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In vitro systems are useful for identifying key metabolic nodes and their threshold effects, but differences in amino acid concentrations in culture media, stimulation conditions, and criteria for polarization assessment may all affect the extrapolatability of the conclusions

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Yes, but precision depends on dose size

Nevertheless, heteroplasmy complicates clinical interpretation, as mutation burdens differ across tissues and tumor subclones, challenging the integration of these data into diagnostic or prognostic models [158]

The distribution can be either diffuse (e.g., ethionine) or zonal (e.g., centrilobular in CCl4