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Fourteen healthy subjects [seven men: age, 29 ± 3 yr; body mass index (BMI), 24.20 ± 0.94 kg/m 2 ; seven women: age, 27 ± 2 yr; BMI, 20.80 ± 0.88 kg/m 2 ; P < 0.05 for BMI] participated in the metabolic studies. In eight healthy subjects (two men and six women; age, 25 ± 1 yr), we determined the effect of water drinking on plasma osmolarity. All subjects were drug-free and nonsmoking. The institutional review board approved all studies, and written informed consent was obtained before study entry.

Subjects did not eat 12.5 h before and did not drink 1.5 h before testing. Three separate studies were conducted. In the first study, we assessed the effect of drinking 500 ml of water on energy expenditure and substrate oxidation rates by using indirect calorimetry. In the second study, we used the microdialysis technique to characterize the effect of drinking 500 ml of water on adipose tissue blood flow and metabolism. In a subgroup, studies were conducted twice, once after ingestion of placebo and once after ingestion of 100 mg of the β-adrenoreceptor blocker metoprolol (Stada Arzneimittel AG, Bad Vilbel, Germany). The medications were ingested in a single-blinded fashion 1 h before water drinking. In the third study, we determined venous plasma osmolarity at baseline and 30 and 60 min after water drinking.

Oxygen uptake and carbon dioxide production were measured by using a respiratory chamber to assess changes in energy expenditure, respiratory quotient (RQ; CO 2 produced/O 2 consumed), and carbohydrate and lipid oxidation rates, respectively. In previous studies in 16 healthy subjects, the maximal spontaneous change in metabolic rate over a 3-h period was 0.2 ± 0.09 kJ/min (3%). Throughout the study, subjects remained seated. After a run-in period of 15 min, resting energy expenditure was determined for 30 min. Then, the subjects ingested 500 ml water (22 C). In a subgroup, we also tested the effect of 500 ml of 37 C warm water. After completion of drinking, measurements were continued for another 90 min.

Microdialysis studies were conducted in the supine position as described previously ( 7 , 8 ). Briefly, one (systemic β-adrenoreceptor blockade) or two (local β-adrenoreceptor blockade) microdialysis probes were inserted into sc adipose tissue at the level of the umbilicus. Before insertion of the probes, the respective area was anesthetized superficially with EMLA cream (AstraZeneca GmbH, Wedel, Germany). On the day with systemic β-adrenoreceptor blockade, the subjects ingested metoprolol (Stada Arzneimittel AG) before testing, and the probe was perfused with Ringer’s solution (Serumwerke Bernburg AG, Bernburg, Germany) supplemented with 50 m m ethanol (B. Braun Melsungen AG, Melsungen, Germany), for monitoring changes in blood flow, and 10 μ m ascorbate (Jenapharm GmbH Co. KG, Jena, Germany). On the day with local β-adrenoreceptor blockade, the subjects ingested placebo before testing, and the perfusate for one microdialysis probe was supplemented with 100 n m of the nonselective β-adrenoreceptor blocker propranolol (Obsidan; ALPHARMA-ISIS, Langenfeld, Germany). CMA/60 microdialysis probes and CMA/102 microdialysis pumps (both from CMA Microdialysis AB, Solna, Sweden) were used. The flow rate was 2 μl/min.

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