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Histological changes in DR include retinal folds, new vessel formation in the optic disk, intraocular hemorrhage, anterior chamber angle, intraretinal neovascularization, mild focal retinal infarcts, and retinal gliosis of the internal plexiform layer (57)
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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 ฮผM [ 13 C 3 , 15 N]-cysteine-NEM, 36.4 ฮผM [ 13 C 2 , 15 N]-GSH-NEM, 10 ฮผM [D 4 ]-Cystine, 0.92 ฮผM [ 13 C 5 , 15 N 2 ]-GSSG, 20 ฮผM [D 4 ]-Hypotaurine, and 20 ฮผM of [ 13 C 2 ]-Taurine at a final concentration of 50 mg/mL followed by incubation on 4 ยฐC for 24 hr