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For both cohorts, animals were deeply anesthetised 90 min later (pentobarbital 100 mg/kg, i.p) and transcardially perfused with cold 0.1 M of phosphate buffer (PB) for 1.5 min followed by 4% of paraformaldehyde (PFA) in PB for 2.5 min

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This is why a physician may look at waist circumference, clothing fit, weekly averages, and symptoms rather than single weigh-ins alone