The subjects ERK signaling inhibitors in the present study were adolescents belonging to the 1993 Pelotas Birth cohort. Pelotas is a medium-sized city in Southern Brazil with a population of approximately 340 thousand. The present study evaluated the 2008 follow-up when subjects were aged 14–15 years (mean 14.3; SD 0.6). During this follow-up, we traced

4325 of the original 5429 subjects, an 82.5% follow-up rate when considering the 147 known deaths. Additional information on the methods of the cohort study can be found elsewhere (Araujo et al., 2010 and Victora et al., 2008). The four behavioral risk factors investigated were defined as follows: a) Smoking: having smoked at least one cigarette in the last 30 days (Malcon et al., 2003). This information was obtained by means of a confidential questionnaire administered to the adolescent. Risk behaviors were coded as a binary variable (presence = 1; absence = 2). Prevalence of multiple risk behaviors was estimated based on the sum of individual behaviors, which generated a score ranging from 0 to 4 (0 = no risk factors; 4 = all four risk factors) based on the distribution observed in the sample. The present analysis was carried out in three stages. First, we analyzed the cluster of risk factors, stratified by sex. Clustering occurs when the observed prevalence of a combination of factors exceeds the expected prevalence for this combination.

Expected prevalence for CHIR99021 a given combination is calculated by multiplying the individual probabilities of each behavior based on their observed occurrence in the survey. Observed/expected (O/E) ratios higher than 1 are indicative of Bay 11-7085 clustering (Galan et al., 2005 and Schuit et al., 2002). The 95% confidence intervals (95%CI) were obtained by binomial exact probability (Daly, 1992). Second, odds ratios (OR) were used to calculate the clustering of two behaviors in the presence of another risk behavior. The OR represents the additional estimate that one behavior may have in relation to the other, and is calculated using the equation below

(Schuit et al., 2002): N11×N00/N10×N01N11×N00/N10×N01where N11 is the number of responders displaying both risk factors, N00 is the number of respondents without any of the risk factors, N10 is the number of respondents displaying only one risk factor, and N01 is the number of respondents displaying the other risk factor. For example, an OR of 1.5 indicates that subjects displaying a given behavior (e.g. physical inactivity) are 1.5 times more likely to display another behavior (e.g. low fruit intake) when compared to those not exposed to the first behavior (physical inactivity). Third, for multivariate analysis, we carried out a Poisson regression with presence of at least three risk behaviors as the outcome and the following demographic variables as exposures: sex (male, female); age in years (14.0–14.4; 14.5–14.9; 15.0–15.