MATERIALS AND METHODS This observational study Ponatinib CAS included 30 male smokers attending a tertiary care hospital; all were from a rural area in South India. The 30 subjects were divided into three groups of 10 each based on their smoking index (SI), as follows: group I: SI < 150, group II: SI 150�C300, and group III: SI > 300. SI was calculated using the formula given by Srinivasan et al.[17]: SI = Numbers of cigarettes/bidis/cigars per day �� total duration of smoking in years An equal number of age-, sex-, diet- and social status-matched nonsmokers formed the control group for the study. None of the individuals were habituated to alcohol or drugs. Ex-smokers and smokers with chromosomal anomalies were excluded from the study. Informed written consent was taken from all participants.

The study was designed in accordance with the Helsinki II declaration and approved by the Institutional Human Ethical Committee. Lymphocyte cultures were set up from heparinized blood, with minor modification of the method of Hungerford,[18] by the addition of chilled fixative at the end of hypotonic treatment. For each person, 50 well-spread metaphase plates, stained with 4% buffered Giemsa, were analyzed for SAs. SAs involve a specific arrangement of acrocentric chromosomes of ��D�� group (chromosomes 13, 14, and 15) and ��G�� group (chromosomes 21 and 22) with their satellites directed towards each other [Figure 1]. For identifying SAs, the criteria used by Hansson were applied;[7] i.e.

, the satellite ends of the associating chromosomes had to be directed towards each other with their longitudinal axes meeting between their short arms, and the distance between the centromeres of associated chromosomes should not exceed the total length of one ��G�� chromosome after excluding its satellite. Odds ratios and the Student’s t-test were used for statistical analysis. Figure 1 Photomicrograph showing satellite associations between the ��DG�� group chromosomes in nonsmokers RESULTS The mean age of the subjects was 38.25, 40, 40.75, and 39.67 years in smoker groups I, II, and III, and in nonsmokers, respectively. The mean frequency of SAs in nonsmokers, calculated using the SI formula, was 29.83 �� 2.88 [Figure 1]; whereas the mean frequency of SAs in smokers belonging to group I, II, and III was 44.25 �� 3.30, 53 �� 3.46, and 67 �� 6.22, respectively [Figures [Figures22�C4]. The frequency of SAs in smoker groups I, II, and III was higher than in Cilengitide the nonsmokers and, moreover, SA was seen to increase with increase in the SI [Table 1].