This experimental strategy enabled to characterize

and quantify the native glycation state of proteins from human plasma and hemolysate (see Section 5.4). Apart from that, predictive analyses intended to evaluate qualitatively and quantitatively the effect of prolonged hyperglycaemia over the glycation profile can be planned. Further studies on the high-risk population of diabetic check details patients should provide new insights about the influence of glycation on molecular and functional networks related to hyperglycemia. For this reason, as described in Section 3, partners will initially focused on islets of Langerhans, insulin-producing cell lines, and blood human samples from diabetes-related cohorts. In subsequent stages the glycation approach will be applied

to target tissues in which hyperglycaemia could promote dysfunctions such as hepatocytes, muscle tissue, neurons, adipose tissue, vascular endothelial cells, retina, kidney, erythrocytes, peripheral blood mononuclear cell (PBMC), platelets, lacrimal fluid, saliva and cell lines associated with the listed primary cells. A complementary phase could be the application of this methodology to animal models in those situations in which it could be required. This project will be an integral part of the new Human Diabetes Proteome KU-60019 Project (HDPP) initiative to generate systems-level knowledge into diabetes-associated cellular changes. Insulin resistance alone does not result in T2DM because hypersecretion of insulin from beta-cells is able to maintain normal glucose homeostasis. However, subsequent decline of insulin secretion will lead to impaired glucose homeostasis and the development of the disease. Islets from diabetic human donors secrete much less insulin in response to glucose even when correcting for total insulin content [31]. These results suggest beta-cell dysfunction

as an early event during diabetes progression prior to beta-cell Rucaparib loss. The beta-cell acts as a fuel sensor. The uptake and metabolism of nutrients in beta-cells is linked to the formation of downstream signals stimulating insulin secretion. This process is known as metabolism-secretion coupling and is tightly linked to mitochondrial function [32]. Mitochondria are not only the site where nutrients are oxidized but the organelle also exports metabolites that are activators of insulin granule exocytosis. This is best studied for the ATP/ADP ratio, which increases as a result of mitochondrial activation. This rise induces the closure of the KATP channel, depolarization of the plasma membrane resulting in calcium influx, which stimulates insulin granule exocytosis. Consistent with the central importance of mitochondria, inhibition of respiration blocks insulin secretion. Furthermore, mitochondrial dysfunction has been observed in islets from individuals with T2DM.