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Purpose: The aim of this study was to examine the activation of the contralateral upper and lower extremities and trunk muscle during ipsilateral upper extremity diagonal isokinetic exercise. Methods: Twenty-one healthy male subjects with no history of shoulder injury undertook ipsilateral diagonal isokinetic exercise at 60, 120, and 180°/sec, utilizing a standard Biodex protocol. Muscle activation amplitudes were measured in the upper trapezius, pectoralis major, biceps brachii, rectus abdominis, external oblique, rectus femoris, adductor longus, and biceps femoris muscles using electromyography. A one-way analysis of variance and paired t-tests were conducted, and the data were analyzed using SPSS, version 21.0. Results: The results revealed no statistically significant interaction between motion and angular velocity and no statistically significant contralateral muscle activation according to angular velocity (p˃0.05). However, they revealed statistically significant contralateral muscle activation according to motion (p˂0.05). Conclusion: These results suggest that the movements involved in contralateral upper extremity diagonal isokinetic exercise can enhance muscle strength in patients affected by stroke, fracture, burns, or arthritis.