Salivary Cortisol Measurements Research Articles

Short-term equine interaction for reducing test anxiety and facilitating coping skill development in college students during examination periods: A preliminary study

Mental health concerns are common among college students and more prevalent during examination periods when stress and anxiety are heightened. While the benefits of equine interaction have been reported, the impact of short-term equine interaction on coping skills valuable for managing test anxiety have yet to be studied. Therefore, the purpose of this preliminary study was to evaluate the impact of short-term equine interaction during the college examination period on development of coping skills and anxiety levels associated with testing, both self-reported and those reflected in physiological measures. Full-time college students engaged in a one-hour equine interaction session held on campus at the onset of final exam week. Participants completed a survey targeting perception of anxiety levels and coping skills. Heart rate and salivary cortisol measurements were collected pre- and post-session. Paired t-tests were performed between pre- and post- session measures and correlations between survey answers and physiological measures determined. Significant reductions were found in all areas of anxiety (Overall Academics P = 0.0003, Final Exam Preparation P = 0.0003, Taking Final Exam P = 0.0003). The reduction in exam-related anxiety was positively correlated to cortisol concentrations (r = 0.69, P = 0.03) with lower concentrations (P = 0.05) and increased heart rate (P = 0.003) post-session. Regarding coping skills, improvements were observed post-session in assertiveness (P = 0.008). Skill development coupled with reduced anxiety supports the potential benefits short-term equine interaction can offer students during the high stress periods, but these findings should be confirmed with more in-depth study.

Performance characteristics of salivary cortisol measurement

Performance characteristics of salivary cortisol measurement

Sex-specific associations between childhood trauma and adult systemic inflammation in daily life.

Sex-specific associations between childhood trauma and adult systemic inflammation in daily life.

The Leuven Prolonged Acute Stress Test (L-PAST): A novel paradigm to induce a subjective and glucocorticoid stress response for at least ninety minutes

Laboratory stress tests typically administer stress acutely, ranging from 3 to 15 minutes. However, everyday stressors usually last longer than ten minutes (e.g., examination stressors, work stressors, and social stressors. Moreover, in some studies, it may be relevant to induce stress for a longer period to affect certain psychological or physiological parameters. To this end, we developed a novel stress test that intends to induce psychosocial stress for 90 minutes. The Leuven Prolonged Acute Stress Test (L-PAST) combines physical (hand immersion in cold water), cognitive (mental arithmetic), and psychosocial (social evaluation and feelings of failure) stress elements of two well-known laboratory stress tests, the Maastricht Acute Stress Test (MAST) and the Montreal Imaging Stress Test (MIST). Fifty healthy women were subjected to both the L-PAST and a sham (control) test in a randomized and counterbalanced manner. The stress response was determined by salivary cortisol measurements and assessment of subjective stress ratings at regular time points during the time preceding the stress period (5 min), the stress period (90 min), and the recovery period (35 min). Cognitive reactivity to failure and subjective pain levels were also assessed at various time points. The childhood trauma questionnaire (CTQ) and the perceived stress scale (PSS) were administered prior to the testing phase. As expected, linear mixed models revealed that the stress response was significantly higher during the L-PAST as indicated by a significant time point by condition interaction effect for both the cortisol response (F(10,450)=12.21, p < 0.0001, ηp2=0.11) and the subjective stress response (F(13,637)=13.98, p < 0.0001, ηp2 = 0.12). Moreover, there was a significant time point by condition interaction effect for cognitive reactivity to failure (F(13,637) = 7.97, p < 0.0001, ηp2 = 0.07) and subjective pain (F(13,637) = 38.52, p < 0.0001, ηp2 = 0.27), indicating that the levels were higher during the L-PAST at most stress induction time points. Lastly, higher CTQ scores were associated with higher subjective pain levels during the L-PAST (F(1,44)=6.05, p = 0.02). Collectively, our results confirm the efficacy of the L-PAST in inducing a prolonged subjective as well as cortisol stress response.

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.

Colorimetric Determination of Salivary Cortisol Levels in Artificial Saliva for the Development of a Portable Colorimetric Sensor (Salitrack)

Mental illnesses, such as clinical depression, have taken an unprecedented toll on society and the economy on a global scale. The relationship between stress management and mental health decline is of utmost significance, especially as most avenues of mental health management remain inaccessible for the majority of the general public, i.e., interview-based, and face-to-face interventions or costly drug-based therapies. Cortisol, the primary stress hormone, regulates the stress response in the human body and, through persistent activation, can lead to chronic stress and mental health deterioration. Thereby, the measurement and evaluation of cortisol within saliva could harness potential developments in management and diagnostic tools to monitor physiological and psychological stress in simple point-of-care applications. The current study aims to determine the concentration of salivary cortisol in spiked artificial saliva samples using blue tetrazolium (BT) dye as a colorimetric indicator. The proposed method showcases the use of the BT dye as an effective method for the rapid measurement of salivary cortisol, with accuracy comparable to the gold-standard method for salivary cortisol analysis, enzyme-linked immunoassays (ELISAs). Finally, a prototype colorimetric sensor has been developed for point-of-care applications of stress monitoring via salivary cortisol measurement.

Is It Possible to Mitigate Fear of Fireworks in Dogs? A Study on the Behavioural and Physiological Effects of a Psychoactive Supplement.

Canine fear of fireworks is a common problem worldwide, with serious implications for the welfare of both dogs and their owners. Therapies for the problem are available, and herbal and nutraceutical agents are increasingly suggested by professionals; nonetheless, studies on their real efficacy in reducing firework fear are lacking. In a randomised, double-blinded, placebo-controlled study, 44 dogs (25 in the "supplement" group and 19 in the "placebo" group) completed a long-term continuous treatment with either a supplement made of tryptophan, valerian, and passiflora or a placebo, including two real exposures to fireworks (on 2020 Christmas and 2021 New Years' Eve, after 42 and 48 days of treatment, respectively). Owners of both groups received the same general environmental management and food/toy offering recommendations for trying with their dogs on those nights. Behavioural (measured by LSSS-Lincoln Sound Sensitivity Scale and PANAS-Positive and Negative Activation scale, as rated by the owners) and stress (measured via salivary cortisol measures) reactions were evaluated. Significantly greater fear decrease (LSSS) was recorded in the "supplement" dogs, as compared to the "placebo" group. Cortisol dosages on New Year's Eve ("noisy" night) were in line with behavioural results; "supplement" dogs showed a smaller increase in the stress response from 22:30 to 00:30 h on New Year's Eve and a greater decrease in their stress response from 02:30 h to 10:30 h on New Year's Day compared to "placebo" dogs. Smaller cortisol levels were also shown by "supplement" dogs than "placebo" dogs on a controlled "quiet night" (27th December). Owners' rates on PANAS remained stable during the whole period of therapy for both groups. The evaluated supplement, a combination of tryptophan, valerian, and passiflora, showed satisfactory results and rare side effects when treating dogs fearful of fireworks.

Occupational Health Challenges for Aviation Workers Amid the Changing Climate: A Narrative Review.

Although there are many forecasts regarding the impact of climate change on the aviation sector, a critical but frequently neglected dimension is the occupational safety risks faced by aviation professionals. This narrative review explores the potential impacts of the changing climate on the health and safety of aviation personnel. Furthermore, we examine the significance of resilience in helping these workers adapt and effectively manage climate-related challenges in their professional lives. Climate change poses increasing threats to the well-being of flight personnel through elevated temperatures, heightened ultraviolet radiation exposure, increased mental workload from extreme weather events, and other psychological stressors. Building resilience through workforce training, planning, and adaptation can reduce vulnerability. In future research, the iterative process of selecting measurement components to gauge the impact of climate change should balance feasibility, relevance for stakeholders, and accurately capturing exposure effects. For instance, while salivary cortisol measures stress biologically, assessments of depression or burnout may provide more nuanced insights on pilot health for industry decision-makers managing climate impacts. In conclusion, a strategic emphasis on enhancing the physical and psychological well-being of the aviation workforce is imperative for facilitating a more efficient adaptation within the sector. This is of paramount importance, considering the critical function that aviation serves in fostering human connectivity. Consequently, it is essential for regulatory bodies and policymakers to prioritize the safeguarding of employee health in the face of climate change challenges.

An LC-MS/MS-based method for the simultaneous quantification of melatonin, cortisol and cortisone in saliva

Disturbances in the diurnal pattern are associated with several clinical and psychological conditions, including depression and fatigue. Salivary sampling for melatonin, cortisol and cortisone provides a non-invasive method for frequent sampling and obtaining biochemical insight into the diurnal pattern of individuals. Therefore, a new liquid chromatography-tandem mass spectrometry-based method for the measurement of salivary melatonin, cortisol and cortisone was developed and validated. The method required 250 μl saliva, used isotope dilution methodology and was based on a liquid-liquid extraction for sample preparation, reversed-phase chromatography and multiple reaction monitoring on a mass spectrometer for quantitation. The lower limits of quantification obtained were 0.010 nmol/L for melatonin, 0.5 nmol/L for cortisol and 1.00 nmol/L for cortisone and the limits of detection were 0.003 nmol/L, 0.15 nmol/L and 0.1 nmol/L respectively. The method imprecision was ≤14% for all measurands, and the method comparison showed highly comparable results with high correlation coefficients (all ≥0.964). Potential interference of cortisol and cortisone by prednisolone was observed and could be detected by chromatogram review. Typical diurnal patterns for melatonin, cortisol and cortisone were observed in the saliva of 20 cancer survivors who collected saliva throughout the day.

Relationships Among Number of Stressors, Perceived Stress, and Salivary Cortisol Levels During the Third Trimester of Pregnancy

ObjectiveTo examine relationships among the number of stressors, perceived stress, and salivary cortisol levels during the third trimester of pregnancy. DesignSecondary analysis of cross-sectional data. SettingParticipants’ homes. ParticipantsWomen during the third trimester of pregnancy (N = 73). MethodsParticipants provided saliva samples at four time points over 2 days for cortisol assay and completed questionnaires to assess stressors and perceived stress. We computed multiple linear regression models to examine the relationships among the number of stressors and perceived stress to cortisol awakening response, diurnal slope, and overall cortisol secretion. We also computed a multiple linear regression model to examine the relationship between perceived stress and the number of stressors. ResultsGreater perceived stress was associated with reduced overall cortisol secretion across the day (β = −0.41, p = .01). The number of stressors was associated with perceived stress (β = 0.48, p = .002) but not salivary cortisol measures. ConclusionElevated perceived stress and the related cortisol alterations that we identified could represent salient targets for enhancing hypothalamic-pituitary-adrenal axis function during the third trimester. Perceived stress may shape the relationship between exposure to stressors and cortisol response during pregnancy. Future research is warranted to confirm study results and to understand the implications for parturition and fetal development.

Preliminary validation of the Stress and Adversity Inventory for Adults (Adult STRAIN) in Dutch and its associations with in‐vivo locus coeruleus integrity

AbstractBackgroundThe Stress and Adversity Inventory (STRAIN) systematically assesses lifetime stressor exposure, but its validity has yet to be established in The Netherlands. This work aims to validate the Dutch translation of STRAIN and its association with key biological correlates: cortisol measures and the locus coeruleus (LC), a brainstem nucleus involved in stress responses and accumulating phosphorylated tau early in adulthood.MethodsEighty‐seven cognitively unimpaired Dutch participants (31‐81 years; ∼45% female; Table 1) completed stress‐ and affective‐related questionnaires, cognitive testing, salivary and plasma cortisol measurements, and 7T‐MRI. The STRAIN was administered after translation into Dutch. LC MRI signal intensity was extracted from an MT‐TFL sequence by normalizing LC signal against the pontine tegmentum (reference region) and creating a sample‐specific LC template. Pearson correlation models assessed associations between STRAIN summary scores (total stressor count and severity), and stress‐related (MIA‐Anxiety, PSS, DASS‐Stress, NEO PI‐R‐Neuroticism), affective (DASS‐Anxiety‐Depression, HDRS), cognitive (WLTDR, LDST, STR3), stress‐related biological (salivary‐, plasma cortisol), and LC measures (Table 1). HDRS models employed Spearman correlations. Age and sex were included as covariates, additionally including education for cognitive measures. Analyses were FDR‐corrected and conducted based on complete cases.ResultsOverall, the sample had relatively low lifetime stressor counts (M = 14.08; Figure 1A). When assessing concurrent validity, the STRAIN was positively associated with other stress measures (r = .21,.36; pFDR &lt;.05; Figure 1B,C). Models including cortisol revealed that less stressor exposure and severity were associated with higher cortisol levels (r = ‐.22,‐.23; pFDR &lt;.05). Affective and cognitive measures were investigated for discriminant validity. STRAIN was not associated with DASS, but was positively associated with HDRS (r = .17,.28; pFDR &lt;.05). Generally, greater stressor exposure was related to poorer cognitive performance (r = .13,‐.19; pFDR &lt;.05), except for a positive association between WLTDR and stressor severity (r = .18; pFDR &lt;.05). No significant associations were found with LC MRI signal intensity (Figure 1B).ConclusionThis study demonstrates preliminary good concurrent and discriminative validity for the Dutch STRAIN translation, even among individuals with low‐to‐moderate self‐reported stress levels. We found no direct associations between LC integrity and lifetime stressors. Future research will investigate associations with other STRAIN domains and potential moderating effects of Alzheimer’s Disease‐related biomarkers on the relationship between cumulative stressor exposure and LC integrity.

Heat inactivation of SARS-CoV 2 enabled the measurement of salivary cortisol during COVID-19 pandemic

Background and aimSalivary cortisol has become an essential tool in the management of cortisol-related disease. In 2020 the sudden outbreak of COVID-19 pandemic caused several concerns about the use of saliva, due to the risk of contamination, and a European consensus further discourage using salivary cortisol. To decrease infectious risk, we handled specimens by applying a heat treatment to inactivate viral particles, further evaluating the impact of the COVID-19 pandemic on the use of salivary cortisol in clinical practice.Material and methodsSaliva samples were exposed for 10 min at 70 °C, then cortisol was measured using LC-MS/MS. The number of salivary cortisol examinations from 2013 to 2022 was extracted from the local electronic database: those performed in 2019, 2020, and 2021 were analyzed and compared with the historical data.ResultsDuring 2020 we observed a decrease of 408 (−20%) examinations (p = 0.05) compared to 2019; especially in salivary cortisol daily rhythm and salivary cortisol/cortisone ratio (respectively reduction of 47% and 88%, p = 0.003 and p = 0.001). Analyzing year 2021 compared with 2020 we reported an increase of 420 examinations (+20%, p = 0.01), with a complete recovery of salivary cortisol measurement (considering 2019: p = 0.71). Major differences were observed between morning salivary cortisol (−20%, p = 0.017), LNSC (−21%, p = 0.012) and salivary cortisol rhythm (−22%, p = 0.056). No Sars-Cov2 infections related to working exposure were reported among laboratory’s employers.ConclusionsWe speculate that the adoption of an appropriate technique to inactivate viral particles in saliva specimens allowed the safety maintenance of salivary collections, also during the Sars-CoV-2 outbreak.

Predicting salivary cortisol and sexual behavior stigma among MSM in the American Men’s Internet Survey 2019

Physiological stress levels in response to sexual behavior stigma among men who have sex with men (MSM) in the United States (US) are understudied. The current study aims to explore the relationship between sexual behavior stigma and salivary cortisol both overall and stratified by race/ethnicity. If such an association exists, it may suggest that sexual behavior stigma can be physiologically measured or indicated by the presence of heightened salivary cortisol. A subsample of 667 MSM participants from the 2019 American Men’s Internet Survey (AMIS; N = 10,129) submitted morning (AM) and evening (PM) saliva cortisol samples using at-home mail-in collection kits. Average daily cortisol and daily cortisol change were calculated; simple linear regressions estimated associations between cortisol measures and sexual behavior stigma characterized in four different ways (ever and recent experience of individual stigma items; average ever and recent experience of three stigma scales: stigma from family and friends, anticipated healthcare stigma, general social stigma). Participants reported a mean age of 36.0 years (SD = 14.9), with most being non-Hispanic white (n = 480, 72.0%), Hispanic (n = 164, 12.3%), or Black/African American (n = 146, 10.9%), and identified as homosexual/gay (n = 562, 84.3%). Reporting ever experiencing healthcare providers gossiping was significantly associated with higher PM cortisol (β = 0.12, p = 0.001) and higher average daily cortisol (β = 0.11, p = 0.004), while reporting ever experiencing police refusing to protect was associated with higher AM cortisol (β = 0.08, p = 0.03) and higher average daily cortisol (β = 0.09, p = 0.02). Recent experiences of stigma were not significant predictors of any measure of cortisol. Measures of salivary cortisol may be used to characterize sexual behavior stigma among MSM populations, however more insight is needed to determine its exact relationship and strength.

Noise level and surgeon stress during thyroidectomy in an endocrine surgery operating room.

Noise in the operating room is an ongoing problem that impacts the outcome of any surgery. Noise as a stressor can produce a startling reaction and activate the fight or flight response of the autonomic and endocrine systems. The psychobiology of stress as assessed by salivary cortisol level is a sensitive measure of allostatic load. This study aims to correlate, both subjectively and objectively, the salivary cortisol levels of the surgeon with noise level measurement in an endocrine surgery operating room (OR). A prospective observational study was conducted in the Endocrine surgery OR of a tertiary care center. We recorded the noise from the shifting in of patients in the OR to shifting out using a digital sound level meter. The operating surgeon (S) provided two salivary cortisol samples (normal salivary cortisol <5 nmol/L), one baseline and another after the procedure. The questionnaire for the assessment of distraction during thyroidectomy was filled in by the S at the end of the procedure. Salivary cortisol levels were analyzed using SLV-4635 (formerly SLV-2930) DRG Instruments GmbH German using the ELISA technique. Statistical analysis was performed using SPSS 22.0. A total of 37 procedures with 74 salivary cortisol samples and 259 questionnaire responses from S were analyzed. All patients with only benign FNAC were operated upon (64.9% colloid). Mean TSH levels were 3.5 ± 6.7 mIU/L. The majority had a solitary thyroid nodule (STN) (25/37, 67.6%). Nineteen patients (51.3%) underwent open hemithyroidectomy, 10 patients total thyroidectomy, and eight patients endoscopic hemithyroidectomy. The mean noise level in the OR was 70 db. The maximum and minimum noise level in the OR was 90.06 and 51.81 dB, respectively. A total of 74 salivary cortisol samples from the S were collected (baseline and post-noise exposure) and mean cortisol levels were recorded. The surgeon was more significantly affected by surrounding noise, especially during critical phases 3 of surgery, mainly, RLN dissection and parathyroid dissection as recorded by their responses in the questionnaire (p = 0.003). The maximum value of post-operative salivary cortisol of surgeon was recorded as 23. 48 ng/mL and the minimum value recorded was 0.49 ng/mL. The difference in baseline cortisol and post-noise exposure cortisol levels of surgeon was found to be significant (p < 0.001). Maximum and mean noise levels were significantly associated with post-noise exposure salivary cortisol elevation in the surgeon (p = 0.032 and 0.014, respectively). The noise levels during RLN dissection were borderline significant with the post-noise exposure salivary cortisol of the surgeon (p = 0.055). Our research is the first such study which has been done to assess noise levels and their effect on thyroidectomy using objective salivary cortisol measurement. It challenges the misconstrued notion that visceral surgeries requiring lesser instruments are not associated with noise-related stress. Noise is a major distraction and the effect of long-term effect on the entire surgical team needs to be studied.

SAT626 The Use Of Salivary Cortisol Measurement By Mass Spectrometry To Identify Low Serum Cortisol Values In Healthy Volunteers Receiving CRN04894, A Melanocortin 2 Receptor (MC2R) Antagonist

Abstract Disclosure: A. Ayala: Employee; Self; Crinetics Pharmaceuticals Inc. C. LaCerte: Employee; Self; Crinetics Pharmaceuticals Inc. R. Luo: Employee; Self; Crinetics Pharmaceuticals Inc. C. Davidson: Employee; Self; Crinetics Pharmaceuticals Inc. J. Wang: Employee; Self; Crinetics Pharmaceuticals Inc. P. Trainer: Employee; Self; Crinetics Pharmaceuticals Inc. A. Krasner: Employee; Self; Crinetics Pharmaceuticals Inc. Clinical trials of cortisol-lowering drug candidates would benefit from user-friendly methods to measure cortisol in an outpatient setting. CRN04894 is a potent, orally bioavailable, MC2R antagonist in development for the treatment of ACTH-dependent Cushing’s syndrome and congenital adrenal hyperplasia. We have previously reported results from a randomized, double-blinded, placebo-controlled (6 active:3 placebo/cohort), multiple (10-day) ascending dose (40 to 80 mg/day) study in healthy volunteers. Declines in basal and ACTH-stimulated serum cortisol levels and a median reduction of 75% in 24-hr urinary-free cortisol (UFC) were seen in the 80 mg cohort. Asymptomatic glucocorticoid deficiency (defined as 08:00 serum cortisol of &amp;lt;5 µg/dL) was the most commonly observed adverse event. Here we report the comparison of salivary cortisol measurements with serum cortisol day curves and 24-hr UFC observed in this study. We evaluated cortisol and ACTH data from the three cohorts dosed once daily (QD) at 22:00 with either 40, 60, or 80 mg. Cortisol (salivary &amp; serum) and ACTH day curves (results reported as the mean of 4 or 5 available samples collected at 22:00, 08:00, 12:00, 16:00 &amp; 20:00) studies were undertaken on days -1, 1, 4, and 9. Mean serum cortisol was moderately correlated with 24-hr UFC:creatinine ratio (ug/g) (r = 0.495) and strongly correlated (r = 0.734) with mean salivary cortisol. There was across all three doses of CRN04894 a consistent magnitude ranking change in measured cortisol analytes: serum (free plus bound) &amp;lt; saliva &amp;lt; urine, with substantial changes being seen within 24 hours of initiation of CRN04894 and no loss of cortisol suppression over 9 days despite median ACTH changes of &amp;gt;1000% from baseline (60 &amp; 80 mg doses). Time-matched individual saliva and serum cortisol values were used (without timepoint or visit restrictions) to explore the relationship between low serum cortisol and salivary levels. A saliva cutoff for cortisol deficiency was optimized for diagnostic accuracy (92.2%) using a serum cortisol value of &amp;lt;5 µg/dL as the reference. Four subjects had 08:00 serum cortisol &amp;lt;5 µg/dL and saliva cortisol data. All four cases were correctly identified as cortisol-deficient using a simultaneous saliva cortisol cutoff of &amp;lt;0.0491 µg/dL. In conclusion, CRN04894 effectively lowered cortisol measured in serum, saliva, and urine in healthy volunteers. Salivary cortisol measured by tandem mass spectrometry correlated well with serum cortisol day curves and may be useful in the diagnosis of biochemical cortisol deficiency in outpatient studies. Future studies will explore the use of salivary glucocorticoid measurement in patients with Cushing’s disease receiving CRN04894. Presentation: Saturday, June 17, 2023

THU011 Cushing’s Disease Diagnosis Challenge With Standard Pituitary MRI: A Clinical Case Report

Abstract Disclosure: E. Kasiri: None. N. Shah: None. Introduction: Cushing’s disease (CD); the most common cause of endogenous Cushing’s syndrome (CS) is mainly caused by an adrenocorticotropin (ACTH)-secreting pituitary adenomas. Although MRI remains the preferred imaging to detect these adenomas, still about 50% of microadenomas may be missed using standard 1.5-Tesla MRI and on the other hand higher resolution MRI with 3T magnets is not widely used. Here we present a case of Cushing’s disease with equivocal 1.5T MRI which made a next step of management rather challenging. Case presentation: A 57-year-old female with history of hypertension, type 2 diabetes mellitus and obesity (BMI: 34.41 kg/m2) presented to our endocrinology clinic to establish care. Her main complaint was difficulty losing weight. She denied taking any steroids or oral contraceptive pill. Vitals showed BP: 124/85 mm Hg, HR: 100 bpm and T: 98.3 °F. Physical exam was significant for moon facies, buffalo hump and abdominal obesity. She was screened for Cushing syndrome with 1mg overnight low dose dexamethasone test which resulted in abnormal cortisol response with a level of 2.1mcg/dl (&amp;lt;1.8 mcg/dl). A 24hr urine free cortisol: 7.3 mcg/dl (&amp;lt;45 mcg/dl) and two late night salivary cortisol measurements resulted in 0.388 and 0.229 mcg/dl (&amp;gt; 3 times upper limit of normal). Other labs showed elevated baseline ACTH :67.4 pg/ ml (7.2 - 63.3 pg/mL), cortisol :19.4 mcg/dl (4.5-23 mcg/dl) and normal DHEAS:1864.1ng/ml (650-3400 ng/mL). We decided to distinguish ectopic vs pituitary source of hypercortisolism by an 8 mg (high dose) dexamethasone suppression test which showed suppressed ACTH &amp;lt;1.5 pg/ml, Cortisol :2.6 mcg/dl (&amp;gt;50% suppression from baseline) suggesting pituitary origin. The standard pituitary 1.5T MRI with and without contrast as next step resulted in questionable microadenoma in the right aspect of pituitary gland and was equivocal. Within the multidisciplinary team decision was made to perform 3T pituitary MRI rather than invasive inferior petrosal sinus sampling (IPSS) which confirmed the pituitary microadenoma of 8.1 mm size with no mass effect and patient is scheduled for transsphenoidal pituitary surgery without needing further testing. Discussion: 3T MRI, with higher magnetic field than 1.5T allows to improve image quality and help with tumor detection in Cushing’s disease. It reportedly has higher sensitivity (80%) and specificity (∼100%) compared to 1.5T MRI which has overall sensitivity and specificity of 71% and 87% respectively. However, it is not the first to order by the clinicians when evaluating the ACTH secreting pituitary adenoma. In the case of equivocal 1.5 T MRI usually the next step is IPSS which is invasive. With availability of Corticotropin Releasing Hormone in the USA being difficult, limited studies of using DDAVP and limitations in finding tertiary centers to perform IPSS, a 3T MRI would be more feasible as a next step to identify a pituitary adenoma. Presentation: Thursday, June 15, 2023

A population-based study of children suggests blunted morning cortisol rhythms are associated with alterations of the systemic inflammatory state

BackgroundIn children, digital media, lifestyle, and the COVID pandemic have impacted sunlight exposure, exercise, and diet patterns - cues that entrain the circadian clock. We hypothesized that low morning cortisol reflects a weak circadian clock, impacting the pro-inflammatory state. The primary objective was to test relationships between diurnal cortisol fluctuations and the inflammatory state in children as a means of providing indirect support for this hypothesis. MethodsThe Cardiovascular Health Intervention Program (CHIP) was a population-based cross-sectional and longitudinal study of circadian health in public elementary school children in Southern Maine, USA (recruitment period 2012–2017). Participants were 689 students in 4th grade (baseline; age=9.2 ± 0.4 years), and 647 students in 5th grade (age=10.5 ± 0.5 years). Nine salivary cortisol measures per child (2 awakening and 1 prior to bed for 3 sequential days) (n = 1336 child phenotype days; n = 7987 cortisol assays), 10 cytokines measured in morning and evening saliva samples (n = 202 child phenotype days), and lipids were measured. Clinical outcomes were blood pressure, weight and height (body mass index [BMI]; BMI = kg/m2), among others. FindingsUpon-waking cortisol levels were 0.28 ± 0.13 µg/dL, 30-minute post-waking 0.33 ± 0.15 µg/dL, and evening 0.08 ± 0.10 µg/dL. Salivary cytokine levels (n = 202) showed interleukins (IL) IL-1β and IL-8 were highest in early morning (upon awakening; AM), and IL-6 and tumor necrosis factor (TNF) TNF-α highest before bed (PM) (IL-1β AM > PM [−4.02 fold; p < 0.001]; IL-8 AM > PM [−1.36 fold; p < 0.001]; IL-6 AM < PM [+1.49 fold; p < 0.001]; TNF-α AM < PM [+1.73 fold; p = 0.03]. Regression modeling showed high morning cortisol was associated with high morning IL-1β (p = 3.82 ×10−6), but low evening IL-1β (p = 6.27 ×10−4). Regression modeling of BMI z-score as the response variable showed the expected significant relationships to high density lipoprotein (HDL) (negative; p < 0.001), mean arterial pressure (positive; p < 0.001), and morning cortisol (negative; p = 0.01) but only weak relationships to either evening cortisol (p = 0.1) or cytokine (positive; p = 0.02; from the model with smallest Rsquared) levels. InterpretationWe provide preliminary data on diurnal fluctuations of inflammatory cytokines in saliva in a population-based cohort of children. Correlation of morning and evening cortisol levels with inflammatory cytokines in the same saliva samples showed that high morning cortisol was associated with high morning IL-1β and low evening IL-1β. Future studies may test the hypothesis that strong diurnal cycling of IL-1β may serve as a homeostatic mechanism keeping the immune system in check, and that low morning cortisol (possible circadian misalignment) may lead to less stringent control of inflammatory networks.

A-030 Indicators Generated from Salivary Cortisol Measurement to Assess Circadian Rhythm: Comparison of Immunoassay and Liquid Chromatography-Tandem Mass Spectrometry

Abstract Background Salivary cortisol is considered a reliable alternative to serum-free cortisol for evaluating hypothalamic-pituitary-adrenal axis rhythmicity. Its advantages include free hormone measurement, non-invasiveness, precision, and cost-effectiveness. Several cortisol indicators in serially collected saliva have been used in research studies to evaluate the circadian rhythm. We aimed to find if the distribution of those indicators is affected by the specific assay platform used. Methods Four-time points of saliva specimens were obtained immediately after awakening, 30 min and 60 min post-awakening, and in the nighttime from 49 subjects (aged between 19 and 25, mean ± SD: 28.9 ± 8.4 years). Salivary cortisol was analyzed by ELISA method using the Cortisol Saliva ELISA kit (IBL, Hamburg, Germany). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was performed on the API 4000 triple quadrupole mass spectrometer coupled with an Agilent 1200 liquid chromatography system. Four cortisol indicators (DCS (diurnal cortisol slope), CARi (cortisol awakening response by an absolute increase within 30 min of awakening), AUCg (area under the curve with respect to ground), and AUCi (AUC with respect to increasing)) were calculated from each patient by the method. Correlation between the two methods was evaluated for overall cortisol concentrations, DCS, CARi, AUCg, and AUCi. Results The cortisol concentrations between the two methods strongly correlated (r = 0.973) but with a mean bias of 3.4 nmol/L (95% CI −3.8 to −2.9). Four cortisol indicators strongly correlated between the two methods, but showed significant differences. DCS data generated from LC-MS/MS measurement was more broadly distributed than those from ELISA. On the contrary, CARi data generated from ELISA measurement dispersed more than those from LC-MS/MS. Conclusion The concentrations of salivary cortisol strongly correlated between ELISA and LC-MS/MS, though considerable proportional bias between the two methods. The cortisol indicators generated from different methods can affect its distribution, therefore possibly affecting its discrimination power.

Effects of a Canine-Assisted Intervention in Perceived and Physiological Stress of Spanish University Students

Abstract Many universities, concerned with finding methods to improve the academic environment and support their students, carry out animal-assisted interventions. This study measured the effect of a canine-assisted intervention program on the stress of first-year university students at the Complutense University of Madrid. A quasi-experimental pretest-posttest design was followed, with a follow-up measure. The instruments used were the Perceived Stress Questionnaire and salivary cortisol measures. The sample was 102 students. The program, called “Compludog,” consisted of one 45-minute session a week for three weeks. The results demonstrated significant improvements in physiological and perceived stress in the posttest and follow-up measure. In addition, analyses revealed that the sociodemographic variables had no influence on the effects of the program. It is concluded that a canine-assisted intervention program is effective. Therefore, Spanish universities should consider this kind of program to support their students and reduce their stress, even during exam time.

Effect of Hatha Yoga and meditation on academic stress in medical students—Clinical trial

IntroductionMedical students use to deal with high levels of academic stress. Hatha yoga and meditation have been found to reduce variables associated with stress. Objective: To evaluate and compare the effect of Hatha yoga and meditation on academic stress in medical students at a private university in Cali, Colombia. Methods and materialsA total of 27 volunteers were randomly divided into two groups: Hatha Yoga(n = 13) and Meditation (n = 14). Each group undertook two weekly one-hour sessions for 14 weeks. Study outcomes were salivary cortisol and perceived academic stress. Anthropometric and clinical variables were additionally measured as characteristics of the study sample. Perceived academic stress was estimated via a structured questionnaire. An intra-group analysis was also conducted to compare the outcomes at baseline and post-intervention within the groups. ResultsAt baseline demographic, clinical, and anthropometric variables were similar between the intervention groups. After the intervention, no differences were found between meditation and hatha yoga groups in salivary cortisol measures (time 0, 15, 30, 45 min, and mean of the 4 measures) and perceived academic stress. However, the intra-group analysis showed that only in Hatha yoga there was a significant increase in cortisol measures and improvement in the scale of perceived academic stress. ConclusionBased on the intra-group but not between-group comparisons of outcomes at baseline and post-intervention, Hatha yoga reduced perceived academic stress despite a concomitant significant increase in cortisol levels. Further trials with larger samples of medical students should be conducted to contrast our findings.