the respective other study group. Of the total of 68 recruited participants 3 scheduled for the dark chocolate group had to stop participation due to technical problems with initial blood sampling (catheter insertion failure (N=2); and catheter occlusion (N=1)) rendering final study samples of 31 subjects in the dark chocolate group and 34 subjects in the placebo chocolate group. Participants abstained for 48h prior to the experimental session from strenuous physical activity and from consumption of substances that contain polyphenols including dark chocolate, red wine, black tea, or apples. In addition, they abstained from consuming milk and milk-containing products as well as coffee (or substances containing caffeine such as coke or energy drinks) and alcohol starting on the evening before the experimental day. They did not consume food and drinks (other than water) in the 2h before reporting to the lab. Participants reported to the lab at 9:50 am where they received a standardized breakfast (1 soya roll, 20g margarine, 30g honey, and water) at 10:00 am. A venous catheter was inserted at 10:45 am followed 45 min later by the first saliva and blood sampling with subsequent administration of 50g of dark or placebo chocolate required to be consumed instantly. Based on previous observations on the kinetics of plasma flavonoid increases following oral intake of dark chocolate, subjects underwent the psychosocial stressor 2 h after chocolate ingestion, when plasma flavonoid levels are expected to peak (,). We presumed that dark chocolate ingestion would induce significantly elevated plasma epicatechin levels during the entire stress experiment since flavonoid plasma levels were found to be still elevated 6 h following oral administration of dark chocolate (). We applied the standard protocol of the widely used Trier Social Stress Test (TSST), which combines a short introduction phase followed by a 3-min preparation phase, a 5-min mock job interview, and a 5-min mental arithmetic task in front of an audience (). The TSST evokes reliable stress hormone increases (,). During recovery from stress, subjects remained seated in a quiet room.
the respective other study group. Of the total of 68 recruited participants 3 scheduled for the dark chocolate group had to stop participation due to technical problems with initial blood sampling (catheter insertion failure (N=2); and catheter occlusion (N=1)) rendering final study samples of 31 subjects in the dark chocolate group and 34 subjects in the placebo chocolate group. Participants abstained for 48h prior to the experimental session from strenuous physical activity and from consumption of substances that contain polyphenols including dark chocolate, red wine, black tea, or apples. In addition, they abstained from consuming milk and milk-containing products as well as coffee (or substances containing caffeine such as coke or energy drinks) and alcohol starting on the evening before the experimental day. They did not consume food and drinks (other than water) in the 2h before reporting to the lab. Participants reported to the lab at 9:50 am where they received a standardized breakfast (1 soya roll, 20g margarine, 30g honey, and water) at 10:00 am. A venous catheter was inserted at 10:45 am followed 45 min later by the first saliva and blood sampling with subsequent administration of 50g of dark or placebo chocolate required to be consumed instantly. Based on previous observations on the kinetics of plasma flavonoid increases following oral intake of dark chocolate, subjects underwent the psychosocial stressor 2 h after chocolate ingestion, when plasma flavonoid levels are expected to peak (,). We presumed that dark chocolate ingestion would induce significantly elevated plasma epicatechin levels during the entire stress experiment since flavonoid plasma levels were found to be still elevated 6 h following oral administration of dark chocolate (). We applied the standard protocol of the widely used Trier Social Stress Test (TSST), which combines a short introduction phase followed by a 3-min preparation phase, a 5-min mock job interview, and a 5-min mental arithmetic task in front of an audience (). The TSST evokes reliable stress hormone increases (,). During recovery from stress, subjects remained seated in a quiet room.
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