Two-step cluster analysis of three phenotypic variables resulted in three significantly different clusters.

<p>Mean scores are represented as percentages of possible maximum points for each measurements. MNWS, Minnesota Nicotine Withdrawal Scale; ZSDS, Zung Self-Rating Depression Scale.</p

Haplotype association tests on C3 phenotypic cluster in GLM and HapScore tests.

<p>Allelic components of the presented haplotypes are rs3787138, rs1044396 and rs3787140 SNPs, respectively. ^a<i>p_model = 0.018</i>; GLM, general linear model.</p

MNWS and ZSDS scores and CO levels of the three phenotypic clusters (C1, C2, C3) identified by two-step cluster analysis.

<p>MNWS, Minnesota Nicotine Withdrawal Scale; ZSDS, Zung Self-Rating Depression Scale; SD, standard deviation.</p

Descriptive characteristics of the study population.

<p>FTND, Fagerström Test for Nicotine Dependence; MNWS, Minnesota Nicotine Withdrawal Scale; COPD, chronic obstructive pulmonary disorder; ZSDS, Zung Self-Rating Depression Scale; SD, standard deviation; MDD, major depressive disorder.</p

Linkage disequilibrium map of the seven investigated SNPs.

<p>1 = rs4522666, 2 = rs6090378, 3 = rs3787138; 4 = rs1044396; 5 = rs3787140, 6 = rs2093107, 7 = rs755203.</p

Frequencies of different haplotypes in phenotypic clusters.

<p>Allelic components of the presented haplotypes are rs3787138, rs1044396 and rs3787140 SNPs, respectively. ^achi-square test indicated a significantly higher frequency of GCC haplotype in C3 compared to non-C3 clusters.</p

Results of risk analysis of haplotype carrying in different phenotypic clusters.

<p>Allelic components of the presented haplotypes are rs3787138, rs1044396 and rs3787140 SNPs, respectively. OR, odds ratio.</p

<i>5-HTTLPR</i>xRLE interaction, with PLINK (left column) and Bayesian (right column) analyses.

<p>LR+: likelihood ratio of emergence of the disease; BSI: Brief Symptom Inventory; BSI-DEP: BSI depression score; BSI-ANX: BSI anxiety score; DEP: lifetime depression; RLE: recent negative life events (in the last year). The three categories of RLE are: low = 0–1, medium = 2, high = 3 or more. Numbers in groups: low RLE: ss = 292, sl = 746, ll = 480; medium RLE: ss = 82, sl = 207, ll = 144; high RLE: ss = 51, sl = 146, ll = 134. Standard errors of means are displayed in case of continuous variables (left column). Right column figures display outlines of posterior distributions of Bayesian Odds Ratios of <i>5-HTTLPR</i> ss versus ll genotype with respect to DEP, BSI-DEP (severe vs. low), and BSI-ANX (severe vs. low). Subsets according to RLE categories (low, medium and high) were analyzed individually. Curve flatness refers to the number of possible models, each with a different odds ratio. An odds ratio greater than one represents a risk for the given phenotype. <b>1A.</b> Logistic regression analysis showed that having the more s alleles increased the risk of DEP with increasing number of RLE. <b>1B.</b> Regarding DEP there is a clear difference between subjects with low RLE (with a Bayesian Odds Ratio close to 1) and subjects with medium or high RLE (where the effect of ss genotype is stronger). <b>1C.</b> As in case of DEP: having the more s alleles also increased BSI-DEP with increasing number of RLE, using linear regression analysis. <b>1D.</b> As in case of DEP: effect of ss genotype on BSI-DEP is negligible in the low RLE group, but higher in the medium, and especially high in the high RLE group. <b>1E.</b> In contrast to depression phenotypes: linear regression analysis showed that carrying the more s alleles increased BSI-ANX without interaction with RLE. <b>1F.</b> In contrast to depression phenotypes, ss genotype represents a risk for BSI-ANX irrespective of RLE group.</p

Bayesian posterior probabilities of relevance of <i>5-HTTLPR</i> for the multivariate phenotype.

<p>RLE was grouped into three categories: low = 0–1, medium = 2, high = 3 or more number of recent negative life events (in the past year). CHA (childhood adversity) was divided into two categories (based on the original three): low = 0–3, medium or high = 4 or more scores. Multivariate phenotype (more accurately describes depression related psychiatric state than one phenotype measure alone) encompasses lifetime depression, BSI depression score and BSI anxiety score. Results are displayed according to CHA and age, in groups differentially exposed to RLE. <b>3A and 3B.</b> Results demonstrate moderate Bayesian probability of relevance of <i>5-HTTLPR</i> in both age groups and CHA groups in those who had 3 or more RLE. <b>3C.</b> In the younger age group (≤30) <i>5-HTTLPR</i> was strongly relevant in those who had medium or high CHA and even moderate number of RLE. <b>3D.</b> In the older age group (>30) <i>5-HTTLPR</i> was strongly relevant in those who had 3 or more RLE, irrespective of CHA.</p

Summary of the genetic association and interaction results using PLINK.

<p>BSI: Brief Symptom Inventory; BSI-DEP: BSI depression score; BSI-ANX: BSI anxiety score; CHA: childhood adversity; DEP: lifetime depression; Pperm: permutated p values; RLE: recent negative life events (in the last year). Additive genetic models were calculated, where <i>5-HTTLPR</i> s allele represents the minor allele. Results are displayed in different groups: total population (all); those up to 30 years (≤30); and those above 30 years (>30). Regression equations (linear regression and beta for BSI-DEP and BSI-ANX scores, and logistic regression and odds ratio for DEP) always involve gender and age as covariates. In case of interaction models, main effect of the respective life event (RLE or CHA) was also covariate in the equation, besides its interaction with <i>5-HTTLPR</i>. And in case of the third model BSA-ANX was also a covariate. Permutated p values were calculated for the nominal p<0.05 results using PLINK—mperm 1000 for main 5HTTLPR effect on anxiety and using the “glmperm” R-package (<a href="http://cran.r-project.org/web/packages/glmperm/index.html" target="_blank">http://cran.r-project.org/web/packages/glmperm/index.html</a>, with 1000 permutations) for the interaction effects on DEP and BSI-DEP.</p><p>The three categories of RLE were: low = 0–1, medium = 2, high = 3 or more number of recent negative life events. The three categories of CHA were: low = 0–3, medium = 4–6, high = 7 or more scores.</p><p>Italics represent trends, and bold represents significant findings.</p><p>Summary of the genetic association and interaction results using PLINK.</p