Seasonal differences in the diurnal pattern of cortisol secretion in healthy participants and those with self-assessed seasonal affective disorder

This study compared the daily pattern of free salivary cortisol secretion in winter and in summer between two groups; participants with self-assessed seasonal affective disorder (SAD) and age- and sex-matched healthy controls. Fifty-two participants completed the study with an equal number in each group. The diurnal pattern of cortisol secretion was assessed across two consecutive weekdays in summer, and two in winter, with conditions being counterbalanced. On each study day participants collected multiple saliva samples in the domestic setting to capture the cortisol awakening response (CAR) and declining levels across the day. In addition, perceived stress, anxiety, depression, state stress and state arousal were assessed using validated questionnaires. There was no evidence for any seasonal changes in psychological data or cortisol pattern for the healthy control population. In summer, self-assessed SAD and control participants had similar psychological and cortisol profiles. In winter however, SAD participants reported greater depression, stress and anxiety, and lower levels of arousal. Furthermore, the CAR was significantly attenuated in SAD participants during winter months. There was no difference in cortisol levels during the rest of the day between controls and SAD participants in winter. In line with the above findings and previous research, there was an inverse relationship between the increase in cortisol following awakening and a measure of seasonality in winter. Furthermore in winter, a general dysphoria construct correlated inversely with the CAR, indicating that participants reporting greater depression, stress and anxiety and lower arousal, exhibited lower CARs. In conclusion, during the shortened photoperiod in winter, the cortisol response to awakening is attenuated in participants with self-assessed SAD in comparison to controls. These findings contribute to the understanding of the physiology of SAD.     

Circadian regulation of cortisol after hippocampal damage in humans.

BACKGROUND: There is substantial evidence that the hippocampus (HC) regulates the activity of the hypothalamic-pituitary-adrenocortical (HPA) axis. Damage to the HC in animals produces a transient alteration in diurnal and stress-related HPA activity. This study was designed to examine the effects of HC damage on basal cortisol secretion in humans. METHODS: Salivary cortisol was measured in 22 patients with HC damage (12 with bilateral damage and 10 with unilateral damage), 7 brain-damaged comparison participants, 10 healthy, age-matched comparison participants, and 6 of the patients' caregivers. Salivary cortisol samples were taken immediately after awakening, 30 min after awakening, at 8:00 am, 11:00 am, 3:00 pm, 6:00 pm, and at bedtime on a single day. Brain-injured patients underwent a structural magnetic resonance imaging scan to examine quantitative volumes of the HC. RESULTS: Both bilateral and unilateral HC damage abolished the cortisol response to awakening documented in the comparison groups. Caregivers of bilateral HC patients showed a reduced response to awakening. The remainder of the circadian pattern was not affected in the HC patients; all groups showed a significant diurnal variation. There was no association between HC volume and cortisol secretion. CONCLUSIONS: Hippocampal damage in humans abolishes the cortisol response to awakening, whereas the remainder of the diurnal cycle is unaffected in these patients. These data suggest a unique role of the HC in the control of basal cortisol secretion.     

Diurnal patterns of salivary cortisol across the adolescent period in healthy females

When examining the diurnal profile of the hormone cortisol in children and adolescents developmental issues are particularly relevant. Previous findings regarding relationships between cortisol secretory activity and reproductive (pubertal) maturation lack clarity and may reflect methodological inconsistencies between studies. This study examined the diurnal cortisol profile across female adolescence, with a particular focus on an obvious and unique marker of development: menarche. In a cross-sectional design, 61 healthy female adolescents aged 9-18 years (mean age 13.89 years, S.D.+/-2.72) collected eight saliva samples per day on two consecutive weekdays. Samples were collected at awakening, 15, 30 and 45min and 3, 6, 9 and 12h post-awakening in order to capture both the cortisol awakening response (CAR) and the subsequent period of decline. Demographic information was recorded and participants also completed the Spielberger State-Trait Anxiety Inventory. Patterns of cortisol secretion exhibited good intra-individual stability across the two sampling days. Participants evidenced a robust diurnal pattern, with cortisol levels peaking approximately 30-45min post-awakening (the CAR) and steadily declining concentrations over the remainder of the day. Differences according to developmental status (in terms of whether or not participants had experienced first menses: menarche) were observed in the time of peak secretion of the CAR, and these distinct patterns could not be accounted for by group differences in demographic, situational or psychological characteristics measured in this study. This effect for the CAR was associated with the onset of menarche alone, unlike cortisol levels over the remainder of the day. For those who had undergone menarche, were older and of greater BMI, cortisol levels remained higher over the day. There was a significant difference in cortisol concentrations at 6h post-awakening between pre- and post-menarche groups. Again, these differences in daytime cortisol secretory activity could not be attributed to situational or psychological factors. Establishing patterns of cortisol secretion in healthy female adolescents provides an important baseline from which to investigate hypothalamic-pituitary-adrenal (HPA) physiology, measured via salivary cortisol, in adolescent populations with known or suspected psychopathology.     

Diurnal salivary cortisol levels in postpartum mothers as a function of infant feeding choice and parity

Daily stress and sleep deprivation can influence the diurnal pattern of cortisol, which normally consists of high morning levels and a gradual decline throughout the day. While most individuals have consistent declining cortisol concentrations over days, others display either flat or inconsistent profiles. Postpartum mothers experience considerable home demands and sleep deprivation, yet, breastfeeding mothers perceive lower stress and reduced negative mood states compared to bottlefeeders. On the other hand, multiparity (having more than one child) is associated with reduced steepness in diurnal cortisol decline. Interestingly, no study to date has investigated the diurnal cortisol pattern and its stability across days in postpartum women as a function of their choice of infant feeding and parity. In this study, we measured salivary cortisol at four different time points during the day, on two non-consecutive days in first-time (primiparous) and second-time (multiparous) mothers at 5-20 weeks postpartum who were exclusively breastfeeding or bottlefeeding, and in non-postpartum mothers of young children (1-6 years). Among multiparous mothers, we found that cortisol levels in those who were bottlefeeding were higher than in breastfeeding mothers at both awakening and 1600 h. This effect remained significant after controlling for individual differences related to infant feeding choice, such as estradiol levels, education and income. No effect of infant feeding choice on cortisol concentrations was observed in primiparous mothers. While a consistent decline across days was common, some mothers presented a flat or inconsistent profile, a profile that was not associated with infant feeding choice or parity. Importantly, mothers with consistent declining profiles had the highest household income. Our findings suggest that although breastfeeding might promote a tighter regulation of diurnal basal cortisol secretion, in particular for multiparous mothers who are likely to be exposed to greater home demands and maternal responsibilities, some aspects of socioeconomic status such as income can also play a significant role in the stability of diurnal cortisol secretion across days.     

Decreased hair cortisol concentrations in generalised anxiety disorder

Previous research examining hypothalamic-pituitary-adrenal (HPA) axis activity in generalised anxiety disorder (GAD) has suggested a general hypercortisolism. These studies have mostly relied on salivary, plasma or urinary assessments, reflecting cortisol secretion over short time periods. The current study utilised the novel method of cortisol assessment in hair to obtain a retrospective index of cortisol secretion over a prolonged period of time. Hair cortisol levels were determined in 15 GAD patients and in 15 age- and gender-matched controls. In addition, participants collected six saliva samples (on awakening, +30 min, 12:00, 16:00, 20:00 h and at bedtime) on two consecutive weekdays for the assessment of the diurnal cortisol profile. Results revealed significantly lower (50-60%) cortisol levels in the first and second 3-cm hair segments of GAD patients compared to those of controls. No significant between-group differences were seen in diurnal cortisol profiles. The hair cortisol findings tentatively suggest that under naturalistic conditions GAD is associated with hypocortisolism. If corroborated by future research, this demonstrates the important qualities of cortisol measurement in hair as an ecologically valid, retrospective index of long-term cortisol secretion and as a marker for psychiatric disorders associated with hypo- or hypercortisolism.     

Association between time of awakening and diurnal cortisol secretory activity

A 12-hour diurnal profile of salivary free cortisol was measured in healthy adults (n=40) on two consecutive days. Samples were collected at timed intervals synchronised to awakening. The mean profile is characterised by a marked increase in cortisol concentration following awakening, peaking after about 30 min, and a subsequent decline over the remainder of the day. Thus two components of the diurnal cycle were examined: a) the first 45 min post-awakening (the awakening cortisol response) and b) the underlying 12 h profile from immediately until 12 h post awakening (but without the awakening response). Both of these components were analysed in two ways such as to provide an indication of overall cortisol concentration and the degree of change in cortisol concentration, i.e. the rise for the awakening response and the diurnal decline. Both components of the cortisol diurnal profile were negatively correlated with awakening time. Thus, those subjects who awoke earliest had higher levels of cortisol over the 45 min following awakening as well as throughout the rest of the day. They also displayed a more marked diurnal decline to be convergent with late awakeners at the end of diurnal measurement, 12 h following awakening. Hence the diurnal cortisol cycle, which is synchronised to awakening, is significantly related to awakening time. These findings support the notion of a close association between suprachiasmatic nucleus (SCN) control of both awakening and cortisol secretory activity.     

Neuroticism and introversion are associated with salivary cortisol patterns in adolescents

Previous studies have yielded equivocal findings on the relationship between personality and cortisol activity. The present study examined associations between personality and cortisol activity in a large, diverse adolescent sample, while partialling the effects of relevant demographic and health-related covariates. A subsample of 230 participants (57% of whom reported elevated neuroticism) was selected from a larger sample of 16-18-year olds involved in a study on risk factors for emotional disorders. Subsample participants completed a battery of personality questionnaires, and saliva collection was requested several months later on three consecutive days at six time points per day, from wakeup to bedtime. Associations between personality and cortisol rhythms were examined using multilevel growth curve modeling. Neuroticism (N) and introversion (I) were significantly and differentially associated with features of diurnal cortisol patterns. Specifically, a significant N x gender interaction was observed, demonstrating flatter cortisol rhythms across the waking day among male participants with higher N. Elevated I, however, was associated with lower cortisol awakening responses for both male and female participants, and higher cortisol at the time of waking for male participants only. The present study supports personality as a significant predictor of diurnal cortisol patterns in late adolescence, after accounting for the effects of demographic and health covariates, and suggests that gender plays a role in moderating associations between personality and cortisol.     

The diurnal patterns of the adrenal steroids cortisol and dehydroepiandrosterone (DHEA) in relation to awakening

The steroid hormones, cortisol and dehydroepiandrosterone (DHEA) are the two main peripheral secretory products of the hypothalamic-pituitary-adrenal stress-neuroendocrine axis. The diurnal pattern of cortisol secretory activity has been well characterised. Various aspects of this pattern have been related to time of awakening, light exposure, psychological dimensions of affect, immune function and systemic health and well-being. DHEA is also an important adrenocortical steroid whose secretory activity has been related to immune function, psychological and health variables. The most pronounced feature of the diurnal cortisol cycle is a burst of secretory activity following awakening with a diurnal decline thereafter. We mapped DHEA secretory activity onto this cycle by measuring both steroids in saliva samples collected at distinct time points over the diurnal cycle, synchronised to awakening. Both steroids, particularly DHEA, showed stability across days of sample collection. A main distinction between cortisol and DHEA was that although DHEA was elevated in post-awakening samples compared with later in the day there was no evidence of an awakening stimulatory burst of DHEA secretory activity. Although DHEA in many respects paralleled cortisol secretory activity there was some dissociation; mean levels were positively but not tightly correlated. The secretory pattern of DHEA is very stable whereas cortisol secretory activity seems more sensitive to day-to-day variability.     

Ageing, physical function, and the diurnal rhythms of cortisol and dehydroepiandrosterone

The present study examined the relationship between ageing, physical function and the diurnal rhythms of cortisol and dehydroepiandrosterone (DHEA). Participants were 36 community dwelling older adults aged between 65 and 86 years old. Salivary cortisol and DHEA were measured over the course of one day: immediately upon awakening, 30 min later, and then 3 h, 6 h, 9 h and 12 h post-awakening. Participants completed the Nottingham extended activities of daily living index, the Berg Balance Scale and their handgrip strength was assessed. Older participants had a significantly higher cortisol area under the curve (AUC), lower overall DHEA levels, lower DHEA AUC, a decreased diurnal slope of decline and increased cortisol:DHEA ratio. Lower diurnal cortisol levels were associated with poorer performance on the Berg Balance Scale and lower handgrip strength, and those with a flattened DHEA diurnal profile reported less independence in carrying out daily tasks. These associations withstood adjustment for age. In conclusion, this study suggests an association between cortisol, DHEA, ageing and physical function.     

Transactions among adolescent trait and state emotion and diurnal and momentary cortisol activity in naturalistic settings

In a community sample of 52 adolescents, multilevel growth curve modeling was utilized to examine whether within-person changes in momentary mood states, and individual differences in trait emotional functioning, were related to adolescent cortisol levels in naturalistic settings. Salivary cortisol levels were measured seven times a day on two typical weekdays in conjunction with diary reports of adolescent mood states. Questionnaire reports of trait emotional functioning (depression, anxiety, and anger) were obtained, as were reports of demographic, developmental, and health control variables. After accounting for the effects of time of day and a wide range of control variables, within-person increases in state negative mood (worry/stress and anger/frustration) were significantly associated with within-person increases in cortisol. When examining trait emotional functioning, adolescents with higher levels of depressive symptoms had slightly lower basal cortisol levels, and adolescents with higher levels of trait anger had a significantly stronger cortisol response to awakening. Several developmental effects were found-adolescents at higher stages of pubertal development had daytime basal cortisol curves that were more elevated, had a steeper diurnal decline, and showed a lesser cortisol awakening response, and cortisol responses to worry/stress increased with age. Cortisol levels were also higher at moments adolescents were alone rather than with others, an effect that declined significantly with age. Cortisol levels were also higher at moments adolescents were alone rather than with others, an effect that declined significantly with age. Results suggest that ongoing transactions occur between adolescents' everyday emotional experiences and their cortisol levels, and that adolescent cortisol activity is modified by age/pubertal stage and by trait emotional functioning.     

Associations of the cortisol awakening response (CAR) with cortical activation asymmetry during the course of an exam stress period

The cortisol awakening response (CAR) is a cortisol rise which is distinct from the circadian rise in hypothalamus-pituitary-adrenal (HPA) axis activity before awakening. The CAR has been shown to be related to experiences of stress and negative affect, and activation of neocortical networks has been suggested as a mechanism. Right-sided cortical activation has been shown to be correlated with negative affect, and an association of electroencephalogram (EEG) asymmetry measures with cortisol secretion has been demonstrated. Therefore, we investigated for associations of the CAR with lateralised trait-like cortical activation and with changes in EEG asymmetry during a putative stressful period. We examined 37 undergraduate students before, during, and after an academic exam period. CARs were measured five times and EEG was measured both about 6 weeks before the beginning of the exams and 1 day before an exam. Trait-like interindividual differences in posterior cortical asymmetry were differentially associated with CARs at different measurement occasions. Participants with greater right centroparietal cortical trait activation showed an increased CAR in anticipation of the exams, whereas all other participants showed an increased CAR in response to the exams. Furthermore, EEG measures taken directly before the exam revealed that greater right frontal cortical activation was related to higher cortisol levels after awakening. The results suggest that lateralised cortical activation moderates CAR changes during the course of a stressful period. Lateralised cortical activation may be an important link between the CAR and health-related variables like experiences of stress and negative affect.     

Abnormal diurnal patterns of salivary α-amylase and cortisol secretion in acute patients with anorexia nervosa

Abstract

Objectives. The evidence that the activity of the sympathetic nervous system (SNS) is decreased in acute anorexia nervosa (AN) is not consistent. Therefore, we aimed to assess the SNS basal activity in malnourished AN patients through the measurement of diurnal salivary levels of α-amylase, whose secretion is regulated by the SNS. As secondary aim, we measured also salivary cortisol. Methods. Eight symptomatic female patients with restrictive AN and eight age-matched healthy women underwent saliva sample collection at awakening and over the day. α-amylase and cortisol were assayed by ELISA method. Results. In both patients and controls, saliva α-amylase levels significantly decreased during 60 min after awakening and then progressively rose towards the afternoon/evening. AN patients exhibited significantly reduced levels of the salivary enzyme with a significant decrease in its overall diurnal secretion and a dysregulated secretory pattern. As compared to control women, AN patients exhibited significantly enhanced levels of salivary cortisol at awakening, an enhanced and advanced cortisol secretion after awakening but no significant change in the overall diurnal secretion of the salivary hormone. Conclusions. These results suggest that the activity of the SNS, evaluated through the assessment of the diurnal secretion of salivary α-amylase, is impaired in the acute phase of AN whereas the cortisol awakening response is enhanced.     

Cortisol secretion and fatigue: Associations in a community based cohort

The association between fatigue and reduced activity in the hypothalamo-pituitary–adrenal (HPA) axis has been described. However the temporal association between fatigue and HPA activity is under debate. We examine whether alterations in cortisol secretion play a role in the development of fatigue or whether changes occur later as a consequence of fatigue in a longitudinal cohort study of 4299 community dwelling adults (mean age 61). Cortisol secretion was measured from saliva samples collected waking, waking + 0.5, 2.5, 8, 12 h and bedtime at phase 7 (2003–2004) of the Whitehall II study. Fatigue was measured at phase 6 (2001), phase 7 and phase 8 (2006) of the Whitehall II study. Three elements of secretion were examined: waking cortisol, the cortisol awakening response and diurnal slope in cortisol secretion. Fatigue was determined using the vitality sub-scale of the Short Form-36. A wide variety of co-variates were measured. We find that fatigue measured at phase 6 was not associated with cortisol secretion at phase 7. At phase 7, low waking cortisol levels and a flat slope in diurnal cortisol secretion were associated with fatigue independently of co-variates. In participants low or free of fatigue at phase 7 low waking cortisol and flatter slope in cortisol secretion were associated with new-onset fatigue at phase 8 (for example, odds ratio for lowest vs. highest tertile of waking cortisol 1.50; 95% confidence intervals, 1.08, 2.09 after adjusting for all co-variates). In conclusion, we find that low waking salivary cortisol and a flat slope in cortisol secretion is associated with fatigue. Cortisol is also associated with future onset of fatigue suggesting that changes in cortisol secretion are etiologic or occur early in the genesis of fatigue.     

Familial influences on basal salivary cortisol in an adult population

To understand the underlying genetic and environmental sources of individual variation in basal cortisol levels, we collected salivary cortisol at awakening and at six fixed time points during the day in adult twins and their singleton siblings. Reported time of awakening was verified with heart rate and body movement recordings. Cortisol data were available for 199 MZ twins, 272 DZ twins and 229 singleton siblings from 309 twin families. No differences in cortisol means and variances were found between twins and singleton siblings. Additionally, the correlations for DZ twins and siblings were not significantly different, indicating generalizability of twin study results to the general population. Genetic model fitting showed heritability for cortisol levels during the awakening period (34% for cortisol level at awakening and 32% for cortisol level at 30 min after awakening) but not for cortisol levels later during the day. The current study shows that, while cortisol levels in the awakening period are influenced by genetic factors, cortisol levels throughout most of the day are not heritable, indicating that future gene finding studies for basal cortisol should focus on the first hour post-awakening.     

Diurnal rhythm of cortisol during late pregnancy: associations with maternal psychological well-being and fetal growth

Maternal psychological functioning during pregnancy affects both maternal and fetal well-being. The hypothalamic-pituitary-adrenal (HPA) axis provides one mechanism through which maternal psychosocial factors may be transduced to the fetus. However, few studies have examined maternal psychological factors or birth outcomes in relation to the diurnal pattern of cortisol across the day. The current study examined maternal psychological well-being, parity status, and birth weight in relation to the maternal cortisol diurnal rhythm in a group of 98 low-risk pregnant women (51 primiparae). At 36 weeks gestation, participants completed both pregnancy-specific and general self-report measures of psychological functioning and provided saliva samples at 8:00, 12:00, and 16:00h on 2 consecutive working days for the assay of cortisol. The expected diurnal decline in salivary cortisol was observed. Higher trait anxiety was associated with a flatter afternoon decline for all mothers. For primiparae, steeper morning cortisol declines were associated with lower infant birth weight. The findings suggest that regulation of the HPA axis may differ by parity status with downstream implications for fetal growth and development.     

Differences in cortisol awakening response on work days and weekends in women and men from the Whitehall II cohort

It is thought that the salivary cortisol awakening response can serve as a reliable marker of hypothalamic-pituitary-adrenocortical activity. Response magnitude might be influenced by stress, but results of studies on work-related stress have been inconclusive. Non-compliance with the sampling schedule is a potential confounding factor that has rarely been controlled. The objective of the study was to determine whether the cortisol awakening response is greater on a work day than on a weekend day, and whether responses vary with gender and socioeconomic position. Compliance was controlled by excluding participants who reported a delay of more than 10 min between waking up and taking the first saliva sample. Data were collected from 196 men and women aged 47 to 59 years drawn from the Whitehall II cohort of British civil servants, with free salivary cortisol sampled immediately after waking up and 30 min later on 2 days. Data on stress, perceived control and happiness over the remainder of the day, and on sleep quality, time of waking, and health behaviour were also obtained. The awakening response was lower in non-compliant participants than in compliant ones, so non-compliant participants were excluded from further analyses. Salivary cortisol levels on waking did not differ by gender or socioeconomic position, or between work and weekend days. However, the cortisol awakening response (defined as the difference between waking and 30 min later) was greater on work than weekend days (mean increases 10.5 and 3.7 nmol/l, P < 0.001). On the work day, women showed larger increases than men (P = 0.011), but there were no gender differences on the weekend day. Across both days, lower socioeconomic position was associated with a larger cortisol awakening response (P = 0.014). Time of waking up was not related to the cortisol awakening response on either day. Participants rated themselves as more stressed, less in control, and less happy over the remainder of the work than weekend day. These results indicate that the cortisol awakening response occurs on both work and non-work days, but that anticipation of the working day is associated with an enhanced response. Cortisol output over the early part of the day may be particularly sensitive to the influence of chronic stress and its anticipation, especially in women.     

Parental adiposity and cortisol awakening responses in young men and women

A heightened cortisol awakening response (CAR) is associated with adiposity in middle-aged men, but the causal significance of this effect is not known. We hypothesised that if disturbance in cortisol secretion is involved in the development of overweight and obesity, then it might be present in normal weight adults at increased risk of obesity on account of parental adiposity. The CAR and cortisol profile over the day were measured in 33 men and 62 women aged 18-25 years. Parental adiposity was assessed with figure ratings derived from the Contour Drawing Rating Scale, and these were correlated with parental self-reported body mass index (BMI) in a subset of participants (r=0.66-0.79). In men, a positive association was observed between the CAR and their judgements of their fathers' adiposity after controlling for age, smoking status, time of waking, and the participants' own BMI; the correlation was 0.56 (P=0.008) for the cortisol increase between waking and 30 min, and 0.47 (P=0.028) for the cortisol area under the curve. The correlation between the CAR and fathers' own reported BMI and figures ratings were also significant. The relationship between parental adiposity and the CAR in women was inconsistent, and the associations between the CAR and opposite gender parental adiposity were not significant. Parental adiposity was not related to cortisol output over the rest of the day or to the slope between waking and evening in either sex. The results of this study suggest that disturbances of cortisol secretion may present before the emergence of heightened adiposity in young men at raised risk for obesity.     

Chronotype and diurnal cortisol profile in working women: differences between work and leisure days

The influence of chronotype on the diurnal profile of salivary cortisol was examined in a sample of 187 healthy women: 21 evening chronotype, 24 morning chronotype and 142 intermediate chronotype. Saliva samples were collected at waking, 30 min post-awakening, at 1000 h, 1200 h, 1500 h, 1700 h and at bedtime on one work and one leisure day. Several components of the diurnal profile were examined including the cortisol awakening response, the total cortisol output and the diurnal profile on both the work and the leisure day, a significant main effect of time emerged (both p<0.01). After adjustment for age, smoking status, self-rated health, time of waking, and sleep problems, no effect of chronotype was evident for cortisol in the evening, the cortisol awakening response, or total cortisol output over the working day. However, on the leisure day, total cortisol output was greater in evening-types than intermediate or morning-types, after adjustment for covariates (p=0.029). The present data indicate that chronotype has a limited impact on the diurnal cortisol profile of healthy women, and may be somewhat impervious to individual preferences for morning or evening activity.     

Cortisol fluctuates with increases and decreases in negative affect

Prior studies have reported cortisol secretion to be primarily influenced by negative affect, but less is known about cortisol activity during states of activation involving increased positive affect and decreased negative affect. On separate days, 30 healthy young men experienced: an activating and humorous video; a speech stressor; and a resting control period. Cortisol was measured in saliva before and after each 30-min mood induction. Positive affect (activation) was increased similarly by both the video and the speech compared to rest (p < .0001). Negative affect increased during the speech and decreased during the video (p < .001). Cortisol increased only during the speech (p < .0001). Following the video, however, cortisol was decreased significantly (p < .0001). Rest day cortisol revealed no differences across periods (p > .1). These results suggest that the hypothalamic-pituitary-adrenocortical axis is a dynamic system influenced by changes in negative affect irrespective of the experience of generalized activation.