Each patient collected one MSC sample between 23.30 and 00.30 h, using the Salivette® sampling method (Sarstedt, Nümbrecht, Germany) (22 (link), 30 (link), 31 (link)).
Patients had a restriction period of 30 min prior to sampling when they were told not to eat, drink, smoke, use snuff, or perform physical exercise, and a period of 60 min prior to sampling when they should avoid brushing their teeth. MSC samples delivered within 1 week from BP measurements were included. The samples were centrifuged and frozen at −25°C until assayed within a year. The Roche Cobas Cortisolassay®, a competitive electrochemiluminescence immunoassay (ECLIA) was used on an Elecsys 2010 immunoanalyser system (Roche Diagnostics) (32 (link)). The intra-coefficient of variation was <3%.
High MSC was defined as ≥9.3 nmol/L (22 (link), 30 (link), 33 (link)), which corresponds to the 83rd percentile in these participants.
Out of 292 patients who provided informed consent to participate, nine subjects were excluded due to the use of systemic corticosteroids, two subjects using topical steroids had very high MSC values (82 and 72 nmol/L) and were therefore excluded as contamination was suspected, and 85 subjects chose not to deliver or failed to deliver proper MSC samples (Fig. 1 ) (22 (link)). Finally, there were proper MSC samples for 196 participants.
Patients had a restriction period of 30 min prior to sampling when they were told not to eat, drink, smoke, use snuff, or perform physical exercise, and a period of 60 min prior to sampling when they should avoid brushing their teeth. MSC samples delivered within 1 week from BP measurements were included. The samples were centrifuged and frozen at −25°C until assayed within a year. The Roche Cobas Cortisolassay®, a competitive electrochemiluminescence immunoassay (ECLIA) was used on an Elecsys 2010 immunoanalyser system (Roche Diagnostics) (32 (link)). The intra-coefficient of variation was <3%.
High MSC was defined as ≥9.3 nmol/L (22 (link), 30 (link), 33 (link)), which corresponds to the 83rd percentile in these participants.
Out of 292 patients who provided informed consent to participate, nine subjects were excluded due to the use of systemic corticosteroids, two subjects using topical steroids had very high MSC values (82 and 72 nmol/L) and were therefore excluded as contamination was suspected, and 85 subjects chose not to deliver or failed to deliver proper MSC samples (