Brachial artery FMD (flow-mediated dilation) is impaired with aging and is associated with an increased risk of CVD (cardiovascular disease). In the present study, we determined whether regular aerobic exercise improves brachial artery FMD in MA/O (middle-aged/older) men and post-menopausal women. In sedentary MA/O adults (age, 55-79 years) without CVD, 8 weeks of brisk walking (6 days/week for approx. 50 min/day; randomized controlled design) increased treadmill time approx. 20% in both MA/O men (n=11) and post-menopausal women (n=15) (P<0.01), without altering body composition or circulating CVD risk factors. Brachial artery FMD increased >50% in the MA/O men (from 4.6±0.6 to 7.1±0.6%; P<0.01), but did not change in the post-menopausal women (5.1±0.8 compared with 5.4±0.7%; P=0.50). No changes occurred in the non-exercising controls. In a separate cross-sectional study (n=167), brachial artery FMD was approx. 50% greater in endurance-exercise-trained (6.4±0.4%; n=45) compared with sedentary (4.3±0.3%; n=60) MA/O men (P<0.001), whereas there were no differences between endurance-trained (5.3±0.7%, n=20) and sedentary (5.6±0.5%, n=42) post-menopausal women (P=0.70). Brachial artery lumen diameter, peak hyperaemic shear rate and endothelium-independent dilation did not differ with exercise intervention or in the endurance exercise compared with sedentary groups. In conclusion, regular aerobic exercise is consistently associated with enhanced brachial artery FMD in MA/O men, but not in post-menopausal women. Some post-menopausal women without CVD may be less responsive to habitual aerobic exercise than MA/O men.