Forearm I/R trial. Study participants reported to the Kronos Longevity
Research Institute Clinical Research Center in the morning after
consuming a light breakfast without caffeinated beverages. Upon arrival,
an i.v. catheter was inserted in the nondominant arm and a baseline
blood sample was collected (234 min). The catheter was kept in the
study participant with a saline drip throughout the trial. A blood
pressure cuff was placed on the same arm, inflated to 200 mm Hg and
kept inflated for 10 min, then released for 2 min. This inflation procedure
was repeated twice more (total time = 34 min). At the 3 time points of
cuff inflation when blood flow was occluded, ~2 mL of heparin flush (10
kUSP/L) was injected into the blood sample tubing to prevent any
clotting at the site. A blood sample was drawn 15 min after the final
release of the blood pressure cuff and then at minutes 30, 60, 120, 180,
and 240 post final cuff deflation.