Blood samples were obtained from the antecubital vein of the forearm via standard venepuncture technique (Vacutainers Systems, Becton–Dickinson). Samples were collected into vacutainers containing Silica (Clot Activator) and stored on ice until centrifugation for 10-min at 1,200 g at 4 °C. Serum aliquots were stored at − 80 °C for subsequent analyses. Commercially available pre-coated high-sensitivity ELISA kits (Thermo Fisher Scientific) were used to determine interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α) and high-sensitivity c-reactive protein (hs-CRP), while commercially available pre-coated standard ELISA kits (Thermo Fisher Scientific) were used to determine tissue plasminogen activator (t-PA) and von Willebrand factor (vWF). IL-6, TNF-α and hs-CRP were analysed as they are implicated in mood disorders [21 (link)] and cognitive functioning [22 (link)]. t-PA and vWF were analysed owing to their association with reductions in CBF [23 (link)]. Kits were stored and utilised according to the manufacturer’s instructions. An automated plate reader (CLARIOstar, BMG LABTECH GmbH, Offenburg, Germany) was used to read the raw absorbance values at the 450 nm wavelength for all assays. Each sample was analysed in duplicate.
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