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Assay MethodsSamples for carnitine analysis were obtained fromsubjects at rest, after an overnight fast. Three bloodsamples (5 ml each) were drawn, the samples centrifugedat 600g for 3 minutes in chilled centrifuge tubes,and plasma aliquots stored at -70° C. Carnitine wasmeasured by a radioenzymatic assay as previouslydescribed.33 Plasma samples were prepared in perchloricacid for the determination of free carnitine andshort-chain acylcarnitines (acyl groups less than 10carbon atoms). Total acid soluble carnitine concentrationrefers to the sum of the free carnitine andshort-chain acylcarnitine concentrations. The ratio ofshort-chain acylcarnitine concentration to total acidsoluble carnitine concentration was also calculatedfrom these measurements. Each assay was performedin duplicate, and the measurements from the threeplasma samples were averaged to determine restingvalues. In three treated subjects, carnitine measurementswere not made (one due to technical problemswith blood drawing and two because they were notfasting at the time of the measurement).
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