Sequence and Seasonal Effects of Salivary Cortisol

Abstract

Assessments of cortisol levels in saliva have been widely used by both researchers and clinicians as an index of adrenal functioning. Quarterly measurements of morning and evening cortisol levels were determined in a longitudinal study of 147 participants (72 women and 75 men) followed for I year each. The analysis of salivary cortisol revealed no significant gender or age differences in the sample. There was a sequence effect in quarterly cortisol values with a progressive decrease in serial measurements, especially notable in the morning values, as well as a seasonal variation in cortisol levels with significantly higher levels found in winter and fall, compared with spring and summer. The findings in this study suggest that repeated saliva sampling and seasonal variation in cortisol levels may independently affect adrenal response and, therefore, need to be accounted for in longitudinal studies.     

Day-to-day variation in saliva cortisol—Relation with sleep, stress and self-rated health

The objective was to examine the day-to-day variation in cortisol among healthy individuals and its relation to the time of saliva sampling, work, stress and fatigue. During 4 consecutive weeks, 14 office workers provided saliva samples (at awakening, 15 min after awakening and at bedtime) and made diary ratings for each day. Results showed a variation in cortisol values between participants but also within individuals. After controlling for the individual differences, results showed that low cortisol levels in the morning were associated with sleepiness at awakening and anxiety, exhaustion, and poor health the day before. High evening levels of cortisol were associated with symptoms of stress and poor self-rated health. Further analysis of the cortisol awakening response (CAR) showed that all participants had a mixture of both a positive and negative responses. During mornings with a negative response participants stayed in bed for a longer time after the initial awakening, which might be a sign of snoozing, thus missing the awakening response.     

Development and validation of an enzyme-linked immunosorbent assay for detection of cortisol in human saliva

Non-invasive measurement of cortisol in saliva is of prime importance as it represents a bioavailable neuroendocrine marker for stress. Therefore, in this study, we developed an enzyme immune assay that was suitable for salivary cortisol measurements. For that purpose, rabbit polyclonal antibody was raised against cortisol-3-CMO:BSA conjugate. The test was based on competition of liquid phase cortisol with conjugated cortisol on the solid phase. Primary antibody was used to bind available sites on the conjugate, which was proportional to numbers of cortisol in liquid phase. Biotinylated secondary anti-rabbit antibody was used to detect primary antibodies by addition of streptavidin peroxidase and substrate, respectively. Color formation was stopped and yellow color was read by a plate-reader spectrophotometer. Additionally, validated test was used to met all validation criteria including. Test developed was used to establish cortisol awakening response (CAR) in saliva samples collected in the morning after awakening (0, 15, 30, and 60^th min) from women (n = 4) and men (n = 4) at 8 or 4 different days, respectively. Diurnal cortisol levels were assessed (n = 8) at after awaking 60 min at morning, 12:00, 19:00, and 22:00 hr. In conclusion, an enzyme immunoassay test was successfully produced, validated and used for cortisol measurement in saliva samples.     

The relationship between long-term job strain and morning and evening saliva cortisol secretion among white-collar workers

The objective of this study was to assess long-term job strain impact on morning and evening salivary cortisol secretion. In all 77 white-collar workers (31% women; sample mean age, 42 years at baseline) volunteered to sample morning (immediately after waking up) and evening (10 p.m.) salivary cortisol for 7 consecutive days. By median split on aggregated self-reported isostrain from three consecutive questionnaires distributed in a period of approximately 3.5 years the participants were classified into a high or low long-term isostrain condition. Regardless of strain condition, there was a significant reduction in morning salivary cortisol secretion from the working week to the weekend, whereas evening salivary cortisol secretion showed no significant variation during the week. Although chronic isostrain did not affect the morning saliva cortisol measures, evening cortisol secretion was significantly elevated in the chronic high isostrain group throughout the whole week. The elevated evening cortisol measures associated with chronic high strain are concordant with the findings in other studies on long-term strain.     

Salivary cortisol and DHEA levels in the Korean population: age-related differences, diurnal rhythm, and correlations with serum levels

PURPOSE: The primary objective of this study was to examine the changes of basal cortisol and DHEA levels present in saliva and serum with age, and to determine the correlation coefficients of steroid concentrations between saliva and serum. The secondary objective was to obtain a standard diurnal rhythm of salivary cortisol and DHEA in the Korean population. MATERIALS AND METHODS: For the first objective, saliva and blood samples were collected between 10 and 11 AM from 359 volunteers ranging from 21 to 69 years old (167 men and 192 women). For the second objective, four saliva samples (post-awakening, 11 AM, 4 PM, and bedtime) were collected throughout a day from 78 volunteers (42 women and 36 men) ranging from 20 to 40 years old. Cortisol and DHEA levels were measured using a radioimmunoassay (RIA). RESULTS: The morning cortisol and DHEA levels, and the age- related steroid decline patterns were similar in both genders. Serum cortisol levels significantly decreased around forty years of age (p < 0.001, when compared with people in their 20s), and linear regression analysis with age showed a significant declining pattern (slope=-2.29, t=-4.297, p < 0.001). However, salivary cortisol levels did not change significantly with age, but showed a tendency towards decline (slope=-0.0078, t=-0.389, p=0.697). The relative cortisol ratio of serum to saliva was 3.4-4.5% and the ratio increased with age (slope=0.051, t=3.61, p < 0.001). DHEA levels also declined with age in saliva (slope=-0.007, t=-3.76, p < 0.001) and serum (slope=-0.197 t=-4.88, p < 0.001). In particular, DHEA levels in saliva and serum did not start to significantly decrease until ages in the 40s, but then decreased significantly further at ages in the 50s (p < 0.001, when compared with the 40s age group) and 60s (p < 0.001, when compared with the 50 age group). The relative DHEA ratio of serum to saliva was similar throughout the ages examined (slop=0.0016, t=0.344, p=0.73). On the other hand, cortisol and DHEA levels in saliva reflected well those in serum (r=0.59 and 0.86, respectively, p < 0.001). The highest salivary cortisol levels appeared just after awakening (about two fold higher than the 11 AM level), decreased throughout the day, and reached the lowest levels at bedtime (p < 0.001, when compared with PM cortisol levels). The highest salivary DHEA levels also appeared after awakening (about 1.5 fold higher than the 11 AM level) and decreased by 11 AM (p < 0.001). DHEA levels did not decrease further until bedtime (p=0.11, when compared with PM DHEA levels). CONCLUSION: This study showed that cortisol and DHEA levels change with age and that the negative slope of DHEA was steeper than that of cortisol in saliva and serum. As the cortisol and DHEA levels in saliva reflected those in serum, the measurement of steroid levels in saliva provide a useful and practical tool to evaluate adrenal functions, which are essential for clinical diagnosis.     

Impact of stress reduction instructions on stress and cortisol levels during pregnancy

This pilot study examined whether giving stress reduction (SR) instructions to pregnant women would be effective in regulating stress, mood, and cortisol levels during pregnancy. Forty-one predominantly low-income Latina women, receiving prenatal services at a public county hospital, completed measures of stress and mood (depressive symptoms, positive and negative affect) and provided morning and evening saliva samples to measure cortisol prior to and after receiving SR instructions. We hypothesized that adherence to these SR instructions would result in lower levels of stress, negative mood states, and cortisol levels when compared to baseline values. Repeated measures ANOVA analyses demonstrated significantly lower levels of stress (P < 0.001), lower symptoms of depression and negative affect (P < 0.001), and lower levels of morning cortisol (P = 0.01) under the SR condition, compared to baseline. Health behaviors that were engaged in during the SR condition and implications for prenatal health interventions are discussed.     

Allopregnanolone serum concentrations and diurnal cortisol secretion in women with premenstrual dysphoric disorder

Most prior studies in patients with premenstrual dysphoric disorder (PMDD) indicate a blunted hypothalamus–pituitary–adrenal axis function. However, the relationship between neuroactive progesterone metabolites, such as allopregnanolone, and hypothalamus–pituitary–adrenal (HPA) axis function in PMDD patients is relatively sparsely studied. The primary aims of this study were to assess diurnal variation in circulating cortisol and low-dose dexamethasone suppression in PMDD patients and healthy controls, and the relationship between these two HPA axis indices and allopregnanolone serum concentrations. Twenty-six women with prospectively defined PMDD and 30 healthy controls were recruited. Participants underwent diurnal sampling for cortisol serum concentrations and a low-dose dexamethasone suppression test. In addition, morning allopregnanolone serum concentrations were determined. There was no difference in diurnal secretion of cortisol and degree of dexamethasone suppression of cortisol between PMDD patients and healthy controls. However, PMDD patients with high allopregnanolone levels displayed blunted nocturnal cortisol levels in comparison with healthy controls who had low allopregnanolone serum concentrations. In women with PMDD, diurnal secretion of cortisol may be influenced by allopregnanolone levels of the luteal phase. This finding may be attributed to timing of blood sampling in the late luteal phase as well as the individual level of allopregnanolone but could potentially explain the discrepancies in results between studies examining HPA axis function in women with PMDD.     

Effects of long-term inhaled corticosteroids on adrenal function in patients with asthma.

BACKGROUND: Inhaled corticosteroids (ICSs) are effective asthma controllers, but long-term use could lead to adverse effects. Objective: To examine adrenal responsiveness of patients with persistent asthma treated with long-term ICSs. METHODS: Morning plasma cortisol levels before and 30 minutes after adrenocorticotropic hormone (ACTH) (1 microg intravenously) stimulation were compared. Primary end points included mean prestimulation and poststimulation cortisol levels; secondary end points included morning cortisol level of 5 microg/dL or less, post-ACTH stimulation cortisol level of 18 microg/dL or less, and/or a net change of 7 microg/dL or less from baseline. RESULTS: A total of 103 asthmatic patients (29 in the triamcinolone acetonide group, 18 in the flunisolide group, 45 in the fluticasone propionate group, and 11 in the oral corticosteroids group [positive controls]) completed the study. Mean daily ICS doses and durations were as follows: triamcinolone acetonide: 448 microg for 36 months; flunisolide: 1,181 microg for 41 months; and fluticasone propionate: 745 microg for 19 months. Eleven of 30 patients taking high-dose ICSs (10 of 28 taking fluticasone propionate and 1 of 2 taking flunisolide) had both low morning cortisol levels and abnormal post-ACTH stimulation cortisol levels. Few patients taking lower doses of any ICS had abnormal results. CONCLUSIONS: Patients who require long-term treatment with high-dose ICSs may have abnormal morning plasma cortisol levels and reduced responsiveness to ACTH stimulation. Careful monitoring of adrenal function should be considered in such patients.     

Patterns of salivary cortisol levels can manifest work stress in emergency care providers

To develop objective assessments of work fatigue, we investigated the patterns of changes in salivary cortisol levels in emergency care providers working extended work shifts. Fourteen subjects, comprising seven physicians and seven physician assistants, provided unstimulated saliva samples at regular intervals over the course of a 24-h work shift and over their subsequent free day. There was a significant time effect, with early morning cortisol levels being significantly attenuated following the work shift. Native diurnal variations varied by gender, with the female subjects manifesting greater cortisol levels. Physicians also had higher cortisol profiles even though their wake–rest cycles were similar to those of the physician assistants. Our results suggest that temporal changes, as well as diurnal similarities, in the salivary cortisol patterns can reflect work-related stress and recovery. In particular, early morning cortisol levels may manifest individual reactivity to work stressors as well as sleep deprivation.     

Relationship between salivary cortisol and depression in adolescent survivors of a major natural disaster

The purpose of this study was to determine the utility of salivary cortisol levels for screening mental states such as depression in adolescents following a natural disaster. We examined the relationship of salivary cortisol levels in adolescent survivors of the 2011 Tohoku Earthquake with the depression subscale of the 28-item General Health Questionnaire (GHQ). Subjects were 63 adolescent survivors (age = 14.29 years ± 0.51) who were administered the GHQ and provided saliva samples thrice daily (morning, afternoon and evening) over the course of 3 days. Based on the GHQ-depression subscores, subjects were divided into low and high depression groups. About 22 % of the subjects were classified into the high symptom group. When data collected over 3 days were used, a significant difference was observed between the two groups in the salivary cortisol levels at the evening time point as well the ratio of the morning/evening levels (p < 0.05). Analyzed by means of receiver-operating characteristic curves, the morning/evening ratios showed a good power in discriminating between subjects with and without depressive symptoms. Our study suggests that repeated measurement of salivary cortisol levels over 3 days has utility in screening for depressive states in adolescents following a natural disaster.     

唾液皮质醇检测在抑郁症诊断中的应用探讨

研究抑郁症患者与健康人早晨唾液皮质醇水平的差异,探讨唾液皮质醇检测对抑郁症诊断的应用价值。早晨收集31例抑郁症患者（患者组）和31名健康人（对照组）的唾液样本,采用酶免疫法（EIA）测定其皮质醇水平;同时以汉密尔顿抑郁量表（24项,HAMD）评定研究对象的抑郁程度。患者组唾液皮质醇水平明显高于对照组（P＆lt;0.01）。唾液皮质醇水平和HAMD评分在对照组中呈正相关（Spearman相关系数=0.663,P＆lt;0.01）;在患者组则无显著相关（P=0.199）。用受试者工作特征（receiver operating characteristic,ROC）曲线分析唾液皮质醇对抑郁症的诊断价值,曲线下面积为0.771（95%可信区间：0.654~0.887）;11.75nmol/L为综合考虑灵敏度和特异性时的最佳诊断阈值,其对应的灵敏度为64.5%,特异性为77.4%;16.73nmol/L是特异性为100%时的诊断阈值,其对应的灵敏度为41.9%。抑郁症患者早晨唾液皮质醇水平比健康人高,唾液皮质醇检测可作为抑郁症诊断的一种辅助手段,但尚不能用于判断患者的抑郁程度。     

Diurnal cycle in salivary cortisol levels in common marmosets

A noninvasive method of saliva sampling was used to assess the diurnal cortisol rhythm from 0900 to 1700 hr in the common marmoset (Callithrix jacchus). The levels of cortisol were highest in the morning and declined significantly over the day. Individual marmosets varied in the magnitude of the cycle, and the greatest individual variability occurred in the morning levels. The decrease in cortisol levels was more rapid after than before the midday feeding period in subordinate marmosets (aged 53-63 months) compared to dominant marmosets (aged 79-80 months), and overall, the levels of cortisol were higher in the subordinate marmosets. We found no effect of sex on cortisol levels across the cycle.     

Seasonal variation of cortisol plasma levels in depressives

There is some evidence for a seasonal variation of plasma cortisol levels after administration of dexamethasone in depressed subjects. This variation is suspected to be associated with similar season-related changes in pre-dexamethasone cortisol levels. Therefore, cortisol plasma levels in depressed subjects before and after dexamethasone administration were investigated. No seasonal variation was found in pre- or post-dexamethasone cortisol levels.     

Effect of hostility on salivary cortisol levels in university students

The influence of hostility on cortisol levels in saliva was investigated in 47 university students (27 males and 20 females). The students were divided into high and low hostility groups by cluster analyses based on their scores on the Buss-Perry Aggression Questionnaire and the Müller Anger Coping Questionnaire. Their saliva was collected twice, in the morning immediately after awakening at a two week interval. On the day before saliva collection, they completed questionnaires about stressful events and their moods in the previous two weeks. The results of the ANOVA showed higher cortisol levels and higher frequencies of stressful events and negative moods in the high hostility group. The effect of hostility on cortisol levels was diminished by controlling for stressful events and negative moods, which suggests that stressful events and negative moods are mediators between hostility and cortisol. The results are discussed in relation to the association between hostility and coronary heart disease.     

Is free plasmatic cortisol measurement useful in intensive care unit?

Serum cortisol free fraction is responsible for its physiological function. Determination of serum total cortisol concentration does not allow accurate evaluation of hypothalamic pituitary adrenal axis when there is a quantitative variation in serum transcortin. Although free cortisol can be assayed by salivary or urinary cortisol measurements, these methods can not be easily applied in intensive care units. Free plasmatic cortisol measurement could provide a better reflect of circulating cortisol in patients with critical illness whom have a decreased concentration of binding proteins and whom adrenal status is an important prognostic factor. Free cortisol can be measured after separation from bound cortisol, calculated with equation based on equilibrium binding or evaluated by cortisol/CBG ratio. In various studies, free plasmatic cortisol allows in critical care patients a better appreciation of adrenal status than total cortisol and be more differential than total cortisol increment after stimulation with synacthen used to diagnose "relative adrenal insufficiency".     

Personality, adrenal steroid hormones, and resilience in maltreated children: a multilevel perspective

In this multilevel investigation, resilience in adaptive functioning among maltreated and nonmaltreated low-income children (N = 677) was examined in relation to the regulation of two stress-responsive adrenal steroid hormones, cortisol and dehydroepiandrosterone (DHEA), as well as the personality constructs of ego resiliency and ego control. Maltreatment status was not related to differences in average levels of morning or afternoon cortisol or DHEA. However, lower morning cortisol was related to higher resilient functioning, but only in nonmaltreated children. In contrast, among physically abused children, high morning cortisol was related to higher resilient functioning. Morning and afternoon DHEA was negatively related to resilient functioning. Although diurnal change in cortisol was not related to resilience, for DHEA, maltreated children with high resilience showed an atypical rise in DHEA from morning to afternoon. Morning and afternoon cortisol/DHEA ratios were positively related to resilient functioning, but did not interact with maltreatment status. Ego resiliency and ego control strongly differentiated maltreated and nonmaltreated children, and the personality variables were substantially predictive of resilience. When considered together, demonstrated effects of personality, cortisol, and DHEA maintained independent contributions in predicting resilience among high-risk youth.     

Low salivary cortisol levels and externalizing behavior problems in youth

Research linking basal cortisol levels with internalizing and externalizing behavior problems in youths has yielded inconsistent results. We hypothesize that the high moment to moment variation in adrenocortical activity requires an analytical strategy that separates variance in cortisol levels attributable to "stable traitlike" versus "state or situationally specific" sources. Early morning saliva samples were obtained from 724 youths (M age = 13.5 years; range = 6-16 years in Year 1) on 2 successive days 1 year apart. Latent state-trait modeling revealed that 70% of the variance in cortisol levels could be attributed to statelike sources, and 28% to traitlike sources. For boys only, higher levels of externalizing problem behaviors were consistently associated with lower cortisol attributable to traitlike sources across 3 years of behavioral assessment. The inverse association between individual differences in cortisol and externalizing problem behavior has previously only been reported in studies of at-risk or clinical groups. The present findings suggest the relationship is a stable phenomenon that spans both normative and atypical child development. Studies are needed to reveal the biosocial mechanisms involved in the establishment and maintenance of this phenomenon, and to decipher whether individual differences in this hormone-behavior link confers risk or resilience.     

Theophylline Concentrations in Serum and Saliva after Oral Microcrystalline and Sustained-Release Preparations in Asthmatics

Theophylline concentrations in saliva have been compared with those in serum in 13 asthmatic patients after single and multiple oral dosage of a microcrystalline theophylline tablet (Nuelin®) and a sustained-release preparation (Nuelin Retard®).
There was wide individual variation in the ratio of serum to saliva concentrations indicating that, even when individual serum to saliva ratios have been established, the use of saliva theophylline measurements alone is unreliable for the routine monitoring of theophylline therapy.
Both preparations produced mean serum theophylline concentrations within the 10–20 mcg/ml range, the sustained-release tablet giving higher morning trough values and smaller trough to peak variation. The preparations were equally and highly bioavailable.
Side effects were reported by three patients, which were the patients with the highest AUC.
The Nuelin Retard preparation appears to provide smooth, sustained serum theophylline concentrations within the therapeutic range and the simple, twice-daily dosage should result in improved patient compliance.     

Concurrent levels of maternal salivary cortisol are unrelated to self-reported psychological measures in low-risk pregnant women

Associations between salivary cortisol and maternal psychological distress and well-being were examined prospectively on 112 women with normally progressing, singleton pregnancies between 24 and 38 weeks gestation. At each of 5 visits, conducted in 3-week intervals, women provided a saliva sample and completed questionnaires measuring trait anxiety, depressive symptoms, pregnancy-specific hassles and uplifts, and psychological well-being. Maternal salivary cortisol was unrelated to psychological measures with the exception of minor associations detected with measures of anxiety and depressive symptoms between 30 and 32 weeks only. Findings indicate that self-reported maternal psychological distress and well-being are not associated with significant variation in maternal salivary cortisol levels during the second half of gestation. This suggests that studies that measure psychological factors in pregnancy but do not measure maternal cortisol should exercise caution in assuming activation of the maternal hypothalamic–pituitary–adrenal axis is the mechanism through which maternal psychological factors are transduced to the fetus.     

Salivary cortisol in women with major depressive disorder under selective serotonin reuptake inhibitors therapy

Depression is one of the widespread diseases whose etiology is still unclear. Dysregulation of the hypothalamic–pituitary–adrenal axis can be the cause of this illness which is concomitant with a high level of cortisol. For this reason, the purpose of the study was to estimate the influence of the selective serotonin reuptake inhibitors (SSRIs) therapy used in monotherapy and polypragmasy on cortisol level in saliva of depressed women. Cortisol was determined in saliva collected from 40 depressed patients treated with SSRIs. HPLC with UV detection was used for quantification of cortisol after its extraction with dichloromethane. For statistical evaluation of the data, the cluster analysis and principal components analysis were used. Results of the study have shown that the SSRIs treatment reduces the cortisol level in saliva. The therapy with sertraline and polypragmasy had a strong influence on suppressing the cortisol secretion. Besides, the amplitude of changes of the cortisol level during the treatment had an impact on the duration of hospitalization. In conclusion, it can be stet that the process of reduction of the cortisol level is multiphasic and that the combination treatment had a stronger influence on suppressing the cortisol secretion than did antidepressants used in monotherapy.