Glycotoxins in cigarette smoke condensate induce fluorescence and covalently cross-link proteins. (A) RNase A was exposed to mainstream cigarette smoke for 0, 5, 8, 24, 48, and 72 h and then assayed for RNase A-AGE-specific fluorescence by measuring emission at 440 nm upon excitation at 370 nm in the presence and absence of aminoguanidine. Circles, RNase A alone; squares, RNase A after incubation with cigarette smoke; triangles, RNase A after incubation with cigarette smoke and 5 mM aminoguanidine; diamonds, RNase A after incubation with cigarette smoke and 50 mM aminoguanidine. (B) RNase A was exposed to mainstream cigarette smoke for 0, 5, 8, 24, 48, and 72 h. To assess the formation of dimers, the samples were subjected to SDS/PAGE under reducing conditions, followed by transfer to nitrocellulose and Western blotting with a rabbit anti-RNase A antibody conjugated to horseradish peroxidase. Circles, RNase A alone; squares, RNase A after incubation with cigarette smoke; triangles, RNase A after incubation with cigarette smoke and 5 mM aminoguanidine; diamonds, RNase A after incubation with cigarette smoke and 50 mM aminoguanidine.