials While the previous set of studies suggest that repeated exposure to exercise may mitigate elevated neurobiological responses to acute exposure to a psychosocial stressor, they remain quasi-experimental in nature, and thus causal, definitive conclusions cannot be drawn. To seek better clarity about whether the HPA-axis can be ‘trained’ to respond differently over time to a psychosocial stressor with repeated bouts of exercise, randomized controlled trials are required. Klaperski and colleagues (2014) exposed office workers to the Trier Social Stress Test-Group (TSST-G), followed by randomization to one of three groups: (1) an exercise intervention group, (2) a relaxation intervention group, or (3) a waitlist control group. Participants in the exercise condition were asked to complete aerobic moderate-intensity exercise for 60 minutes, two times/week for 12 weeks, whereas those in the relaxation condition were taught relaxation techniques like muscle relaxation, and breathing techniques, also completing two 60-minute sessions/week for 12 weeks. Both groups had one supervised-group session per week and one session was conducted individually. Control group participants did not receive any exercise or relaxation program. Following the 12 weeks, all participants were exposed to the TSST-G a second time. The results showed no significant group differences from baseline to completion of the trial in cortisol levels at the first sampling point (i.e., baseline values pre-stressor exposure) 10 of the TSST-G between all 3 groups. However, cortisol reactivity to the TSST-G between the exercise and the control group was significantly different (p = .01), but not between the exercise and relaxation groups (p = 0.44). With regard to within group analyses, both the exercise and the relaxation groups had significantly lower cortisol reactivity following their 12-week programs compared to pre-program initiation. 1.4.3 Cross-stressor adaptation through single bouts of exercise The pattern emerging from the literature suggests that those who partake in regular physical activity or have higher levels of physical fitness demonstrate a dampened response to psychological stressors. Fewer studies have investigated the effects of a single bout of exercise on a subsequent exposure to a psychosocial stressor. Zschucke and colleagues (2015) recruited 40 men [n=20 highly trained athletes; men who trained at high intensity 3+ times a week at an endurance sport) and n=20 sedentary men who did not exercise at all (< once per week)] and randomized them into either a stretching condition or a moderate-to-vigorous 30-minute treadmill walk/run at 60-70% their VO2max, corresponding to vigorous exercise prescriptions according to the American College of Sports Medicine (2018). Both groups then had a 90-minute recovery period before completing an adapted version of the Montreal Imaging Stress Task (MIST) while undergoing a functional magnetic resonance imaging (fMRI) scan. Saliva samples were also repeatedly taken to assess cortisol reactivity to the MIST. Their findings showed a significant reduction in the treatment group’s cortisol response to the MIST compared to the control group. The results also demonstrated an overall reduced cortisol response to the MIST in those with higher aerobic fitness levels (Zschucke et al., 2015). Another study conducted by Wood and colleagues (2018) examined the impact of a single bout of exercise before a subsequent psychosocial stressor with 164 female participants 11 (mean ± SD age: 20.1 ± 3.4 years). Seventy-five of the 164 (48.2%) females took part in a 1.9 mile (30-minute) moderate-intensity group urban walk prior to completing a group-based version of the TSST (TSST-G), whereas the other 89 participants were only exposed to the TSST-G. Wood and colleagues (2018) results did not demonstrate any significant differences in cortisol secretion between the exercise and control groups when exposed to the psychological stressor. Significant differences were seen between the “fit” participants compared to the “unfit” participants regardless of their randomization. “Fit” participants demonstrated a lower cortisol response to the subsequent psychological stressor. These results support the findings of quasi-experiments that explored the effects of habitual exercise on the reactivity to a psychological stressor but do not support the hypothesis that a single bout of exercise alters HPA reactivity (Rimmele et al., 2007, 2009; Traustadóttir et al., 2005). Wunsch and colleagues also explored the effects of acute and habitual exercise on reactivity to a psychosocial stressor (2019). The study included 84 male participants who were randomized into either completing a single bout of exercise or a control stretching condition. Fifty percent of the sample was classified as meeting recommendations for regular weekly physical activity (i.e., at least 150 mins of aerobic exercise per week during the previous 6-month period prior to the study) and the other 50% was considered inactive (i.e., reported no regular activity during the previous 6 months). Participants randomized to the exercise condition completed a 30-minute ergometer bicycling session at 70% of their individual maximal load capacity (Watts/kg established by a prior bicycle ergometer procedure). Those randomized to the stretching condition completed a 30-minute stretching session. Immediately following completed of either the bouts of exercise or stretching protocols, all participants completed the TSST-G and remained in the lab for an additional 65-minute recovery period (Wunsch et al., 2019). Salivary 12 cortisol samples were taken at seven-time points throughout the procedure (pre/post-exercise bout, pre/post-TSST-G, and 10, 30, and 65-minutes into recovery). Results indicated that both habitual exercise and a single bout of exercise were associated with different cortisol responses to the acute stressor. Specifically, habitual exercise was associated with a significantly less steep rise in cortisol and a significantly lower peak cortisol level when compared to the inactive participants. Participants who completed the single bout of exercise prior to the stressor, when compared to those who did not, also demonstrated a less steep rise in cortisol. Habitual exercise, however, did not moderate the effects of a single bout of exercise on cortisol release (Wunsch et al., 2019). In another study, Caplin and colleagues (2021) measured the cortisol response after exposure to varying intensities of a single bout of exercise and a subsequent acute psychological stressor induced 45 minutes later in a non-athlete population. The study was a randomized-controlled experimental design consisting of 83 untrained men, aged 21 on average, who were randomized into three separate exercise intensity groups: light (30% of their heart rate reserve; HRR), moderate (50% HRR), and vigorous (70% HRR). These designated intensities were established based on the American College of Sports Medicine guidelines (2018). Participants completed a single bout of exercise with their TSST 45 minutes post-exercise. Participants remained seated in the lab for 70 additional minutes post-stressor. Saliva was sampled before and after the TSST, and then 5 additional times following its completion. Their results demonstrated different trajectories of cortisol output in response to the TSST, dependent on the intensity of the exercise bout. Specifically, they demonstrated that completion of a single bout of vigorous exercise dampened the HPA-axis stress response when compared to lesser exercise intensities. When compared to the moderate and light exercise groups, those in the vigorous group had 13 comparatively higher levels of cortisol at entry to the TSST, showed dampened cortisol reactivity to the stressor, and had a faster return to baseline levels during the recovery period (Caplin et al., 2021). Additionally, when comparing the moderate and light groups, the moderate group demonstrated a significantly slower increase rate and deceleration (Caplin et al., 2021). 1.5 Limitations with the Literature While previous research has established that exercise at moderate and vigorous intensities may mitigate HPA reactivity to a subsequent psychosocial stressor, the studies have some important limitations that the current study seeks to address. Specifically, previous studies demarcated light, moderate, and vigorous intensities based on zones prescribed for exercise engagement from the Guidelines for Exercise Testing and Prescription Manual from the American College of Sports Medicine for the general population (2018). These intensity zones are drawn from percentages of heart rate reserve (Zuhl, 2020), which do not consider the variability found in individual metabolic boundaries (Gaskill, et al 2001). Percent of HRR or maximal load capacity does not represent true physiological demarcations of significant metabolic and functional changes within the cardiovascular and respiratory systems that occur during a bout of exercise. Demarcating intensity zones based on the ventilatory thresholds is mechanistically more precise, when considering previous research on the effects of VT1 on metabolic and functional changes, and in our case, cortisol release. 1.6 The Current Study The purpose of the current study is to build on the work by Caplin et al. (2021), by establishing intensity zones based on physiologically meaningful cut offs. Specifically, by using each participant’s ventilatory thresholds to determine individualized light, moderate, and vigorous intensity zones prior to randomization, we can better examine the effects of exercise 14 intensity on cortisol reactivity to and recovery from a subsequent laboratory induced psychosocial stressor. To achieve this, participants complete a modified maximal cardiopulmonary exercise test (CPET), adapted from the Bruce Protocol to determine their individual ventilatory thresholds. Participants then return to the lab within a 2 to 14-day window to complete the experiment, whereby they will be randomized to their intensity zone based on their CPET results. They will then complete a 30-minute bout of exercise, followed by a 45-minute recovery phase. Then, participants complete the Trier Social Stress Test, followed by a 70-minute recovery phase. During this second day experimental protocol, participants provide 12 saliva samples to help understand the impact of different intensities of a single bout of exercise on HPA reactivity to the TSST. 1.7 Hypothesis Hypothesis: Vigorous intensity, defined as the midway point between the first and second ventilatory thresholds, will dampen the cortisol trajectory following a subsequent psychological stressor, compared to moderate (defined as being 10% below the VT1) and light (set at 1.7km/hour) intensities. Moderate intensity will be evidenced to dampen the cortisol trajectory as compared to the light intensity, but to a lesser extent than vigorous. 15 Chapter 2: Methodology, Study Protocol, and Planned Analysis The methodology reported in this section encompasses the parent study, of which my thesis is a part. The purpose of this thesis is to examine the trajectory of cortisol reactivity to and recovery following a psychological stressor that follows a single bout of exercise at varying intensities. The parent study expands on prior evidence by investigating the extent to which a single bout of exercise, performed at light, moderate, or vigorous intensities based on individualized physiological indicators of intensity, alters varied cardiovascular, neurobiological, and psychological responses to a subsequently induced psychosocial stressor. The literature has established that a relationship can be found between the HPA-axis cortisol response to both exercise and psychological stress; specifically, fitness levels and single bouts of exercise at specific intensities can dampen the cortisol response to a psychological stressor (Caplin et al., 2021; Rimmele et al., 2007, 2009; Traustadóttir et al., 2005; Zschucke et al., 2015; Wunsch et al., 2019). However, why this relationship exists between exercise and stress regarding the cortisol response is unknown. The parent study is seeking to understand the extent to which blunted cortisol responses seen after exposure to a psychological stressor following a single bout of exercise at moderate or vigorous intensities are simply a function of the negative feedback loop (whereby cortisol release during exercise when VT1 is surpassed feeds back onto the HPA-axis thus suppressing further release), or are part of a larger array of psychological (i.e. affective, cognitive) and physiological (e.g. cardiovascular) changes that occur in tandem, as should be expected by the cross-stressor adaption hypothesis. 2.1 Participants 2.1.1 Inclusion Criteria 16 The target population is healthy adults between 18 and 35 years of age who are free of any heart or lung conditions that could impede their ability to perform vigorous exercise. We include both biological males and females in our sample compared to our previous study which only focused on males (Caplin et al., 2021). We seek to recruit 84 participants: 42 of each sex. Individuals who are currently undergoing hormone therapy or have lived experience as a transgender person will be included in the study as well, in attempts to broaden the research participation for populations that have been historically underrepresented. 2.1.2 Exclusion Criteria Potential participants with any medical condition affecting the heart or lungs, or that would prohibit their capacity to perform strenuous exercise, are excluded from the study. Individuals are also ineligible if they have been diagnosed with any psychiatric disorders, specifically if they are currently or have recently been treated with any exogenous glucocorticoids (i.e., medications that impact cortisol release) or have any chronic illnesses or are taking any medications that impact hormone production. Individuals currently using or have recently used the following medications, will be excluded: corticosteroids, beta-blockers, beta-2 agonists, psychotropics, angiotensin- converting enzyme inhibitors, ketoconazole, metyrapone (treatment of Cushing’s syndrome), or hormonal contraceptives. Females on hormonal contraceptives are excluded from the study due to the blunting of cortisol production with contraceptives (Kirschbaum, et al., 1999). However, a potential participant can be deemed eligible if they previously have stopped taking hormonal contraceptives for a minimum of 6 months prior to scheduling. Individuals suffering from substance abuse or addiction are excluded. Individuals who smoke (including vaping) are excluded from the study as well. Nicotine, like caffeine/alcohol, is 17 a stimulant and can interfere with the activation of the HPA-axis, and adrenocorticotropic hormone and cortisol production (Kirschbaum et al 1993; Rohleder & Kirschbaum, 2006). Persons are ineligible if they are classified as an elite athlete, defined by Swann, Moran, and Piggott as anyone participating in sport at the regional level or above (2015), due to the literature suggesting that elite athletes have a blunted response to the Tier Social Stress Test (Rimmele, et al., 2007). Finally, non-English speaking individuals unable to understand potential risks that may be associated with the study or who may not understand questionnaires and procedures will also be excluded from participating (see Appendix C). 2.1.3 Recruitment Recruitment occurs on the UBC Vancouver campus through posters, advertisement stands, and in-class recruitment (see Appendix A.1-A.4). Advertisements are also posted on social media pages that are directed to UBC staff and student audiences. All recruitment ads prompt potential participants to visit the study website at www.eaststudy.ca where they can get more information about the study, review basic eligibility criteria, and book a phone screener appointment with our study coordinator, if interested. Participants are compensated for their time with a fitness report from their Cardiopulmonary Exercise Test and either with an $80 cash stipend or 6.5 Human Study Pool (HSP) course credits that can be applied to UBC Psychology courses. The UBC Psychology HSP is a program connected to the UBC Psychology undergraduate program that provides an avenue for students to be introduced to the research process by offering opportunities for them to take part in studies while receiving psychology course credits for their commitment. EAST has a recruitment advertisement on the HSP SONA portal which is accessible to all psychology students. On the site, a summary of the study is provided, and individuals are directed to the 18 EAST website and prompted to book a phone screening consultation with our research staff. Based on the allotted time required to participate in the study as well as the fact that the study is in person and not online, our study provides 6.5 HSP credits to those who complete both days. 2.1.4 Screening Procedures All interested participants are required to complete an in-depth screening process (see Appendix C & K.4) to ensure interest and eligibility. In this phone interview, participants are asked demographic questions like age (in years), sex assigned at birth, gender identity, and lived experience as a trans person, as well as questions regarding their medical history and current health status. Participants are also asked to complete the Physical Activity Readiness Questionnaire (PAR-Q+; see Appendix K.4; Warburton et al., 2011), and if any of their responses to the screening criteria are positive, they are required to consult a medical provider to receive approval to engage in any intensity exercise as required by our study. The Leisure-Time Activity Categorical Item (L-CAT; Appendix K.5; Kiernan et al., 2013) is used to record their regular engagement in exercise in their everyday lives. All interested and eligible females are included in the study, though their participation in the experimental day is limited to when they are in their luteal phase of their menstrual cycle. 2.1.5 Menstrual Phase Identification Endocrinological differences between the sexes require consideration when examining how the body responds to physical and psychological stressors. One major difference between the sexes is the menstrual cycle (McNulty et al. 2020). Two of the main hormones that fluctuate throughout this cycle are estrogen and progesterone (Davis & Hackney 2017; Landgren et al., 1980; Owen, 1975). These fluctuations have been broken down into measurable and predictable phases within the cycle and are largely classified into the following three phases, the follicular 19 phase (low estrogen and progesterone), the ovulatory phase (high estrogen and low progesterone), and the mid-luteal phase (high estrogen and progesterone) (de Jonge et al., 2019). These hormonal fluctuations have interactive effects on a wide range of biological systems (Davis & Hackney, 2017; Chrousos et al, 1998; Ansdell et al., 2019), including physical performance (McNulty et al., 2020) and physiological responses to psychosocial stressors (Kirschbaum et al., 1999). In terms of exercise performance, McNulty et al.’s (2020) systematic review and meta-analysis demonstrated that there is a small decrease in performance when females are in their follicular phase compared to the luteal phase. In terms of reactivity to psychosocial stressors, females in their follicular phase have apparent blunting of their cortisol responses to the TSST, as compared to females in their luteal phase or compared to men (Kirschbaum et al., 1999). To account for this hormonal response, female participants of the study must be within their luteal phase during the time of the Day 2 session. This will be tracked via a self-report measure. When scheduled for their lab sessions, female participants will be asked to count backwards 14 days since their last onset of their menses and will be scheduled within that 14–28-day period. Participants are also asked to contact the study coordinator if they menstruate before their scheduled Day 2 visit, in which case it will be rescheduled for the following cycle. 2.2 Study Design This study is a true-experimental, between-group design with three levels, using stratified randomization to group 84 participants (42 females and 42 males) into one of the following three levels: light, moderate, and vigorous exercise. During their first assessment day, all participants undergo a modified maximal cardiopulmonary exercise test (CPET), adapted from the Bruce Protocol, to determine their ventilatory thresholds and establish their light, moderate, and 20 vigorous intensity zones. On a subsequent experimental day, participants are randomized to complete a single bout of exercise for 30 minutes in one of these zones, followed by the TSST 45 minutes later. The targeted outcome for this project is salivary cortisol. The study takes place over two in-lab sessions located at the Fitness, Aging, and STress (FAST) Lab on the campus of the University of British Columbia. These visits will be referred to as ‘Day 1’ and ‘Day 2’, during which participants are randomized to their respective condition. 2.3 Measures My thesis will exclusively examine salivary cortisol levels prior to and following the TSST as the outcome. Secondary outcomes measured during or following the TSST that are included in the parent study include blood pressure [monitored with a Spot Vital Signs® LXi Device (Masimo Set, Welch Allyn blood pressure cuff)], physiological indicators of threat and challenge (i.e., heart rate, heart rate variability, cardiac output, total peripheral resistance, monitored with the BIOPACTM), affective arousal (Felt Arousal Scale, see Appendix K.9; Svebak & Murgatroyd, 1985), affective valence (Feeling Scale, see Appendix K.10; Hardy & Rejeski, 1998), Negative and Positive Affect (PANAS-X, see Appendix K.11; Watson & Clark, 1994), and cognitive appraisals [i.e., appraisals of threat and challenge (see Appendix K.12; Mendes et al., 2007)], though will not be discussed since they do not pertain to my thesis. Only variables included in this master’s thesis will be reviewed in greater detail. 2.3.1 Primary Outcome Measure 2.3.1.1 Salivary Free Cortisol Salivary cortisol is the unbound, active, and functional portion of cortisol compared to blood cortisol (total bound and unbound cortisol; Bozovic et al., 2013; O’Connor & Corrigan, 1987) and is considered a reliable measure of the HPA-axis response to both acute psychological and physiological stressors while maintaining a reasonable correlation with free cortisol found in 21 blood (Bozovic et al., 2013; Vining et al., 1983). Salivary cortisol collection methods also provide numerous advantages compared to plasma and serum. For example, salivary cortisol collection methods (saturation of specialized material) provide a relatively stress-free, non-invasive way of collection, and does not require medical training or professionals to administer sample collection, making it the suggested cortisol measure method for stress research (Hellhammer et al., 2009). In the current study, twelve saliva samples are taken on Day 2 (prior to and following the exercise bout as well as the TSST) with the Sarstedt Salivette®, with the cotton Salivette placed into the participant’s mouth for 60 seconds, chewing for the entire duration before maneuvering it back into the tube and sealing the cap (see Figure 1). Once the participants samples are collected, they are stored at -20º C in the FAST Lab’s storage freezer before being shipped to Dresden, Germany for processing at Dresden Lab service, the laboratory of Clemens Kirschbaum for analyzing. More details about the timing of the samples are provided in Section 2.1.9. Saliva samples were stored and stored at -20 degrees Celsius in the Dresden lab until analysis. After thawing, samples were centrifuged at 3,000 rpm for 5 min, which resulted in a clear supernatant of low viscosity. Salivary concentrations were measured using commercially available chemiluminescence immunoassay with high sensitivity (Tecan - IBL International, Hamburg, Germany; catalogue number R62111). The intra- and inter-assay coefficients of variance were below 9% (2.0% and 3.0%, respectively), and determined acceptable. 22 Figure 1. Salivette® Instructions for use (Sarstedt, Rommeldsorf, Germany) 2.3.2 Secondary Outcome Measure 2.3.2.1 Physical and Mental Health Indicators To describe the sample, I report participants’ resting blood pressure on Day 2 prior to completing the exercise protocol. I also report the sample’s (and SD) perceived stress levels, as well as depressive symptomatology from the survey they completed on Day 2 prior to completing the exercise protocol. 2.3.2.2 Blood Pressure Blood pressure (BP) refers to the amount of force the heart requires to pump blood through the cardiovascular system and is measured via systolic (pressure in the arteries during the heartbeat) and diastolic (pressure in the arteries at rest/between heartbeats) (Centre for Disease Control and Prevention, 2021). According to the American College of Sport Medicine (2018) a healthy resting BP should read <120/80 which should be measured after a 5-minute restful sitting period where the back is supported in a chair, feet are planted on a hard surface, and arms are resting around the level of the heart (American College of Sports Medicine, 2018). Due to the nature of the maximal and submaximal exercise performances required during the lab visits, BP is recorded multiple times during the baseline protocol (see Figures 2 & 3) via the Spot Vital Signs® LXi Device (Masimo Set, Welch Allyn) and participants are ineligible to complete the exercise tasks if their resting BP is >140/90 to mitigate potential cardiac events (American College of Sports Medicine, 2018). 23 2.3.2.3 Depression Scale The Center for Epidemiologic Studies Short Depression Scale (see Appendix K.6; Eaton et al., 2004) is a 10-question scale with a 4-point rating system ranging from ‘rarely or none of the time’ to ‘all of the time’ and asks specific questions regarding how the individual has felt over the last week. Internal consistency among the items was high (Cronbach a=0.91). 2.3.2.4 Perceived Stress The Perceived Stress Scale (PSS, see Appendix K.7; Cohen et al., 1994) is a 10-item scale designed to measure how individuals are currently appraising life events as stressful by capturing their experiences from the previous month. Internal consistency among the items was good (Cronbach a=0.82). 2.4 Procedures 2.4.1 First Assessment Day (Day 1). Day 1 takes approximately 90 minutes to complete and is held between the hours of 9 am and 12 pm. This assessment day includes a verbal review and signing of the consent form (see Appendix E), assessment of demographic factors via a QualtricsTM survey (see Appendix K.1-K.4), as well as anthropometrics (height in meters, weight in kilograms), systolic and diastolic blood pressure, and heart rate are measured. Participants are fitted with the BiopacTM and 3 sensors for measurement of continuous heart rate. Participants are familiarized with the Rating of Perceived Exertion (see Appendix K.8; Borg, 1985), Felt Arousal Scale (see Appendix K.9; Svebak & Murgatroyd, 1985), and Feeling Scale (see Appendix K. 10; Hardy & Rejeski, 1989). Participants are then fitted with the mask for gas exchange analysis with the COSMED metabolic cart. Following this, a modified maximal cardiopulmonary exercise test (CPET), adapted from the Bruce Protocol, is completed. The modified Bruce Protocol has been determined as a highly 24 reliable and valid indicator of cardiorespiratory fitness (Kaminsky & Whaley, 1998). The modified Bruce Protocol lasts a maximum of 21 minutes and involves incrementally increasing speed and grade every 20 seconds. The original Bruce Protocol had an incremental ramp increase every three minutes allowing measurement of overall cardiorespiratory fitness. However, it did not allow for measurement of specific incremental changes in gas exchange that occurs at a more granular level. Thus, the modified version of the Bruce Protocol (BSU Bruce Ramp Protocol, Kaminksy and Whaley, 1998) was created in 1998 to predict maximal oxygen uptake and the ventilatory thresholds (Kaminksy and Whaley, 1998). The Feeling Scale (Hardy & Rejeski, 1998), Felt Arousal Scale (Svebak & Murgatroyd, 1985), and the Rating of Perceived Exertion (Borg, 1985) are asked every 2 minutes during the test, until completion. Once the test is complete, an exercise team member removes the VO2 mask, and the participants start a 3-minute walking recovery period. Once completed, participants continue the recovery period seated, have their blood pressure taken three times to ensure proper recovery is taking place, and are asked the Feeling Scale (Hardy & Rejeski, 1998) and Felt Arousal Scale (Svebak & Murgatroyd, 1985) one final time before finishing the study day. All equipment is removed at this point. See Figure 2, for the first assessment day timeline and see Appendix I.1 for the Day 1 data collection sheet. 25 Figure 2: First Assessment Day Timeline 2.4.3 Cardiopulmonary Exercise Test (CPET) – Prescribed Intensities via Ventilatory Thresholds COSMED Quark CPET software produces a Breath-by-Breath analysis generating multiple visuals, including graphs comparing VO2 (mL/min) with VCO2 (mL/min). The VO2max is autodetected by the computer generated VO2 max graph, confirmed by the team. While VT1 and VT2 boundaries are also autodetected by the software, it is confirmed by a team member in the lab. Each participant’s individual boundaries are referenced when determining the intensity at which they will exercise, when randomized to either one of the three conditions: light (LICT), moderate (MICT), and vigorous intensity continuous training (VICT). If randomized into the LITC, participants stay at a 1.7 mph walking pace with no incline. If randomized to MICT, participants maintain a ventilation rate on Day 2 that corresponds to just below 10% of their oxygen uptake of their VT1 determined on Day 1, and VICT will maintain a ventilation rate on Day 2 midway between their VT1 and VT2 determined on Day 1. Since this process does rely 26 on human input, the team has done multiple checks to limit potential bias when finding these points. 2.4.3 Second Assessment Day (Day 2): Experimental Procedures Day 2 – the experimental day – takes approximately 270 minutes to complete and is held between the hours of 12 pm and 5 pm. The Day 2 protocol includes a 30-minute bout of exercise completed at one of the randomized intensity levels, based on their Day 1 results for their ventilatory thresholds. The exercise bout is followed by a post-exercise recovery period of 45 minutes, a psychosocial stress task (the Trier Social Stress Test; see Appendix J; Kirschbaum et al., 1993), and a post-stressor recovery period of 70 minutes. The two days of assessments must take place between 48 hours and 14 days of each other. Please refer to Figure 3 for the Day 2 timeline breakdown and Appendix I.2 for protocols. 2.4.3.1 Screening and Consent Upon arrival to the lab on Day 2, participants review the consent form (see Appendix E) with study personnel and complete a series of preliminary questionnaires (see Appendix K.2-K.4) to ensure no changes in their health status or capacity to exercise have occurred. Following that, participants are asked to complete the following questionnaires: the Centre for Epidemiologic Studies Depression Revised Scale (see Appendix K.6; Eaton et al., 2004), and the Perceived Stress Scale (see Appendix K.7; Cohen et al., 1983). Participants are also refamiliarized with the Rating of Perceived Exertion Scale (Borg, 1985). Felt Arousal Scale (Svebak & Murgatroyd, 1985), and the Feeling Scale (Hardy & Rejeski, 1989). Following this, participants change into activewear and are fitted with the Biopac, with a total of 11 electrodes (see Appendix M.2) to measure heart rate, heart rate variability, cardiac output, and total peripherical resistance (for physiological assessment of threat and challenge; Blascovitch & 27 Mendes, 2000). Height and weight are recorded again, and participants are fitted with the mask for gas exchange analysis during the exercise protocol using the COSMED metabolic cart. 2.4.3.2 Baseline Once participants are fitted, the baseline period consists of a 3-minute standing period, a 10-minute seated portion, and an additional 3-minute standing period during which multiple blood pressures are recorded and participants answer the Feeling Scale (Hardy & Rejeski, 1998), Felt Arousal Scale (Svebak & Murgatroyd, 1985), and an adapted version of the Positive and Negative Affect Scale (see Appendix K.12; Watson, & Clark, 1994). 2.4.3.3.Single Bout of Exercise Following the 16-minute baseline period, the first saliva sample is taken. Then, participants are informed to which exercise intensity condition (light, moderate, or vigorous) they are randomized (see Appendix L.2) and complete the Feeling Scale (Hardy & Rejeski, 1998), Felt Arousal Scale (Svebak & Murgatroyd, 1985), an adapted version of the Positive and Negative Affect Scale (Watson, & Clark, 1994), and the Threat and Challenge Appraisals Scale (see Appendix K.12; Mendes et al., 2007). Blood pressure is also taken during this time. To establish the individualized targeted VO2 at which they perform their 30-minute exercise activity, information on the ventilatory thresholds from Day 1’s maximal CPET are utilized. If randomized into the LICT, participants stay at a 1.7 mph walking pace for 30 minutes with no incline. In consultation with exercise physiologist, Dr. Stacey Hutton, we have determined that MICT can be set at 10% below their oxygen uptake of their VT1. If the participants are randomized into the MICT condition, the first five minutes is a period of slow incremental adjustment to their targeted VO2 and participants hold that level of intensity for the remaining 25 minutes. Targeted VO2 for VICT is prescribed midway between their VT1 and 28 VT2, and participants are slowly increased to this intensity for the first seven minutes, and then continue at this intensity for 23 minutes, totaling 30 minutes. During the ramp up portions for the MICT and VICT groups, the treadmill is set to an incline of 6.5% and a team member gradually increases the speed to hit the predetermined threshold numbers at the appropriate time (5 or 7 minutes for MICT and VICT, respectively). Since there are expected fluctuations within the ventilatory output of every individual, they are kept within a 5% range on either side of their ideal threshold. To keep them within that determined zone, the team member adjusts the speed of the treadmill as needed. During the duration of the exercise component, another team member is stationed beside the treadmill monitoring the participants and asks participants to respond to the Feeling Scale (Hardy & Rejeski, 1998), Felt Arousal Scale (Svebak & Murgatroyd, 1985), and Rating of Perceived Exertion (Borg, 1985) every two minutes for the duration of the exercise. For more details regarding exercise protocol see Appendix L.2. 2.4.3.3 Exercise Recovery As soon as the exercise protocol is finished, participants enter the 45-minute exercise recovery phase. Participants start a 3-minute standing cooldown during which the VO2 mask stays on and two blood pressure readings are taken a minute apart. At this time, they repeat the Feeling Scale (Hardy & Rejeski, 1998), Felt Arousal Scale (Svebak & Murgatroyd, 1985), Rating of Perceived Exertion (Borg, 1985), an adapted version of the Positive Affect and Negative Affect Scale (Watson, & Clark, 1994), and Threat and Challenge Appraisals Scale (Mendes et al., 2007). Once complete, the mask is removed, and they begin a 2-minute walking recovery period and take their second saliva sample. After the treadmill cooldown, participants start a 40-minute seated portion of the 45-minute exercise recovery period. They are given the option to change out of their active wear if needed before watching the first few episodes of 29 Season 4 “How It’s Made” on YouTubeTM. The third saliva sample is taken 20 minutes into the recovery period, preceded by the Feeling Scale (Hardy & Rejeski, 1998) and Felt Arousal Scale (Svebak & Murgatroyd, 1985). The fourth cortisol sample takes place 45 minutes following the completion of the exercise bout, preceded by the Feeling Scale (Hardy & Rejeski, 1998) and Felt Arousal Scale (Svebak & Murgatroyd, 1985). Once the exercise-recovery period is complete, participants start the pre-TSST procedures and provide their fifth saliva before starting the Trier Social Stress Test (TSST) protocol. 2.4.3.5 Psychosocial Stress Task: The Trier Social Stress Test The TSST is considered the gold standard psychosocial stressor to induce physiological stress, due to its ability to raise heart rate, blood pressure, and cortisol levels and produce a threatened response (Allen et al., 2017; Kirschbaum et al., 1993). The TSST includes both social-evaluative threat and uncontrollability components which are crucial for a significant cortisol response (Dickerson and Kemeny, 2004). Upon starting the protocol, participants are given the instructions of the task by a team member, who explains that they will be completing a speech to a panel of judges in the role of the interviewee in a fake job interview. Participants are also informed of a second task that will be given to them by the panel after the completion of the interview. Participants are informed that the judges are trained in behavioural analysis and will take notes and are told that their session will be video recorded for later voice and behaviour analysis. This multi-method approach of pairing an audience-based-evaluative performance (interviewing in front of a panel of judges) with a recorded component (i.e., video camera) is argued to provide larger effect sizes for cortisol response compared to other variations of the 30 TSST protocol where peer-evaluations and recordings were absent and targets the critical social evaluative threat variable (Dickerson & Kemeny, 2004). Once the instructions are delivered, participants transition into a 5-minute preparation period where they stand behind the podium equipped with a pen and paper. During this preparation phase participants complete another blood pressure measurement, the Threat and Challenge Appraisals Scale (Mendes et al., 2007) and provide their sixth saliva sample. Following this, participants complete the 5-minute mock-interview with a panel of judges and a 5-minute mental arithmetic task before providing their seventh saliva sample. The total TSST time is 17 minutes. For a review of the timing and completion of different questionnaires throughout and following the TSST, please see Figure 3 and Appendix J. 2.4.3.6 Psychosocial Stress Task Recovery Following the completion of the TSST, participants then enter the stress-recovery period that lasts 70 minutes, during which the participants are seated and continue to watch “How It’s Made.” Ten minutes into recovery they are asked to complete the Thoughts questionnaire (see Appendix K.13) and provide their eighth cortisol sample. The following four saliva samples (9-12) are taken at 20-, 30-, 50-, and 70- minutes post-TSST completion. 2.4.3.7 Debrief Protocol Upon completion of the Day 2 protocol participants are informed about the purpose of the TSST, an explanation for its deception, and a breakdown of the specific measures that were taken (see Appendix F and G). Participants are then introduced to the TSST judges, who are no longer wearing the white lab coats and are more affable. Participants are debriefed on the purpose of the study, as well as provided UBC Counseling service information before reviewing their CPET results from their first lab visit. 31 2.4.3.8 Cash Honorarium Protocol Participants who sign up for the cash reimbursement are given the prorated $20 for the completion of the first visit and $60 for the completion of the second visit ($80 total). A receipt is created for both the participant and the research team (see Appendix H.1). 2.4.3.9 HSP Honorarium Protocol Participants who sign up for the study through HSP receive their first 1.5 credits immediately following the completion of their Day 1 (see Appendix H.2), and then receive the remaining 5 credits during the debrief protocol (see Appendix G.2). Participants are also given a Take-home Debrief Form (see Appendix G.3) that outlines the study parameters. Participants will also need to complete a brief online review worksheet (see Appendix G.2) as per SONA requirements. 2.4.4 Additional Protocol Information During the Day 1 lab visit, participants can drink water throughout the session, apart from when wearing the VO2 mask. Day 2 does include a water restriction, which participants are made aware of in the consent form, due to the collection of cortisol saliva samples. The water restriction occurs as soon as baseline begins but they are provided a couple of ounces of water immediately following their second saliva sample (after the exercise) and once more after their third saliva sample. This is to control for the potential dilution of the sample. However, in the case of an emergency, water is not withheld from the participant. During both visits, but especially the second visit, participants are prohibited from bringing their phones and any exercise gadgets/technology into the main area of the lab once the protocols begin. They are asked to leave all devices including smart watches (many of which have a texting function) with their belongings in the greeting room. 32 Figure 3: Second Assessment Day Timeline 2.4.5 Mitigating Risks The associated risks in this study are primarily regarding the maximal and submaximal exercise testing. However, the risks that have been reported by the American College of Sports Medicine are associated with clinical populations, thus the risk for healthy individuals is significantly lower (Goodman et al., 2011). Our screening protocol has been designed to control for this with a thorough list of health questions accompanied by the PARQ+ (see Appendix C, K.2, and K.4; exclusion criteria, section 2.1.2). In addition to the screening process, the protocol does contain multiple safety precautions to minimize all potential risks. Participants are never left in the lab alone; a team member is always present to monitor their physical states. At least one (if not all) scheduled research team members is certified in cardiopulmonary resuscitation (CPR) and automated external defibrillator (AED) use and the lab is equipped with a first aid kit and working AED and stethoscope. The treadmill and COSMED cart are both equipped with an emergency stop button and an emergency 33 stop string is always attached to the participant when on the moving treadmill. Participants’ heart rates are closely monitored throughout the visit via the BIOPAC ECG transmitter on the BIOPAC computer stationed in the lab. Participants who are randomized into the vigorous intensity group maintain a threshold halfway between their VT1 and VT2; however, if a participant is not able to sustain this threshold for the duration of the CPET protocol, they can be dropped to just above their VT1 threshold and still be considered vigorous intensity. 34 Chapter 3: Data Analytic Approach 3.1 Power Calculations Given that our primary goal is to examine the differences between vigorous intensity exercise with both moderate and light exercise, we calculated the effect sizes for cortisol differences from Caplin et al., 2021 for each time point following the TSST (15, 25, 35, 45, 65, 85 minutes following the start of the TSST) for VICT compared to MICT and VICT compared to LICT. We then averaged the effect sizes (0.67) and calculated the required sample size based on this estimated effect size using Optimal design (Duan et al., 2015). Based on these data, as well as 80% power, alpha = 0.05, a required sample size of 74 was required for the parent study. As we also expect to covary for the effect of one additional factor (i.e., sex) in the parent study, we added ten more participants to the sample size (i.e., N = 84). These analyses, surrounding sex differences, will not be investigated due to the limitations surrounding the current thesis sample size at the time of analysis (see section 4.1). 3.2 Statistical Analysis Descriptive statistics for demographic factors and baseline physical fitness (i.e., VO2, VT1, VT2, body mass index, resting blood pressure, etc.) were reported for all participants, as either mean and standard deviation for all continuous variables or number and percent of the sample for all categorical variables. Growth curve models with restricted maximum likelihood were developed to examine trajectories of cortisol reactivity to and recovery from the TSST. Growth curve models can estimate multiple components of the stress response including the estimated intercept, slope, and curvature to provide a more nuanced examination of the estimated baseline, reactivity, peak, and recovery of the cortisol response to the TSST following different intensities of exercise. 35 Multilevel models, including growth curve models, also allow for robust handling of skewed and missing data (Raudenbush, 2002; Singer, et al., 2003). First, an unconditional means model (i.e., no predictors in the model) was created to determine the proportion of unexplained within- and between-person variance in cortisol output (See Equation 2). 𝐶𝑜𝑟𝑡𝑖𝑠𝑜𝑙!"#$!! = b& + 𝜀 (Eq. 3) Second, a Level-1 growth model including the fixed effect of time and time-squared (see Equation 3) was analyzed to test the shape of the cortisol response as a function of time across all participants. 𝐶𝑜𝑟𝑡𝑖𝑠𝑜𝑙!"#$!! = b& + b' ∗ 𝑇𝑖𝑚𝑒 + b( ∗ 𝑇𝑖𝑚𝑒( + 𝜀 (Eq. 4) Third, a Level-2 growth model was used to determine direct effects of exercise groups, with the vigorous intensity exercise group as the referent. 𝐶𝑜𝑟𝑡𝑖𝑠𝑜𝑙!"#$!! = b& + b'(𝑇𝑖𝑚𝑒) + b((𝑇𝑖𝑚𝑒() + b)(𝐿𝐼𝐶𝑇) + b*(𝑀𝐼𝐶𝑇) + b+(𝑇𝑖𝑚𝑒 ∗ 𝐿𝐼𝐶𝑇) + b,(𝐿𝐼𝐶𝑇 ∗ 𝑇𝑖𝑚𝑒() + b-(𝑀𝐼𝐶𝑇 ∗ 𝑇𝑖𝑚𝑒) + b.(𝑀𝐼𝐶𝑇 ∗ 𝑇𝑖𝑚𝑒() + 𝜀 (Eq. 5) The estimated time of cortisol secretion peaks were determined for all three groups, with the following equation. 𝐶𝑜𝑟𝑡𝑖𝑠𝑜𝑙 /$01 = −𝛽!234$(2 ∗ 𝛽56#7$) (Eq. 6) Primary analysis compares the vigorous group’s estimated intercept, slope, and curvature to both the light and moderate groups, with a follow-up analysis comparing the moderate to the 36 light and vigorous groups’ estimates to determine the effects between the moderate and light group only. All analyses and visualizations were created with R statistical software version (Version 4.3.2; R Core Team, 2023). Data cleaning and figures used functions from the tidyverse suite of packages (Wickham et al., 2019) as well as the wesanderson (Ram & Wickham, 2023) and ggpubr packages (Kassambara, 2023). Multilevel models were created and interpreted with lme4 (Bates et al., 2015), lmerTest (Kuznetsova et al., 2017), broom.mixed (Bolker & Robinson, 2022) and performance (Lüdecke et al., 2021) packages. Two dummy variables were created from the three conditions, with vigorous-intensity continuous training (VICT) as the referent group to compare the moderate (MICT) and light (LICT) conditions to the vigorous group. A follow-up analysis was completed in which light condition was the referent group to test for any differences with the moderate group. 3.3 Hypotheses A review of the hypotheses are as follows: Vigorous intensity will significantly dampen the cortisol trajectory following a subsequent psychological stressor, compared to moderate and light intensities. Moderate intensity will be evidenced to dampen the cortisol trajectory as compared to the light intensity, but to a lesser extent than vigorous intensity. 37 Chapter 4: Results The data presented in this section are part of an ongoing parent study examining the physiological (e.g., cortisol, heart rate, blood pressure, cardiac output, total peripheral resistance) and psychological responses (e.g., affect, cognitive appraisal) to a stressor. However, these analyses are outside the scope of this thesis. The data presented in this section include only cortisol reactivity to and recovery from the induction of a psychological stressor following a single bout of exercise at different intensities. 4.1 Participants At the time of analysis, 42 of the parent study’s 84 participants’ saliva samples were collected and assayed for cortisol concentrations. Mean age of the sample was 23.95 (SD = 4.85), with the majority being males (62%, n = 26). Self-reported ethnic origin was collected: 52.4% of the sample was of Asian descent (n = 22); an additional 19% (n = 8) was Caucasian; 7.14% (n = 3) of Middle Eastern descent; 4.76% (n = 2) of South East Asian descent; 2.38% (n = 1) of each of the following: Latin American descent, of South Asian descent, African-American descent, Eurasian descent, Middle Eastern/Caucasian descent, South American descent; and 2.38% (n = 1) unidentified. Participants were randomized via stratified randomization into three exercise groups: light (LICT; n = 9), moderate (MICT; n = 16), and vigorous (VICT; n = 17). The light group consisted of 67% (n=6) Asian participants, and 11% (n=1) Caucasian, South-East Asian, and Eurasian. The moderate group consisted of 56% (n=9) Asian participants, 13% (n=2) Caucasian, 0.06% (n=1) South-East Asian, Latin American, South Asian, South American and one undisclosed participant. The vigorous group consisted of 41% (n=7) Asian participants, 29% (n=5) Caucasian, 18% (n=3) Middle Eastern, 0.06% (n=1) African American and Middle 38 Easter/Caucasian. Twenty-six participants reported being undergraduate students, 13 reported being graduate students, one reported being employed and one was undisclosed. Table 1 presents means and standard deviations for BMI, VO2max, VT1, and VT2, CES-D, PSS, L-CAT, resting blood pressure (systolic and diastolic), room temperature and humidity in each group. All participants completed data collection between the hours of 12 pm and 5:30 pm at the University of British Columbia in the Fitness, Aging, and Stress Lab. Table 1: Demographic Statistics Overall (N = 42) LICT (N = 9) MICT (N = 16) VICT (N=17) p BMI 23.54 (3.73) 23.95 (3.37) 24.88 (4.55) 22.09 (2.53) ns VT1 20.27 (3.05) 20.99 (1.90) 19.34 (2.96) 20.77 (3.52) ns VT2 35.08 (7.04) 36.72 (8.33) 31.42 (5.37) 37.65 (6.60) < 0.05A VO2Max 39.43 (7.91) 41.00 (8.23) 35.71 (6.34) 42.11 (8.13) 0.05A Depressive Symptoms 11.68 (9.93) 12.74 (14.48) 11.06 (7.26) 11.71 (9.86) ns Psychological Stress 15.69 (5.50) 15.78 (7.22) 16.81 (5.26) 14.59 (4.80) ns Leisure Time Physical Activity 3.74 (1.15) 4.11 (1.05) 3.25 (1.13) 4 (1.12) ns Resting Systolic BP 108.16 (10.93) 111.56 (11.38) 109.75 (10.29) 105.06 (11.15) ns Resting Diastolic BP 64.71 (6.42) 66.44 (9.33) 65.28 (5.92) 63.35 (5.33) ns Room Temperature 26.5 (2.9) 26.34 (2.96) 25.35 (1.21) 27.73 (3.65) ns Room Humidity 24.98 (7.1) 24.75 (5.87) 26.81 (8.85) 23.25 (5.46) ns Note: All categorical data were analyzed via Chi-squared. A An Omnibus ANOVA found that there were between-group differences. Tukey’s HSD suggested that there was a significant difference between the VICT and MICT group, but no significant difference between either group and LICT. ANOVAs revealed there were no significant between group differences in BMI (MBMI = 23.54, SD = 3.73), the first ventilatory threshold (MVT1 = 20.27, SD = 3.05), resting systolic and diastolic blood pressure (MSYS = 108.16, SD = 10.93; MDIA = 64.71, SD = 6.42), depressive symptoms (M= 11.68, SD= 9.93), perceptions of psychological stress (M = 15.69, SD = 5.50), or 39 leisure time physical activity (M = 3.74, SD = 1.15). However, there were differences between the VICT and MICT group for age (MMICT = 26.50, SDMICT = 5.70; MVICT = 22.24, SDVICT = 3.63), ventilatory threshold 2 (MMICT = 31.42, SDMICT = 5.37; MVICT = 37.65, SDVICT = 6.60), and VO2max (MMICT = 35.71, SDMICT = 6.34; MVICT = 42.11, SDVICT = 8.13). See Table 1 for the group, sample means, and standard deviations. 4.2 Protocol Adherence All participants completed their exercise intensity as prescribed. However, one participant in the moderate group had to be held at an average speed of 1.4 mph at a 6.5% to stay within their prescribed threshold. 4.3 Salivary Cortisol Assays Descriptive statistics (Mean and SD) for cortisol concentrations prior to and following the TSST are presented in Table 2. Table 2: Cortisol Descriptive Statistics Time Overall Light Moderate Vigorous 0 2.19 (0.93) 2.19 (0.82) 1.5 (0.49) 2.83 (0.87) 5 2.11 (0.88) 2.18 (0.78) 1.44 (0.41) 2.7 (0.82) 20 3.66 (2.08) 5.21 (3.18) 2.79 (1.14) 3.65 (1.64) 30 5.34 (3.55) 7.81 (4.46) 5.47 (3.68) 3.91 (2.02) 40 4.22 (2.55) 6.5 (3.37) 4.13 (2.37) 3.11 (1.2) 50 3.42 (2.07) 5.31 (3.24) 3.15 (1.47) 2.69 (1.02) 70 2.7 (1.35) 3.57 (1.49) 2.46 (0.88) 2.47 (1.52) 90 2.24 (1.07) 2.92 (1.25) 2.05 (0.77) 2.07 (1.13) 4.5 Unconditional Means Model An unconditional means model (i.e., no predictors in the model) was created to determine the proportion of unexplained within- and between-person variance cortisol output (See Equation 1). The interclass correlation for the unconditional means model (see Table 3) was 0.354, 40 suggesting that 35% of the variance can be explained by differences between individuals and 65% occurred at the within-person level. Table 3: Unconditional Means Model Unconditional Level-1: Time(2) Level-2: Full Model Rand: Intercept (SD) 0.34 0.35 0.33 Rand: Residual (SD) 0.46 0.37 0.33 R2 Conditional 0.35 0.57 0.66 R2 Marginal 0.00 0.19 0.34 Chi squared to prev. model * * Notes: Unconditional, Level-1 (time-only), and Level-2 (including covariates) Growth Model Comparisons *p<.001 4.6 Level-1 Growth Model; cortisol response as a function of time The Level-1 growth model includes the fixed effect of time and time-squared (see section 3.2, Equation 4). The trajectory of cortisol over time followed a curvilinear shape. The estimated intercept for log-transformed cortisol at baseline was 0.68 (SE = 0.069, 95%CI = [.543, 0.817], p < 0.05 ), with an estimated rate of increase at 0.027 (SE = 0.002, 95%CI = [0.023, 0.032], p < 0.05 ) per minute, and an estimated curvature of -0.0003 (SE = 2.5e-5 , 95%CI = [-0.0004, -0.0003], p = 0.05). A chi-square test suggested that this model had better fit than the unconditional growth model (χ = 120.91, df = 2, p < 0.05). Compared to the residuals of the unconditional means model, this model reduced variance by 19%. 4.7 Level-2 Growth Model; Cortisol response between groups The final model added the direct effects of exercise groups (2 dummy coded variables with VICT as the referent group), in addition to the interactions for groups with time, as well as groups with time2. Figure 4 depicts the estimated trajectories for each group, utilizing each group’s estimated intercepts, slopes, and curvatures. The model accounted for 11% additional variance compared to the conditional model with just time and time-squared included. As seen in 41 Table 4, the estimated log-cortisol intercept for VICT (b = 1.01) was significantly larger than for MICT (b = -0.688, SE = 0.143, CI = [-0.9734, -0.402], p < 0.001), but did not show a significant difference when compared to LICT (b = -0.302, SE = 0.169, CI = [-0.641, 0.036], p = 0.079). Estimated log-cortisol intercept for MICT was significantly different from LICT (b = 0.39, SE = 0.17, CI = [0.0435, 0.7273], p<0.05). The rate of increase in cortisol for VICT (b = 0.007, SE =0.003, CI = [0.001, 0.014], p= 0.028) was significantly slower compared to both LICT (b = 0.036, SE =0.006, CI = [0.025, 0.047], p<0.001), and MICT (b = 0.032, SE =0.005, CI = [0.023, 0.041], p<0.001). VICT (b = -0.0001, SE =0.0004, CI = [-0.0002, -0.0001], p<0.001) also demonstrated a significant deceleration compared to LICT (b = -.00003, SE = 0.0001, CI = [-0.0004, -0.0002], p<0.001) and MICT (b = -0.0003, SE = 0.0001, CI = [-0.0004, -0.0002], p< 0.001). Results indicated that MICT and LICT did not have significantly different slopes (b = 0.004, SE =0.006, CI = [-0.008, 0.015], p= 0.525). MICT did not have a significant deceleration compared to LICT (b = 0.000, SE = 0.0001, CI = [-0.0002, -0.0001], p = 0.431). See Figure 4 for individual and group trajectories. The estimated times at which cortisol peaked were calculated for each group (see section 3.2, Equation 6), and were as follows: LICT = 46.53, MICT = 47.53, and VICT = 26.85 minutes. 42 Figure 4: Group and Individual Trajectories of Cortisol for Low, Moderate, and High Intensity Groups Note: Bold line indicates the average trajectory per group, while the thin lines represent each individual within the group. Table 4: Coefficients with Alternate Group Coding (i.e., different referent groups) Model 3: Low Comp Model 3: Mod Comp Model 3: Vig Comp Intercept 0.70 (0.14)* 0.32 (0.1)* 1.01 (0.1)* Time 0.04 (0.005)* 0.04 (0.003)* 0.01 (0.003)* Time2 -0.00046 (0.00005)* -0.00041 (0.00004)* -0.00013 (0.00004)* Low 0.39 (0.17)* -0.30 (0.17) Mod -0.39 (0.17)* -0.688 (0.14)* Vig 0.302 (0.17) 0.688 (0.14)* Low*Time 0.004 (0.01) 0.036 (0.01)* Low*Time2 -0.00005 (0.0001) -0.00033 (0.0001)* Mod*Time -0.004 (0.01) 0.032 (0.005)* Mod*Time2 0.00005 (0.0001) -0.00028 (0.0001)* Vig*Time -0.036 (0.006)* -0.032 (0.005)* Vig*Time2 0.00033 (0.0001)* 0.28 001)* 43 Note: “Low Comparison” compares both moderate and vigorous. “Moderate Comparison” compares both the Low and Vigorous group to the Moderate group. “Vigorous Comparison” compares those in the low and moderate groups to those in the Vigorous. * p < 0.05 4.8 Missing Data One participant was missing the temperature and humidity readings from their visit. Two participants missed a single item on the PSS, and one participant missed a single item on the CES-D. We used random-forest imputation to estimate these values for baseline demographics. 44 Chapter 5: Discussion In this thesis, I aimed to examine the effects of varying intensities of a single bout of exercise on cortisol trajectories following exposure to a subsequent psychological stressor. The prescription of the intensities for the bouts of exercise were marked by physiologically meaningful cut-points for each participant. Although the analyses were completed on half the required sample size based on the power analysis for the parent study, significant effects were observed. In support of the first hypothesis, a growth curve model analysis demonstrated that participants who exercised at vigorous intensities had a dampened cortisol response to the TSST, such that their rise was slower and, consequently, their deceleration was slower when compared to participants in the moderate and light intensity groups. On the other hand, and in contrast to the second hypothesis, no differences in cortisol trajectories were observed between the light and moderate groups, apart from the fact that those in the moderate group had lower estimated starting points prior to starting the TSST. Thus, these results partially support our hypotheses. 5.1 Glucocorticoid Negative Feedback Loop Previous studies have shown mixed findings when individuals completed a stressor task following either a bout of moderate-to-vigorous exercise when compared to either a series of light stretches (Zschucke et al., 2015; Wunsch et al., 2019) or no movement at all (Wood et al., 2018). In Zschucke and colleagues’ (2015) study, a group of 20 highly trained men, defined as men who trained at least three times a week for an endurance sport, and 20 sedentary men, defined as men who exercised less than once per week (or not at all), were randomized to either jog or run for 30 minutes at 60-70% of their VO2max (determined from a previous CPET) or a placebo exercise (light stretching without considerable elevation of heart rate) before exposing them to a psychological stressor. They demonstrated reduced cortisol reactivity in response to the 45 stressor task in those who exercise prior to the stressor compared to those who did not. These effects were independent of their reported typical exercising levels. Similarly, Wunsch et al. (2019) found significantly dampened cortisol responses to a psychosocial stressor in men who exercised prior on an ergometer at 70% of their maximal load (Watts/kg) when compared to those in a group of men who completed a series of light stretches. On the other hand, Wood and colleagues (2018) exposed 164 women to the TSST-G immediately following randomization to either participate in a group-based, 30-minute moderate-intensity group walk in an urban park (N=75) or to a group that did not complete any exercise at all. Wood and colleagues’ results revealed no differences in cortisol responses following the TSST-G between the two groups. It is apparent from these studies that perhaps intensity of the bout of exercise matters. More specifically, how different exercise intensities are defined could be influencing the interpretation of results in past studies. 5.2 Demarcating exercise intensity levels Zschucke and colleagues (2015) defined their exercise bout as moderate intensity, but prescribed at a level that potentially surpassed the first ventilatory threshold (participants held 60-70% of their VO2max). It is noteworthy that exercise at this intensity is typically labeled vigorous intensity by ACSM, not moderate intensity; prior research suggests that the first ventilatory threshold is typically surpassed around 50-60% of one’s VO2max (Hansen et al., 2003). Additionally, Zschucke and colleagues (2015) confirmed via functional magnetic resonance imaging that the mediating effects were a product of the activation of the negative feedback loop which is activated when cortisol levels are released (i.e., due to heightened energy demands, or threatened stress appraisals). Prior evidence does suggest that cortisol can be released in response to moderate intensity (just below the first ventilatory threshold); however, 46 duration of the bout (i.e., minimum 90 minutes) is the determining factor (Athanasious et al., 2022). Wunsch and colleagues also reported their exercise intensity as moderate-to-high while having their participant execute the exercise protocol at 70% of their maximal load (Watts/kg) on an ergometer for 30 minutes (2019). However, demarcating specific intensity levels based on metabolic needs is challenging due to ventilatory data not being captured. Woods and colleagues (2018) prescribed a 30-minute moderate intensity bout defined as 55% to 70% of maximum heart rate which was analyzed after the completion of the exercise. The randomized exercise cohort completed a group urban walk at a group-selected walking pace. No participants ran or jogged making it very unlikely that anyone surpassed the first ventilatory threshold for most of the 30-minutes. Interestingly, no dampening cortisol effects were found between the exercise and the control group during or after the exposure to the psychological stressor. Caplin and colleagues (2021) sought to reveal the impact of exercise bout intensity on cortisol responses to the TSST, and prescribed intensity zones based on ACSM guidelines, benchmarking light exercise at 30% of their CPET-determined heart rate reserve (HRR), moderate at 50% HRR, and vigorous at 70% HRR. Caplin et al.’s study demonstrated significant dampened cortisol responses to the TSST when participants completed a bout of exercise at vigorous intensity when compared to moderate and light intensities, and dampened responses at moderate intensity when compared to light intensity. Our findings partially support Caplin and colleagues’ (2021) study that different trajectories of cortisol output in response to a psychological stressor is dependent on the intensity of the exercise bout. Significant dampening of the cortisol response for the vigorous group was demonstrated in both studies when compared to the moderate and light groups. However, the current study failed to find a significant 47 difference between the moderate and light groups when utilizing individualized designation for intensity levels. The notion that both moderate and vigorous exercise dampen cortisol release to a subsequent heterotypical stressor found by Caplin et al (2021), Zschucke et al (2015), and Wunsch et al (2019) have been argued to provide support to the cross-stressor adaptation hypothesis (Sothmann et al., 1996). As a reminder, the cross-stressor adaptation hypothesis suggests that the neurobiological response to a homotypic stressor (the same type of stressor) is habituated to repeated exposure of the stressor (i.e., regular engagement in exercise reduces the stress response reactivity to the exercise and elicits a faster recovery), whereas heterotypic stressors (dissimilar in nature) can produce either habituated or more extreme responses after exposure (Sothmann, 2006; Sothmann et al., 1996). However, the current trial’s results suggest otherwise, that in fact the mechanism to explain the effects of a single bout exercise on cortisol trajectories to a subsequent stressor is merely the negative feedback loop when the first ventilatory threshold is surpassed, if the current results hold with the full sample. Cortisol has been shown to be primarily released when individuals surpass the ventilatory threshold due to its increased energy demands, requiring glucose to be created for continued activity through cortisol’s conversion of proteins and lipids into glucose, a process known as gluconeogenesis (Del Corral et al., 1998). As part of the process, the release of cortisol from the adrenal glands leads to activation of glucocorticoid receptors in the hypothalamus and pituitary glands, terminating production and release of their HPA-axis associated hormones and consequently the release of cortisol from the adrenal glands, in what is known as a negative feedback loop (Gunner & Quevedo, 2006; Herman, 2022). 48 Participants in the moderate and light intensity groups in the current study, purposefully did not surpass their ventilatory thresholds within the 30-minute bout. Consequently, with the lack of cortisol release, dampening of HPA axis activity following exercise through negative feedback was not initiated, and thus cortisol release to the TSST was perhaps able to continue uninhibited. On the other hand, in the current study, vigorous activity purposefully surpassed the ventilatory threshold, initiating cortisol release and the negative feedback loop, thus dampening the cortisol release to the TSST. This hypothesis likely reveals why Zschucke and colleagues (2015) and Wunsch and colleagues (2019) both reported dampening effects on cortisol release in response to a psychological stressor for their exercise groups compared to their control groups, whereas Wood and colleagues did not find any dampening effects between their two intervention groups (2018). Additionally, while Caplin and colleagues found a statistically significant dampening of the cortisol response to the TSST following moderate exercise compared to light, it is possible that participants in the moderate group did surpass their ventilatory threshold while exercising at 50% of their heart rate reserve for 30 minutes. However, ventilatory data were not collected and HRR is not equivalent to VO2 max (Swain & Leutholtz, 1997), making it reasonable to postulate that the dampened cortisol trajectory for the vigorous group in the current study could simply be explained by the negative feedback loop. 5.3 Understanding the stress response To determine whether the results from the current study, when contrasted with previous studies, simply reveals that dampened cortisol responses to the TSST are a result of the HPA-axis negative feedback loop or experimental evidence in support of the cross-stressor adaptation hypothesis, it is important to examine the potential role of exercise on mitigating more than just 49 the cortisol response to a subsequent psychosocial stressor. The parent study within which the current study is embedded is designed to investigate the effects of different intensities of exercise on a wider range of outcomes, including affective and cognitive responses to the psychosocial stressor as well as other physiological responses, including heart rate variability, cardiac output, and total peripheral resistance. It has been established that specific physiological responses to acute psychological stressors are determined by a cognitive appraisal process that consciously or unconsciously determines if an individual has the psychological or behavioural resources to meet the demands of the stressor that results in a threat or challenge appraisal (Blascovich et al., 2004; Lazarus & Folkman, 1984). Depending on whether the person appraises the stressor as a threat or challenge, different coping strategies are deployed, and different emotional responses are experienced (Lazarus & Folkman, 1984). For example, negative emotions such as anxiety and fear, as well emotional regulation strategies like rumination or distraction are suggested to be heightened during a threatened response, whereas the appraisal of a stressor as a challenge is associated with more positive emotions and problem-solving strategies (Blascovich & Tomaka, 1996; Lazarus & Folkman, 1984). Physiologically, work by Blascovich and colleagues (Blascovich et al., 2004, Blascovich & Tomaka, 1996) has demonstrated that challenge responses are associated with the SAM-p release of epinephrine (i.e., adrenaline) and norepinephrine (i.e., noradrenaline) into the bloodstream, increasing heart rate and cardiac output and decreasing total peripheral resistance (Cannon, 1929; McEwen, 2008; Behnke & Kaczmarek, 2018). Threat responses activate the HPA axis resulting in the release of cortisol, which limits cardiac output and increases total peripheral resistance (Behnke & Kaczmarek, 2018; Blascovich et al., 2004). These different, yet 50 specific, physiological reactions highlight the important implications of the psychological and cognitive appraisal process. Interestingly there are a limited number of studies that explore the effects of single bouts of exercise on the response to a psychological stressor on such a wide range of psychological and physiological responses. In a systematic review currently in preparation, Hadwen and colleagues have determined that 20 studies have explored the effects of a single bout of exercise on a range of outcomes following a psychosocial stressor. The review indicates that significant effects of exercise on psychological and physiological responses are, in large part, mostly when studies examined the effects of a single bout of exercise on cortisol reactivity to and recovery from the psychosocial stressors. Specifically, six of the 20 articles (30%) explored the effects of a single bout of exercise on cortisol secretion. Of those six, five (83%) demonstrated significant dampened cortisol responses to psychosocial stressors following exercise compared to no exercise, noting that higher intensities and longer bouts elicited large dampening cortisol output in response to the psychological stressors compared to lower intensities and shorter bouts. On the other hand, much fewer studies demonstrated significant effects on other outcomes. For example, effects of a single bout of exercise on heart rate and blood pressure elevations following a psychosocial stressor were inconsistent. Only two out of eight studies (25%), demonstrated faster returns to baseline in systolic BP after the exposure to psychological stressors when followed by single bouts of exercise when compared to no exercise. One out of eight studies (13%) saw similar return to baseline in diastolic BP after the psychological stressor, and 67% (four out of six studies) saw significant results in heart rate (HR) recovery. However, significant results surrounding heart rate recovery were mixed, only finding effects of a single bout of exercise on HR recovery from the psychological stressor in adolescent sample not young adults. One study 51 also reported elevated HR levels in the exercise groups during the recovery from the subsequent stressor; however, the exposure to the psychological stressor occurred immediately after the completion of the exercise bout (Chafin, 2007). No studies demonstrated significant effects of exercise on heart rate variability responses to a stressor. Additionally, none of the 20 studies investigated total peripheral resistance and only one study investigated cardiac output. As mentioned, cardiac output and total peripheral resistance, when combined into a ratio score, are considered to represent either a threat or challenge response to a psychosocial stressor as stated by the Biopsychosocial Model of Threat and Challenge (Behnke & Kaczmarek, 2018; Blascovich & Tomaka, 1996). In terms of psychological responses to the stressor, seven of the 20 studies (35%) in the systematic review (Hadwen et al., in preparation) measured different concepts associated with stress perceptions and affective responses (e.g., perceived stress, stressor difficulty, etc.) after the exposure to the psychological stressor. Of those studies, six (86%) had participants rate perceived stress (via a single-item Likert Scales or Visual Analogue Scales), and only one (17%) reported that a single bout of exercise at self-selected intensity significantly lowered stress perceptions in an adolescent sample (Lambiase et al. 2010). Interestingly, no studies using young adult samples or prescribed moderate-to-vigorous exercise intensities found significant results. Stress difficulty was measured in two studies with differing sample characteristics and measurement tools. One study with an adolescent sample measured stress difficulty via a 5- point Liket-Scale (Aladro-Gonzalvo et al., 2019), while the other study with young adults used a 7-point Likert-Scale (Paine et al., 2013). Neither produced significant differences regarding preceptions of stress difficulty in either the exercise or control groups. Only one study examined stress appraisals in young adults, reporting that the control group precieved the psychological 52 stressor as more threatening than the exercise group; however, the findings were not significant (Jacquart et al. 2020). Rumination, mood, and affect have also been explored in seven of the 20 studies in Hadwen and colleagues’ (in preparation) review. Findings for these outcomes are also inconclusive. Four of these studies explored the effects of a single bout of exercise on rumination following induction of the TSST, and none reported significant effects (Bernstein et al., 2020; Bernstein & McNally, 2018; Chafin, 2007). Notably, exercise was demonstrated to be a significant moderator of the association between rumination and negative affect and self-criticism (Bernstein et al., 2020; Bernstein & McNally, 2018). Additionally, no evidence was found to support the idea that a single bout of exercise modifies the effects of a psychosocial stressor on affective or mood states (Hadwen et al., in preparation). Hadwen and colleagues concluded that the inconsistencies in study design and measurement procedures make firm conclusions difficult to make. In summary, effects of a single bout of exercise seem to have few consistent findings across a wide range of psychological and physiological outcomes, except for cortisol reactivity. This suggests that it is possible that the dampened cortisol response to psychosocial stressors following vigorous intensity exercise compared to no exercise is simply negative feedback, and not evidence of any immediate cross-stressor adaptation. The application of the cross-stressor adaptation hypothesis to the role of exercise in mitigating responses to psychosocial stressors, however, is premised on the notion that repeated exposure to exercise should lead to mitigated reactivity and accelerated recovery from the stressor. Mücke and colleagues’ (2018) conducted a systematic review of fourteen studies that examined the differences between regular exercisers and/or more fit individuals on a wide range of physiological and psychological responses to the TSST compared to those who are physically 53 less active or less fit. In their review, 58% (seven of twelve studies) demonstrated reduced cortisol reactivity in those who exercise regularly or are more fit compared to those who are less active or fit. Forty-four percent (4 of 9) demonstrated lower heart rate reactivity in the exercisers and more fit adults compared to the lower active and less fit adults. In terms of psychological factors, 50% of the 4 studies demonstrated significantly higher levels of calmness and lower levels of state anxiety following the stressor in those who exercised regularly or were designated as fit, compared to those who were not regular exercisers or were of lower fitness. Similarly, 60% of studies demonstrated better positive mood states following the stressor in those who exercised or were more fit, compared to those who were not. The systematic review did, however, conclude a potential dose-response relationship, whereby high levels of regular physical activity seem to have stronger effects than moderate physical activity levels when compared to a physically inactive lifestyle. Taken together, results from Mücke and colleagues’ systematic review that there is some, but inconclusive, evidence that engaging in regular physical activity reduces physiological and psychological reactivity to psychosocial stressors. When considering the findings from the current study, Hadwen and colleagues systematic review that is currently in preparation, and Mücke and colleagues’ review, it is clear that more research is required to extend this area of research, including more consistent study designs that can be replicated across labs, wider ranges of physiological and psychological outcomes that best represent models of challenge and threat, and the determination of intensity zones based on metabolically relevant needs. 5.4 Future Directions The literature has demonstrated that a single bout of moderate-to-vigorous exercise dampens the cortisol reactivity to and recovery from a subsequent psychological stressor. 54 However, previous studies have utilized parameters to demarcate intensity zones that are not directly associated with clear biological mechanisms. Therefore, future studies should seek to better standardize intensity zones with metabolically meaningful cut-off points (i.e., ventilatory thresholds) to better understand the negative feedback loop and the cross-stressor adaptation hypothesis. Additionally, the totality of the effects of a single bout of exercise, specifically regarding the cross-stressor adaptation hypothesis, remain inconclusive due to many the inconsistencies within research protocols. To address these issues future studies should consider the standardization of protocols and the inclusion of cardiovascular and psychological measures associated with the stress response. Future studies should also expand their populations to include equal numbers of females and males to determine if sex differences do exist within this literature. Previous research that has explored the impacts of habitual exercise on cortisol reactivity following the exposure to psychosocial stressors in females found reduced reactivity compared to non-exercisers (Klaperski et al., 2013), similar to studies completed in males (Rimmele et al., 2007; 2009). However, sex differences between females and males in the effects of a single bout of exercise on stressor reactivity still needs to be explored to avoid speculations on the generalization that males and females would demonstrate the same effects. Women have historically been underrepresented or excluded in this area of research. The majority of studies reviewed here have excluded females from cortisol research based on the fluctuation of its levels throughout the menstrual cycle (Caplin et al., 2021; Wunsch et al., 2019). Kirschbaum and colleagues (1999) explored how the different menstrual phases impacted HPA-axis regulation after exposure to the TSST and found that women in their luteal phase had higher levels of salivary free cortisol compared to women in their follicular phase and women on 55 contraceptives. The study also looked at sex differences and found that even though men, overall, tended to have higher cortisol reactivity when exposed to stressful stimuli, only women in the luteal phase showed comparable. Therefore, it can be speculated that potential sex differences could be found when looking at HPA axis reactivity to stressors that are dissimilar in nature due to the fluctuation in cortisol levels during different menstrual phases compared to the relatively consistent levels in males. The current study did not explore sex differences due to lack of female participants during the time of analysis. However, the parent study is exploring sex differences between males and females within the luteal phase of their menstrual cycle and excluding those using hormonal contraceptives. Exploring the relationship between exercise and the stress response during all phases of the menstrual cycle, as well as including those on hormonal contraceptives, can perhaps provide a more comprehensive and generalizable understanding of the complex relationship between the menstrual cycle and the HPA-axis response to stressors following engagement in exercise or not. 5.5 Limitations The current report is limited by its incomplete sample size when compared to the required sample size required by the power analysis calculation. The study is also limited by the fact that I did not analyze results for the other collected psychological or physiological collected measures, nor did I explore sex differences within the presented sample due to there being a small number of females. However, the final sample will be exploring potential sex differences between the groups to determine the extent to which exercise intensity impacts the stress response to psychosocial stressors. 56 Regarding generalizability, our study’s sample included a healthy and young population (majority being university students), excluding anyone who was above the age of 35 and those who had any health conditions that would impede their ability to exercise as well as those who with any diagnosed psychological disorder. Future studies should include more diverse samples including older populations and adolescents, diverse occupational backgrounds (i.e., shift workers, office workers, etc.), and difference socioeconomic status ranges. 5.6 Conclusion The prolonged activation of the stress response can have catastrophic consequences on one’s overall wellbeing, as well as mental and physical health (Kivimaki & Steptoe, 2018; Miller et al., 2007; Kopp & Réthelvi, 2004; Cohen et al., 2007; O’Connor et al, 2021) presenting a critical need to investigate ways to mitigate the negative consequences of the stress response. Long-term physical activity and exercise have been demonstrated to alter the HPA axis stress response when exposed to acute stressors in a majority of studies (Mücke et al., 2018). 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American College of Sports Medicine. https://www.acsm.org/docs/default-source/files-for-resource-library/exercise-intensity-infographic.pdf 69 Appendices Appendix A Recruitment Advertisements A.1.1 Advertisement Poster 70 A.1.2 Advertisement Slide 71 A.1.3 In-Class Advertisement Script EAST In class announcement advertising Hello everyone, My name is <name> of the School of Kinesiology at UBC and I’m here to recruit participants for our current study – called “the EAST study”. First, I am going to tell you what the study is all about and then you are very welcome to ask any kind of questions that may come to your mind. Finally, I will tell you what to do, if you are interested in taking part in the study. In brief, we are interested in how different intensities of exercise impacts our responses to social interactions. The EAST-study will consist of two sessions: The first lasting 90 minutes and the second lasting 4 hours and 30 minutes. Sessions will be scheduled at least 3 days apart, so that both sessions are completed within 14 days. (1) The first session will last approximately 90 minutes. You will be asked to fill out some brief questionnaires and review procedures and the consent form. Then you will be fitted with a Biopac monitor and gas exchange mask to test multiple variables like heart rate, and respiratory rate before starting a light warmup. From there you will be asked to run on a treadmill as long as you can until you feel you can work no harder, followed by a 5-minute low-intensity recovery period and 5-minute sitting period. (2) The second session will last approximately 4 hours and 30 minutes and follow very similar procedures. You will be fitted with a Biopac monitor and gas exchange mask to measure heart rate, respiratory and more before starting a light warmup. From there you run for 30 minutes at a fixed intensity that is lower than the maximum reached during the first visit and followed by a 5-minute low-intensity recovery period and sitting period. Following this you will be asked to participate in a short social interaction. What will you gain from this study? First, like most participants in studies, you’ll feel great that you’re helping scientific discoveries! Second, you will receive a cash honorarium to reimburse you for your time. Or you can find us on SONA and receive HSP credits! Additionally, at the end of the study, you will also get your personal results from the fitness testing, which will give you an objective measure of your aerobic fitness. If you are interested in the results of the EAST study, we would be happy to send them to you via email, as soon as our study is finished and published. Who can take part in our study? Participants will be fully screened by a member of the study team but broadly speaking: Participants are eligible if they: • Are between the ages of 18-35 • Are free of any medical conditions that would prohibit their capacity to perform strenuous exercise. • Are free of psychiatric disorders, Cushing’s syndrome, Addison’s Disease • Do not currently smoke/vape or use medications/hormone supplements that would affect your cardiovascular system (full list available during scheduling) • Are not currently (or within the last 12 months) involved in organized competitive sport. • If female, are not currently taking hormonal contraceptives. The study will take place in the Medical Sciences Block C, room number 108, 2176 Health Sciences Mall at UBC campus (http://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3). It should be noted that your academic standing in this class or any other class will not be affected by your decision to participate or withdraw from the study. You can scan the code on the screen to access our website or find us at eaststudy.ca. There you can find more information about the study and book an appointment to finish screening with our study coordinator! Or you can find us on SONA and enter the study through that system. Looking forward to seeing you again! Thanks for your time, Bye! 72 A.1.4 HSP Advertisement Study Title: EAST Study Researcher: Hana Hicks, Luke Peddie, Dr. Eli Puterman, Dr. Frances Chen Description: In brief, we are interested in how different intensities of exercise impacts our responses to social interactions. The EAST-study will consist of two sessions: (1) The first session will last approximately 90 minutes. You will be asked to fill out some brief questionnaires and review procedures and the consent form. Then you will be fitted with a Biopac monitor and gas exchange mask to test heart rate, heart rate variability, cardiac output, total peripheral resistance, and respiratory rate before starting a light warmup. From there you will be asked to run on a treadmill as long as you can until you feel you can work no harder, followed by a 5-minute low-intensity recovery period and 5-minute sitting period. (2) The second session will last approximately 4 hours and 30 minutes and follow very similar procedures. You will be fitted with a Biopac monitor and gas exchange mask to test heart rate, heart rate variability, cardiac output, total peripheral resistance, and respiratory rate before starting a light warmup. From there you run for 30 minutes at a fixed intensity that is lower than the maximum reached during the first visit and followed by a 5-minute low-intensity recovery period and sitting period. Following this you will be asked to participate in a short social interaction. You will also be asked to provide of Eligibility: Participants will be fully screened by a member of the study team. Please let them know if you have any questions about the following criterial. Participants are eligible if they: • Are between the ages of 18-35 • Are free of any medical conditions that would prohibit their capacity to perform strenuous exercise • Are free of psychiatric disorders, Cushing’s syndrome, Addison’s Disease • Do not currently smoke/vape or use medications/hormone supplements that would affect your cardiovascular system (full list available during scheduling) • Are not currently (or within the last 12 months) involved in organized competitive sport. • If female, are not currently taking hormonal contraceptive. Location: The study will take place in the Medical Sciences Block C, room number 108, 2176 Health Sciences Mall at UBC campus (hLp://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3). Contact Information: eaststudy.ca Reimbursement/Time: Participants will be provided with 6.5 HSP credits and a complimentary fitness assessment, pro-rated if participants do not complete both study sessions. Study End Date: Ongoing. Version 3 | 01/24/2024 Page 1 of 1 73 A.1.5 Initial Contact Email Hello <name>, Thank you for your interest in participating in the EAST study and for booking a screening and scheduling phone call. Please review the following information prior to the 15-minute call. We are happy to answer any questions that you may have by email or during the initial call. Here is a link to our website for you to review the study in more detail: eaststudy.ca <add link to website here> Please find attached the study Information and Consent Form. Please read these documents carefully and if you have any questions, we would be happy to answer any questions concerning the consent form during the eligibility screening phone call. We have also included the link to our website: eaststudy.ca In brief, we are interested in how different intensities of exercise impacts our responses to stressful experiences. The EAST-study will consist of two sessions: (1) The first session will last approximately 90 minutes. You will be asked to fill out some brief questionnaires and review procedures and consent form. Then you will be fitted with a Biopac monitor and gas exchange mask in order to test heart rate, heart rate variablity, cardiac output, and total peripheral resistance and respiratory rate before starting a light warmup. From there you will be asked to run on a treadmill as long as you can until you feel you can work no harder, followed by a 5-minute low-intensity recovery period and 5-minute sitting period. (2) The second session will last approximately 4 hours and 30 minutes and follow very similar procedures. You will be fitted with a Biopac monitor and gas exchange mask in order to test heartrate, heart rate variability, cardiac output, total peripheral resistance and respiratory rate before starting a light warmup. From there you run for 30 minutes at a fixed intensity that is lower than the maximum reached during the first visit and followed by a 5-minute low-intensity recovery period and sitting period. Following this you will be asked to participate in a short social interaction, similar to a mock-job interview. You will also be asked to provide a total of 12 saliva samples and complete some brief questionnaires and a final rest period for 70 minutes. At the end of the study, you will receive either HSP credits or cash to reimburse you for your time. Your method of compensation will be confirmed during your phone call. You will also receive a personalized fitness assessment. If you are interested in becoming more active, we will be happy to offer you a list of tips for increasing and maintaining an active lifestyle. The study will take place in the Medical Sciences Block C, room number 108, 2176 Health Sciences Mall at UBC campus (http://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3). Thank you again for scheduling your initial phone call. Please refer to the confirmation email you received after booking for details regarding your appointment. If you are determined to be eligible and agree to participate during the call, we will make appointments for the exercise sessions as described above. We are also happy to answer any questions you have by email or during this phone call. I am a graduate student at UBC and will be your first point of contact 74 concerning the EAST study. The Principal Investigator of the Study is Dr. Eli Puterman, Assistant Professor in the School of Kinesiology. I look forward to speaking with you, <insert name> 75 A.1.6 Phone Screener Phone Screener Hello <name>, my name is <researcher name> and I am calling you regarding the EAST research study that you showed interest in participating in. Is now still a good time to have a 10-15-minute conversation about the study? Thank you again for your interest in participating in the EAST study and for making an appointment online. I understand you were sent a copy of the study consent form, have you had the opportunity to review it? If you have any questions about it, please let me know. I’m going to review some study details, and then you can tell me if you are still interested in participating. In brief, we are interested in how different intensities of exercise impacts our responses to social interactions. The EAST-study will consist of two sessions: (1) The first session will last approximately 90 minutes. You will be asked to fill out some brief questionnaires and review procedures and consent form. Then you will be fitted with a Biopac monitor and gas exchange mask in order to test heart rate, heart rate variability, cardiac output, total peripheral resistance and respiratory rate before starting a light warmup. For the Biopac, we will place multiple electrodes on the front and back of your torso and will ask you to wear a strap to hold the electrodes and monitor in place. From there you will be asked to run on a treadmill as long as you can until you feel you can work no harder, followed by a 5-minute low-intensity recovery period and 5-minute sitting period. (2) The second session will last approximately 4 hours and 30 minutes and will follow very similar procedures. You will be fitted with a Biopac monitor and gas exchange mask in order to test heart rate, heart rate variability, cardiac output, total peripheral resistance and respiratory rate before starting a light warmup. From there you run for 30 minutes at a fixed intensity that is lower than the maximum reached during the first visit, followed by a 5-minute low-intensity recovery period, and a sitting period. Following this you will be asked to participate in a short social interaction, similar to a mock-job interview. You will also be asked to provide a total of 12 saliva samples and complete some brief questionnaires and a final rest period for 70 minutes. Do you have any questions? If YES: <answer questions> Note here the given information: ______________________________________________________________________________ _____________________________________________________________________________________________________________________ Are you are still interested in participating? If YES: Before we schedule your visits I just need to ask you a few questions to confirm you are eligible to participate and also a few questions to get general information about you: Screening Questions Does qualify Does NOT qualify 1. How old are you? Note the precise age here: __________________________ o 18-35 o <18 o >35 76 2. What was your sex assigned at birth? Note here: __________________________ o Male, Female, Intersex o N/A 3. If female: a. Is your average menstrual cycle between 21-35 days? b. Are you currently using hormonal contraceptives? o Yes o No o No o Yes 4. What is your gender identity? Note here: __________________________ o All o N/A 5. Do you have lived experience as a trans person? a. Note here: __________________________ b. If yes, are you currently on any hormone therapy? (Make note) o Yes o No o N/A 6. Have you been diagnosed with any psychiatric disorder(s)? (e.g. depression, anxiety disorder or substance abuse) - if yes: - are you still experiencing these issues? o No o Yes 7. Do you have Cushing Syndrome, Addison’s Disease or adrenal insufficiency? o No o Yes 8. Are you currently taking or have you recently taken any of the following medications or supplements: corticosteroids, beta-blockers, psychotropics, angiotensin-converting enzyme inhibitors, ketoconazole, metyrapone, and hormonal contraceptives? - if yes: which medication, for how long and when was it last taken: o No o Yes 9. Are you a Smoker (i.e., cigarettes, marijuana, and/or vapes)? o No o Yes 10. Are you currently or have you recently (within the past year) participated in sport at a regional, university, or semi-professional league? o No o Yes 11. Have you ever participated in a study where you had to prepare a presentation for a panel of judges? o No o Yes 12. Has your doctor ever said that you have a heart condition OR high blood pressure? o No o Yes 13. Do you feel pain in your chest at rest, during your daily activities of living, OR when you do physical activity? o No o Yes 77 14. Do you lose balance because of dizziness OR have you lost consciousness in the last 12 months? Please answer NO if your dizziness was associated with over-breathing (including during vigorous exercise). o No o Yes 15. Have you ever been diagnosed with another chronic medical condition (other than heart disease or high blood pressure)? PLEASE LIST CONDITION(S) HERE: o No o Yes 16. Are you currently taking prescribed medications for a chronic medical condition? PLEASE LIST CONDITION(S) AND MEDICATIONS HERE: o No o Yes 17. Do you currently have (or have had within the past 12 months) a bone, joint, or soft tissue (muscle, ligament, or tendon) problem that could be made worse by becoming more physically active? Please answer NO if you had a problem in the pastor presently, but it does not limit your current ability to be physically active. PLEASE LIST CONDITION(S) HERE: o No o Yes 18. Has your doctor ever said that you should only do medically supervised physical activity? o No o Yes 19. Are you fully vaccinated against COVID-19? Please note that you will be required to provide proof of vaccination and photo identification. o Yes o No 20. Are you able to read and understand English at the level necessary to fully understand what you will be required to do in the study and the risks associated? o No o Yes [If answered yes to questions 7-13] Thank you for your responses. You may still be eligible for this study but I will have to ask that you complete a short questionnaire about your health, which I will send you via email. [If participant has time to complete on the phone, stay on line; if not, schedule a call back] [If does not qualify] Ok. Unfortunately, you are not eligible for this study. We thank you for your interest and your time. [go down to the recruitment below!] [If qualifies] You qualify for the study. Before we get started with scheduling, here is some additional information: as mentioned in the consent form, this study does involve capturing variables like heart rate, heart rate variability, and total peripheral resistance. These measures will be captured with a Biopac motor and electrodes which will be adhered to the skin around your chest, neck and back area. For them to be properly adhered to the skin body hair may need to be trimmed by the research staff at 78 the site of contact with an electric trimmer. This will only be the case if the hair is preventing a proper connection. Further, the research assistant placing the electrodes will be gender matched to you. If identifies as non-binary: We currently do not have any research assistants on our team who identify as non-binary. At this time, I want to check that you are comfortable with the placing of electrodes, and if so, which gender of research assistant you would prefer to place them? Last, before we schedule you in can you please confirm how you heard about this study? [If through Human Subject Pool] Great. Can you please share with me your SONA ID so that we have it on file to allocate credits once your testing has concluded (make note)? You will be allocated 1 credit after completion of the first testing day and 4.5 credits after completion of the second testing day. [If not through Human Subject Pool]. Great, thank you for confirming (make note so that provided cash honorarium instead of credits on SONA). We would like to schedule you for two appointments. The first visit can be scheduled in the morning or afternoon and the second visit needs to be scheduled in the afternoon. *For female assigned at birth: The second visit needs to take place at a specific phase of your menstrual cycle and within two weeks of the pre-study day. To do that, could you please take a minute to identify the day that your most recent menses/period started? Bleeding Onset: ___________________ TAKE A MOMENT TO WRITE DOWN 14-28 DAYS FROM THAT DAY. Thank you. It appears that your second visit needs to take place between <date1> and <date2>. FIND A TIME THAT WORKS FOR SECOND VISIT. JUDGES CANT BE FROM DAY 1 TAKE A MOMENT TO WRITE DOWN WHAT DATES DAY 1 CAN BE BETWEEN (UP TO 14 DAYS BEFORE BUT NOT WITHIN 3). Your first visit can occur anytime within fourteen days of <date1>. Would any of the following dates or times work? *For male assigned at birth or intersex: Would any of the following dates or times work? <dates and times> [if scheduling successful] Ok then, I will send you a short reminder email with the date and time. I’ll also send you the “information and consent form” by email. Please read it carefully and let us know, if you have any questions. At our first meeting on <date> we are happy to answer all of your questions concerning the consent form. Please make sure to read preparation instructions listed in the email carefully, as certain clothing cannot be worn for the electrodes. Please feel free to contact us by email if you have any questions you want to ask before our first meeting. At 79 the first meeting, we will also review the consent form together before you sign it, if you so choose to. The study will take place in the Medical Sciences Block C, 2176 Health Sciences Mall at UBC campus, just across from Woodward Library. Do you require directions? If yes, refer to the link in the first email response: (http://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3) [if scheduling not successful] Ok. Well if anything changes please feel free contact me to set something up. You can reach me or another lab coordinator at kin.fastlab@ubc.ca, Monday through Friday! The FAST lab is also conducting various other projects. If you are interested in participating in these other opportunities, may we store your contact information so that we can contact you in the future? [if yes] àadd to recruitment list Ok, Thanks for your time and have a good day, goodbye. Note: • Questions number 8–14 are the seven General Health Questions of the “Physical Activity Readiness Questionnaire for Everyone” (Warburton et al., 2021) 80 A.1.7 Reminder Emails Reminder Email Dear <name>, Thank you for taking part in the EAST study! Please review the Information and Consent Form carefully and let us know if you have any questions. At our first meeting on <date>we are happy to answer all of your questions concerning the consent form. Please feel free to respond to this email or visit our website if you have any questions you want to ask before our first meeting. Your first visit will be on <date>, from<time-time>. Your second visit will take place on <date>, from<time-time>. Do you use an online or paper calendar? If so, can you please enter these dates now. If you need to cancel, please provide us with 24 hours’ notice. All meetings will be located at the FAST lab, Medical Sciences Block C, 2176 Health Sciences Mall at UBC campus. If your appointment falls on a weekend, the building will be locked so one of our team members will meet you at the front entrance. (http://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3). If you still have any questions, please feel free to contact us at any time: website: eaststudy.ca For all meetings we ask that you bring with you: • We will provide a specialized starp that will be worn under your active wear. • Please bring clothing appropriate for exercise, running shoes and a towel (there will be the possibility to take a shower after the study), in addition to your normal daily attire. • Please note on both visits' electrodes will be secured to your torso (three on your vist fisit and eleven on your second). • If necessary, we may need to use a hand trimmer to remove hair on the neck and back to secure the electrodes properly to the skin. • If wearing a sports bra, if possible, try to not wear one with a high razor back. • Please try to avoid wearing shirts made of synthetic materials as it may interfere with our HR monitors. • If possible, please bring your own water bottle. Also, please consider the following instructions: • In the days prior to and on the day of your appointments, please make sure to hydrate well • Please do not exercise vigorously within 24 hours of your visits • Do not eat large meals 2 hours before coming to the lab for your visits • Please try to limit the consumption of any foods with live cultures at all on the day of your 2nd appointment. This includes milk, yogurts, yeast products, etc. • We also ask that you do not drink coffee up to two hours prior to your meetings. • Please abstain from alcohol on the day of your assessments. • If you are feeling sick or have developed symptoms, please contact us as soon as possible to reschedule your appointment. 81 If these instructions are not fulfilled, you will be asked to reschedule your appointment; so, please make sure to follow the instructions listed above. Thank you very much for your time and cooperation, we look forward to seeing you soon! [Name] Study Coordinator 82 Second Reminder Email Dear <name>, Thank you for taking part in the first session of the EAST study! At our second meeting on <date> we are happy to answer any other questions you may have. Or please feel free to contact us by phone if you have any questions you want to ask before our second meeting. Your second visit will take place on <date>, from<time-time>. Do you use an online or paper calendar? If so, can you please enter these dates now. If you need to cancel, please provide us with 24 hours’ notice. All meetings will be located at the FAST lab, Medical Sciences Block C, 2176 Health Sciences Mall at UBC campus. If your appointment falls on a weekend, the building will be locked so one of our team members will meet you at the front entrance. (http://www.maps.ubc.ca/PROD/index_detail.php?locat1=523-3). If you still have any questions, please feel free to contact us at any time: For all meetings we ask that you bring with you: • Spare clothing appropriate for exercise, running shoes and a towel (there will be the possibility to take a shower after exercise), in addition to your normal daily attire. • Please try to avoid wearing shirts made of synthetic materials as it may interfere with our HR monitors. • If possible, please bring your own water bottle. • Proof of COVID-19 vaccination and photo identification • Completed COVID-19 Health Check (https://bc.thrive.health/covid19/en) Also, please consider the following instructions: • In the days prior to and on the day of your appointments, please make sure to hydrate well • Please do not exercise vigorously within 24 hours of your visits • Do not eat large meals 2 hours before coming to the lab for your visits • Please try to limit the consumption of any foods with live cultures at all on the day of your 2nd appointment. This includes milk, yogurts, yeast products, etc. • We also ask that you do not drink coffee up to two hours prior to your meetings. • Please abstain from alcohol on the day of your assessments. • If you are feeling sick or have developed symptoms, please contact us as soon as possible to reschedule your appointment. If you need to reschedule or cancel please try to give us a minimum of 48 hours notice via email. If your period begins prior to your second testing session, please email us as soon as possible as we will need to reschedule. If these instructions are not fulfilled, you will be asked to reschedule your appointment; so, please make sure to follow the instructions listed above. We will remind you of this information by phone the day before the meeting. Thank you very much for your time and cooperation, we look forward to seeing you soon! <attach Information & Consent form> [Name]: Study Coordinator 83 Appendix B Consent Forms B.1 Cash Honorarium Consent Form 84 85 86 87 88 89 90 91 92 93 94 B.2 HSP Consent Form 95 96 97 98 99 100 101 102 103 104 105 Appendix C Deception Form DECEPTION FORM Some types of research can be carried out only if the participants do not know the true purpose of the research in advance. However, because such studies involve a potential breach of trust, they should not be entered into without sober reflection on the necessity for using deception and its potential impact on participants, once disclosed. Researchers intending to conduct studies involving deception will need to ensure that their research meets the conditions outlined in the TCPS 2 Article 3.7A.* a) The research involves no more than minimal risk to participants b) The lack of participant consent is unlikely to adversely affect the welfare of the participant c) It is impossible or impracticable to carry out the research and to answer the research question properly, given the research design, if prior consent of participants is required d) There is a plan to provide a debriefing and the opportunity to withdraw consent and data As noted in the TCPS2, debriefing is an important mechanism in maintaining the participant’s trust in the research community and should be proportional to the sensitivity of the issue studied. Where minor deception has been used, often the debriefing can be a simple candid disclosure. However, in more sensitive research, researchers should provide a full explanation of why participants were misled, why it was necessary and express their concern about the welfare of participants. Re-establishing trust that might have been lost is an important dimension of the debriefing process and researchers should be sensitive to participants’ needs, feelings, reactions and concerns. If your study involves deception, please (1) complete the next page; (2) delete this instruction page before saving; (3) attach the next page to your ethics application. *References in the TCPS 2 (2018) To access the print version (PDF): https://ethics.gc.ca/eng/documents/tcps2-2018-en-interactive-final.pdf. Page numbers refer to the print pages. PRE1 website Print version Article 3.7A http://pre.ethics.gc.ca/eng/tcps2-eptc2_2018_chapter3-chapitre3.html#7a Alterations to Consent Requirements | see page 37 Article 3.7B http://pre.ethics.gc.ca/eng/tcps2-eptc2_2018_chapter3-chapitre3.html#7b Debriefing in the Context of Alterations to Consent Requirements | see page 40 Exceptions to the requirement to debrief | see page 42 1 The Interagency Advisory Panel on Research Ethics (PRE) 106 Use as much space as needed. 1. Deception undermines informed consent. Describe: a. why you believe deception is necessary to achieve your research objectives (i.e. that the research cannot be done in any other way) b. why you believe that the benefits of the research outweigh the costs to participants. (A) One major objective of this study is to induce a stress response in the study participants using a mild-to- moderate stressor. The TSST is the gold-standard of psychosocial stress protocols to date (Kirschbaum et al., 1993), and has been used in a plethora of studies in the fields of health, social and clinical psychology (over 800 published studies with the TSST in title or abstract). In studies using the TSST to induce stress, a small degree of deception is necessary to avoid participants developing anticipatory coping behaviors to manage the stressor. Implicit to its very nature, psychosocial stress is characterized by events that are uncontrollable and socially evaluative (Dickerson and Kemeny, 2004). If participants are fully informed to the TSST protocol upon consent, these two essential features become null, as participants may anticipate and prepare to cope with the forthcoming stressor. Since, in this study, we are interested in the question of whether exercise mitigates acute cortisol responses to stressful events, it is important that the induction of mild-to-moderate stress remains uncompromised. The stress signal must be adequately strong to warrant a hormonal stress response and subsequent detection in the saliva. Our study does inform participants that they will be completing a short social interaction (which is referring to the TSST) to make sure participants are aware of the additional component of the study without disclosing the purpose or nature of the test to ensure the stress induction is effective. Various studies have used milder stress protocols, all of which result in stronger noise to signal ratios, increased variance in participants’ coping, and sometimes, no cortisol response at all (Pruessner et al., 2008; Zschucke et al., 2015). Using these less reliable or invalidated protocols would be remiss, compromising the quality and subsequent impact of the research for informing better healthcare. In stress research, the use of standardized protocols, such as the TSST, are necessary so that researchers can compare and cross-reference their respective work. (B) It is our belief that the benefits of this research for improving healthcare far outweigh the minimal, short-term cost, to participants. When participants are debriefed on the purpose of the TSST, they generally recover quickly, experiencing a sense of relief and ease that they were not actually being evaluated on their performance. Fortunately, the TSST only induces mild-to- moderate stress, and is thus unlikely to cause threat beyond that which occurs when stressful events occur in everyday life. The study population will be health men and women, and unlikely to experience abnormal reactions to stress beyond the normal responses. Participants will also be glad to receive a monetary honorarium, or HSP credits, and a fitness evaluation (valued at $100) for their time and effort. Also, we will provide them with resources directing 107 them to UBC’s counselling services, which they may follow up on at their own discretion. In the long term, findings from this study may inform individuals, who actually experience stress in everyday life, how to use physical activity in order to cope better. Given the high rates of stress in everyday life reported by Canadians (approximately 25% report high stress according to Health Canada), our research will shed light on the benefits of exercise to many Canadians. 2. Outline the anticipated impacts of your use of deception on the participants, once they have learned of it. There are a few potential outcomes that may ensue from participants learning of the deception used in this experiment. On the one hand, participants may experience some mild resentment towards the researcher. We consider this outcome unlikely since the deception used is subtle and short-lived: participants will be informed that they will have a social interactions; similar to that of a mock-job interview. They will not be informed, however, that there is a panel of judges or that the judges provide no socially validating cues. The participants will also be unaware of a math-task at the end of the TSST, but this component only lasts 5 minutes. Additionally, the rest of the study, including exercise protocols and all measures, will be fully disclosed to the participants. On the other hand, because of the stressful nature of the task at hand, participants may actually feel a strong sense of relief and ease to find out that they were not actually being judged on their performance. Participants may receive great comfort in learning that what they initially believed to be a mock-job interview, assessing their quality and performance, was not actually evaluating them at all. While learning that mild-stress was purposefully induced may cause participants some initial indignation, we believe that once they are exposed to the friendliness, and moral intention of our research staff, they will regain a sense of security and ease. Another potential impact on participants from learning about the deception, is an initial sense of surprise and confusion: participants may want to know why stress was induced. For this reason, our team will go beyond simply communicating the nature of the deception, and will provide a complete and thorough explanation of the studies purpose and objectives. Participants will be informed on the impact that our study will have on future healthcare, and will be commended for their important contribution. The honorariums that we provide will also contribute to mitigating any adverse reactions to learning of the deception 3. Describe how you will debrief participants at the end of the study. Confirm that participants will be asked whether they want to withdraw their data. Debriefing will occur following the final rest period. First, participants will receive an initial debriefing regarding the purpose of the TSST. During this debriefing, the lead will inform that participant that they will meet the judges, who will expose themselves as researchers. In a friendly and comforting manner, they will explain that they purposefully provided no validating cues during the public presentation. The lead experimenter will then explain what the TSST is and why it was used. Participants will be reassured that they can now relax and that no other stressors will ensue for the remainder of the experiment. They will be told to relax and be proud of the time they have committed to our study. At the end of this recovery phase, the second phase of the debriefing will take place. During this time, participants will be notified of the purpose and objectives of the study, i.e. why stress was induced and what 108 exactly was measured (cortisol, mood and cognitions). Participants will also be informed about how this study might influence future healthcare. They will be thanked and reassured that they have made a significant contribution to the purpose of advancing science in the field of stress and health. Participants will be informed on the types of impacts that stress can have in people’s lives. We will offer them resources for UBC’s counselling services that they can use at their own discretion. Participants will then be handed the results of their fitness test from the first study appoint (separate day from the TSST). The lead experimenter will review the results with them, informing them of several markers of their current fitness status. Lastly, the lead experimenter will provide the participant with a $80 cash honorarium or grant the full 5.5 HSP credits. If the participant is receiving cash they will sign 2 receipts: one for them two keep, and another for the experimenter. The participants will be thanked for all their time and effort and will be guided to the exit. 4. In very limited circumstances, when the debriefing is not possible, practicable or appropriate, it may be acceptable to waive the debriefing requirement. A very strong argument is required before the BREB will approve the use of deception without a debriefing. If you are proposing to not debrief participants, please provide your justification, and include: a. How the participant population will learn about the study b. Whether any representatives of the study population were consulted about the study design and questions. We will always be debriefing the participants. 109 Appendix D Debriefing Protocol D.1 Cash Debriefing Protocol Debriefing Protocol – Section 1 EAST Debriefing Protocol Each participant will be debriefed at the very end of the study to clarify all forms of deception that were used in the TSST portion of the second assessment and to explain the purpose of the entire protocol. Debriefing for TSST: 1. Hi <<participant name>> 2. Standardized debriefing on the purpose of the TSST and the specific measures that were taken: I know that this task may have felt quite stressful, but I would like to reassure you that it was all part of a very safe stress protocol that is often used in the lab. The judges that evaluated you are really just lab members; they were told to provide no validating cues while you gave your presentation. In fact, they were told to be totally non-reactive during your speech. They are really quite nice people, and are simply members of our lab. They haven’t evaluated you in any way. All notes that they took regarding your performance were fake. We are sorry that we couldn’t tell you about the backwards counting component of the task; this surprising portion was meant to catch you off guard. The protocol is referred to as the Trier Social Stress Task (TSST) and its purpose is to induce a psychosocial stressor using a public speaking and arithmetic task. Your performance was not being recorded, nor did we evaluate anything that took place during the task. It was simply used to induce stress. Don’t worry. The study is over now, and you will no longer have to worry about any stressors. You should be very proud of the time and effort that you have committed to this study and all of us here at the FAST Lab commend you for that. Debriefing following entire protocol: 3. Thank you very much for taking part in this study! 4. Standardized debriefing on the purpose of the study and the specific measures that were taken: “In doing this study we hoped to gain insight into the physiological and psychological mechanisms that influence how exercise modifies one’s response to a stressful event. 110 It is well known, that the intensity of a single bout of exercise influences psychological and physiological responses to exercise. For example, the greater the intensity, the more one is likely to experience a dampening on their stress system. Moreover, intense exercise is associated with a hormonal stress response. For example, cortisol, a hormone in our body, rises in response to stress and also in response to hard exercise. We want to have a closer look at the associations between your responses to different intensity exercises and your reactions to a stressful event. You were randomly assigned to the _____________-intensity exercise group. That means that you performed today at a heart rate that corresponds to roughly 30% (LICT – controlled walking speed of 1.7-2 Mph) or 10% (MICT) below the oxygen uptake corresponding to VT1 or halfway between VT1 and VT2 (VICT). At this intensity, we have asked you to rate your feelings, your arousal and your perceived exertion across multiple time points measured before, during and after exercise, and during and following a subsequent stressor, the TSST. The saliva samples were taken to measure the changes in cortisol levels over time. By integrating the data from all participants we can have a closer look at the interplay of these measures and determine in more detail how exercise influence one’s responses to stress. We ask that you do not talk about the specifics of this experiment outside of our lab, especially components related to the stress-task. This is mainly to prevent potential participants from developing expectations or biases prior to engaging in our protocol. Do you still have any questions about this? If there are some questions that come to your mind at a later date, please feel free to contact us at any time (email or phone us).” If you have any concerns about the study you may contact the Clinical Research Ethics Board at UBC which has reviewed this study and is responsible for helping to ensure that the rights of research participants are upheld. Please contact Pia Ganz at 604-875-4149 if you have any concerns. 5. Mentioning UBC Counselling service, mental health support “In our study we asked you to fill in some questionnaires about your levels of stress. We also used a stress induction protocol. Experiencing these events regularly in everyday life can have a significant negative impact, physically and mentally. We don’t know, if this directly applies to you, but here is a contact form for UBC counselling services that you may use at your discretion. Feel free to contact them at any time you like.” 6. Participant will receive personal results of CPET test and will be able to ask questions about it. Hand over personal CPET-results: printed from cosmed computer and placed in an envelope 7. Participant will receive reimbursement ($80) in cash. Participant has to sign two receipts (one for participant and one for us) 111 112 Debriefing Protocol – Section 2 UBC Mental Health Support at Counselling Services A variety of mental health resources and professionals are available through Counselling Services. Counselling Services can work with you to assess your wellness needs, and identify the level of care that's right for you – from self-directed programs and tools to workshops, group programs, and individual therapy – to help you have the best university experience possible. For more information please refer to the resources below: Website: https://students.ubc.ca/health/counselling-services Make an appointment: Counselling Services - Operating hours Brock Hall Room 1040, 1874 East Mall Vancouver, BC V6T 1Z1 Canada Day Hours of Operation Monday 8:30 am - 4:30 pm Tuesday 8:30 am - 4:30 pm Wednesday 8:30 am - 7:00 pm Thursday 10:30 am - 4:30 pm Friday 8:30 am - 4:30 pm Saturday Closed Sunday Closed 113 D.2 HSP Debrief Protocol Section 1 - EAST Debriefing Protocol Each participant will be debriefed at the very end of the study to clarify all forms of deception that were used in the TSST portion of the second assessment and to explain the purpose of the entire protocol. Debriefing for TSST: 8. Preparation to meet the judges 9. Standardized debriefing on the purpose of the TSST and the specific measures that were taken: I know that this task may have felt quite stressful, but I would like to reassure you that it was all part of a very safe stress protocol that is often used in the lab. The judges that evaluated you are really just lab members; they were told to provide no validating cues while you gave your presentation. In fact, they were told to be totally non-reactive during your speech. They are really quite nice people, and are simply members of our lab. They haven’t evaluated you in any way. All notes that they took regarding your performance were fake. We are sorry that we couldn’t tell you about the backwards counting component of the task; this surprising portion was meant to catch you off guard. The protocol is referred to as the Trier Social Stress Task (TSST) and its purpose is to induce a psychosocial stressor using a public speaking and arithmetic task. Your performance was not being recorded, nor did we evaluate anything that took place during the task. It was simply used to induce stress. Don’t worry. The study is over now, and you will no longer have to worry about any stressors. You should be very proud of the time and effort that you have committed to this study and all of us here at the FAST Lab commend you for that. Debriefing following entire protocol: 10. Thank you very much for taking part in this study! 11. Standardized debriefing on the purpose of the study and the specific measures that were taken: This study is a randomized-controlled experimental design and its purpose is to test the effect of different intensities, demarcated using the ventilatory thresholds from a cardiorespiratory exercise test, and examine how they may relate to the neurobiological reactivity to and recovery from a subsequent acute laboratory induced psychosocial stressor (i.e., public speaking task). In doing this study we hoped to gain insight into the physiological and psychological mechanisms that influence how exercise intensity (independent variable) modifies one’s response (dependent variable) to a stressful event. 114 It is well known, that the intensity of a single bout of exercise influences psychological and physiological responses to exercise. For example, the greater the intensity, the more one is likely to experience a dampening on their stress system. Moreover, intense exercise is associated with a hormonal stress response. For example, cortisol, a hormone in our body, rises in response to stress and also in response to hard exercise. We want to have a closer look at the associations between your responses to different intensity exercises and your reactions to a stressful event. You were randomly assigned to the _____________-intensity exercise group. That means that you performed today at a heart rate that corresponds to 30% (LICT – controlled walking pace of 1.7-2 Mph ) or 10% (Moderate intensity exercise) below the oxygen uptake corresponding to VT1, and halfway below VT2 (Vigorous intensity exercise). At this intensity, we have asked you to rate your feelings, your arousal and your perceived exertion across multiple time points measured before, during and after exercise, and during and following a subsequent stressor, the TSST. The saliva samples were taken to measure the changes in cortisol levels over time. By integrating the data from all participants, we can have a closer look at the interplay of these measures and determine in more detail how exercise influence one’s responses to stress. We ask that you do not talk about the specifics of this experiment outside of our lab, especially components related to the stress-task. This is mainly to prevent potential participants from developing expectations or biases prior to engaging in our protocol. Do you still have any questions about this? If there are some questions that come to your mind at a later date, please feel free to contact us at any time (email or phone us).” If you have any concerns about the study you may contact the Clinical Research Ethics Board at UBC which has reviewed this study and is responsible for helping to ensure that the rights of research participants are upheld. Please contact Pia Ganz at 604-875-4149 if you have any concerns. 12. UBC Counselling service, mental health support In our study we asked you to fill in some questionnaires about your levels of stress. We also used a stress induction protocol. Experiencing these events regularly in everyday life can have a significant negative impact, physically and mentally. We don’t know, if this directly applies to you, but here is a contact form for UBC counselling services that you may use at your discretion. Feel free to contact them at any time you like. 13. Review personal results of CPET test Participants are reminded they can ask questions about personal CPET-results: printed from COSMED computer. 14. Participant will receive HSP credits upon completion of the study. Participants will receive 1 credit per hour with a max of 6.5 credits and they will be uploaded into your HSP account after the completion of Day 2. 115 Section 2 - UBC Mental Health Support at Counselling Services A variety of mental health resources and professionals are available through Counselling Services. Counselling Services can work with you to assess your wellness needs, and identify the level of care that's right for you – from self-directed programs and tools to workshops, group programs, and individual therapy – to help you have the best university experience possible. For more information please refer to the resources below: Website: https://students.ubc.ca/health/counselling-services Make an appointment: Counselling Services - Operating hours Brock Hall Room 1040, 1874 East Mall Vancouver, BC V6T 1Z1 Canada Day Hours of Operation Monday 8:30 am - 4:30 pm Tuesday 8:30 am - 4:30 pm Wednesday 8:30 am - 7:00 pm Thursday 10:30 am - 4:30 pm Friday 8:30 am - 4:30 pm Saturday Closed Sunday Closed 116 Section 3 - STUDY REVIEW WORKSHEET Thank you for participating in this study. The research portion of the session is now over, and the research assistant will next spend some time telling you more about this study to help clarify the larger goals of the research. The following review worksheet is designed by the Psychology Department Subject Pool to help you follow along as you learn more about this study and the nature of scientific inquiry in psychology. Although completing this worksheet is voluntary, your responses will help maximize what you learn from your experience as a participant in the Department Subject Pool. Responses can also help researchers refine their procedures and allow the Psychology Department to recognize study review sessions that participants find most educational. Your responses are completely anonymous, will not be used for research purposes or have any bearing on receiving credit for your participation. 1. What was the main purpose and/or broader implication of the current study? 2. This study relies primarily on an experimental or correlational design (circle one)? 3. The primary independent or predictor variable in this study is: _______________________ 4. The primary dependent or outcome variable in this study is: ________________________ 5. The primary hypothesis being tested is: 6. If you were going to do a study like this, what would you change/add/modify? 7. If this study was published in an Introductory Psychology textbook, it would be in a chapter on (circle at least one): Biological Psychology Development Sensation/Perception Learning/Memory Motivation/Emotion Stress/Health Psychological Disorders Personality Quantitative Methods Cognition/Language Social Psychology 8. How clear and informative was the debriefing for this study? not at all 1 2 3 4 5 extremely 9. How interesting and engaging was the debriefing for this study? not at all 1 2 3 4 5 extremely 10. Were you orally debriefed by the researcher? yes no TO BE COMPLETED BY STUDY RESEARCH ASSISTANT Study Name:_______________________________________________________________ Date of Participation: __________________ Principal Investigator (Core Faculty): ___________________________________________ Research Assistant:__________________________________________________________ RA Signature: ______________________________________________________________ Please return this completed form to the RA who conducted your session. 117 D.3 HSP Take-home Debrief Form School of Kinesiology Faculty of Education Room 104-2176 Health Sciences Mall Medical Sciences Block C Vancouver B.C. Canada, V6T 1Z3 Research Title: Exercise and Stress 2.0 (EAST 2.0) The main purpose of this study is to test how different exercise intensities, defined by the ventilatory thresholds from a cardiorespiratory exercise test, influence people’s subsequent reactivity to and recovery from an acute psychosocial stressor (the Trier Social Stress Test; TSST). The TSST consists of a mock job interview and a mental arithmetic component and is the gold standard in reliable laboratory-induced stressors due to its ability to raise cortisol levels and produce a stress response. Throughout your visit, we measured your psychological responses through questionnaires and physical responses with saliva samples (which will examine the hormone cortisol), your heart rate, heart rate variability, and total prereferral resistance and cardiac output with the Biopac. Finally, we measured your ventilatory data with the V2 mask. This study is a randomized-controlled experimental design and hopes to gain insight into the physiological and psychological mechanisms that influence how exercise intensity (independent variable) modifies one’s response (dependent variable) to a stressful event. Our primary hypothesis is that vigorous intensity exercise will dampen the cortisol trajectory following a subsequent psychological stressor, compared to light and moderate exercise. Contact Information If you have any questions or desire for further information about the study please contact the lead graduate students on this study, Hana Hicks, and Luke Peddie. You can also contact the Principal Investigator, Dr. Eli Puterman. 118 Appendix E Receipts E.1 Cash Receipt 119 E.2 HSP Receipt 120 Appendix F Data Collection Forms F.1 Data Collection Forms Day 1 Participant ID: __________________ Lead: __________________ Tester: __________________ Threshold of participant: ______________________________ Baseline – Exercise prep - Exercise period – CD/Exercise recovery Temperature: ______________Humidity: _____________ Ideal Time Record time (timer or day) Blood Pressure/Exercise Group RPE FS FAS Pre-arrival Duties Temp and Hum recorded COSMED calibration COSMED prep Participant file Biopac ready to go -turn on computer -electrodes -devices - tape Questionnaires Time: Participant Arrival Sit in room 108 Notify participants to ask questions throughout ~0 min Time: Review consent form and procedures ~2 min Participant completes: Current health status: - PAR-Q+ Responsibility checklist Demographic questionnaire ~ 8 min Manual Questionnaires Instructions: RPE FS FAS ~11 min Study Prep + fitting Participant clothes Uses washroom ~13 min Time: Record Participant Height Weight ~15min 121 Ideal Time Record time (timer or day) Blood Pressure/Exercise Group RPE FS FAS Fit Biopac -3 EGC on front Fit V2 Mask ~ 17min Time: F1 Baseline (20-min) Stand for 3 min ~ 30 min Time: *Start Timer SBP: __ DBP:___ SBP:__ DBP:___ F1 F2 Blood Pressure (Seated) *can put strap on but don’t start blood pressure measurements until 5 mins in ~33 Time: Blood Pressure ~BP takes 30 seconds and then wait a minute in between ~38 min SBP: ___ DBP: ___ Blood Pressure ~40min SBP: ___ DBP: ___ Blood Pressure ~42 min SBP: ___ DBP: ___ F2 F1 Stand (3-mins) *have participant keep feet on floor markers and look at a painting* ~43 min SBP: __ DBP:___ SBP:__ DBP:___ F1 Exercise Prep V2mask ~45min 122 Ideal Time Record time (timer or day) Blood Pressure/Exercise Group RPE FS FAS Safety protocol F3 Standing (3-mins) ~50 min Time: SBP: ___ DBP: ___ SBP: ___ DBP: ___ F3 F4 Submaximal Exercise Modified Bruce Protocol 21 min max ~53 min (0) Exercise Start time: Timer: Participant Check-in ~55 min (2) Participant Check-in ~57 min (4) Participant Check-in ~59 min (6) Participant Check-in ~61 min (8) Participant Check-in ~63 min (10) Participant Check-in ~65 min (12) Participant Check-in ~67 min (14) Participant Check-in ~69 min (16) Participant Check-in ~71 min (18) Participant Check-in ~73 min (20) 123 Ideal Time Record time (timer or day) Blood Pressure/Exercise Group RPE FS FAS End of Exercise -Have participant grab bars and step on the side of the treadmill ~74 min (21) Time: Timer: F4 *once they are done* F5 Recovery: Take off mask Walking 3 mins – 1.7 mph ~ 75min (0) Recovery Start time: Timer: F5 F6 Standing (2-min) ~78 min (3) F6 F7 Seated (3-min) Bp ~81 min (5) Timer: SBP: ___ DBP: ___ BP (3-min) ~84 min (8) Timer: SBP: ___ DBP: ___ End of Recovery ~86 min (10) Time: F7 Debriefing protocol: - Thank you - Reminder email - Send signed consent form ~90 min Time: |---FS, FAS, RPE every 2 mins of exercising ---| 124 Notes: 125 F.2 Data Collection Forms Day 2 Group randomization: __________________ Participant ID: __________________ Lead: __________________ Tester: __________________ Threshold of participant: ______________________________ Baseline – Exercise prep - Exercise period – CD/Exercise recovery – Pre-TSST/TSST – Stress recovery Saliva Sample Temperature: ______________Humidity: ______________ Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Pre-arrival Duties Temp and Hum recorded COSMED calibration COSMED prep Participant file Biopac ready to go -turn on computer -electrodes -devices - tape Load online/iPad questionnaires 12x Salivettes Labelled Time: Participant Arrival Sit in room 108 Notify participants to ask questions throughout ~0 min Time: Review consent form and procedures Water restriction Fill water bottle ~2 min Participant completes: Current health status: - PAR-Q+ Responsibility checklist PSS Anxiety Inventory Depression Revised Scale ~ 8 min Manual Questionnaires Instructions: RPE FS FAS Saliva sample procedure ~13 min 126 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Study Prep + fitting Participant clothes - change into Biopac shirt Uses washroom ~15 Time: Record Participant Height Weight ~25min Fit Biopac -3 EGC on front -8 electordes on neck and back **get participant to look straight ahead** Fit V2 Mask *Water restriction starts now ~ 27 min Time: F1 Baseline (20-min ish) Stand in Biopac(turn on) for 3 min *have participant keep feet on floor markers and look at a painting* ~ 35 min (0) Time: *Start timer SBP: __ DBP:__ SBP: __ DBP:___ F1 F2 Blood Pressure (Seated) *can put strap on but don’t start blood pressure measurements until 5 mins ~38 min (3) Time: Blood Pressure (Seated) ~43min (8) SBP: ___ DBP: ___ Blood Pressure ~44 min (9) SBP: ___ DBP: ___ Blood Pressure ~45 min (10) SBP: ___ DBP: ___ F2 F1 127 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Stand (3-mins) *have participant keep feet on floor markers and look at a painting* ~46 min (11) Timer: Time of sample 1: SBP: ___ DBP: ___ SBP: ___ DBP: ___ 1 F1 Exercise Prep Put mask on Stand on treadmill Safety protocol *nothing is being measured here ~49min F3 Standing (3-mins) ~59 min Time: SBP: ___ DBP: ___ SBP: ___ DBP: ___ 2 2 F3 F4 Submaximal Exercise First 5-7 minutes will be ramping up to exercise group (MICT, VICT) LICT stays at 1.7 Mph Con’t until 30-minute mark ~62 min (0) Exercise Start time: Timer: Participant reaches ~64 min (2) Participant Check-in ~66 min (4) Participant reaches *MICT Reached just below (V1) ~67 min (5) Participant Check-in ~68 min (6) Participant reaches *VICT Reached ~69 min (7) 128 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Participant Check-in ~70 min (8) Participant Check-in ~72 min (10) Participant Check-in ~74 min (12) Participant Check-in ~76 min (14) Participant Check-in ~78 min (16) Participant Check-in ~80 min (18) Participant Check-in ~82 min (20) Participant Check-in ~84 min (22) Participant Check-in ~86 min (24) Participant Check-in ~88 min (26) Participant Check-in ~90 min (28) End of Exercise -Have participant grab bars and step on the side of the treadmill ~92 min (30) Time: Timer: F4 F5 CD - Standing (3-min) Rest *Mask stays on Actual start of recovery *take two bp readings 1 minute apart ~92 min (0) SBP: ___ DBP: ___ SBP: ___ DBP: ___ 3 3 129 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C F5 Cooldown Begins (2-min) Change treadmill to 1.7 mph *Take mask off* Can have 1-2 oz of water immediately AFTER saliva sample ~95 min Time: Timer: Time of sample 2: F6 Exercise Recovery (45-min) Optional clothing change Seated for duration of recovery How it’s Made ~97 min (5) Recovery Start time: Timer: Check-in (20-min in) * Can have 1-2 oz of water immediately AFTER saliva sample ~112 min (20) Time of sample 3: End of Recovery (45-min) ~137 min (45) Time: Time of sample 4: F6 F1 Pre-TSST Begins (3-min) Standing BP -NA – BP Questionnaires about upcoming task on computer Saliva sample at end of pre-tsst ~137 min Time: Time of sample: 5: SBP: ___ DBP: ___ SBP: ___ DBP: ___ 4 F1 TSST Instructions Walk into room ~139 min Time: 130 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Give instructions F7 TSST -preparation Cort sample and BP#1 Leave room to get the iPad (0.30 -1.0 min) Return to take sample and give iPad to do questionnaire *Record BP#1 Leave TSST room Return after 4 min Return & take BP#2 Leave with ipad ~142 min (0) Time: Time of sample 6: SBP: ___ DBP: ___ SBP: ___ DBP: ___ 5 F7 F8 TSST ~147 min Time: F8 F9 TSST End Stay standing *blood pressure after they exit the TSST room while doing questionnaires on computer ~160 min Time: Time of sample 7: SBP: ___ DBP: ___ SBP: ___ DBP: ___ 6 6 F9 F11 Beginning of Stress-Recovery (70 mins) ~165 Time: GET PARTICIPANT TO FILL OUT THOUGHTS QUESTIONNAIRE (found in iPad) Saliva sample #8 AFTER questionnaire ~175 min (10) Timer (after questionnaire): Time of sample 8: 131 Ideal Time Record time (timer or day) Saliva Sample Blood Pressure/Exercise Group RPE FS FAS NA T/C Saliva Sample #9 ~185 min (20) Timer: Time of sample 9: Saliva Sample #10 ~195 min (30) Timer: Time of sample 10: Saliva Sample #11 ~215 min (50) Timer: Time of sample 11: Recovery Period End Stand for 3 mins before final sample 12 ~235 min (70) Timer: Time of sample 12: 7 F11 Debriefing protocol: - Thank you - Debriefing - UBC counselling service - CPET results - Honorarium ~240 min Time: Ending time second assessment day ~250 132 Notes: 133 Appendix G Trier Social Stress Test Script EAST Public Speech Task [Participant is led into TSST room and asked to step up to line and face judges] [Experimenter Record Polar Time immediately before instructions] The task is introduced to the participant by the lead experimenter: “Your task in this experiment is the following: Please imagine that you had applied for a job and have been invited for an interview. In contrast to a real interview, however, you are supposed to give a talk, in which you are to convince the panel in five minutes why you think that you would be the best candidate for this position. Please note that you will be recorded by a camera and a microphone for later voice and behavioural analysis. These members are trained in behavioural analysis and will take notes during your talk. You should try to leave the best possible impression, and assume the role of the applicant for the duration of the talk as best as you can. The members of the panel will reserve the right to ask follow-up questions in case of uncertainties to receive all necessary information from you. Following your talk you will then be given a second task by the panel, which will only be explained to you by the panel. Do you have any questions?" [If yes, the lead experimenter should respond] We are now going to complete some blood pressures and take a saliva sample. Afterwards you’ll be given an ipad questionnaire to fill out about this task. Then, you may take some notes to prepare, which you must not use during your talk. Then, we will take another blood pressure measurement before your talk begins. If you have no further questions, please stand behind the podium. [Strap BP cuff to participant, give them the sample] [Leave the room] [Grab the iPad with the survey loaded and re-enter to collect the sample – roughly 30 sec-1 min] [Return] [Record BP, take sample and give the iPad to participant] [Leave] [Return at 4 minutes to take final BP reading] [Take iPad and Leave] 134 Interview Task (5 min Prep + 5 min Talk & Qs) After 5 minutes of participant preparation (the final BP has been taken and the Lead has left the room), the Chairperson/Judge says: “Please step up to the line.” Wait for participant to step forward, then turn on camera “Begin your talk." If participant talks for longer than 3 minutes, then interrupt. “That’s enough, now please tell us why you think you’re better qualified than the other applicants” If participant stops before 3 minutes, wait for about 20s and then say: “You still have some time remaining, please continue.” [If participant does not continue after another 10 seconds, start asking questions.] Some research participants use their school-knowledge to distract from their own person. In that case the chair should certainly intervene, for example by saying "We believe you that you know how to execute a market analysis, but we would be more interested to find out why you were so involved in or drawn to this area.") Questions to ask the research participant during the “job interview” – good questions highlighted in red • Why do you think you are especially well-qualified for this task? • What do your family/ friends especially appreciate about you? • What do you appreciate about your friends? • You just pointed out that you were especially good at…, what other characteristics qualify you? • You just spoke about…, what exactly do you then think about…? • Please complete the following sentence: “I am the best in…” • Please list your strengths! • Please list your weaknesses! • What kind of leadership qualities do you have? • What do you think about teamwork? • Where do you see your position in a team? • What can you constructively add to a team? • You just mentioned that you really appreciate teamwork. What do you think about people who like to work independently? • What do your employees appreciate about you most? • Would you be willing to work overtime without compensation? • Would you be willing to work on the weekends if necessary? • What kind of qualities to you expect from your co-workers? • Under what circumstances would you be willing to take responsibility for your employees’ mistakes? • Would you lie in order to gain an advantage? • What do you think about the saying “Everybody determines his own luck” Arithmetic Task (5 min) This part of the test should take five minutes maximum, unless the participant has reached "0" before that. 135 Chairperson/Judge 1 says: “We now want you to complete a mental arithmetic task. Please count aloud backwards from 2023 in increments of 17. Please calculate as quickly and correctly as possible. If you miscalculate, we will point out your mistake and you have to start over again. Do you have any questions?” Answers: 2023 2006 1989 1972 1955 1938 1921 1904 1887 1870 1853 1836 1819 1802 1785 1768 1751 1734 1717 1700 1683 1666 1649 1632 1615 1598 1581 1564 1547 1530 1513 1496 1479 1462 1445 1428 1411 1394 1377 1360 1343 1326 1309 1292 1275 1258 1241 1224 1207 1190 1173 1156 1139 1122 1105 1088 1071 1054 1037 1020 1003 986 969 952 935 918 901 884 867 850 833 816 799 782 765 748 731 714 697 680 663 646 629 612 595 578 561 544 527 510 493 476 459 442 425 408 391 374 357 340 323 306 289 272 255 238 221 204 187 170 153 136 119 102 85 68 51 34 17 0 If the participant miscalculates: "That was incorrect. Please begin at 2023." If they make more than three mistakes. “Please start from (new number) counting down in increments of 17.” If the participant does not miscalculate after several increments: "Please try to calculate faster." If the participant is fidgeting, looking at the floor or ceiling, or avoiding eye-contact: “Please keep your hands by your sides.” or… “Please look straight ahead” or… “Please look into the camera as you calculate.” Saliva Sample #3 Chairperson/Judge 1 says: “That’s enough. Please take this clipboard and marker and step back to the line. Without revealing your responses, mark how you feel on the laminated questionnaires” [Judge 2 hands clipboard and open marker] “We will now take a saliva sample. Please take this salivette and begin to chew for 1 minute.” [Judge 2 hands them salivette; Time approximately 1 minute]. [After chewing for 1 minute] Your minute is up. Please spit the swab back into the tube. Bring the test tube and the clipboard to the experimenter in the other room.” END OF TASK Around 70 minutes after the end of the TSST, the experimenter brings the panel judges into the waiting room. Judges should greet the participant, assure him that he did a great job in the interview, shake hands, explain that their role was to stress him, and can make friendly small talk for a minute before leaving. In other words, feel free to smile and act naturally now! The participant is likely to feel relieved when the judges explain that they were playing a role. 136 Appendix H Questionnaires H.1 Demographic Questionnaires Demographic Questionnaire What is your sex assigned at birth? How would you best describe your gender identity? 1. How would you best describe your ethnic origin o Caucasian o African American o Native American o Asian o Middle eastern o Other ethnic group: ___________________ o Prefer not to answer 2. What is your current occupation o Employed o Unemployed o Retired o Other: ____________________ o Student If yes: o Undergraduate o Graduate o Prefer not to answer 3. How tall are you? <actual measures will be taken by research personnel> 4. What is your weight? <actual measures will be taken by research personnel> _______________________ 137 H.2 Current Health Status 138 H.3 Pre-Exercise Responsibility Checklist Day number? o 1. Pre-study Day o 2. Experimental Day Have you… Yes No Exercised in the last 24 hours? o o Had caffeine within the last 2 hours? o o Eaten a large meal in the last 2 hours? o o Had any alcohol today? o o Hydrated well over the last 2 days? o o 139 H.4 Physical Activity Readiness Questionnaire for Everyone (PAR-Q+) 140 141 142 143 H.5 L-CAT (Leisure Time Categorical Activity Scale) During the past month, which statement best describes the kinds of physical activity you usually did? Do not include the time you spent working at a job. Please read all six statements before selecting one. 1. I did not do much physical activity. I mostly did things like watching television, reading, playing cards, or playing computer games. Only occasionally, no more than once or twice a month, did I do anything more active such as going for a walk or playing tennis. 2. Once or twice a week, I did moderate activities such as getting outdoors on the weekends for an easy walk or stroll. Or once or twice a week, I did chores around the house such as sweeping floors or vacuuming. 3. About three times a week, I did moderate activities such as brisk walking, swimming, or riding a bike for about 15-20 minutes each time. Or about once a week, I did moderately difficult chores such as ranking or mowing the lawn for about 45-60 minutes. Or about once a week, I played sports such as softball, basketball, or soccer for about 45-60 minutes. 4. Almost daily, that is five or more times a week, I did moderate activities such as brisk walking, swimming, or riding a bike for 30 minutes or more each time. Or about once a week, I did moderate difficult chores or played sports for 2 hours or more. 5. About three times a week, I did vigorous activities such as running or riding hard on a bike for 30 minutes or more each time. 6. Almost daily, that is, five or more times a week, I did vigorous activities such as running or riding hard on a bike for 30 minutes or more each time. 144 H.6 Centre for Epidemiology Studies – Depression Scale – Revised 145 H.7 Perceived Stress Scale (PSS) 146 H.8 Rating of Perceived Exertion (RPE-Scale) Ratings of Perceived Exertion Scale (RPE, Borg, 1985) When doing physical activity, we want you to rate your perception of exertion. This feeling should reflect how heavy and strenuous the exercise feels to you, combining all sensations and feelings should reflect how heavy and strenuous the exercise feels to you, combining all sensations and feelings of physical stress, effort, and fatigue. Do not concern yourself with any one factor such as leg pain for shortness of breath but try to focus on your total feeling of exertion. Try to appraise your feeling of exertion as honestly as possible, without thinking about what the actual physical load is. Your own feelings of effort and exertion is important, not how it compares to others. Look at the rating scale below while you are engaging in activity; choose the number from below that best describes your level of exertion. [Ask participant: “How hard are you working?”] 6 No exertion at all 7 Extremely light 8 9 Very light 10 11 Light 12 13 Somewhat hard 14 15 Hard 16 17 Very hard 18 19 Extremely hard 20 Maximal exertion 147 H.9 Felt Arousal Scale (FAS) Felt Arousal Scale (FAS, Svebak & Murgatroyd, 1985) Estimate here how aroused you actually feel. Do this by pointing at the appropriate number. By “arousal” we meant how “worked-up” you feel. You might experience high arousal in one of a varity of ways, for example as excitement or anxiety or anger. Low arousal might also be experienced by you in one of a number of different ways, for example as relaxation or boredom or calmness. [Ask participant: “How aroused do you actually feel?”] 6 High arousal 5 4 3 2 1 Low arousal 148 H.10 Feeling Scale The Feeling Scale (FS, Hardy & Rejeski, 1989) While participating in exercise, it is common to experience changes in mood. Some individuals find exercise pleasurable, whereas others find it to be unpleasant. Additionally, feeling may fluctuate across time. That is, one might feel good and bad a number of times during exercise. Scientists have developed this scale to measure such responses. [Ask participant: “How do you feel?”] +5 Very good +4 +3 Good +2 +1 Fairly good 0 Neutral -1 Fairly bad -2 -3 Bad -4 -5 Very bad 149 H.11 Negative & Positive Affect Questionnaire (PANAS-X) Negative and Positive Affect Schedule (PANAS-X, Watson & Clark, 1994) This scale consists of a number of words that describe different feelings and emotions. Read each item and then indicate your answer by circling the number. Indicate to what extent you feel this way right now, that is, at the present moment. Very slightly or not at all A little Moderately Quite a bit Extremely 1. Excited 0 1 2 3 4 2. Dissatisfied 0 1 2 3 4 3. Afraid 0 1 2 3 4 4. Calm 0 1 2 3 4 5. Strong 0 1 2 3 4 6. Blue 0 1 2 3 4 7. Attentive 0 1 2 3 4 8. Distressed 0 1 2 3 4 9. Upset 0 1 2 3 4 10. Active 0 1 2 3 4 11. Interested 0 1 2 3 4 12. Ashamed 0 1 2 3 4 13. Hostile 0 1 2 3 4 14. At ease 0 1 2 3 4 15. Proud 0 1 2 3 4 16. Guilty 0 1 2 3 4 17. Inspired 0 1 2 3 4 18. Alert 0 1 2 3 4 19. Scared 0 1 2 3 4 20. Alone 0 1 2 3 4 21. Sad 0 1 2 3 4 22. Enthusiastic 0 1 2 3 4 23. Jittery 0 1 2 3 4 24. Relaxed 0 1 2 3 4 25. Nervous 0 1 2 3 4 26. Irritable 0 1 2 3 4 27. Determined 0 1 2 3 4 150 H.12 Task Appraisal Questionnaires Task Appraisal Questionnaire (Pre-Task) Please indicate by circling a number after each statement to indicate how you are feeling right now regarding the task you are about to complete. Strongly Strongly Disagree Neutral Agree 1. The upcoming task is very demanding. 1 2 3 4 5 6 7 2. I am very uncertain about how I will perform during the upcoming task. 1 2 3 4 5 6 7 3. The upcoming task will take a lot of effort to complete. 1 2 3 4 5 6 7 4. The upcoming task is very stressful. 1 2 3 4 5 6 7 5. I have the abilities to perform the upcoming task successfully. 1 2 3 4 5 6 7 6. It is very important to me that I perform well this task. 1 2 3 4 5 6 7 7. I’m the kind of person who does well in these types of situations. 1 2 3 4 5 6 7 8. A poor performance on this task would be very distressing for me. 1 2 3 4 5 6 7 9. I expect to perform well on this task. 1 2 3 4 5 6 7 10. I view the upcoming task as a positive challenge. 1 2 3 4 5 6 7 11. I think the upcoming task represents a threat to me. 1 2 3 4 5 6 7 12. I feel as if I am in complete control of my performance 1 2 3 4 5 6 7 151 Task Appraisal Questionnaire (Post-Task) Please indicate by circling a number after each statement to indicate how you are feeling right now regarding the task you have just completed. Strongly Strongly Disagree Neutral Agree 1. The task was very demanding. 1 2 3 4 5 6 7 2. I am uncertain about how I performed. 1 2 3 4 5 6 7 3. I exerted a lot of effort during the task. 1 2 3 4 5 6 7 4. The task was very stressful. 1 2 3 4 5 6 7 5. I felt that I had the abilities to perform well in the task. 1 2 3 4 5 6 7 6. It was very important to me that I performed well this task. 1 2 3 4 5 6 7 7. I believe I performed well on the task. 1 2 3 4 5 6 7 8. I felt that the task challenged me in a positive way. 1 2 3 4 5 6 7 9. I felt threatened by the task. 1 2 3 4 5 6 7 10. I felt in complete control during the task. 1 2 3 4 5 6 7 152 H.13 Thoughts Questionnaire Please rate each statement as to how often you have thought about that aspect in the time since you gave your speech. Never Not Often Sometimes Often Very Often 1. My speech was good 0 1 2 3 4 2. I could have done much better 0 1 2 3 4 3. How anxious I felt 0 1 2 3 4 4. I should have chosen a different topic 0 1 2 3 4 5. If my blushing/sweating/dry mouth/shaking was obvious 0 1 2 3 4 6. How well I handled it 0 1 2 3 4 7. How bad my speech was 0 1 2 3 4 8. I made a fool of myself 0 1 2 3 4 9. How much I enjoy these situations 0 1 2 3 4 10. How I always do badly in this type of situation 0 1 2 3 4 11. I must have looked stupid 0 1 2 3 4 12. How smoothly it all went 0 1 2 3 4 13. How self-conscious I felt 0 1 2 3 4 14. What a failure I was 0 1 2 3 4 15. That I chose an interesting topic 0 1 2 3 4 16. How many mistakes I made 0 1 2 3 4 17. How confident I felt 0 1 2 3 4 18. I came across as self-assured 0 1 2 3 4 19. How awkward I felt 0 1 2 3 4 20. That I was at my best 0 1 2 3 4 21. How fast my heart was pounding 0 1 2 3 4 22. I didn’t make a good impression 0 1 2 3 4 23. Other aspects of the speech task 0 1 2 3 4 24. The overall speech task situation 0 1 2 3 4 153 Appendix I Pre-exercise Explanation I.1 Full Pre-exercise Explanation Day 1 FULL PRE-EXERCISE EXPLANATION DAY 1 Purpose/Participant goal • You are going to complete a maximal graded exercise test, called the BSU Bruce protocol. • The treadmill is going to increase its speed and grade of the treadmill continuously until the end • Your goal is to walk, jog, and run for as long as you can safely continue. • Once you think you are 10-15 seconds from hitting your capacity to continue the exercise safely, please raise your hand to notify the research team but continue exercising • Once you have hit your max please immediately grab onto the side bars and jump feet onto the stationary sides • The research team will immediately put the treadmill into “recovery mode” • Do not touch red button or pull the cord (those are only for emergencies) • Keep feet on the sides until the research team instructs you that it is safe to “remount” the treadmill pad Mask/Hand signals • You are wearing the V2 mask so we can capture ventilatory data. Feel free to speak and ask questions until the test begins. • Once you start moving, we are going to ask that you don’t speak. Please respond to the questionnaires and our questions by pointing, giving a thumbs up, or using hand signals. • However, in case of emergency, or if you are feeling lightheaded, dizzy, short of breath, nauseous or any symptoms greater than those you would feel during normal exercise, please speak and let us know immediately. Treadmill safety • Your safety is our number 1 priority. A couple of things to be careful of: • Like all treadmills this treadmill has moving pad that is between two stationary sections. Please be aware of your footing to not accidently step on the stationary sides while running • You will be attached to the COSMED computer with the V2 mask. It is important to be aware of where you are on the treadmill to avoid pulling on the cords. o The research team will also inform you to step closer to the top of the treadmill if you start to move back o Tip: it can be helpful to use the bars as a guide to know your placement on the treadmill by tapping the back of your hands on them *demo this* Emergency safety protocol • Again, if you begin to have chest pain or feel lightheaded, dizzy, short of breath, nauseous or any symptoms greater than those you would feel during normal exercise, please speak and let us know immediately. • We will terminate the test and manage whatever symptoms you have. • You are attached to a red safety clip that will immediately stop the treadmill when disconnected 154 • There is also an emergency stop button (big red button) on the treadmill that will have the same effect • There is also an emergency stop button on the COSMED computer which will be monitored by the research team Explanation of tester roles • Throughout the test, both testers will be monitoring your safety as well as doing a number of tasks • Tester 1 will primarily be monitoring your metabolic data and making sure the computer is recording good data. • Tester 2 will ask you to respond to the 3 questionnaires every 2 minutes, please point to indicate your response. They will say your response aloud, please give a thumbs up to confirm. (Practice this prior to test). Questionnaire Demo • I am going to ask you to respond to these 3 questionnaires every 2 minutes. I will hold the questionnaire here and ask you to point to your response. o I will verbally confirm the number with you, please respond with a thumbs up to confirm (again, please don’t speak). To end the protocol reminder • Your goal is to continue the protocol for as long as you can • Once you are 15-10 seconds from hitting your capacity to continue the exercise, please raise your hand to notify the research team and keep going • Once you have hit your max please immediately grab onto the side bars and jump feet onto the stationary sides Are you ready? INSTRUCTIONS FOR TESTERS DURING EXERCISE • This is a max test therefore you need to check-in with them continuously and cheer them on (I.e. “You got this!” “Keep going!” “You're doing great!” “Dig deep here!” etc.) • Ask all three questionnaires every 2 minutes • Always use the same order (RPE, FS, FAS) • Use the exact wording on the questionnaires • Following each 3 responses, ask for thumbs up or a nod to confirm they are doing okay • Make sure to monitor their placement on the treadmill – tell them to move up if they start to slip back on the pad (need to be always on the upper half of pad to avoid pulling the cord) 155 I.2 Full Pre-exercise Explanation Day 2 FULL PRE-EXERCISE EXPLANATION DAY 2 Treadmill safety • Your safety is our number 1 priority. A couple of things to be careful of: • Like all treadmills this treadmill has moving pad that is between two stationary sections. Please be aware of your footing to not accidently step on the stationary sides while running • You will be attached to the COSMED computer with the V2 mask. It is important to be aware of where you are on the treadmill to avoid pulling on the cords. o The research team will also inform you to step closer to the top of the treadmill if you start to move back o Tip: it can be helpful to use the bars as a guide to know your placement on the treadmill by tapping the back of your hands on them *demo this* Mask/Hand signals • You are wearing the V2 mask so we can capture ventilatory data. Feel free to speak and ask questions until the test begins. • Once you start moving, we are going to ask that you don’t speak. Please respond to the questionnaires and our questions by pointing, giving a thumbs up, or using hand signals. Emergency safety protocol • If you begin to have chest pain or feel lightheaded, dizzy, short of breath, nauseous or any symptoms greater than those you would feel during normal exercise, please speak and let us know immediately. • We will terminate the test and manage whatever symptoms you have. • You are attached to a red safety clip that will immediately stop the treadmill when disconnected • There is also an emergency stop button (big red button) on the treadmill that will have the same effect • There is also an emergency stop button on the COSMED computer which will be monitored by the research team Purpose/Participant goal • You have been randomized into the _____ exercise group *If Light read this* • The treadmill will be set to and stay at a 1.7 mph walking pace and you will exercise at that pace for 30 minutes *If Moderate read this* • The treadmill will be risen to a grade of 6.5 degrees • Using your data from Day 1 we have found your first ventilatory threshold (VT1) and will use the first 5 minutes of the exercise to ramp you up to just below this threshold and keep you there for the remaining 25 minutes 156 *If Vigorous read this* • The treadmill will be risen to a grade of 6.5 degrees • Using your data from Day 1 we have found your two ventilatory thresholds (VT1 and VT2) and will use the first 7 minutes of the exercise protocol to ramp you up to a pace that keeps your VT in the middle of these two thresholds • You will be held at this threshold marker for the remaining 23 minutes *Read for all* • At the end of the 30 minutes of exercising the treadmill will automatically go into recovery mode • The research team will instruct you to grab onto the bars and jump feet onto stationary sides • You will immediately start a 3-minute standing rest before taking a saliva sample and going into a 2 minute walking- recovery Explanation of tester roles • Throughout the test, both testers will be monitoring your safety as well as doing a number of tasks • Tester 1 will primarily be monitoring your metabolic data and making sure the computer is recording good data. • Tester 2 will ask you to respond to the 3 questionnaires every 2 minutes, please point to indicate your response. They will say your response aloud, please give a thumbs up to confirm. (Practice this prior to test). Questionnaire Demo • I am going to ask you to respond to these 3 questionnaires every 2 minutes. I will hold the questionnaire here and ask you to point to your response. o I will verbally confirm the number with you, please respond with a thumbs up to confirm (again, please don’t speak). To end the protocol reminder • At the end of the 30 minutes of exercising the treadmill will automatically go into recovery mode • The research team will instruct you to grab onto the bars and jump feet onto stationary sides • You will immediately start a 3-minute standing rest before taking a saliva sample and going into a 2-minute walking- recovery We will start will 3-minute standing baseline and the research team will inform you before the exercise portion is going to start – please don’t speak Are you ready? *starts 3-minute baseline* … after 3 minute mark… start trreadmill 157 INSTRUCTIONS FOR TESTERS DURING EXERCISE • This is a max test therefore you need to check-in with them continuously and cheer them on (I.e. “You got this!” “Keep going!” “You're doing great!” “Dig deep here!” etc.) • Ask all three questionnaires every 2 minutes • Always use the same order (RPE, FS, FAS) • Use the exact wording on the questionnaires • Following each 3 responses, ask for thumbs up or a nod to confirm they are doing okay • Make sure to monitor their placement on the treadmill – tell them to move up if they start to slip back on the pad (need to be always on the upper half of pad to avoid pulling the cord) 158 Appendix J Lab Manual Forms J.1 COSMED Cheat-Sheet Turn on button (green light) before opening Cosmed Omnia Calibration: Gas calibration  metabolic ergo  start  follow prompted instructions from computer  open valve  you want between 5-7 (turn the black nob until you get to 6 then leave it)  plug in the small aux to the main computer before hitting start  leave it alone while it is calibrating  there shouldn’t be any red, if so check the back again and make sure the valve is open properly and there is gas  restart the calibration  if still red … could be sampling line (replace as a last resort) Flowmeter calibration hook up flowmeter (clear tube, turbine, black piece connected to cart)  do six strokes (computer counts for you)  want the numbers to be between -.1 and .1 (it is okay if it is exactly -.1 or .1) *You must calibrate every single participant To turn off the gas once the data collection is complete: Turn black knob until it is loose  release the tube in the back and put it back in  open and close the valve to release access gas (dial should be showing 0)  then turn off COSMED computer (green light should be off) Creating a New Profile (Day 1) Database  “New subject” First name: EAST2 Last name: ID# Gender: female/male Ethnicity: Other Real birthday Height and weight: you collect this data before putting on biopac New Test  Metabolic  Cardio Pulmonary  Breath by breath Day 1 Bruce Protocol (Max Test) Once you hit start a settings page will pop up, select: Type: Max Data filtering: none Ergometer: COSMED treadmill Mph Protocol: BRUCE 159 port: same Everything else same Notes for Bruce Protocol: • Once you hit *start* and *rec* the treadmill will do all the work • You just make sure the data is being collected properly and watch the participant • Participant works until they feel as though they can’t anymore or until 21 minutes (whichever is first) • Once they indicate that they can no longer continue you will click *recovery* and the treadmill will automatically go into recovery mode o Do NOT hit *stop* unless it is an emergency because it will stop the test Day 2 New Test (for Leads – in-depth form to follow)  Find participant in database  Select participant  edit if necessary  New Test  Metabolic  Cardio Pulmonary  Breath by breath Once you hit start a settings page will pop up, select: Type: Submax Data filtering: Rolling Time Average Ergometer: COSMED treadmill Mph Protocol: none port: same Everything else same 160 J.2 BIOPAC Set-up Prepare setup • Tray • Abrasive Pad prepared • 11 electrodes (or 3 + 4 paired electrodes) • 11 Pieces of tape pre-ripped • Measuring Tape • Cords unpacked and prepared for participant ECG Setup Red – Positive: Find Bone Landmark on L-side below heart (last rib White – Negative: Find Bony Landmark on R-side above heart (Clavicle) Black – Ground: Find Bony Landmark on L-side above heart (Clavicle) 161 NICO Setup Electrodes: White – Impedence Stimulus: Place on either side of spinous process, below where the neck folds when participant looks up. Do not touch electrodes together. White – Voltage: Place on either side of spinous process, below the electrodes above. Do not touch electrodes together. Red – Voltage: Landmark 25cm below bottom edge of White – Voltage and equally space the electrodes to the side Red – Impedence Return: Place directly below Red – Voltage, do not touch electrodes together. Wiring (Outside – In Principle): Connect White – Impedence to Red Impedence Connect White – Voltage to Red - Voltage
Thesis/Dissertation
eng
Kinesiology
Vancouver : University of British Columbia Library
Attribution-NonCommercial-NoDerivatives 4.0 International
Graduate
Puterman, Eli
Effects of exercise intensities on cortisol release following the induction of a psychological stressor
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