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For experiments FOXO4-DRI was dissolved in DMEM to generate a 2.5 mM stock solution

This superoxocobalamin can ultimately oxygenate NO to generate nitrate and nitrite through a peroxynitrite pathway.51,52 In essence then, cobalamin may operate as a catalyst in the presence of O 2 and reducing agents (e.g., ascorbic acid, which was recently reported to treat vasoplegia to inactivate or suppress NO signalling).53 Nevertheless, there remains ongoing debate regarding whether nitrite and nitrate can be considered functionally as stores of NOthat is, whether nitrate or nitrite can be reduced to release NO at a later time point.48 Nitric oxide synthase is responsible for the production of NO and exists in at least three forms known today (an inducible form and two calmodulin-dependent formsnNOS and eNOS).52 Inducible NOS is expressed in vascular smooth muscle cells and cardiac myocytes.54 Inflammatory mediators (interferon gamma and tumor necrosis factor ) activate the expression of inducible NOS.52 Nitric oxide causes vasodilatation by inducing smooth muscle-soluble guanylate cyclase to increase the production of cyclic guanosine monophosphate.32,48,54,55 This results in a reduction of intracellular calcium concentrations, leading to smooth muscle relaxation.54,55 As noted previously, V-B12-induced hypertension is suggested to arise from NO scavenging

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