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ts were recruited for this study. Entire blood L-685,458 samples were collected from 360 individuals with CVD from St.Thomas Hospital, Kerala, India. Diagnosis of CVD was based on physical examination and Doppler ultrasound test. CVD resulting from obstructions like neoplasm had been excluded in the study. Differential diagnosis was performed by an experienced vascular surgeon and presence of distichiasis was ruled out by an ophthalmologist. Individuals with kind two diabetes mellitus have been also excluded considering the fact that genetic variants of FoxC2 happen to be reported to result in susceptibility to diabetes mellitus. Blood samples were collected from age and gender matched 352 wholesome controls with no recognized loved ones history for CVD. For tissue level expression evaluation, varicose vein tissue samples were collected from 22 patients admitted for treatment of CVD by operative remedies at Kempegowda Institute of Healthcare Sciences, Bangalore, India. Saphenous handle vein samples from 20 sufferers who underwent coronary artery bypass graft surgery at Sri Jayadeva Institute for Cardiovascular Sciences & Research, Bangalore, India were also collected for the study. Complete blood samples have been also collected from these 22 sufferers and 20 controls for sequencing assays. Relevant data regarding the clinical characteristics of patients were collected from healthcare records of the hospitals participating in the study. Variables Family history Bleeding Thrombophlebitis Cellulitis LL oedema Pigmentation Ulceration CEAP Class two 3 4 5 6 N = 382 n 257 29 3 5 89 185 56 48 11 223 73 27 Data evaluation Demographic data of all study participants and information regarding symptoms for instance pain, itching and throbbing sensation in legs and clinical signs for example hemorrhage, lower limb oedema, Percentages were taken from the column totals. doi:10.1371/journal.pone.0090682.t002 FoxC2 in Chronic Venous Disease a b Genotypes c.-350G.T GG GT TT GT/TT c.-512C.T c CC CT TT CT/TT c.-1538A.G c AA AG GG AG/GG c Individuals n Controls n OR P-value AOR 342 37 3 40 325 46 1 47 1 0.76 2.85 0.81 0.353 0.72 69 209 104 313 118 170 84 254 1 2.1 two.12 2.11 ,0.001 2.37 2.44 two.08 240 100 42 142 280 90 two 92 1 1.3 24.5 1.8 ,0.001 1.22 25.58 1.8 Percentages had been taken in the column totals. Chi-square test for measure of association was used to derive p value. aOdds ratio and 95% confidence intervals of individual polymorphisms. b Adjusted odds ratio and 95% confidence intervals is obtained adjusting for age group and sex in multiple logistic regression model. c Polymorphism previously reported in the Entrez single nucleotide polymorphism database. doi:10.1371/journal.pone.0090682.t003 hyperpigmentation, thrombophlebitis, cellulitis and ulceration have been collected for each patient from healthcare records. Loved ones history, occupational and lifestyle data were collected to examine their influence in aggravating disease manifestation. Disease phenotypes have been categorized according to CEAP classification system. Varicose veins without odema or pigmentation were classified under C2. Only two.9% of all our patients had been in CEAP Class 3 in which varicose vein with oedema alone are found. The patients in this study had been mostly from CEAP Class 4, 5 and 6 who presented various clinical signs like pigmentation, ulceration along with oedema due to CVD.