Between-run quality control sample coefficients of variation (%) for the principal plasma index assays were: plasma phosphorus, 2.3; calcium, 2.7; alkaline phosphatase, 2.6; creatinine,

6.0; albumin, 7.8; antichymotrypsin, 8.0; parathyroid hormone, 8.3; and 25(OH)D, 15.0. Ethics and approvals The study was conducted according to the guidelines laid down in the Declaration of Helsinki, and all procedures involving human BIBW2992 manufacturer subjects were approved by the Local Research Ethics Committees representing each of the 80 postcode sectors used. The protocol was also approved by the Ethical Committee of the MRC Dunn Nutrition Unit (of which the Micronutrient Status Laboratory is now part of MRC Human Nutrition Research) in Cambridge. Written informed consent was obtained from all subjects. Follow-up mortality study The present study included 1,054 participants comprising 538 men and 516 women with partial or complete data available for the analyses of interest here, all of whom agreed to be flagged on the National Register of Births and Deaths and whose status (i.e. as still alive

or registered as having died) was known unequivocally in September 2008. No exclusions, other than those resulting BMS202 datasheet from willingness to participate or the availability of blood samples, were imposed, and there was no evidence of sampling bias. Because of missing values (principally due to incomplete consent availability for the blood sampling), the analyses of the blood biomarker Rabusertib order variables are typically based on a subset of 800–900 participants and of 555 for the parathyroid hormone dataset. Mortality outcomes were obtained from the National Health and Service (NHS) register of deaths, up to September 2008. Statistical analyses Cox proportional hazards models were used, with years of survival as the time scale, to estimate the Lck risk of all-cause mortality. The data were censored to September 2008 in participants who survived. The proportional hazards assumption

was examined by comparing the cumulative hazard plots, grouped as exposure; no appreciable violations were observed. Standardised values (z-scores) were used for each of the explanatory variables, thus expressing the hazard ratios per standard deviation rather than per measurement unit, achieving an enhanced conformity between indices. Adjustment was made for potential confounders, including age and sex, in all models. Multivariable Cox regression model was used to test the independent effect of nutrient status indices or nutrient intake estimates after adjustments for acute phase indicators, functional and anthropometric measures. Since relationships between indices, rather than estimates of prevalence were of interest, the weighting factors used in the Survey Report [5] were not used here. All tests of statistical significance were based on two-sided probability; P < 0.05 was deemed significant.