Calibration plots.

<p>Calibration plots of ACC/AHA risk score before (left side) and after (middle) recalibration and of ESC SCORE (right side). Part A shows calibration in the pooled sample of KORA and HNR. Part B shows calibration by cohort (KORA: filled circles, HNR: triangles). The solid line indicates perfect calibration of the risk score. Depicted are mean estimated risk versus mean observed frequency per decile of estimated risk with axes ranging from 0 to 0.5 (50%).</p

Basic description of study populations after applying exclusion criteria according to the ACC/AHA risk score.

<p>Basic description of study populations after applying exclusion criteria according to the ACC/AHA risk score.</p

Flow chart of study populations.

<p>Flow chart of the KORA and Heinz Nixdorf Recall study populations showing exclusion criteria in accordance with the ACC/AHA risk score.</p

ROC curves.

<p>ROC curves of ACC/AHA risk score before (left side) and after (middle) recalibration and of ESC SCORE (right side). Part A illustrates performance in the pooled sample of KORA and HNR, while part B shows performance separately for KORA (solid) and HNR (dashed). C = C-statistic [95% confidence interval].</p

Modulation of insulin degrading enzyme activity and liver cell proliferation

<p>Diabetes mellitus type 2 (T2DM), insulin therapy, and hyperinsulinemia are independent risk factors of liver cancer. Recently, the use of a novel inhibitor of insulin degrading enzyme (IDE) was proposed as a new therapeutic strategy in T2DM. However, IDE inhibition might stimulate liver cell proliferation via increased intracellular insulin concentration. The aim of this study was to characterize effects of inhibition of IDE activity in HepG2 hepatoma cells and to analyze liver specific expression of <i>IDE</i> in subjects with T2DM. HepG2 cells were treated with 10 nM insulin for 24 h with or without inhibition of IDE activity using <i>IDE</i> RNAi, and cell transcriptome and proliferation rate were analyzed. Human liver samples (n = 22) were used for the gene expression profiling by microarrays. In HepG2 cells, <i>IDE</i> knockdown changed expression of genes involved in cell cycle and apoptosis pathways. Proliferation rate was lower in <i>IDE</i> knockdown cells than in controls. Microarray analysis revealed the decrease of hepatic <i>IDE</i> expression in subjects with T2DM accompanied by the downregulation of the p53-dependent genes <i>FAS</i> and <i>CCNG2</i>, but not by the upregulation of proliferation markers <i>MKI67, MCM2</i> and <i>PCNA</i>. Similar results were found in the liver microarray dataset from GEO Profiles database. In conclusion, <i>IDE</i> expression is decreased in liver of subjects with T2DM which is accompanied by the dysregulation of p53 pathway. Prolonged use of IDE inhibitors for T2DM treatment should be carefully tested in animal studies regarding its potential effect on hepatic tumorigenesis.</p

Explained variance after internal validation.

<p>Explained variance after internal validation.</p

Calibration graphs (in deciles) of the prediction model after internal validation.

<p>(A) For men, in the development dataset (The Hoorn Study, The KORA S4/F4 Study and the Inter99 Study combined). (B) For men, in the development dataset, stratified per cohort: Hoorn Study (dots), KORA F4/S4 Study (squares), Inter99 Study (triangles). (C) For women, in the development dataset (The Hoorn Study, The KORA S4/F4 Study and the Inter99 Study combined). (D) For women, in the development dataset, stratified per cohort: Hoorn Study (dots), KORA F4/S4 Study (squares), Inter99 Study (triangles). (E) For men, in the external validation dataset (The METSIM Study). The diagonal line indicates perfect calibration.</p

Linear regression model predicting HbA1c levels after 6 years.

<p>Linear regression model predicting HbA1c levels after 6 years.</p

Baseline characteristics of the 4 datasets.

<p>Baseline characteristics of the 4 datasets.</p