2.4.1. Sample Collection

A portion of whole heparinized blood was separated for the comet assay; the remaining blood was centrifuged at 2000× g for 10 min in a refrigerated centrifuge.

Frozen tissues were homogenized with a lysis buffer (Cell Lysis Buffer 2; Bio-Techne-R&D Systems, Minneapolis, MN, USA) supplemented with a cocktail of protease and phosphatase inhibitor (Protease Inhibitor Cocktail I, Bio-Techne-Tocris, Minneapolis, MN, USA) at a weight:volume ratio of 1:2, using a tissue homogenizer (Ultra-Turrax model T25; Ika Labortechnik, Staufen, Germany). The homogenate of each sample was centrifuged at 10,000× g for 20 min at 4 °C. The supernatant was collected for the biochemical assays with the exception of mitochondrial ALDH2 activity assay.

A portion of whole heparinized blood was centrifuged (3000 rpm, 4 °C) for plasma separation.