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In a previous study, we developed and validated a new driving posture focused on biomechanical loads for physical fatigue reduction in static long-term sitting. In this study, the posture was evaluated in dynamic long-term driving condition by qualitative and quantitative measurements. The results showed physical fatigue of the new posture was halved in comparison with the one of the conventional posture in same car by subjective evaluations. Physiological indices had same tendency with subjective evaluations. From the results, we extracted seven physiological indices as good measures of physical fatigue while driving. Therefore, fatigue reduction of the new posture was qualitatively validated by physiological measurements.