Platelet aggregation test

We assessed ADP‐induced platelet aggregation by light transmittance aggregometry (PAP‐8E platelet aggregation profiler, Bio/Data Corporation, Horsham, PA, USA) at baseline and after 7 days of intervention. Citrate‐anticoagulated blood (0.32%) was centrifuged (Rotina 420R, Hettich Lab Technology, Tuttlingen, Germany) for 15 min at 180 g to obtain platelet‐rich plasma. Platelet‐poor plasma was prepared by 10 min centrifugation at 1550 g. Experiments were performed at 37°C under stirring conditions. Thrombin receptor‐activated peptide (TRAP; final concentration 15 μmol l⁻¹, Bachem, Bubendorf, Switzerland) was used to induce maximum platelet aggregation (100%). ADP and arachidonic acid were used to initiate the platelet aggregation in the test, final concentrations of ADP (0.1, 0.2, 0.5, 1.0, 2.0 μmol l⁻¹, Sigma‐Aldrich, St. Louis, MO, USA); arachidonic acid (2 mmol l⁻¹, Sigma‐Aldrich, St. Louis, MO, USA). Aggregations were performed with and without the addition of phosphocreatine (CrP 5 mmol l⁻¹ final concentration; Sigma‐Aldrich, St. Louis, MO, USA).