2. Family controls comprised unaffected relatives as defined in this manner, and spouses. Spouses were recruited to increase sample size, reduce residual confounding from unmeasured environmental factors shared with HBM cases and who, as a function of their genetic independence, would be unlikely to share common polygenic influences over BMD. Recruitment ran from September 2008 until April 2010. All participants were clinically assessed by one doctor

using a standardised structured history and examination questionnaire, after which DXA scans were performed for relatives and spouses, using local GE Lunar Inc. Madison, WI, USA) DXA systems applying manufacturer’s standard scan and positioning protocols, and weight and routine height measurements were recorded. Body mass index (BMI) was calculated as weight (kilograms)/height (metres)2. Current and historical physical activity data were collected from HBM cases and family Alectinib in vivo controls

by questionnaire (including the validated international physical activity questionnaire [IPAQ] [6], [7], [8] and [9]). Participants were excluded if under 18 years of age, pregnant or unable to provide written informed consent for any reason. The Hertfordshire Cohort Study is a population based cohort study tracing 42,974 men and women born in Hertfordshire during 1931–1939 and still living there during the period 1998–2003. Individuals were traced using the NHS BMS754807 central registry at Southport and the Hertfordshire Family Health Service Association. Full details of the study design have previously been reported [10]. A planned subsample of 6099 individuals were invited to participate in a clinical study and 3225 (53%) men and women aged 60–75 years were recruited and completed home interviews [10]. In 2004 and 2005 a subgroup (from East Hertfordshire)

were followed up and 322 men (65%) and 321 women (69%) re-attended, completed lifestyle questionnaires which included questions concerning medical history including fractures, smoking and alcohol consumption. Height was measured to the nearest 0.1 cm using a Harpenden pocket stadiometer and weight to the nearest 0.1 kg using floor scales, at the time of pQCT assessment [11]. pQCT scans were performed at the distal and mid-shaft of the Adenosine tibia (4 and 66% from the distal endplate) in the non-dominant lower limb using a Stratec XCT2000L (Stratec Medizintechnik, Pforzheim, Germany); voxel size 0.5 mm, CT speed 30 mm/s, XCT software version 5.50d. A reference line at the distal endplate was determined from initial frontal scout view. Cortical bone was defined using a threshold above 650 mg/cm3 (optimal for bone geometry [12]). Trabecular bone was identified by elimination of cortical bone and therefore trabecular bone mineral density (tBMD) was defined as a density < 650 mg/cm3.