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Sample preparation was conducted using a proprietary (Metabolon, Durham, North Carolina, USA) series of organic and aqueous extractions to remove the protein fraction while allowing maximum recovery of small molecules

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Thus, changes in levels of Fos labeling in the PVH observed in mice lacking robust leptin signaling are most likely due to alterations in the strength of these viscerosensory afferents

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Criteria commonly require documented body mass index consistent with the labeled population, documented weight-related comorbid conditions, and evidence of a prior structured attempt at diet and physical activity