, 2011). Some of those compounds are known carcinogens, such as B(a)P, a PAH and 4-(N-methylnitrosamino)-1-(3-pyridinyl)-1-butanone (NNK), a tobacco-specific N-nitrosamine. Both compounds are classified by IARC as “carcinogenic to humans” (Group 1) ( IARC, 2012a and IARC, 2012b). Testing of complex mixtures is problematic as a small number of particular components can mask the effects of others, especially if they elicit high cytotoxicities. In addition, components can also act synergistically or act competitively, so the testing

of mixtures can only give a global picture Veliparib datasheet obscured by these factors. Fractionation of the different smoke components can assist in the determination of what the key toxic drivers in smoke may be. A global

picture can also be used to compare smoke from different tobaccos, which may have different toxicities. There are different mechanisms by which cigarette smoke carcinogens interact with DNA (Hecht, 1999). DNA adduct formation and oxidative DNA damage are mechanisms known to generate DSBs by acting directly on the DNA. Cigarette smoke has also shown to have aneugenic activity (Van et al., 2008), an indirect-acting mechanism of genotoxicity. However, no single compound present in cigarette smoke has been classified as an aneugenic compound. Currently, the genotoxic potential of cigarette smoke is measured mostly using methods focusing on fixed DNA damage after acute exposures to different forms GSK2118436 Calpain of cigarette smoke (DeMarini et al., 2008, Schramke et al., 2006, Van et al., 2008, Wolz et al., 2002, Nakayama et al., 1985 and Johnson et al., 2009). There are

also multiple clinical studies focusing on the genotoxic effects of cigarette smoke in humans (Hruba et al., 2010; Choudhury et al., 2008 and Mondal et al., 2010). However, these clinical studies fall out of the scope of this review. Recent reviews described the different physical forms of cigarette smoke used in in vitro testing ( Table 4) and the history of the collection of tobacco smoke for toxicology testing ( Breheny et al., 2011 and Johnson et al., 2009). De Marini conducted a detailed review of the genotoxicity of tobacco smoke and tobacco smoke condensate (DeMarini, 2004). Overall, cigarette smoke condensate (CSC) and cigarette smoke total particulate matter (TPM) have been the main testing forms of cigarette smoke in vitro. The use of CSC or TPM for in vitro genotoxicity testing has the advantage that test material can be prepared as a concentrated stock solution in a compatible solvent (usually DMSO) and applied at a relatively high top concentration in a range of in vitro test systems, thus maximizing the potential to detect and quantify a genotoxic effect. Resultant data can be normalized on a per milligram tar, per cigarette or per milligram nicotine basis, facilitating product comparisons ( DeMarini et al., 2008).