Background: The purpose of the present study was to compare endothelial function in lean (body mass index [BMI] = 18.0-24.9 kg/m(2)); overweight (BMI = 25-29.9 kg/m(2)); and obese (BMI >= 30 kg/m(2)), healthy, eumenorrheic women. Methods: Eighteen lean, 22 overweight, and 19 obese eumenorrheic middle-aged women were studied. Vascular structure and function were assessed via non-invasive ultrasound imaging of the carotid and brachial arteries. Body composition, blood pressure, fasting blood lipids, glucose, and insulin also were measured. Results: The groups demonstrated significantly (p < 0.001) different mean values for total body, lean body, and fat masses. The obese group demonstrated significantly (p < 0.05) elevated fasting glucose and insulin levels and lower high-density lipoprotein levels as compared to the lean group. The over-weight group also demonstrated elevated fasting glucose levels as compared to the lean group (p < 0.05) with no significant difference from the obese group. Only systolic blood pressure differed among the three groups, being elevated in the obese group compared to the lean group (P < 0.05). The obese group demonstrated significantly (p < 0.05) elevated carotid artery lumen diameter, carotid artery wall cross-sectional area, and brachial artery lumen diameter with significantly (p < 0.05) lower flow-mediated dilation as compared to the lean group. The overweight group demonstrated elevated carotid artery wall cross-sectional area and brachial artery lumen diameter as well as lower flow-mediated dilation as compared to the lean group (p < 0.05). Conclusions: The results of this study support the hypothesis that carotid artery wall cross-sectional area is elevated and flow-mediated dilation reduced in overweight and obese eumennorheic women as compared to lean counterparts in relation to BMI classification.