None of the individual compounds led to any TNF- secretion from the macrophage-like cells (data not shown). 4. Age groups should be bound to proteins to exert an inflammatory reaction. These Amfr findings display that diet Age groups directly stimulate the inflammatory response of human being innate immune cells and help us define the risk of regular usage of AGE-rich food products on human being health. Keywords: advanced glycation endproducts, swelling, food, macrophages, Maillard reaction 1. Intro During heating of food products comprising protein and carbohydrates, diet advanced glycation endproducts (Age groups) are created via the Maillard reaction [1]. Age groups are greatly responsible for the flavor and taste of food, and increase palatability. In the Maillard reaction, the carbonyl group of a reducing sugars (sugars with a free anomeric carbon [2]) reacts with an amino moiety of an amino acid leading to reactive intermediates, the so-called Amadori products. These Amadori products then form different Age groups [3]. Age groups comprise a large body of different molecules [3]. Examples of well-studied Age groups are: N()-carboxymethyllysine (CML), carboxyethyl-lysine (CEL), methylglyoxal-derived hydroimidazolone (MG-H1), pentosidine, and acrylamide. Even though quantification of Age groups in food products is frequently carried out by antibody-based techniques [4,5], liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the preferred technique to accurately determine and quantify individual Age groups PF-05231023 [1,6,7,8]. Physiological effects of exposure to diet Age groups are not yet fully recognized. Endogenously created Age groups in diabetic patients, however, have been shown to induce an inflammatory reaction and contribute to the onset of cardiovascular diseases, such as atherosclerosis and diabetic cardiomyopathy [9]. The inflammatory effect seen in these situations is definitely proposed to be caused by the binding of Age groups to the specific receptor for advanced glycation endproducts (RAGE). RAGE activates many enzymes and protein complexes, one of PF-05231023 which is definitely nuclear element kappa-light-chain-enhancer of triggered B cells (NF-B) [10]. Even though swelling caused by endogenous Age groups has been thoroughly analyzed, information within the generation of swelling by diet Age groups is definitely scarce. As said, many of the studies on the effects of Age groups are carried out with endogenously created Age groups, by heating bovine serum albumin or human being serum albumin with different sugars at 37 C for a number of hours, mimicking the endogenous scenario. This difference having a home cooking scenario, in which diet proteins and sugars are heated to higher temps, can lead to a different array of molecules, resulting in different immunological results. In addition to endogenous Age groups, some individual Age groups have been investigated. Acrylamide is definitely a well-studied compound, but only with regards to its genotoxic and carcinogenic effects [11,12]. To our knowledge, no study has been published within the inflammatory effect of acrylamide. A definite cause-effect relationship between Age groups and inflammation has also not be founded yet and the available results have been questioned as being caused by endotoxins rather than Age groups [13,14]. To our knowledge, we are the first to PF-05231023 investigate the inflammatory effect of diet Age groups inside a home-cooking environment and the first to investigate the inflammatory effect of acrylamide. The present study aimed at investigating the effects of diet Age groups within the inflammatory response of human being cells of the innate immune system. To mimic a home cooking situation, only dietary protein and sugars were used and heated to a heat of 100 C. The formation of dietary Age groups over time was quantified by LC-MS/MS and the presence of endotoxin was excluded. We also examined the effect of individual Age groups and whether the observed inflammatory effect was induced by activation of RAGE. 2. Materials and Methods 2.1. Chemicals and Reagents Casein from PF-05231023 bovine milk, -lactose monohydrate, NaOH, sodium-phospate, 2-mercaptoethanol, and thiazolyl blue tetrazolium bromide (MTT) were from Sigma-Aldrich (Saint Louis, MO USA). D-glucose, glutamate, fetal bovine serum (FBS), Dulbecccos Phospate-Buffered Saline (DPBS) were from Gibco (Thermo Scientific, Waltham, MA, USA). Analytical requirements of CML (>99%), CEL (>95%), MG-H1 (>93%), and pentosidine (>99%), as well as the deuterium labelled internal requirements CML-d2, CEL-d4, and MG-H1-d3, were from Polypeptide (Strasbourg, France). Boric acid (99.5%), chloroform (99.5%), nonafluoropentanoic acid (NFPA; 99%), sodium hydroxide (98%), sodium borohydride.