 Endocrinological parameters Salivary cortisol levels were measured at T1, after T2 and T3. The saliva samples were collected using a standardized salivette and frozen at -20 until analysis in the laboratory. After thawing, salivary fluids were centrifugated to precipitate mucins and cortisol was assayed in the supernatant with a solid-phase radioimmunoassay, wherein 125-I labeled cortisol competes for a fixed time with cortisol in the biological sample The Coat-A-Count Cortisol, Siemens, Los Angeles, CA. 