Each patient collected one MSC sample between 23.30 and 00.30 h in their home within 1 week after recruitment, using the Salivette® sampling method (Sarstedt, Nümbrecht, Germany) [17 (link), 26 (link), 32 (link)]. Patients had a restriction period of 30 min prior to sampling when they were told not to eat, drink, smoke, use snuff, nor to perform physical exercise, and a period of 60 min prior to sampling when they should avoid brushing their teeth [17 (link), 26 (link)]. The samples were centrifuged and frozen at − 25 centigrades until assayed within a year. The Roche Cobas Cortisolassay immunoassay (ECLIA) was used on an Elecsys 2010 immunoanalyser system (Roche Diagnostics, Mannheim, Germany) [17 (link), 26 (link), 33 (link)]. The detection limit was 1.9 nmol/l. The intra-assay coefficient of variation was < 3%. Low MSC was defined as < 3.7 nmol/l; intermediate MSC as ≥ 3.7 to < 6.7 nmol/l; and high MSC as ≥ 6.7 nmol/l).
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