Melatonin, a pineal hormone synthesized at night, is critical for the synchronization of circadian and seasonal rhythms, being a key regulator of energy metabolism in many animal species. Although studies in humans are lacking, several reports, mainly on hibernating animals, demonstrated that melatonin supplementation and a short photoperiod increase brown adipose tissue (BAT) mass. The present proof-of-concept study is the first, to our knowledge, to evaluate BAT in patients with melatonin deficiency (radiotherapy or surgical removal of pineal gland) before and after daily melatonin (3 mg) replacement for 3 months. All four studied patients presented increased BAT volume and activity measured by positron emission tomography-MRI. We also found an improvement in total cholesterol and triglyceride blood levels without significant effects on body weight, liver fat, and HDL and LDL levels. Albeit not statistically significant, fasting insulin levels and HOMA of insulin resistance decreased in all four patients. The present results show that oral melatonin replacement increases BAT volume and activity and improves blood lipid levels in patients with melatonin deficiency, suggesting that melatonin is a possible BAT activator. Future studies are warranted because hypomelatoninemia is usually present in aging and appears as a result of light-at-night exposure and/or the use of b-blocker drugs.Melatonin, a pineal hormone synthesized and released at night that plays a critical role in the synchronization of circadian and seasonal rhythms, has been studied as a key regulator of energy metabolism in many animal species for a long time (1,2). Pinealectomized rats show increased body weight gain and metabolic disturbances that are prevented by daily melatonin supplementation at night, without decreasing energy intake (1-5). This suggests that melatonin also regulates energy metabolism by its action on energy expenditure, possibly as related to the activation of brown adipose tissue (