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For the global metabolite profiling in serum, the metabolites in 10 L of serum were extracted by the 390 L of pre-chilled 82% MeOH (80 C) followed by 15 min incubation at 80 C

Real-time quantitative polymerase chain reaction Total RNA was extracted from cells using the Trizol reagent (Takara Bio Inc, Japan) according to the manufacturers instructions

Investigators noted that the restoration of melatonin rhythms in aged animals was accompanied by improvements in certain biomarkers of biological aging, suggesting a circadian and endocrine component to epithalon's putative anti-aging activity

[1] Endocrine Axis Models: Used in laboratory models to explore growth hormone axis regulation under controlled experimental conditions