Stress stimuli and altered cellular metabolism might trigger an increase in ROS production and a depletion of antioxidant defense systems, resulting in a defective mitochondrial respiratory chain, mutation in mtDNA, and altered Ca 2+ homeostasis
Dimethyl fumarate (DMF) and its metabolite monomethyl fumarate (MMF) modulate some signaling proteins activity and intracellular concentration, such as the nuclear factor (erythroid-derived 2)-like 2 (Nrf2), nuclear factor-kappa B (Nf-B), and cyclic adenosine monophosphate
Additionally, when the aqueous layer is compromised, the use of artificial tear drops and secretagogues, such as diquafosol sodium eye drops, is recommended to maintain ocular surface lubrication and dilute inflammatory cytokine concentrations
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For the quantification of sulfur metabolites in the tissue samples, the tissue metabolites were extracted and derivatized with NEM in ice-cold extraction solvent (80% MeOH: 20% H 2 O containing 25 mM NEM and 10 mM ammonium formate, pH=7.0) which includes stable isotope labeled internal standards (20 M [ 13 C 3 , 15 N]-cysteine-NEM, 36.4 M [ 13 C 2 , 15 N]-GSH-NEM, 10 M [D 4 ]-Cystine, 0.92 M [ 13 C 5 , 15 N 2 ]-GSSG, 20 M [D 4 ]-Hypotaurine, and 20 M of [ 13 C 2 ]-Taurine at a final concentration of 50 mg/mL followed by incubation on 4 C for 24 hr