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Objective biomarkers like rising malondialdehyde and inflammatory cytokines (IL-6, TNF-alpha) become measurable around 1014 days post-discontinuation

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ELISA for detection of anti-DNP antibodies ELISA 96-well plates (Fisher Scientific) were coated at 4 C overnight with 100 l of 5 g/ml BSA-DNP (Invitrogen) dissolved in coating buffer (1.0 L deionized water containing 8.4 g NaHCO 3 and 3.56 g Na 2 CO 3 , pH: 9.5)