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Angiotensin-converting enzyme (ACE) is known to play an important role in the pathobiology of human essential hypertension. Similarly, cis-unsaturated fatty acids, prostaglandins, and free radicals are believed to play a role in the control of blood pressure. It was observed that all the cis-unsaturated fatty acids tested can inhibit ACE activity to a significant degree. On the other hand, prostaglandins and free radicals, superoxide anion, hydrogen peroxide, and hydroxyl radical did not show significant inhibitory effects on ACE activity in vitro. But, the nitric oxide donor sodium nitroprusside showed a potent inhibitory action on ACE activity, suggesting that one of the possible mechanism(s) by which nitric oxide can bring about its anti-hypertensive action might be by modulating ACE activity in addition to its direct vasodilator action. These results indicate that there is a close interaction among ACE activity, cis-unsaturated fatty acids, and nitric oxide, which may have relevance to the pathobiology of human essential hypertension.