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Objectives: We systematically reviewed published studies looking at the relationship between total serum homocysteine (tHcy) and subclinical markers of atherosclerosis, such as carotid intimal-medial thickness (C-IMT), coronary artery calcium (CAC) and ankle-brachial index (ABI) in asymptomatic individuals. We analyzed these studies to examine this relationship as well as to guide future avenues of investigation by identifying studies that will help in the inclusion of tHcy levels in current guidelines on atherosclerotic disease management. Background: tHcy has been known to be associated with clinically evident atherosclerotic disease. However, tHcy is not incorporated in current guidelines for diagnosis of subclinical disease in high-risk asymptomatic individuals. Methods: We searched online databases (e.g., PUBMED, MEDLINE) for published articles assessing the relationship between tHcy and C-IMT, CAC and ABI. We limited the studies to asymptomatic populations, and excluded any study including symptomatic patients or individuals with a history of coronary, peripheral or cerebrovascular disease. Results: A systemic review of 19 articles revealed a significant association between elevated levels of tHcy and subclinical markers of atherosclerosis in asymptomatic individuals in most studies. Mean tHcy levels were consistently found to be higher in men compared to women. A total of 12 studies showed a significant association between tHcy and other risk markers of atherosclerosis, even after adjusting for age, sex and conventional risk factors. There were seven studies in unique populations that showed no significant relationship. We also observed that studies lowering baseline tHcy levels did not lead to an improvement in C-IMT, CAC or ABI scores. This might indicate that tHcy has a stronger role as a marker of atherosclerotic disease than as a risk factor for the same. Conclusions: Based on our review, we conclude that there is a significant association between the subclinical atherosclerotic process and tHcy, and it shows potential as a cheap marker for risk stratification of asymptomatic patients. However, future studies further elucidating this association and elaborating the exact role of tHcy in the atherosclerotic disease process are required. The results of these studies suggest the incorporation of plasma tHcy levels in future risk reduction protocols for identification of individuals at higher risk of atherosclerotic events, and thus to categorize them for more aggressive treatment with established preventive and therapeutic measures. Clin Chem Lab Med 2007;45:1667–77. [ABSTRACT FROM AUTHOR]