sity lipoprotein; BMD: bone mineral density; LS: lumbar spine; FN: femoral neck; TH: total hip; VF: vertebral fractures. doi:10.1371/journal.pone.0111703.t001 3 DKK1 and Cardiovascular Disease in Type 2 Diabetes tion. Fasting plasma glucose, glycated hemoglobin, calcium, phosphorus and creatinine were measured using standard automated laboratory techniques. Glomerular filtration rate was calculated by MDRD/CKD-EPI formula. High-density lipoprotein, low-density lipoprotein cholesterol and triglycerides were measured by standard biochemical methods. Dyslipidemia was defined according to third report of the expert panel on detection, evaluation, and treatment of high blood cholesterol in adults criteria or current treatment with statins. Calciotropic hormones PubMed ID: measurement included serum intact parathormone and 25-hydroxyvitamin D developed by Biomedica according to the manufacturer’s instructions. The Biomedica DKK-1 ELISA MedChemExpress TMS detects free DKK-1. Intra-assay and interassay variability were of 7% and 9%, respectively. DKK1 measurements are reported throughout in pg/ml after conversion from pMol/L. 1 pMol/l is equivalent to 28,68 pg/ml. carotid bifurcation was determined between the near and far walls of the carotid bifurcation on the right and left sides. Each part was measured from views of both longitudinal and vertical sections at the bifurcation. If a discrepancy was observed in the measured values between longitudinal and vertical sections, the smaller value was selected to avoid overestimation. BIFIMT was defined as the mean of the measurements from the right and left sides. The value for each side was obtained from the mean of 10 wall measurements. The BIFIMT, measured in millimeters, was considered pathological if it was $0.9 mm, and was considered to be carotid atherosclerosis if the BIFIMT was $ 1.2 mm or 50% greater than the BIFIMT in the adjacent area. Plaques were identified as calcified by findings of bright white echoes on sonography. A single trained sonographer performed the study in all subjects. Statistical analysis Data for continuous variables are presented as mean 6 standard deviation. Data for categorical variables are presented as numbers and/or percentages. KolmogorovSmirnov test was used to test the normality of distribution of continuous variables. The association between continuous variables was described by Pearson’s or Spearman’s correlation coefficients. The comparison of categorical variables among groups was performed using Chi-square test or Fisher test. PubMed ID: The comparison of continuous variables among groups was performed using unpaired Student’s t test or Mann-Whitney test. To determine the independent variables which correlated with DKK1, the parameters that correlate significantly in univariate analysis and other which are biologically linked to DKK1 were tested in multiple backward model linear regression analysis. Multiple backward model of logistic regression analysis was performed to identify DKK1 as independent predictor for CVD in T2DM patients. The model included established atherosclerotic risk factors. CVD-defining parameters were not included in the multiple logistic regression model. The usefulness of serum DKK1 as a marker of high risk of atherosclerotic disease in T2DM was Bone density measurement and vertebral fractures assessment Bone mineral density at lumbar spine L2L4, femoral neck and total hip was performed in all patients by dualenergy X-ray absorptiometry using Hologic QDR 4500 densitometer

Wt animals also had higher apoptosis rates than the corresponding ko mice

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