A single site, three cohort, age-stratified, cross-sectional study was conducted in the US in subjects who were exclusive cigarette smokers (n=60), exclusive moist snuff consumers (n=48), and non-consumers of tobacco (n=60) to identify potential CVD-related endpoints that differed among the three cohorts. Enrolled subjects were generally healthy, adult males (ages 26-49) who were free of clinically significant health problems, measured ≥70% of predicted for FEV1 (spirometry), and were willing to undergo all study procedures. Tobacco-use cohorts provided their usual brand (UB cigarettes or moist snuff) for use during an overnight clinical confinement. On Day 1, a 45-minute product abstention period was followed by a UB tobacco product "challenge" appropriate to the cohort. A 10-12 hour overnight "fast" from food/drink and all tobacco products preceded the start of Day 2 procedures. Clinical endpoints, measured 15-minutes Post-challenge on Day 1 and/or Fasting on Day 2, included tobacco- (and potentially CVD-) related biomarker evaluations of spot-urine and blood samples; physiological assessments (flow-mediated dilation, carotid intima-media thickness, ankle brachial index, spirometry and expired CO); and self-reported product use, nicotine dependence and diet/health status measures. Cohort-specific inclusion/exclusion criteria were well-defined to create exclusive use groups with the expectation that the biomarkers of exposure and effect could differentiate the three cohorts.In this study, comparison of the three cohorts revealed that, within each tobacco-use cohort, comparable product usage among the different age groups was reported. No significant differences in the Fagerström nicotine dependence scores were observed within or between the tobacco-use cohorts. Overall, the subjects rated themselves as healthier than the population norm on the health questionnaire (SF-36v2^TM), with no significant age effects observed and only one significant difference between the tobacco cohorts and non-consumers.