FAO was measured with 3H-oleic acid (American Radiolabeled Chemicals) and quantified using tritiated water released per centimeter length of intestine. Briefly, the colon or duodenum (first 5 cm from the stomach) was isolated and then flushed with oxygenated PBS to remove waste. The intestinal segment was everted on a gavage needle and filled with PBS and ligated at both ends to form a sac. The explant was incubated in a buffer containing 3H-oleic acid and 100 mM carnitine for 4 hours at 37°C. At each hour, a 400 μL aliquot of assay buffer was removed from each sample and replaced with 400 μL of fresh assay buffer. The collected aliquot was quenched with 400 μL of 10% (w/v) trichloroacetic acid and centrifuged at 6800 g for 10 minutes. The supernatant was neutralized with 6 N sodium hydroxide solution and applied to columns containing AGX-1 resin (BioRad). The flow-through containing the unretained sample and the fraction eluted with 2 mL of water were pooled. Then, 10 mL of scintillation fluid was added and radioactivity measured on a scintillation counter. For the butyrate studies, explants were incubated with either 10 mM sodium chloride or 10 mM sodium butyrate.

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