Monitoring adrenal insufficiency through salivary steroids: a pilot study.

BACKGROUND: Various glucocorticoid replacement therapies (GRTs) are available for adrenal insufficiency (AI). However, their effectiveness in restoring glucocorticoid rhythm and exposure lacks adequate biochemical markers. We described the diurnal salivary cortisol (SalF) and cortisone (SalE) rhythm among different GRTs and analysed the associations between saliva-derived parameters and life quality questionnaires. METHODS: Control subjects (CSs, n = 28) and AI patients receiving hydrocortisone (HC, n = 9), cortisone acetate (CA, n = 23), and dual-release hydrocortisone once (DRHC-od, n = 10) and twice a day (DRHC-td, n = 6) collected 9 saliva samples from 07:00 to 23:00. Patients compiled Pittsburgh Sleep Quality Index, Hospital Anxiety and Depression Scale, and Addison disease-specific quality-of-life questionnaires. SalE and SalF were measured by liquid chromatography-mass spectrometry. Exposure was monitored using SalE for HC and DRHC and SalF for CA. Area under the curve (AUC) was computed. Different GRTs were compared by Z-scores calculated from saliva-derived parameters. Questionnaire results predictors were evaluated with multiple regression analysis. RESULTS: Compared with controls, all GRTs resulted in glucocorticoid overexposure in the morning. Hydrocortisone, CA, and DRHC-td caused overexposure also in afternoon and evening. Compared with other treatments, CA determined increased Z-score-07:00 (P < .001), DRHC-td determined increased Z-score-AUC07:00→14:00 (P = .007), and DRHC-od induced lower Z-score-AUC14:00→23:00 (P = .015). Z-scores-AUC14:00→16:00 ≥ .619 best predicted questionnaire scores. CONCLUSIONS: None of the GRTs mimics normal glucocorticoid rhythmicity and exposure. SalE, SalF, and Z-score may be useful markers for monitoring and comparing different GRTs. Excess glucocorticoid in early afternoon best associated with depressive symptoms and worse life and sleep quality.

Classic and 11-oxygenated androgens in serum and saliva across adulthood: a cross-sectional study analyzing the impact of age, body mass index, and diurnal and menstrual cycle variation.

OBJECTIVE: 11-oxygenated androgens significantly contribute to the circulating androgen pool. Understanding the physiological variation of 11-oxygenated androgens and their determinants is essential for clinical interpretation, for example, in androgen excess conditions. We quantified classic and 11-oxygenated androgens in serum and saliva across the adult age and body mass index (BMI) range, also analyzing diurnal and menstrual cycle-dependent variation. DESIGN: Cross-sectional. Morning serum samples were collected from 290 healthy volunteers (125 men, 22-95 years; 165 women, 21-91 years). Morning saliva samples were collected by a sub-group (51 women and 32 men). Diurnal saliva profiles were collected by 13 men. Twelve women collected diurnal saliva profiles and morning saliva samples on 7 consecutive days during both follicular and luteal menstrual cycle phases. METHODS: Serum and salivary steroids were quantified by liquid chromatography-tandem mass spectrometry profiling assays. RESULTS: Serum classic androgens decreased with age-adjusted BMI, for example, %change kg/m2 for 5α-dihydrotestosterone: men -5.54% (95% confidence interval (CI) -8.10 to -2.98) and women -1.62% (95%CI -3.16 to -0.08). By contrast, 11-oxygenated androgens increased with BMI, for example, %change kg/m2 for 11-ketotestosterone: men 3.05% (95%CI 0.08-6.03) and women 1.68% (95%CI -0.44 to 3.79). Conversely, classic androgens decreased with age in both men and women, while 11-oxygenated androgens did not. Salivary androgens showed a diurnal pattern in men and in the follicular phase in women; in the luteal phase, only 11-oxygenated androgens showed diurnal variation. CONCLUSIONS: Classic androgens decrease while active 11-oxygenated androgens increase with increasing BMI, pointing toward the importance of adipose tissue mass for the activation of 11-oxygenated androgens. Classic but not 11-oxygenated androgens decline with age.

Comparative analysis of the effects of collection methods on salivary steroids.

BACKGROUND: Steroid hormone test for saliva was a promising area of research, however the impact of different collection methods on salivary steroids was underexplored so far. This study was designed to compare the effects of different collection methods (unstimulated or stimulated by chewing paraffin, forepart or midstream) on salivary flow rate, concentrations and secretion rates of steroids in saliva. METHODS: Whole-saliva samples were collected from 10 systemically and orally healthy participants, whose forepart and midstream segments of saliva were collected under unstimulated and stimulated conditions, with the salivary flow rate of each sample recorded. The concentrations and secretion rates of salivary steroids including testosterone, dehydroepiandrosterone (DHEA) and progesterone were measured by ELISA, with the multiple of change calculated. RESULTS: The results indicated mechanical stimulation used in collection of saliva samples could affect concentrations and secretion rates of steroids, whereas forepart and midstream segments had little differences in levels of salivary steroids, which effects could be partly influenced by individual specificity. The asynchronism in change of secretion rate of steroids with that of salivary flow rate might play an important role during this course. CONCLUSION: Based on these findings, we suggested to use the same collection method throughout one analytical study on salivary steroids or in longitudinal observations to ensure the comparability of the saliva samples collected.

Assessment of correlations and concentrations of salivary and plasma steroids, testicular morphometry, and semen quality in different climatic conditions in goats.

The current study aimed to investigate whether steroids (testosterone, estradiol, and cortisol) in the saliva of goats reflects their concentrations in the plasma. Also, it aimed at ascertaining, for the first time, the effect of changes in climatic conditions (spring versus summer) on the aforementioned steroids, testicular volume, testicular echotexture (pixel intensity; PI, and integrated density; ID), and semen quality in goats. Saliva and plasma samples were collected from 7 male Shiba goats weekly in spring and summer for measurement of testosterone (T; ng/ml), estradiol (E2; pg/ml), and cortisol (ng/ml) using radioimmunoassay. The changes in testicular volumes (TV/ml) and echogenicity were monitored weekly using ultrasonographic assessments concomitantly with subjective evaluations of semen parameters. Results revealed a highly significant positive correlation (r = 0.72; P < 0.0001) between plasma and salivary cortisol. Mild significant positive correlation was found between salivary and plasma T levels (r = 0.31; P < 0.002). However, low non-significant negative correlation was reported between salivary and plasma E2 levels (r = -0.24; P > 0.05). Higher levels of salivary cortisol were found in summer than in the spring (3.8 ± 0.6 ng/ml versus 1.4 ± 0.3 ng/ml; P < 0.001). However, no differences in levels of T, E2, and plasma cortisol were recorded between spring and summer (P > 0.05). Values of testicular PI and ID were significantly (P < 0.001) higher in the summer (94.5 ± 4.9, 258,388 ± 13,190, respectively) than in the spring (74.0 ± 2.1, 200,922 ± 5704, respectively). Meanwhile, TV and semen parameters did not significantly differ between spring and summer. In conclusion, saliva can be considered as an alternative biological fluid for the measurement of cortisol and T, but not suitable for E2. Climatic conditions significantly impact the levels of cortisol in saliva and testicular echogenicity in goats.

Chair-based exercise programs in institutionalized older women: Salivary steroid hormones, disabilities and frailty changes.

PURPOSE: Many people experience aging-related losses in different physical domains, which leads to a condition often called physical frailty (PF). The aim of this study was to analyse the effects of two different, 28-weeks, class chair-exercise protocols on salivary steroid hormones (SH), PF, and functional disabilities (FD) in frail older women. METHODS: A sample of older frail individuals (n = 60, 817.84 years) participated in the study and were divided into three groups: chair elastic-band muscle strength exercises (CSE), n = 20), chair-multimodal exercise (CME, n = 21) and a control non-exercise group (CGne, n = 19). Both exercise programs consisted of 45 min of supervised chair-based exercise group classes, carried out 3 times/week. CME participants performed a progressive training using walking, mobility and body weight resistance exercises. The CSE participants exercised using an elastic-band system of progressive exercises. Both CSE and CME followed a circuit training protocol. The controls did not change their usual lifestyle. The indicators of PF, FD and SH concentrations were analyzed before and after the intervention. RESULTS: Both exercise programs diminished the PF status showing significant time and time versus treatment interactions (p < .01). An increase in the CME group, between baseline and 14-weeks, and in the CSE group, after 28 weeks, for Testosterone concentrations was observed (p < .01). Dehydroepiandrosterone (DHEA) increased after 28-weeks in the CME group and decreased in the CGne after the same period (p < .05). Both exercise programs decreased the negative scores of several FD domains, specially fear of falling that showed significant effects with time (p < .01), and time vs intervention (p < .05). CONCLUSION: Both chair-exercise based programs were effective in stimulating positive changes in physical health and in steroid hormone responses, especially in DHEA. The control group did show a negative trend towards an increased PF status and decreased levels of SH. It is crucial for public health to identify the main factors associated with Functional Disability and Physical Frailty that underlie the development of new methods for complementary therapies, such as the use of low doses of hormonal supplementation combined with long-term exercise interventions.

Testosterone, androstenedione, cortisol and cortisone levels in human unstimulated, stimulated and parotid saliva.

BACKGROUND: Recently, measurements of steroids like testosterone, androstenedione, cortisol and cortisone in saliva are more and more applied in diagnostics and scientific studies. This is mainly due to the simple and non-invasive collection of saliva. We aimed to evaluate the optimal way to collect saliva for steroid hormone measurement. METHODS: We investigated in twenty volunteers whether there is a difference between steroid hormone concentrations in unstimulated and stimulated saliva collected while chewing, using cotton and synthetic Salivettes®, citric acid or chewing gum. Furthermore, total unstimulated saliva was compared to parotid gland saliva. Testosterone, androstenedione, cortisol and cortisone were measured using Liquid-Chromatography Tandem Mass Spectrometry (LC-MS/MS). RESULTS: Salivary testosterone, androstenedione and cortisol concentrations were unaffected by stimulation upon mouth and tongue movements, cortisone levels were on average 16% lower. Concentrations of all hormones were lower in parotid gland saliva compared to total unstimulated saliva (on average 51%, 26%, 66% and 49% lower, for testosterone, androstenedione, cortisol and cortisone, respectively). Concentrations of testosterone as well as androstenedione were lower when using synthetic Salivettes® (58% and 41%, respectively) and were higher when using cotton Salivettes® (217% and 46%, respectively). Cortisol levels in saliva were unaffected by using Salivettes®. However, cortisol and testosterone levels were higher in with chewing gum stimulated saliva (16% and 55%, respectively). Cortisone concentrations were lower in all types of stimulations (on average 25%-35%). CONCLUSION: The way saliva is collected should be considered when analysing and interpreting salivary hormone concentrations. We advocate unstimulated saliva collection in simple polypropylene tubes for all steroid measurements.