lower phosphate levels (4.25 (3.7, 5.3)) vs (4.86 (4.0, 5.6)) mg/dl, respectively, p= 0.008), albumin 50 (3.8 (3.5, 4.0)) vs (3.9 (3.6, 4.2)) g/dL, respectively, p= 0.001). Lower BGP levels were associated 51 with aortic calcification (p< 0.001), iliac calcification (p=0.042) and vertebral fractures (p=0.023). 52The regression model showed that smoking is associated with a significant reduction of total BGP 53 levels by about 18% (p=0.01). 54Conclusion: This is the first clinical study in a haemodialysis population which identifies cigarette 55 smoking as a potential inhibitor of BGP activity, a protective agent in bone and vascular health. 56 Smoking is a risk factor for osteoporosis [6] and vascular calcification [7]. Nicotine and non-71 nicotine tobacco smoke components have been shown to depress osteoblast activity. Gao et al. 72 showed that smoke exposure inhibited bone formation and increased bone resorption. Lumbar spine 73 and femur Bone Mass Density (BMD) was lower in 4-month smoke-exposed female rats than 74 controls. However, there was no significant difference in serum BGP levels between smoke-75 exposed rats and controls [8]. 76The objective of this secondary analysis of the vitamin K Italian (VIKI) study, an observational 77 study designed to assess the prevalence of vitamin K deficiency in haemodialysis patients, was to 78 assess the association between cigarette smoking and BGP levels in an end stage renal disease 79 (ESRD) population. We hypothesized that BGP levels would be lower in smokers than never 80 smokers, and that lower BGP levels would be related to fractures and severity of prevalent vascular 81 calcification. 82 83 5
PATIENTS AND METHODS 84This study is a secondary analysis of the VIKI study, involving 18 dialysis centres in Italy [9]. All 85 the local ethics committees approved the study, which was conducted according to the regulations 86 in force related to observational studies. Approval dates ranged from July 14, 2008 to October 26, 87 2009. Patient enrolment took place between November 2008 and November 2009, and follow-up to 88 assess vital status was performed in December 2011. We included adult patients of both genders 89 who had been on haemodialysis for >1 year, provided that they gave their informed consent, in 90 writing, for the use of their medical records for the study. We excluded patients who had a life 91 expectancy <6 months, cancer (with the exception of basal cell carcinoma), coagulation disorders, 92 or conditions that according to the investigator could interfere with the study outcome. Study 93 subjects were administered a questionnaire to ascertain smoking status (i.e. current, past or never 94 smoker). 95
Laboratory determination 96
Parathyroid hormone (PTH) 97The method for quantitative determination of PTH in serum was the automated LIAISON ® N-98 Tact ® PTH Assay 310910 (DiaSorin Inc., Stillwater MN, USA), a direct, 2-site, sandwich-type 99 chemiluminescence immunoassay (CLIA) carried out on the LIAISON ® (DiaSorin Inc., Stillwater 100 MN, USA) ...