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16 Microsomal membrane fractions of CYP- and FMO-overexpressing cell lines were prepared by differential centrifugation as previously described, with protein concentrations estimated using the BCA assay, as per the manufacturers recommendations

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In contrast, an individual who must have a double espresso each morning to wake up but who experiences no negative health consequences of doing so could be described as being caffeine dependent, but they are not addicted in the clinical definition

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