Hypotheses and fundamental study design characteristics for evaluating potential reduced-risk tobacco products. Part I: Heuristic.

The risk-reducing effect of a potential reduced-risk tobacco product (PRRP) can be investigated conceptually in a long-term, prospective study of disease risks among cigarette smokers who switch to a PRRP and in appropriate comparison groups. Our objective was to provide guidance for establishing the fundamental design characteristics of a study intended to (1) determine if switching to a PRRP reduces the risk of lung cancer (LC) compared with continued cigarette smoking, and (2) compare, using a non-inferiority approach, the reduction in LC risk among smokers who switched to a PRRP to the reduction in risk among smokers who quit smoking entirely. Using standard statistical methods applied to published data on LC incidence after smoking cessation, we show that the sample size and duration required for a study designed to evaluate the potential for LC risk reduction for an already marketed PRRP, compared with continued smoking, varies depending on the LC risk-reducing effectiveness of the PRRP, from a 5-year study with 8000-30,000 subjects to a 15-year study with <5000 to 10,000 subjects. To assess non-inferiority to quitting, the required sample size tends to be about 10 times greater, again depending on the effectiveness of the PRRP.

Cigarette smoke exposure promotes arterial thrombosis and vessel remodeling after vascular injury in apolipoprotein E-deficient mice.

BACKGROUND: Cigarette smoking is a major risk factor for the development of cardiovascular disease. However, in terms of the vessel wall, the underlying pathomechanisms of cigarette smoking are incompletely understood, partly due to a lack of adequate in vivo models. METHODS: Apolipoprotein E-deficient mice were exposed to filtered air (sham) or to cigarette mainstream smoke at a total particulate matter (TPM) concentration of 600 microg/l for 1, 2, 3, or 4 h, for 5 days/week. After exposure for 10 +/- 1 weeks, arterial thrombosis and neointima formation at the carotid artery were induced using 10% ferric chloride. RESULTS: Mice exposed to mainstream smoke exhibited shortened time to thrombotic occlusion (p < 0.01) and lower vascular patency rates (p < 0.001). Morphometric and immunohistochemical analysis of neointimal lesions demonstrated that mainstream smoke exposure increased the amount of alpha-actin-positive smooth muscle cells (p < 0.05) and dose-dependently increased the intima-to-media ratio (p < 0.05). Additional analysis of smooth muscle cells in vitro suggested that 10 microg TPM/ml increased cell proliferation without affecting viability or apoptosis, whereas higher concentrations (100 and 500 microg TPM/ml) appeared to be cytotoxic. CONCLUSIONS: Taken together, these findings suggest that cigarette smoking promotes arterial thrombosis and modulates the size and composition of neointimal lesions after arterial injury in apolipoprotein E-deficient mice.

Fibrinolytic balance of the arterial wall: pulmonary artery displays increased fibrinolytic potential compared with aorta.

Classic pulmonary thromboembolism research has documented that large pulmonary thromboemboli lyse spontaneously, suggesting potent fibrinolytic activity in human pulmonary artery (Pa). This concept conflicts with published animal studies in which the proximal Pa was reported to be devoid of tissue plasminogen activator (t-PA) expression. The current study used in situ hybridization protocols to demonstrate t-PA expression in samples of human main Pa (n = 30). Real-time PCR was used to demonstrate quantitatively that the levels of t-PA transcripts were higher than those of its primary regulator [ie, plasminogen activator-inhibitor 1 (PAI-1)] in the Pa samples. Immunologic and functional assays extended these observations by demonstrating that levels of t-PA antigen were higher than PAI-1 antigen, which resulted in the detection of free t-PA activity. This contrasted with the fibrinolytic balance of matched samples of aorta (n = 6) in which PAI-1 transcripts and antigen values were higher than the corresponding t-PA values, and only M(r) 110 kDa t-PA-PAI-1 complexes could be detected in functional assays. To assess the relative fibrinolytic contribution of the endothelial cell layer, Pa endothelial cells and aortic endothelial cells were scraped and propagated in culture for 20 +/- 6 days. Pa endothelial cell-conditioned media revealed increased t-PA/PAI-1 antigen ratios. Taken together, our data indicate that the balance between t-PA and PAI-1 is shifted in human main Pa to favor net PA activity.