Diurnal Salivary Cortisol Profiles Research Articles

Comparative analysis of salivary cortisol measurements using different assay methods in relation to serum-free cortisol measurement

ObjectivesSalivary cortisol reflects the biologically active form of serum cortisol, offering a noninvasive evaluation method for the diurnal rhythm of the hypothalamic-pituitary-adrenal (HPA) axis. While liquid chromatography-tandem mass spectrometry (LC-MS/MS) is known for its specificity, immunoassays (IA) are commonly used because of their simplicity. This study aimed to assess the performance of salivary cortisol measurement using both IA and LC-MS/MS in comparison to serum-free cortisol measurement. MethodsAssay results for 188 saliva and 94 serum samples from 47 participants were analyzed. Salivary samples collected at different time points were analyzed using IA and LC-MS/MS. Serum samples were analyzed for cortisol, cortisol-binding globulin, and free cortisol. The statistical analyses included correlations and method comparisons. ResultsThe diurnal salivary cortisol profiles exhibited a comparable circadian rhythm pattern; however, the concentrations measured using IA were consistently higher than those measured using LC-MS/MS. The correlation analysis revealed robust associations among salivary cortisol (IA), salivary cortisol (LC-MS/MS), and serum-free cortisol levels (LC-MS/MS). However, the method comparison revealed a systematic bias between IA and LC-MS/MS in salivary cortisol measurement. ConclusionsThis study contributes to the ongoing debate on assay techniques by affirming the suitability of IA and LC-MS/MS for salivary cortisol measurement to assess dynamic changes in HPA axis activity. The identified systematic bias emphasizes the importance of selecting methods based on specific research or clinical requirements.

Serial Diurnal Salivary Cortisol Profiles in 667 pregnant women - Association with Cardiometabolic Complications.

Maternal obesity, hypertensive pregnancy disorders and gestational diabetes (GDM) are linked to an increased risk of negative offspring health outcomes. This association may be mediated by maternal hypothalamic-pituitary-adrenal axis (HPA axis) activity, resulting in elevated maternal cortisol levels and fetal exposure, but evidence remains scarce. We examined (1) maternal diurnal cortisol profiles longitudinally across gestation, and (2) explored associations with maternal cardiometabolic complications. Women in the InTraUterine sampling in early pregnancy (ITU) study (n=667) provided seven salivary cortisol samples from awakening to bedtime up to three times during pregnancy (median gestational week 19.3, 25.7, and 38.1, n=9,356 samples). Changes in cortisol awakening response and diurnal slope (indicative of HPA-axis activity) and their associations with maternal body mass index (BMI), hypertensive pregnancy disorders and GDM were examined using linear mixed models. The cortisol awakening response declined in in 60%-67% of women, and the diurnal slope attenuated from early to late pregnancy (b = 0.006, p = .001). Higher BMI was associated with less decline in cortisol awakening response (b= 0.031, p = .0004), and less attenuation in diurnal slope from early to late pregnancy (b = -0.001, p = .006). Hypertensive pregnancy disorders and GDM were not significantly associated with diurnal cortisol profiles. The attenuation in cortisol awakening response and diurnal slope support HPA-axis hypo-responsivity during pregnancy. Less attenuation of both markers in women with a higher BMI may indicate reduced adaption of the HPA-axis to pregnancy, presenting a mechanistic link to offspring health outcomes.

Online, low-volume meditation does not alter immune-related biomarkers

ObjectivesPrior studies of mindfulness meditation have demonstrated anti-inflammatory and immunoregulatory effects but whether meditation courses delivered online can exert similar effects is poorly understood. Barriers to large scale implementation of traditional mindfulness meditation programs has created an increased interest in the effect of less time- and resource-intensive online meditation courses. The purpose of this study was to determine whether a 6-week online mindfulness program with low time demands on nurses would lead to changes in gene expression, cytokine profiles, telomerase activity, and cortisol profiles. MethodsThis was a randomized, parallel pilot study comparing an online mindfulness-based stress management program to an active control group from December 2018 to May 2019. Healthy nurses with above average levels of perceived stress were randomized to receive a 6-week online mindfulness-based stress management program including ≥5 min daily meditation practice or listen to relaxing music for ≥5 min daily as the control arm. Blood samples were collected at baseline and after 6 weeks, and various self-reported measures of stress, physical and emotional health were collected at baseline, after 6 weeks, and after 12 weeks. Whole transcriptome mRNA sequencing of whole blood at baseline and after 6 weeks was performed along with measurement of plasma IL-6, IL-8, IL-10, TNF-α, and IFN-γ. Peripheral blood mononuclear cells were isolated, and telomerase activity was measured. Diurnal salivary cortisol profiles were assessed at baseline and after 6 weeks. The primary outcome was change over time in a pre-determined set of 53 genes representative of the immune-related changes seen with stress, which was analyzed using a mixed linear model. Secondary outcomes included all other self-reported measures and biomarkers mentioned above. ResultsA total of 61 nurses were randomized, with 52 having sufficient data to include in the final analysis. After 6 weeks, nurses in the control group reported significant reductions in stress as measured by the Perceived Stress Scale while those in the mindfulness group did not. However, after 12 weeks, the mindfulness group also showed a significant reduction in stress. When compared to the control group, no significant changes in RNA gene expression or any other biomarkers were observed in the nurses who participated in the mindfulness program. ConclusionsOur study found that this brief online mindfulness-based intervention was effective in reducing stress in nurses, albeit with a delayed effect compared to listening to relaxing music. Regarding immunoregulatory effects, there were no significant differences between treatment and control groups in transcriptomic or other tested biomarkers of immune function. This study provides evidence for a floor effect of mindfulness on transcriptional and circulating biomarkers of immune function.

Diurnal Salivary Cortisol Profiles in Patients with Cushing's Syndrome.

Quantification of salivary cortisol is one of the highly sensitive and specific screening parameters for Cushing's syndrome (CS). However, only late-night salivary cortisol is part of the standard screening procedure. In this study, we aimed to analyze salivary cortisol day profiles in patients with different types of CS to test whether specific patterns might be relevant for diagnosis and subtyping. Among 428 patients including those with confirmed Cushing's syndrome (N=111, of those 75 with Cushing's disease, 27 patients with adrenal CS and nine patients with ectopic CS), autonomous cortisol secretion (N=39) or exclusion of CS (control group, N=278) salivary cortisol was measured five times a day. At each of the five time points, salivary cortisol was significantly higher in patients with CS compared to the control group (p≤0.001). Using the entire profile instead of one single salivary cortisol at 11 p.m. improved diagnostic accuracy (85 vs. 91%) slightly. Patients with ACTH-dependent CS had higher salivary cortisol levels than patients with adrenal CS. Also, morning cortisol was significantly higher in patients with ectopic CS than in patients with Cushing's disease (p=0.04). Nevertheless, there was a strong overlap between diurnal profiles, and the diagnostic yield for subtyping was low. The study results show that using diurnal salivary cortisol profiles for CS diagnosis results in a limited increase in diagnostic accuracy. With significant differences between Cushing subtypes, cortisol profiles are not useful in everyday clinical practice for subtyping of CS.

Exploring associations between diurnal cortisol, stress, coping and psychopathology in adolescents and young adults with 22q11.2 deletion syndrome

Background22q11.2 deletion syndrome (22q11DS) is a neurogenetic condition associated to a high risk for psychiatric disorders, including psychosis. Individuals with 22q11DS are thought to experience increased levels of chronic stress, which could lead to alterations in hypothalamic-pituitary-adrenocortical (HPA)-axis functioning. In the current study, we investigated for the first time diurnal salivary cortisol profiles in adolescents and young adults with 22q11DS as well as their link with stress exposure, coping strategies and psychopathology, including psychotic symptoms. MethodsSalivary cortisol was collected from adolescents and young adults with 22q11DS (n = 30, age = 19.7) and matched healthy controls (HC; n = 36, age = 18.5) six times a day for two days. Exposure to stressful life events, including peer victimization, coping strategies and general psychopathology were assessed with questionnaires. Psychotic symptoms and psychiatric comorbidities were evaluated with clinical interviews. ResultsWe observed similar daily levels and diurnal profiles of salivary cortisol in adolescents and young adults with 22q11DS compared to HCs. However, participants with 22q11DS reported less frequent exposure to stress than HCs. In 22q11DS, we observed a significant association between the use of non-adaptive coping strategies and the severity of psychotics symptoms. Cortisol level was not associated to severity of psychotic symptoms, but elevated cortisol awakening response (CAR) was found in participants with 22q11DS with higher levels of general psychopathology. ConclusionsOur results do not support earlier propositions of altered HPA-axis functioning in 22q11DS but highlight the need to further investigate diurnal cortisol as an indicator of HPA-axis functioning and its link with (earlier) stress exposure and psychopathology in this population. Interventions should target the development of adaptive coping skills in preventing psychosis in 22q11DS.

Impact of coping strategies on perceived stress, depression, and cortisol profiles among gynecologic cancer patients.

We explored associations between problem-focused, emotional processing, and emotional expression coping strategies and markers of stress including perceived stress, depressive symptoms, and diurnal cortisol profiles among women with gynecologic cancer. Problem-focused coping was associated with less perceived stress, fewer depressive symptoms, and more rhythmic diurnal salivary cortisol profiles. Emotional processing was associated with lower perceived stress and fewer depressive symptoms. Emotional expression was associated with fewer depressive symptoms and elevated diurnal mean and evening cortisol levels. Results point to key differences in coping strategies. In this sample, only problem-focused coping was linked with adaptive differences in both psychological and physiological stress measures.

Low-Grade Inflammation and Ambulatory Cortisol in Adolescents: Interaction Between Interviewer-Rated Versus Self-Rated Acute Stress and Chronic Stress.

To determine whether the association between self-rated or interviewer-rated recent acute stress exposures and low-grade inflammation and daily cortisol production in adolescents is moderated by chronic stress ratings. Acute and chronic stress exposures were assessed in 261 adolescents aged 13 to 16 years using a semistructured life stress interview. The negative impact of acute stressors was independently rated by both adolescents (self-rated) and interviewers (interviewer-rated). Markers of inflammation (interleukin (IL)-6, IL-1ra, C-reactive protein) were measured from peripheral blood samples obtained via antecubital venipuncture. Participants collected 4 saliva samples at home on each of 6 consecutive days for the analysis of diurnal salivary cortisol profiles. There were no main effects of acute stressors (self- and interviewer-rated) and chronic family or peer stress on adolescent inflammation markers and cortisol (p values > .10). However, the interaction between interviewer-rated acute stress and chronic family stress was significantly associated with adolescent inflammation markers (IL-6, IL-1ra). Specifically, as chronic family stress increased, the association between acute stressor impact (interviewer-rated) and inflammation markers became more positive (IL-6 (B = .054, SE = .023, p = .022); IL-1ra (B = .030, SE = .014, p = .034)). Interactions between self-rated acute stress and chronic family stress were not associated with any biological measures (p values > .10). Interactions between acute stressor impact (both self- and interviewer-rated) and chronic peer stress were also not significantly associated with any biological measures (p values > .05). Among adolescents, interviewer-based ratings of acute stressor impact may allow for better prediction of health-relevant inflammation markers than adolescents' own ratings.

The CIRCORT database: Reference ranges and seasonal changes in diurnal salivary cortisol derived from a meta-dataset comprised of 15 field studies

Diurnal salivary cortisol profiles are valuable indicators of adrenocortical functioning in epidemiological research and clinical practice. However, normative reference values derived from a large number of participants and across a wide age range are still missing.To fill this gap, data were compiled from 15 independently conducted field studies with a total of 104,623 salivary cortisol samples obtained from 18,698 unselected individuals (mean age: 48.3 years, age range: 0.5–98.5 years, 39% females). Besides providing a descriptive analysis of the complete dataset, we also performed mixed-effects growth curve modeling of diurnal salivary cortisol (i.e., 1–16h after awakening). Cortisol decreased significantly across the day and was influenced by both, age and sex. Intriguingly, we also found a pronounced impact of sampling season with elevated diurnal cortisol in spring and decreased levels in autumn. However, the majority of variance was accounted for by between-participant and between-study variance components. Based on these analyses, reference ranges (LC/MS–MS calibrated) for cortisol concentrations in saliva were derived for different times across the day, with more specific reference ranges generated for males and females in different age categories. This integrative summary provides important reference values on salivary cortisol to aid basic scientists and clinicians in interpreting deviations from the normal diurnal cycle.

No alterations in diurnal cortisol profiles before and during the treatment in patients with stress-related exhaustion.

Several theories have emerged in recent years suggesting that neuroendocrinological alterations, mainly changes in cortisol, could be of importance with respect to the link between chronic stress and disease. This study investigated possible deviations in the diurnal cortisol profiles of patients with clinically diagnosed stress-related exhaustion (exhaustion disorder - ED) compared with healthy controls. Salivary cortisol samples taken at home in the morning directly after waking up, 30 min later, and in the evening were compared between ED patients (N = 122; 25% men) and healthy controls (N = 98; 44% men). Follow-up measurements were performed after 6 months (79 patients) and 12 months (68 patients) of the treatment. There were no clear differences in diurnal salivary cortisol profiles between the patients and healthy controls. Moreover, salivary cortisol levels and diurnal profiles did not change significantly during the treatment in the patient group. There was some indication of a smaller cortisol awakening response in the male patients compared to the male controls, but the difference appeared to be mainly related to the antidepressant use. Diurnal salivary cortisol profiles, at least as measured in this study, give a rather poor reflection of the prolonged stress exposure experienced by patients with ED. Such basal salivary cortisol measurements do not seem suitable as biomarkers for stress-related conditions such as ED or burnout, or as an aid to assess the effects of prolonged stress load in a routine clinical practice.

Preterm delivery as a predictor of diurnal cortisol profiles in adulthood: evidence from Cebu, Philippines.

Fetal exposure to elevated maternal cortisol can permanently modify hypothalamic-pituitary-adrenal (HPA) axis function, and thereby have long-term health impacts. Maternal cortisol steadily increases throughout normal pregnancy, but is abnormally high in preterm deliveries (<37 weeks). Prematurity remains a widespread public health problem, yet little is known about its potential long-term effects on adult HPA function. Here we test the hypothesis that diurnal cortisol profiles measured in young adulthood will vary based upon an individual's preterm status. Diurnal salivary cortisol profiles, a marker of HPA-axis function, were measured in 1,403 young adults (ages 21-23 years) participating in the Cebu Longitudinal Health and Nutrition Survey, located in Metropolitan Cebu City, Philippines. Males who had been born preterm exhibited lower morning cortisol and non-significantly elevated evening cortisol, resulting in a more adverse, flatter rate of decline across the day. In contrast, there were no significant differences by preterm status in cortisol measured at any time of day in females. These findings point to potential long-term effects of having been born preterm on adult HPA-axis function, and add to evidence from this and other populations for sex differences in the biological and health impacts of prenatal stress exposure.

Salivary cortisol in pregnant women suffering from blood and injection phobia

Stress and/or anxiety during pregnancy affect maternal and fetal well-being and can cause premature delivery and postnatal pathology in the child. Women suffering from phobias related to blood and injections are prone to high levels of stress, including anxiety and sometimes panic attacks, during pregnancy. Cortisol is amongst the mediators through which the neurohormonal expressions of maternal psychological factors may be transduced to the fetus. The aim of this study was to investigate whether pregnant women suffering from blood and injection phobia have raised cortisol levels or are characterized by unusual diurnal salivary cortisol profiles compared with healthy controls. The sample consisted of 110 pregnant women with blood and injection phobia and 110 pregnant healthy controls. Both groups provided morning and evening saliva samples in weeks25 and 36 for the assay of cortisol. In gestational week25, when blood was drawn for the mandatory blood testing, extra blood was taken to analyze corticotrophin-releasing factor, adrenocorticotropic hormone, and cortisol in serum. The diurnal decline in salivary cortisol as well as increased cortisol levels were observed during pregnancy. Pregnant women suffering from blood and injection phobia had a higher output of cortisol compared with women without the phobia (F = 6.25, df = 1, p = 0.014), but no marked difference in the diurnal cortisol rhythm was found between groups. Our findings indicate that untreated blood and injection phobia during pregnancy increases cortisol concentrations. Blood and injection phobia is treatable, and cognitive behavioral therapy can be used. Women with blood and injection phobia during pregnancy therefore need to be recognized and offered treatment without delay in early pregnancy.

The ratio of cortisol/DHEA in chronic fatigue syndrome

Introduction: Hypoactivity of the hypothalamic-pituitary-adrenal axis (HPA axis) has been one of the most consistent biological findings in chronic fatigue syndrome (CFS). However, it has been hypothesised that the adrenal steroid dehydroepiandrosterone (DHEA) may counteract the effects of cortisol in the brain (Kroboth et al., 2003) and that consequently a true measure of the “net” effect of cortisol on the brain is better obtained by calculating the ratio of cortisol to DHEA. Studies have not attempted to measure this pre and post Cognitive Behavioural Therapy (CBT), an effective treatment for CFS (Whiting et al., 2000) also recommended byNational UK guidelines. Methods: Basal (0900 h) plasma levels of cortisol, DHEA and their ratio were measured in 41 patients with CFS without depression and in 28 matched controls. In 33 of the above patients this ratio was repeated following a 6 month course of CBT. Furthermore diurnal salivary cortisol profiles were measured at 0800, 1200, 1600 and 2000 h pre- and post-CBT in all 41 patients. Results: The plasma ratio of cortisol/DHEA (mean±SD) did not differ between patients and controls (5.91±0.46 vs 5.86±0.46, p=0.97). Although there was no difference comparing the 0900 h plasma cortisol/DHEA ratio before and after CBT, 0900 h plasma DHEA was significantly lower after CBT (34.7±18.4 vs. 31.2± 21.0 nmol/l, p=0.001). While total daily salivary cortisol output from 0800–2000 h (measured as the area under the curve) increased significantly from 68.2±19.6 to 79.4±21.8 nmol/l h (t=2.62, p<0.05) after CBT. Conclusions: The combined novel findings of increased total daily salivary cortisol output and reduced plasma DHEA suggest that ‘functional’ cortisol is increased following CBT for CFS. We suggest that there disavantages in using single plasma levels and that further clarity will be obtained in future research by measuring both cortisol and DHEA concurrently in saliva throughout the day.

High Prevalence of Central Adrenal Insufficiency in Patients with Prader-Willi Syndrome

The annual death rate of Prader-Willi syndrome (PWS) patients is very high (3%). Many of these deaths are sudden and unexplained. Because most deaths occur during moderate infections and PWS patients suffer from various hypothalamic insufficiencies, we investigated whether PWS patients suffer from central adrenal insufficiency (CAI) during stressful conditions. Overnight single-dose metyrapone tests were performed. Metyrapone (30 mg/kg) was administered at 2330 h. At 0400, 0600, and 0730 h, ACTH, 11-deoxycortisol, cortisol, and glucose levels were measured. Diurnal salivary cortisol profiles were assessed on a different day at wake-up, 30 min after wake-up, at 1400 h, and at 2000 h. The study was conducted in a pediatric intensive care unit. Patients included 25 randomly selected PWS patients. Patients were considered as having CAI when ACTH levels remained below 33 pmol/liter at 0730 h. Median (interquartile range) age was 9.7 (6.8-13.6) yr. Fifteen patients (60%) had an insufficient ACTH response (CAI, P < 0.001). There was no significant difference in age, gender, genotype, and body mass index SD score between patients with CAI and those without. Morning salivary cortisol levels and diurnal profiles were normal in all children, suggesting that CAI becomes apparent only during stressful conditions. Strikingly, 60% of our PWS patients had CAI. The high percentage of CAI in PWS patients might explain the high rate of sudden death in these patients, particularly during infection-related stress. Based on our data, one should consider treatment with hydrocortisone during acute illness in PWS patients unless CAI has recently been ruled out with a metyrapone test.

Exploration of basal diurnal salivary cortisol profiles in middle-aged adults: Associations with sleep quality and metabolic parameters

The use of saliva samples is a practical and feasible method to explore basal diurnal cortisol profiles in free-living research. This study explores a number of psychological and physiological characteristics in relation to the observed pattern of salivary cortisol activity over a 12-h period with particular emphasis on sleep. Basal diurnal cortisol profiles were examined in a sample of 147 volunteers (mean age 46.21+/-7.18 years). Profiles were constructed for each volunteer and explored in terms of the area under the curve (AUC) of the cortisol-awakening response with samples obtained immediately upon waking (0, 15, 30 and 45 min post waking) and at 3, 6, 9 and 12h post waking to assess diurnal decline. Diurnal mean of cortisol was based on the mean of cortisol at time points 3, 6, 9 and 12h post waking. Psychological measures of perceived stress and sleep were collected with concurrent biological assessment of fasting plasma glucose, insulin, blood lipids and inflammatory markers. Blunted cortisol profiles, characterised by a reduced AUC, were observed in the majority (78%) of a middle-aged sample and were associated with significantly poorer sleep quality and significantly greater waist-hip ratio (WHR). Blunted cortisol profiles were further associated with a tendency to exhibit a less favourable metabolic profile. These findings suggest that reduced cortisol secretion post waking may serve as an additional marker of psychological and biological vulnerability to adverse health outcomes in middle-aged adults.

Cortisol secretory activity in older people in relation to positive and negative well-being

Secretion of the hormone cortisol, a physiological correlate of affect, has been studied mostly in relation to negative states, especially stress. By contrast, policy initiatives aimed at older populations now routinely emphasise well-being and a 'positive ageing' perspective. In this study, we examined diurnal salivary cortisol profiles from 50 active seniors recruited into a wider community research project (mean age 74 years; 34 F/16 M). Participants' wrist activity was continuously monitored by actimeters in their homes over a 48 h period. During this time two diurnal cycles of cortisol data were collected (8 samples per day); with actimeter data providing a compliance check in regard to timing of self-administered saliva collections. Prior to the trial, participants had completed the GHQ-30 which was scored separately to yield both positive and negative well-being scores which matched closely normative data from over 6000 cases in a large survey. Our data suggest that positive and negative psychological well-being are quite strongly and inversely correlated. However, neither on their own was associated with basal levels of cortisol. Rather, for cortisol secretion in the 45-min period following awakening, but not during the rest of the day, we found a significant interaction between positive and negative well-being (p<0.024). Further analysis of this interaction showed that among participants low on negative well-being, higher positive well-being was significantly associated with lower cortisol; equally, among participants high on positive well-being, lower negative well-being was significantly associated with lower cortisol. Thus, a powerful synergy seemed to be operating in this early morning period such that cortisol secretion was 27% lower in participants with both higher-than-average positive well-being and lower-than-average negative well-being (comprising 34% of the sample). We conclude that cortisol secretion in the first 45 min following awakening is distinct from the rest of the day and most able to discriminate well-being states.

Predicting Posttraumatic Stress Symptoms From Pretraumatic Risk Factors: A 2-Year Prospective Follow-Up Study in Firefighters

Most studies focusing on risk factors for posttraumatic stress disorder (PTSD) have used retrospective study designs. Only a small number of studies have prospectively examined risk factors in the immediate aftermath of trauma exposure in predicting PTSD symptoms. The purpose of this study was to identify predictive risk factors for posttraumatic stress symptoms and comorbid psychopathological symptoms present during the time before exposure to traumatic stress in a high-risk population. Forty-three professional firefighters were assessed immediately after basic training (baseline) and at 6, 9, 12, and 24 months after entry into firefighter service. Subjects were screened for psychopathological symptoms, including symptoms of PTSD, depression, and anxiety. Subjects were also characterized with regard to personality traits such as self-efficacy, hostility, and alexithymia. Neuroendocrine activity was assessed by examination of awakening and diurnal salivary cortisol profiles and 24-hour urinary catecholamine excretion. Multiple linear regression analysis was used to analyze posttraumatic stress symptoms at 24-month follow-up as a function of pretraumatic characteristics. A high level of hostility and a low level of self-efficacy at baseline accounted for 42% of the variance in posttraumatic stress symptoms after 2 years. Subjects who had both risk factors at baseline showed a significant increase in measures of PTSD symptoms, depression, anxiety, general psychological morbidity, global symptom severity, and alexithymia during the 2-year period. Biological characteristics were not predictive of the development of psychopathological symptoms. These results suggest that specific personality traits may constitute markers of vulnerability to the development of psychopathological symptoms after trauma exposure. Early identification of preexisting risk factors is needed to provide effective prevention and intervention for individuals who are at risk of developing trauma-related disorders.