2.4. Oxidative Stress
BLOOD, SEPARATION, OXIDATIVE STRESS
Blood samples were collected during the study visits in BD Vacutainer CPT mononuclear cell preparation tubes that contained blood separation media comprising a thixotropic polyester gel and a Ficoll Hypaque solution. The tubes were centrifuged to isolate live peripheral blood mononuclear cells (PBMC), and oxidative stress was measured using a fluorogenic cell-permeant probe CellROX Orange (Thermo Fisher, Waltham, MA, USA). Fluorescence upon oxidation by reactive oxygen species was quantified on a BD Accuri C6 flow cytometer using absorption/emission maxima of 545/565 nm and presented as relative fluorescent units (RFUs).