Cortisol Measurements Research Articles

Wearable and Implantable Cortisol-Sensing Electronics for Stress Monitoring.

Cortisol is a steroid hormone that is released from the body in response to stress. Although a moderate level of cortisol secretion can help the body maintain homeostasis, excessive secretion can cause various diseases, such as depression and anxiety. Conventional methods for cortisol measurement undergo procedures that limit continuous monitoring, typically collecting samples of bodily fluids, followed by separate analysis in a laboratory setting that takes several hours. Thus, recent studies demonstrate wearable, miniaturized sensors integrated with electronic modules that enable wireless real-time analysis. Here, the primary focus is on wearable and implantable electronic devices that continuously measure cortisol concentration. Diverse types of cortisol-sensing techniques, such as antibody-, DNA-aptamer-, and molecularly imprinted polymer-based sensors, as well as wearable and implantable devices that aim to continuously monitor cortisol in a minimally invasive fashion are discussed. In addition to the cortisol monitors that directly measure stress levels, other schemes that indirectly measure stress, such as electrophysiological signals and sweat are also summarized. Finally, the challenges and future directions in stress monitoring and management electronics are reviewed.

Psychobiological effects of an eHealth psychoeducational intervention to informal caregivers of persons with dementia: a pilot study during the COVID-19 pandemic in Italy

BackgroundThe workload associated with caring for a person with dementia (PwD) could negatively affect informal caregivers’ physical and mental health. According to the recent literature, there is a need for studies testing the implementation of affordable and accessible interventions for improving caregivers’ well-being.AimsThis study aimed to explore the feasibility and effectiveness of an 8 week eHealth psychoeducation intervention held during the COVID-19 pandemic in Italy in reducing the psychological burden and neuroendocrine markers of stress in caregivers of PwD.MethodsForty-one informal caregivers of PwD completed the eHealth psychoeducation intervention. Self-reported (i.e., caregiver burden, anxiety symptoms, depressive symptoms, and caregiver self-efficacy) and cortisol measurements were collected before and after the intervention.ResultsFollowing the intervention, the caregivers’ self-efficacy regarding the ability to respond to disruptive behaviours improved (t = − 2.817, p = 0.007), anxiety and burden levels decreased (state anxiety: t = 3.170, p = 0.003; trait anxiety: t = 2.327, p = 0.025; caregiver burden: t = 2.290, p = 0.027), while depressive symptoms and cortisol levels did not change significantly. Correlation analyses showed that the increase in self-efficacy was positively associated with the improvement of caregiver burden from pre- to post-intervention (r = 0.386, p = 0.014). The intervention had a low rate of dropout (n = 1, due to the patient’s death) and high levels of appreciation.DiscussionThe positive evidence and participation rate support the feasibility and effectiveness of the proposed eHealth psychoeducational intervention to meet the need for knowledge of disease management and possibly reduce detrimental effects on caregivers’ psychological well-being.ConclusionFurther placebo-controlled trials are needed to test the generalizability and specificity of our results.

Forests are chill: The interplay between thermal comfort and mental wellbeing

As global warming and urbanisation intensify unabated, a growing share of the human population is exposed to dangerous heat levels. Trees and forests can effectively mitigate such heat alongside numerous health co-benefits like improved mental wellbeing. Yet, which forest types are objectively and subjectively coolest to humans, and how thermal and mental wellbeing interact, remain understudied. We surveyed 223 participants in peri-urban forests with varying biodiversity levels in Austria, Belgium and Germany. Using microclimate sensors, questionnaires and saliva cortisol measures, we monitored intra-individual changes in thermal and mental states from non-forest baseline to forest conditions. Forests reduced daytime modified Physiologically Equivalent Temperature (mPET; an indicator for perceived temperature) by an average of 9.2 °C. High diversity forests were the coolest, likely due to their higher stand density. Forests also lowered thermal sensation votes, with only 1 % of participants feeling ‘warm’ or ‘hot’ compared to 34 % under baseline conditions. Despite the desire for a temperature increase among 47 % participants under cool forest conditions, approximately two-thirds still reported feeling very comfortable, in contrast to only one-third under baseline conditions. Even at a constant perceived temperature, participants were 2.7 times more likely to feel warmer under baseline conditions compared to forests. A forest-induced psychological effect may underlie these discrepancies, as supported by significant improvements in positive and negative affect (emotional state), state anxiety and perceived stress observed in forests. Additionally, thermal and mental wellbeing were significantly correlated, indicating that forest environments might foster a synergy in wellbeing benefits.

Relationship between wool cortisol, wool quality indices of Australian Merino rams and climatic variables in Tasmania

IntroductionAustralia is the world’s largest producer and exporter of fine wool. External factors (i.e., non-genetic) that affect wool phenotype need to be identified and managed to minimize any negative effects on wool quality. Climatic stress is one external event that can affect wool phenotype, other stressors include shearing, nutrition, disease, pregnancy, and lambing. This study focuses on the relationship between Saxon Merino ram wool phenotype and wool cortisol. The hypotheses had two parts (i) wool cortisol levels would vary seasonally and variations would correlate with wool phenotype (ii) that these seasonal variations would also correlate with thermal indices.MethodsCortisol levels in Saxon Merino ram wool samples were determined using a cortisol enzyme-immunoassay (EIA) using polyclonal cortisol antibody. The wool cortisol results were correlated for significance with thermal indices including Temperature Humidity Index [THI] and Chill Index [CI] obtained from the Australian Bureau of Meteorology.ResultsPart 1 of hypothesis one was supported with wool cortisol showing significant seasonal variation. Part 2 of hypothesis 1 was not supported, with no statistically significant causality present between total wool cortisol levels and the price index of Saxon Merino ram wool. However, there was significant causality between wool cortisol and coarse edge measurement (CEM) of wool. Hypothesis 2, the proposal that thermal indices would impact on wool cortisol was also not supported.DiscussionIt can be concluded that although conclusive causality between total wool cortisol and wool phenotype quality could not be ascertained, there were indirect effects of cortisol levels on wool phenotypes with significant effects on the CEM. The CEM of Merino Saxon ram wool can be manipulated by assessing and managing cortisol levels during the growth cycles of Merino wool, through appropriate nutrition and husbandry procedures stress could be managed.

Physiology of human-horse interactions during substance withdrawal within psychotherapy participants

Abstract Psychotherapy incorporating equine interaction (PIE) is emerging as an effective treatment for substance use disorder (SUD); however, research concerning physiological impacts of PIE during substance withdrawal is lacking. This study investigated impacts of PIE on salivary cortisol concentrations and heart rates in SUD patients during withdrawal. Heart rate and cortisol concentrations were also measured in horses to investigate potential human-horse coupling during PIE. Saliva samples and heart rates were collected from SUD patients (n = 18) and their therapy horses (n = 4) prior to the introduction of the horse and following equine interaction within a residential psychotherapy program during the substance withdrawal period. Without the presence of the horse, the equine environment during the first week of withdrawal produced lower () cortisol and heart rate measures than found in the equine interaction for the SUD patients. Human heart rates, however, decreased () in the second week in response to the equine interaction. A strong negative correlation (r = −0.9, ) was found within the changes in human and horse cortisol concentrations during week two as human cortisol concentrations decreased while horse cortisol concentrations increased. Results indicate equine interaction during psychotherapy is more effective in the second week than the first at mitigating stress for withdrawing residential SUD treatment program patients and the equid environment, even without the presence of a horse, can positively impact stress parameters in withdrawing SUD patients during the first week of treatment.

The utility of a baseline morning cortisol measurement, in the screening for adrenal insufficiency amongst non-critically ill hospital inpatients

BackgroundSerum cortisol levels vary according to the daily circadian cycle, with peak levels seen in the morning. There is evidence in ambulatory patients that a morning cortisol level can predict an adequate cortisol response to Synacthen (tetracosactide) stimulation. AimThe aim of this study is to determine the utility of baseline analysis of the serum cortisol levels in the morning “baseline morning cortisol”, determined using a newer immunoassay, in the screening for adrenal insufficiency amongst non-critically ill hospital inpatients. MethodsThis is a retrospective cross-sectional cohort study. Results64 adult inpatients had undergone a short Synacthen test (SST) (measurement of serum cortisol levels 30 and 60 min after administration of tetracosactide) during the study period. 17 patients returned an abnormal SST result. The measured level of cortisol in the morning correlated to both the 30 min and 60 min stimulated cortisol values (r = 0.612, p < 0.001 and r = 0.639, p < 0.001). After inspecting the receiver operating characteristic curve, a cortisol concentration of 200nmmol/L measured in the morning was selected as a threshold for predicting the SST result. Using this cut off, sensitivity for predicting a normal SST was 100 %, specificity 56 %, positive predictive value 45 % and negative predictive value 100 %. ConclusionA morning cortisol measurement, determined using a newer immunoassay, is sufficient in most cases to screen for adrenal insufficiency amongst non-critically ill hospital inpatients. Use of clinical judgement in conjunction with single morning cortisol measurement is likely to reduce the need for SST testing amongst inpatients.

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.

Concentration levels of selected hormones in judokas and the extent of their changes during a special performance test at different ambient temperatures

BackgroundThere is little scientific literature available on the diversity of physiological responses of judokas to anaerobic interval exercises in warm environments. Understanding the dynamics of changes in the concentration of selected hormones during a special endurance test at different ambient temperatures may have significant practical value, as it provides an opportunity for optimal programming and monitoring of the training process. So, the main aim of the research was to survey interval anaerobic exercises in different ambient temperatures on Concentration levels of selected hormones in judokas.Methods15 judokas athletes (age: 20.65 ± 2.03 years; body height: 178.00 ± 6.31 cm; body mass: 76.26 ± 12.57 kg; training experience: 12.1 ± 1.57 years) volunteered for the study. The judokas performed five sequences (each lasting 7.20 min) of pulsatile exercises on a cycle ergometer and hand ergometer in a thermoclimatic chamber at temperatures of 21 ± 0.5 °C and 31 ± 0.5 °C. The exercises were different from typical interval exercises, with varying times, upper and lower limb loads, and were followed by a 15-minute break after each sequence. Total duration of the experiment, including the five sequences of pulsating exercise and four 15-minute rest breaks between each exercise sequence, amounted to 96 min and 20 s. The workload was increased by 20 W for the lower limb tests and 12 W for the upper limb tests every 2 min. Biochemical measurements of testosterone (T), cortisol (C), growth hormone (HGH), adrenocorticotropic hormone (ACTH), follicle-stimulating hormone (FSH), adrenaline (E), noradrenaline (NE), and β-endorphin (β-end)were performed using the enzyme-linked immunosorbent assay (ELISA) method on blood samples taken before and after five series of pulsatile exercises, at 1, 24, and 48 h.ResultsPulsatile exercise at ambient temperatures of 21 and 31 °C resulted in a decrease in body weight of the studied athletes (p < 0.05) and significantly reduced body volume and plasma volume after training (p < 0.05). The concentration of HGH, testosterone, cortisol and NE showed a statistically significant difference after the end of the series of pulsating exercises at both temperatures (p < 0.05) and did not significantly affect the concentration of ACTH, FSH and adrenaline concentration.ConclusionsAn increase in the concentration of growth hormone, cortisol and NE was observed after doing the work at both 21 and 31 °C ambient temperature. Physical exertion in both ambient temperatures contributed to a statistically significant decrease in testosterone concentration. Based on the obtained research results, it can be concluded that physical activity in various thermal conditions of the external environment activates the hormonal response to varying degrees, with the direction of changes depending on the external thermal factor.

Attachment in close relationships and glycemic outcomes in children with type 1 diabetes

BackgroundOur aim was to determine whether child attachment to parents, parent attachment style, and morning cortisol levels were related to diabetes outcomes measured by average glycated hemoglobin (HbA1c), HbA1c variability over 4 years and time in range (TIR) in children with type 1 diabetes (T1D).Research design and methods101 children with T1D and one of their parents were assessed at baseline for child attachment (Child Attachment Interview; CAI) and parent attachment (Relationship Structures Questionnaire; ECR-RS). Serum samples were collected for cortisol measurements before the interviews. HbA1c levels were measured during a 4-year follow-up period at regular 3-monthly visits, and data for TIR were exported from blood glucose measuring devices. Multivariate linear regression models were constructed to identify independent predictors of glycemic outcomes.ResultsMore girls than boys exhibited secure attachment to their mothers. The results of the regression models showed that securely attached girls (CAI) had higher average HbA1c than did insecurely attached girls (B = −0.64, p = 0.03). In boys, the more insecure the parent's attachment style, the worse the child's glycemic outcome: the higher the average Hb1Ac (B = 0.51, p = 0.005), the higher the HbA1c variability (B = 0.017, p = 0.011), and the lower the TIR (B = −8.543, p = 0.002).ConclusionsAttachment in close relationships is associated with glycemic outcomes in children with T1D, and we observed significant differences between sexes. A sex- and attachment-specific approach is recommended when treating children with less favorable glycemic outcomes. Special attention and tailored support should be offered to securely attached girls in transferring responsibility for diabetes care and at least to male children of insecurely attached parents to prevent suboptimal glycemic control. Further studies in larger samples and more daily cortisol measurements may help us better understand the links between stress response, attachment and T1D.

Characteristics of very low birthweight infants who have cortisol measurements taken and associations with neonatal acute kidney injury.

Characteristics of very low birthweight infants who have cortisol measurements taken and associations with neonatal acute kidney injury.

Comprehensive assessment of cortisol and cortisol metabolites provides insight into the complex relationship between HPA axis function and BMI

Aims/objectivesAlthough considerable effort, both experimental and theoretical, has been directed towards understanding the relationship between the HPA axis and weight regulation, no true consensus exists in the literature as to the nature of the relationship. The aim of this study was to explore potential correlations between BMI and measures of cortisol and cortisol metabolites using dried urine and saliva sampling in a large sample of individuals with BMIs ranging from underweight to obese. Materials and methodsA cohort of patients with data available from urinary and/or salivary measures of cortisol and cortisol metabolites who met inclusion criteria was extracted from the database of a commercial clinical laboratory. Pearson correlation coefficients were used to determine associations between variables; Student's t-test and one-way ANOVA were used to examine differences between groups, and the Jonckheere-Terpstra trend test was used to assess for trends by BMI category. A multivariable linear regression model was created to determine which variables explained the largest amounts of variance in BMI. ResultsA significant correlation was observed between the urinary cortisol metabolites and BMI (P < 0.0001). In addition, cortisol metabolites were associated with changes in BMI over time. No significant correlation was observed between urinary free cortisol and BMI, and correlations observed between BMI and other variables, with the exception of age, were either weak or not statistically significant. ConclusionsThe data presented in this study suggest that cortisol metabolism is a key component of weight regulation and that cortisol metabolite concentrations may potentially serve as informative biomarkers to characterize the relationship between the HPA axis and changes in BMI. The implications of this affect both clinical practice and the research and development of both prevention and treatment strategies aimed at either decreasing or increasing BMI.

Hyponatremia due to adrenal insufficiency after a Mamushi bite: a case report

BackgroundMamushi bites are the most common venomous snake bites in Japan, with known complications including rhabdomyolysis and acute kidney injury; however, adrenal insufficiency as a result of snake bites has not been previously reported. We report a case of empty sella with transient adrenal insufficiency during hospitalization for a Mamushi bite.Case presentationAn 84-year-old man was admitted to our hospital with a Mamushi bite on the right fifth finger. Serum sodium (Na) level remained in the normal range. On the ninth day of admission, he developed hyponatremia, with a serum Na level of 114 mEq/L and serum cortisol level of 4.0 μg/dL (reference value 4.5–21.1 μg/dL). His serum Na level was restored within the normal range after administration of corticosteroids with 3% NaCl solution. Both rapid adrenocorticotrophin and corticotropin-releasing hormone loading tests showed low cortisol response. Based on the results of the hormone loading tests, a diagnosis of pituitary adrenal insufficiency was made. Contrast-enhanced pituitary magnetic resonance imaging (MRI) showed primary empty sella. After discontinuation of corticosteroids, the hyponatremia did not recur, and the patient was discharged on the 24th day of hospitalization. After discharge, the patient visited an outpatient clinic, but hyponatremia recurrence was not observed.ConclusionsThis is the first report of hyponatremia due to pituitary adrenal insufficiency during hospitalization for a Mamushi bite in a patient with empty sella. When hyponatremia occurs during hospitalization for a Mamushi bite, cortisol measurement, hormone loading test, and head MRI should be performed to search for pituitary lesions because of the possibility of adrenal insufficiency caused by snake venom.

Adrenocortical function in patients with Single Large Scale Mitochondrial DNA Deletions: a retrospective single centre cohort study.

Single Large Scale Mitochondrial DNA Deletions (SLSMDs), Pearson Syndrome (PS) and Kearns-Sayre Syndrome (KSS), are systemic diseases with multiple endocrine abnormalities. The adrenocortical function has not been systematically investigated with a few anecdotal reports of overt adrenal insufficiency (AI). The study aimed to assess the adrenocortical function in a large cohort of SLSMDs. A retrospective monocentric longitudinal study involved a cohort of 18 SLSMDs patients. Adrenocortical function was evaluated by baseline adrenocorticotrophic hormone (ACTH) and cortisol measurements and by high- (HDT) and low-dose (LDT) ACTH stimulation tests and compared with 92 healthy controls (HC). Baseline adrenocortical function was impaired in 39% of patients and by the end of the study, 66% of PS and 25% of KSS showed an insufficient increase after ACTH stimulation, with cortisol deficiency due to primary AI in most PS and subclinical AI in KSS. Symptomatic AI was recorded in 44% of patients. Peak cortisol levels after ACTH stimulation tests were significantly lower in patients than in HC (P < .0001), with a more reduced response to LDT vs HDT (P < .05). Our study highlights that cortisol deficiency due to primary AI represents a relevant part of the clinical spectrum in SLSMDs, with more severe impairment in PS than in KSS. Basal and after-stimulus assessment of adrenocortical axis should be early and regularly investigated to identify any degree of adrenocortical dysfunction. The study allowed the elaboration of a diagnostic process designed for the diagnosis, treatment, and follow-up of adrenocortical abnormalities in SLSMDs.

Effects of temperature and feeding regime on cortisol concentrations in scales of Atlantic salmon post-smolts

Anadromous fish are vulnerable to anthropogenic and environmental stressors including pollution, rising temperatures, and changes in food availability. Knowledge of how fish respond to specific stressors aid our understanding of population declines and inform predictions of how populations will react to future environmental change. There is increasing interest in using cortisol measurements from fish scales to measure chronic stress in fish, however the effects of increased metabolic rate on scale cortisol dynamics must also be considered. This study examines the effect of temperature and feeding conditions on scale cortisol in Atlantic salmon (Salmo salar). Post-smolts were subjected to three different temperatures (6, 10.5 and 15 °C) and four feeding/starvation treatments over a 12-week period. Females held at 6 and 15 °C had significantly higher scale cortisol levels than those held at 10.5 °C, while rearing temperature had no effect on scale cortisol in males. The increase in scale cortisol at 6 °C indicated that temperature related differences were not driven solely by metabolic rate. A two-week starvation period produced an increase in scale cortisol in males and females held at 10.5 °C but not at 6 °C or 15 °C. The study demonstrates that scale cortisol fluctuations can be detected in a low amount (∼10 mg) of Atlantic salmon scales for monitoring of the physiological stress response. Scale cortisol shows potential for monitoring physiological responses during the marine phase in Atlantic salmon. However, the influence of environmental stressors on scale cortisol needs to be better understood, with consideration for sex-specific and interactive effects.

THU021 Mysterious Resolution Of Cushing’s Disease

Abstract Disclosure: D.R. Whiteman: None. T. Hoang: None. V. Del Signore: None. P.V. Shah: None. Cushing’s disease is a unique problem to the endocrinologist. Clinical acumen is needed to guide correct therapy. We describe a case of Cushing’s disease in a young reproductive female with abnormal menses and obesity. Post-operative surgical pathology did not confirm her disease, but her menses normalized and biochemical evidence of disease did not persist. Case Presentation: A 24 year old female presented with irregular menses since menarche. She had 2-3 menstrual cycles per year prior to oral contraceptive pills (OCPs). She stopped OCPs and failed to conceive for &amp;gt; 1 year. Other symptoms included chronic fatigue, increased weight gain, abdominal striae, and hirsutism on her chin, arms, abdomen, and nipples. Prior evaluation showed positive progesterone withdrawal. Endocrine work-up included low dose 1mg dexamethasone suppression (LDST), total testosterone, FSH, LH, estradiol, sex hormone binding globulin, 17-hydroxyprogesterone, DHEA-sulfate, IGF-1, TSH, free T4, prolactin, and hemoglobin A1c. Work-up only positive for LDST: cortisol of 3.8 ug/dL. Additional screening tests included: 24 hour urine cortisol collection of 32 ug/24hr but volume not reported and late-night salivary cortisol (LNSC) of &amp;gt;0.1 ug/dL. Three more LNSC tests resulted as 0.212, 0.739, and 0.154 ug/dl. Confirmatory testing with 8mg Dexamethasone suppression test was positive: cortisol of 0.4 ug/dL. ACTH was 21.6 pg/mL, consistent with ACTH dependent disease. A pituitary MRI with and without contrast was negative for pituitary adenoma. Next, she completed inferior petrosal sinus sampling (IPSS) that lateralized to the left sella. Her central to peripheral ACTH ratio was &amp;gt; 2 and lateralized to the left with a ratio &amp;gt; 1.4. She had transsphenoidal pituitary resection and postoperative cortisol initially dropped to 5.5 ug/dL, but increased to 13.3 ug/dL prior to discharge. No glucocorticoid replacement required. Surgical pathology demonstrated non-neoplastic pituitary tissue with no pathologic change. Post-operative IPSS revealed all cortisol levels &amp;lt; 2.0 ug/dL. Two post-operative AM cortisol measurements were 5.4 ug/dL and 10.8 ug/dL suggestive of no remission. Interestingly, 2 months post-operatively repeat screening tests were negative: LDST cortisol of 0.6 ug/dL, salivary cortisol &amp;lt;0.010 ug/dL, and 24 hour urine cortisol 18 ug/24 hour. Furthermore, menses returned post-operatively. Discussion: Missing a diagnosis of Cushing’s disease results in increased morbidity. The diagnostic process can be both complex and invasive. Localization of the adenoma and resection are needed for cure. Standard practice dictates confirmation with pathology and biochemical remission. Interestingly, initial cortisol levels did not suggest remission, but later biochemical testing ruled out ongoing Cushing’s disease. Due to inadequate tissue data, true etiology of disease and remission remains a mystery. Presentation: Thursday, June 15, 2023

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

THU315 Sleep And Circadian Regulation, And Intrinsically Photosensitive Retinal Ganglion Cells Function In Diabetic Retinopathy

Abstract Disclosure: S. Reutrakul: None. J.J. McAnany: None. J.C. Park: None. F.Y. Chau: None. K.K. Danielson: None. P. Bharati: None. S. Pannain: None. E. Van Cauter: None. E.C. Hanlon: None. Background: Diabetic retinopathy (DR) is associated with a dysfunction of intrinsically photosensitive retinal ganglion cells (ipRGCs), a crucial component of the entrainment of the circadian system and of the photic entrainment of melatonin secretion. We explored ipRGC function, sleep, 24-h melatonin and cortisol profiles in patients with and without DR. Methods: Type 2 diabetes patients with DR (n=10) and without DR (n=7,) participated. ipRGC function was inferred from the post illumination pupil response (PIPR). Sleep was assessed by 14-day actigraphy. Serum was sampled at hourly intervals for 24hrs for melatonin and cortisol measurement. Results: Mean (SD) age (53.8 (8.1) vs (54.0 (8.6) yr) and mean A1C (7.5 (1.7) vs. 7.1 (0.9)%) did not differ between DR and no-DR patients. PIPR was significantly smaller in DR than no-DR patients (0.14 (0.16) vs. 0.33 (0.10), p=0.001). Nightly sleep duration and efficiency were similar, but sleep variability (standard deviation of daily sleep duration over 7 nights) tended to be larger in DR than no-DR patients (84.1 (29.8) vs. 59.5 (21.8) minutes), p=0.08. Twenty four-hour serum melatonin output was significantly lower in DR than no-DR patients (103.1 (95.4) pg/ml vs. 231.1 (131.3) pg/ml, p=0.034). All no-DR patients exhibited the classical 24-h profile of melatonin with consistently low levels during the day time and a clear nocturnal rise peaking around the middle of the night. In contrast, a normal melatonin rhythm was observed only in 3 of the 10 patients with DR. Six DR patients had 24-h mean melatonin level under 3 pg/ml, five without any evident day-night pattern. One DR patient had a shift in melatonin timing. There was a significant correlation between PIPR and 24-h melatonin output, r= 0.726, p&amp;lt;0.001, but not with sleep variability. In contrast, mean 24-h cortisol profiles and levels, however, were not significantly different between the two groups (mean levels DR 6.18 (1.23) vs no-DR 6.18 (0.73) μg/dL), Conclusion: DR is associated with ipRCGs dysfunction, irregular sleep, and with major disruptions of the 24-hour melatonin rhythm while cortisol rhythm was maintained, suggesting circadian dysfunction in those with type 2 diabetes and retinopathy. Whether melatonin supplementation is beneficial in type 2 diabetes with DR should be explored. Presentation: Thursday, June 15, 2023

Technical validation and a comparison of two methods to quantify individual levels of glucocorticoids in Alpine marmot hair

Quantification of cortisol concentration in hair has become a promising conservation tool for non-invasive monitoring of “stress” in wild populations, yet this method needs to be carefully validated for each species. The goals of the study were:•Immunologically validate two methods (study 1 and 2 respectively) to extract and quantify cortisol in the hair of wild Alpine marmots.•Compare the amount of cortisol extracted from hair samples using two methods i.e. cut into fine pieces (study 1) and hair samples pulverized using a ball mill (study 2).•Determine the extent to which methods in study 2 could provide individual specific hair cortisol (HC) measures when samples were taken from the same body location. Within and between individual variations in HC levels were examined from multiple hair samples from 14 subjects in study 2. We evaluated if inter-individual variations in HC levels could be explained by sex and age.At least twice the amount of cortisol was obtained per g/hair when samples were pulverized in a ball mill prior to extraction compared to when cut into pieces. Our methods demonstrated intra-individual consistency in HC at a given time point: inter-individual variation in HC was three times larger than within individual variance. Sex and age did not impact HC levels.

THU623 Leuprolide Treatment Of Mild Cortisol Secretion (MACS) Due To Primary Bilateral Macronodular Hyperplasia (PBMAH) Expressing Aberrant LH Receptors

Abstract Disclosure: L. Branchaud-Croisetière: None. M. St-Jean: Speaker; Self; Recordati rare disease, GlaxoSmithKline. Stock Owner; Self; Spruce bioscience. Cushing syndrome (CS) is caused by PBMAH in &amp;lt;2% of cases. Treatment is mostly surgical with unilateral or bilateral adrenalectomy, but in some cases, identification of aberrant receptors can allow the use of targeted medical therapies. Only a few cases of adrenal CS associated with an aberrant LH/hCG receptors that were treated with GnRH agonist were described. In December 2020, we saw a 72-year-old woman for bilateral adrenal nodules and an ACTH independent hypercortisolism. Her past medical history includes depression, insomnia, and osteoporosis with multiples fragility fractures. She was taking citalopram, calcium, vitamin D and alendronate was added in 2021. In the follow-up she developed dyslipidemia and hypertension. There were no sign of CS on physical examination. In 2020, a non-contrast thoracic scan was done and showed a right adrenal nodule of 3x1,4 cm and a left adrenal nodule of 3 x 2 cm both with densities &amp;lt; 10 HU and no atrophy of the adrenal limbs. In August 2021, a FDG PET scan showed an SUV of 3,4 on the right side and 3,7 on the left side. Morning cortisol after 1 mg DST (done twice) were 122 and 133 nmol/L (N &amp;lt;50). Aldosterone to renin ratio and 17-OHP were normal. In September 2020, a 4 mg IV dexamethasone suppression test was done and cortisol was 189 nmol/L at 9h am on day 2. Between 2020 and 2021, ACTH (3 values) were between 1,3-1,7 pmol/L (N 0-10 pmol/L) and DHEAS (3 values) between 1-1,6 (N 0,95-11,67 mol/L). 24h UFC (7 values) varied from 280,6 to 600,6 nmol/d (N 153-789 nmol/d). LNSC (8 values) were normal &amp;lt;2 to 3 nmol/L (N &amp;lt;5) except for one value at 6 nmol/L. In February 2022, systematic research for aberrant receptors was done. The cortisol have showed an ACTH-independent increased of 242% following 100 mcg IV of LHRH. There was also a significant ACTH-independent increase after 10 IU of vasopressin (+ 66%) and 10 mg of maxeran PO (+ 98%). There was no significant increase in cortisol after stimulation with glucagon 1mg IV and the upright posture test. We couldn’t obtain cortisol measures after stimulation with cortrosyn and the mixed-meal test because of issues with samples. Treatment with Leuprolide 3,75mg IM monthly was started in April 2022 and the dose was increased to 7,5 mg in August 2022 because of partial response observed initially. There was a response with an increase in ACTH from &amp;lt;1,1 to 2,6 pmol/L until December 2022. Since January 2023, ACTH (3 values) have decreased between 1,8-2 pmol/L. In January 2023, the morning cortisol after 1 mg DST was 104 nmol/L and 24h UFC were 161 and 306 nmol/d and two LNSC were &amp;lt;2 nmol/L. However, with the recent decrease in ACTH, we suspect an escape to Leuprolide. In summary, this is a rare case of MACS due to PBMAH expressing aberrant LH receptors with initial response to Leuprolide. We are waiting for the results of genetic testing. The patient could eventually benefit from a left unilateral adrenalectomy if escape from medical treatment is confirmed. Presentation: Thursday, June 15, 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