Study Design

For gene expression analysis, groups of three male rats received acetaminophen (APAP, 99% pure, Sigma, St. Louis, MO) as a suspension in 0.5% aqueous ethyl cellulose (USP/FCC grade; Fisher Scientific Company, St. Louis, MO) by gavage. Doses utilized in this study were 0 (vehicle only), 50, 150 and 1500 mg kg-1 BW-1. The animals were sacrificed (see below) at 6, 24 or 48 h after dosing.

For ATP measurements, groups of two male rats were dosed with 0 (vehicle only), 50, 150 or 1500 mg kg-1 BW-1 APAP and sacrificed after 3, 6, 24 or 48 h.

The entire study was replicated for a biological confirmation of the results. In the case of the gene expression analysis, the data presented are primarily derived from one of the two replicates, for ATP measurements the data is presented as average of both replicates.

We performed the studies according to the guidelines in the NIH Guide for the Care and Use of Laboratory Animals (12). An approved Animal Study Protocol was on file prior to initiation of the study.

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