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Postulated mechanisms of metformin action, including AMPK activation and complex I inhibition, have been demonstrated in the setting of a consistent pleiotropic activity of this drug, suggesting that several molecular targets could be involved (LaMoia and Shulman, 2021)

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Traditional reversibility assays are based on regained enzymatic activity after rapid dilution [73] or washout [34], or on detection of released unbound inhibitor upon protein denaturation/digestion [55] or chasing with a competitive irreversible ABP [74]