The aim of the present study was to investigate associations between spontaneous meal initiations and blood glucose dynamics in overweight male subjects in negative energy balance. In a randomized crossover design, fifteen overweight male subjects (BMI 28.6 (SD 1.8 kg/m2) participated in three treatments, each of which consisted of 2 weeks consuming a low-energy diet followed by a test of voluntary food ingestion in the absence of time-related cues. The low-energy diet consisted of three daily meals (947 kJ) which were either semi-solid with or without 2.5 g guar gum, or solid, and a dinner of subject's own choice. During the time-blinded test, on the first, second, and third meal initiation subjects ingested a low-energy meal corresponding to that used during the preceding weeks. Changes in blood glucose were monitored on-line. Associations between spontaneous meal initiations and blood glucose dynamics were determined using the chi2 test. No difference was found between treatments in the occurrence of postabsorptive and postprandial declines in blood glucose or in associations between meal initiations and blood glucose dynamics. Postprandial dynamic blood glucose declines were associated with meal initiation (chi2 26 8, P<0.00 1), but postabsorptive and postprandial transient declines were not. In overweight subjects, the usual association between transient declines and spontaneous meal initiation was completely absent in negative energy balance.