In a previous study, the authors reported that the 24-h rhythms of pituitary and adrenal hormones—that is, thyrotropin (TSH), prolactin (PRL), growth hormone, and cortisol—adapted only partially in a group of permanent night workers. However, the real impact of circadian rhythm alterations on the health and well-being of subjects is still unclear. In this study, the authors focus on an ergonomic field and address questions of adaptation of these hormones during the usual day sleep time (0700-1500 h) and during the usual night work time (2200-0600 h) in permanent night workers. Eleven night workers, working a night schedule for at least 2 years, submitted to a high-frequency blood sampling procedure (10 min) and to electroencephalographic recordings during sleep. The endocrine profiles of night workers were compared to those of day-active subjects studied during their usual sleep-wake schedule. During usual day sleep, despite an adapted sleep structure, cortisol levels among night workers were abnormally enhanced, whereas the TSH decreased in comparison to the plateau observed among day-active subjects. During usual work time, some hormonal disturbances persisted, in particular concerning cortisol and PRL (two hormones known to reflect the level of activation). Among night workers, the work time was associated with the quiescent period of cortisol secretion normally occurring during the first hours of sleep, and with a transient PRL increase. These results revealed altered hormonal profiles during the sleep time of night workers that do not result in an altered sleep pattern. The nocturnal work time, which requires a high level physical and mental performance, is associated with some endocrine alterations reflecting an eventual phase of hypovigilance.