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Two experiments are reported which investigated whether male and female operator duration estimation and subjective workload followed conventional circadian fluctuation. In the first experiment, twenty-four subjects performed a filled time-estimation task in a constant blacked-out, noise-reduced environment at 0800, 1200, 1600, and 2000 h. In the second experiment, twelve subjects performed an unfilled time estimation task in similar conditions at 0900, 1400, and 1900 h. At the termination of all experimental sessions, participants completed the NASA TLX workload assessment questionnaire as a measure of perceived mental workload. Results indicated that while physiological response followed an expected pattern, estimations of duration and subjective perception of workload showed no significant effects for time-of-day. In each of the experiments, however, there were significant differences in durational estimates and mental workload response depending upon the gender of the participant. Results are taken to support the assertion that subjective workload is responsive largely to task-related factors and indicates the important differences that may be expected due to operator gender.