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We have therefore undertaken a retrospective analysis of repeat SSTs performed in patients with potentially reversible causes of AI to determine if there are features of the SST results (basal, 30-minute, or delta cortisol) that might both guide a strategy for repeat testing and in addition help to identify groups of patients in whom HPA axis function is likely (or unlikely) to be restored. Materials and Methods Patient selection Difficulties in interpretation of the baseline serum cortisol and the response to Synacthen may be encountered when patients are on steroid therapy. Please note that prednisolone produces a significant positive interference in the cortisol assay used in this laboratory. Cho et al. investigated [4] over 200 healthy individuals to define the normal thresholds for serum cortisol levels upon stimulation in dynamic studies; however, they used an RIA instead of the traditional fluorometric assay, used in earlier studies. [1] Following an ITT, the 95 th percentile of the peak serum cortisol was 15 μg/dL (414 nmol/L), which was proposed to be the reference level for healthy volunteers. The study participants also underwent either a low dose (1 μg) or standard dose (250 μg) SST. All those who underwent the low dose SST had serum cortisol level >18 μg/dL (497 nmol/L) while those who underwent the standard-dose SST had serum cortisol ≥20 μg/dL (550 nmol/L). The use of this threshold as the standard dose SST was therefore suggested. [4] They also measured cortisol levels with 2 different RIAs and found that the results correlated with each other. Proposed flow chart for the use of SST in patients with potentially reversible causes of AI. *Random morning cortisol was measured between 9 and 12 am and at least 18 h after the last dose of glucocorticoid.

A 30-minute serum cortisol level following ACTH injection might lead to a false positive diagnosis of adrenal insufficiency as some patients cross the threshold cortisol levels only at 60 minutes. This practice can lead to a lifelong exposure to unnecessary treatment with exogenous steroids that can have harmful effects, including increased healthcare costs related to over-prescribing, monitoring, physician time, and enormous emotional and psychological impacts on patients. With patients in the 30-minute SST only protocol for cortisol measurement with an inadequate response, a repeat test should be done for up to 60 minutes if a robust basal to 30-minute cortisol change was observed. We reminded the clinicians of their responses to the standard dose SST (250 mcg cosyntropin injection) to enable comparison with our previous study. 1 We excluded pediatric clinicians from the survey.Data collection and curation: Hadeel Aljamei, Lama Amer, Muhammad Sohaib Khan, Eman Alrajhi, Anhar Alnassar, Reem Alahmed, Mohammed Abufarhaneh, Fayha Farraj Abothenain, Dina Mahmoud Ahmad Aljayar. the expiry of the 12 months from the creation of the short SST for antisocial behaviour (18 months in cases where an extension notice has been served following the creation of the short SST),

In patients with SAI, the 30-minute cortisol response post-SST was the best indicator of future (and more prompt) adrenal recovery, but accuracy of predicting future recovery of adrenal function was further enhanced by combining the 30-minute cortisol SST value with the baseline cortisol (which can be taken as being equivalent to a random morning cortisol). In the TAI group it was the delta cortisol across the SST (the difference between the baseline and 30-minute sample) that was the best predictor for future adrenal recovery. Combining the delta cortisol threshold with a subsequent random cortisol (above or below 200 nmol/L) helped to refine the predictive ability of the test. As our study was a retrospective analysis, we collected data on all protocols adopted by the clinicians. We defined a normal response as a stimulated cortisol value ≥550 nmol/L achieved at 30 or 60 minutes or at both time points. An abnormal response referred to a stimulated cortisol value <550 nmol/L. Primary adrenal insufficiency was defined when the patient had an inadequate response (ie, cortisol <550 nmol/L with corresponding elevated ACTH levels when ACTH results were available). Secondary adrenal insufficiency was defined by an inadequate response (ie, cortisol <550 nmol/L with corresponding low ACTH levels when ACTH results were available). 3.2 Statistical analysis Failure to meet the above criteria indicates probable Addison's disease or very marked adrenal atrophy secondary to prolonged absence of ACTH stimulation. Further tests are required to differentiate between the two.

A binomial logistic regression was performed on the whole cohort to ascertain the effects of selected variables on the likelihood that participants will show recovery at the subsequent test. Six variables were inserted into the model: age, sex, 30-minute cortisol, basal cortisol of the subsequent test, use of steroid medication, and different assay used. Linearity of the continuous variables with respect to the logit of the dependent variable was assessed via the Box-Tidwell (1962) procedure. A Bonferroni correction was applied using all six terms in the model resulting in statistical significance being accepted when P< 0.008. Based on this assessment, all continuous independent variables were found to be linearly related to the logit of the dependent variable. Data are expressed as median with 95% CI assuming a normal distribution.

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