Carboxymethyllysine Concentration Research Articles

Determination of AGE-CML Level in Commercially Available Sports Bar in Italy.

The process of bar creation involves improving the texture of the product to increase its palatability, which can be further induced by various physical or chemical changes during storage, such as sugar crystallization and molecular migrations in which Maillard's reaction occurs, forming the N-epsilon- (carboxymethyl) lysine (CML) adduct. In this study, we aimed to assess (the CML) adduct used in commercial bars today as meal substitutes or for athletic or sports purposes. The adduct CML is an advanced glycation end-product (AGEs) found in the human body (serum) and foods. It is the significant ligand for the Receptor for Advanced Glycation End Products (RAGE), resulting in chronic inflammation upon CML activation. Additionally, it aimed to assess the amount of AGEs-CML in various energy bars available on the Italian market. CML OxiSelect ™ ELISA was used to assess the quantity of CML bars. The amount of AGE-CML was assessed in commercially available energy bars. According to the ELISA analysis, CML concentrations per g protein in all the tested energy bars varied from 138,42 to 1387,54 μg/gr per bar and from 461,41 to 3970,46 μg/gr per 100 gr of product, which depends on the quantity of protein. The amount per gram of protein is relatively uniform (with a variation of about 10%), and when compared to other foods, it is positioned in a medium-low range.

Formation of AGEs in Penaeus vannamei fried with high oleic acid sunflower oil.

Here, we investigated the effects of frying process on the formation of advanced glycation end products (AGEs) in shrimps using Penaeus vannamei as the raw material. The results showed that the oil, malondialdehyde, fluorescent AGEs, carboxymethyl lysine (CML), methylglyoxal hydroimidazolone (MG-H1) and the outer layer carboxyethyl lysine (CEL) content was higher in the fried shrimps than that in the raw unfried shrimps. The outer layer CML, CEL and inner CEL, MG-H1 values all reached the maximum after the first batch of frying (22.43mg/kg, 304.24mg/kg, 83.76mg/kg, and 169.42mg/kg respectively). However, fluorescent AGEs and MG-H1 of the outer layer reached the maximum after the fifth and fourth batches of frying (1230.0 AU/g and 341.63mg/kg). Malondialdehyde, fluorescent AGEs, CML, MG-H1, and CEL concentration in the fried shrimps firstly increased and then decreased to stabilization with more frying batches, with higher content in the outer layer of fried shrimps.

Correlation between serum advanced glycation end products and dietary intake of advanced glycation end products estimated from home cooking and food frequency questionnaires

Background & aimsTo our knowledge the association between dietary advanced glycation end-products (dAGEs) and cardiometabolic disease is limited. Our aim was to examine the association between dAGEs and serum concentration of carboxymethyl-lysine (CML) or soluble receptor advanced glycation end-products (sRAGEs), and to assess the difference on dAGEs and circulating AGEs according to lifestyle and biochemical measures. Methods and results52 overweight or obese adults diagnosed with type 2 diabetes were included in this cross-sectional analysis. dAGEs were estimated from a Food Frequency Questionnaire (FFQ) or from a FFQ + Home Cooking Frequency Questionnaire (HCFQ). Serum concentrations of CML and sRAGEs were measured by ELISA. Correlation tests were used to analyze the association between dAGEs derived from the FFQ or FFQ + HCFQ and concentrations of CML or sRAGEs. Demographic characteristics, lifestyle factors and biochemical measures were analyzed according to sRAGEs and dAGEs using student t-test and ANCOVA.A significant inverse association was found between serum sRAGEs and dAGEs estimated using the FFQ + HCFQ (r = −0.36, p = 0.010), whereas no association was found for dAGEs derived from the FFQ alone. No association was observed between CML and dAGEs. dAGEs intake estimated from the FFQ + HCFQ was significantly higher among younger and male participants, and in those with higher BMI, higher Hb1Ac levels, longer time with type 2 diabetes, lower adherence to Mediterranean diet, and higher use of culinary techniques that generate more AGEs (all p values p < 0.05). ConclusionsThese results show knowledge on culinary techniques is relevant to derive the association between dAGEs intake and cardiometabolic risk factors.

Serum carboxymethyllysine concentration is associated with erosive hand osteoarthritis

Carboxymethyllysine (CML) and homocitrulline (HCit) are the products of two non-enzymatic post-translational modifications of protein, a process related to age. We investigated whether serum CML and HCit concentrations were associated with hand osteoarthritis (HOA), especially erosive HOA. Serum CML and HCit were measured by using liquid chromatography coupled with tandem mass spectrometry at inclusion in 386 patients included in the DIGItal Cohort Design (DIGICOD) cohort. We investigated whether serum CML and/or HCit concentrations were associated with erosive HOA or with HOA clinical and radiological features. Moreover, we compared the tissular concentrations of CML and HCit in OA and non-OA cartilage from proximal interphalangeal and metacarpo-phalangeal (MCP) joints from human cadaveric donors. Median (IQR) serum CML concentration was lower in patients with erosive HOA than those with non-erosive HOA (178.7 [157.1-208.8] vs 194.7 [168.9-217.1]μmol/mol Lys, P=0.002), but median HCit concentration did not differ between the groups (193.9 [162.9-232.0] vs 193.9 [155.9-224.6]μmol/mol Lys). Cartilage HCit and CML concentrations were not correlated with clinical features. Serum CML concentration was higher in OA than non-OA MCPs (7.0 vs 4.0mmol/mol Lys, P=0.01). Serum CML concentration was lower in erosive HOA than non-erosive HOA, and cartilage CML concentration was higher in OA than non-OA cartilage. These results encourage further studies to test whether serum CML could be a new prognostic biomarker in HOA.

Lactococcus lactis KF140 Reduces Dietary Absorption of Nε - (Carboxymethyl)lysine in Rats and Humans via β-Galactosidase Activity.

Background and AimsExcessive intake of advanced glycation end products (AGEs), which are formed in foods cooked at high temperatures for long periods of time, has negative health effects, such as inflammatory responses and oxidative stress. Nε-(Carboxymethyl)lysine (CML) is one of the major dietary AGEs. Given their generally recognized as safe status and probiotic functionalities, lactic acid bacteria may be ideal supplements for blocking intestinal absorption of food toxicants. However, the protective effects of lactic acid bacteria against dietary AGEs have not been fully elucidated.Materials and MethodsWe investigated the effect of treatment with Lactococcus lactis KF140 (LL-KF140), which was isolated from kimchi, on the levels and toxicokinetics of CML. The CML reduction efficacies of the Lactococcus lactis KF140 (LL-KF140), which was isolated from kimchi, were conducted by in vitro test for reducing CML concentration of the casein-lactose reaction product (CLRP) and in vivo test for reducing serum CML level of LL-KF140 administered rats at 2.0 × 108 CFU/kg for14 days. In addition, 12 volunteers consuming LL-KF140 at 2.0 × 109 CFU/1.5 g for 26 days were determined blood CML concentration and compared with that before intake a Parmesan cheese.ResultsAdministration of LL-KF140 reduced serum CML levels and hepatic CML absorption in rats that were fed a CML-enriched product. In a human trial, the intake of LL-KF140 prevented increases in the serum levels of CML and alanine aminotransferase after consumption of a CML-rich cheese. LL-KF140 was determined to presence in feces through metagenome analysis. Furthermore, β-galactosidase, one of the L. lactis-produced enzymes, inhibited the absorption of CML and reduced the levels of this AGE, which suggests an indirect inhibitory effect of LL-KF140. This study is the first to demonstrate that an L. lactis strain and its related enzyme contribute to the reduction of dietary absorption of CML.

Nɛ-(carboxymethyl)lysine, Available Lysine, and Volatile Compound Profile of Biscuits Enriched with Grape by-Product During Storage.

We investigated the changes in Nɛ-(carboxymethyl)lysine (CML) and available lysine content, antioxidant properties, volatiles, and oxidation products of biscuits enriched with grape by-product (GP), stored for six months under a modified atmosphere of 0%/30%/70% O2/CO2/N2 and in air. Fresh GP-formulated biscuits showed lower concentrations of CML (89%), available lysine (40%), and pyrazines (75%), but higher antioxidant capacities (~ sixfold), furans (12-fold), and lipid-derived compounds (three-fold) than the control. Although ~ 15% higher losses of Maillard-type volatiles were identified in the air atmosphere during storage, lipid oxidation was ~ 30% less pronounced in the modified atmosphere. A significant correlation of 0.994 between the reduction in CML and the available lysine suggest further CML reactions with the ɛ-NH2 group of amino acids. Significant correlations (of -0.550 to -0.980) between oxidation products, antioxidant capacities, and changes in CML content during storage suggest that these parameters might be involved in the CML elimination mechanism.

Advanced glycation end products, leukocyte telomere length, and mitochondrial DNA copy number in patients with coronary artery disease and alterations of glucose homeostasis: From the GENOCOR study

Background and aimAlterations of glucose homeostasis can increase advanced glycation end products (AGEs) that exacerbate vascular inflammatory disease and may increase vascular senescence and aging. This study examined the relationships between carboxymethyl-lysine (CML) and soluble receptor for AGEs (sRAGE) with leukocyte telomere length (LTL) and mitochondrial DNA copy number (mtDNAcn), as cell aging biomarkers, in patients with established coronary artery disease (CAD). Methods and resultsWe studied 459 patients with CAD further categorized as having normal glucose homeostasis (NG, n = 253), pre-diabetes (preT2D, n = 85), or diabetes (T2D, n = 121). All patients were followed up for the occurrence of major adverse cardiovascular events (MACEs). Plasma concentrations of sRAGE and CML were measured by ELISA. mtDNAcn and LTL were measured by qRT-PCR. CML levels were significantly higher in patients with preT2D (p < 0.007) or T2D (p < 0.003) compared with those with NG. mtDNAcn resulted lower in T2D vs preT2D (p = 0.04). At multivariate Cox proportional hazard analysis, short LTL (HR: 2.89; 95% CI: 1.11–10.1; p = 0.04) and high levels of sRAGE (HR: 2.20; 95% CI: 1.01–5.14; p = 0.04) were associated with an increased risk for MACEs in patients with preT2D and T2D, respectively. T2D patients with both short LTL and high sRAGE levels had the highest risk of MACEs (HR: 3.11; 95% CI: 1.11–9.92; p = 0.04). ConclusionsHigh levels of sRAGE and short LTL were associated with an increased risk of MACEs, especially in patients with diabetes, supporting the usefulness of both biomarkers of glycemic impairment and aging in predicting cardiovascular outcomes in patients with CAD.

Advanced Glycation End-Products (AGEs) and Their Soluble Receptor (sRAGE) in Women Suffering from Systemic Lupus Erythematosus (SLE).

Systemic lupus erythematosus (SLE) is characterized by abnormal action of the immune system and a state of chronic inflammation. The disease can cause life-threatening complications. Neoepitopes arising from interdependent glycation and oxidation processes might be an element of SLE pathology. The groups included in the study were 31 female SLE patients and 26 healthy female volunteers (the control group). Blood serum samples were obtained to evaluate concentrations of advanced glycation end-products (AGEs), carboxymethyllysine (CML), carboxyethyllysine (CEL), pentosidine, and a soluble form of the receptor for advanced glycation end-products (sRAGE). Compared to a healthy control group, the SLE patients exhibited a higher concentration of AGEs and a lower concentration of sRAGE in serum. There were no statistically significant differences in serum CML, CEL, and pentosidine concentrations between the groups. Therefore, SLE patients could be at risk of intensified glycation process and activation of the proinflammatory receptor for advanced glycation end-products (RAGE), which could potentially worsen the disease course; however, it is not clear which compounds contribute to the increased concentration of AGEs in the blood. Additionally, information about the cigarette smoking and alcohol consumption of the study participants was obtained.

The impact of a modified carbohydrate formula, and its constituents, on glycaemic control and inflammatory markers: A nested mechanistic sub-study.

Hyperglycaemia occurs frequently in the critically ill. Dietary intake of advanced glycation end-products (AGEs), specifically Nε-(carboxymethyl)lysine (CML), may exacerbate hyperglycaemia through perturbation of insulin sensitivity. The present study aimed to determine whether the use of nutritional formulae, with varying AGE loads, affects the amount of insulin administered and inflammation. Exclusively tube fed patients (n = 35) were randomised to receive Nutrison Protein Plus Multifibre®, Diason® or Glucerna Select®. Insulin administration was standardised according to protocol based on blood glucose (<10 mmolL-1 ). Samples were obtained at randomisation and 48 h later. AGEs in nutritional formula, plasma and urine were measured using mass spectrometry. Plasma inflammatory markers were measured using an enzyme-linked immunosorbent assay and multiplex bead-based assays. AGE concentrations of CML in nutritional formulae were greatest with delivery of Nutrison Protein Plus® (mean [SD]; 6335pmolmol-1 [2436]) compared to Diason® (4836pmolmol-1 [1849]) and Glucerna Select® (4493pmolmol-1 [1829pmolmol-1 ]) despite patients receiving similar amounts of energy (median [interquartile range]; 12 MJ [8.2-13.7 MJ], 11.5 MJ [8.3-14.5 MJ], 11.5 MJ [8.3-14.5 MJ]). More insulin was administered with Nutrison Protein Plus® (2.47unitsh-1 [95% confidence interval (CI)=1.57-3.37unitsh-1 ]) compared to Diason® (1.06unitsh-1 [95% CI=0.24-1.89unitsh-1 ]) or Glucerna Select® (1.11unitsh-1 [95% CI=0.25-1.97unitsh-1 ]; p = 0.04). Blood glucose concentrations were similar. There were associations between greater insulin administration and reductions in circulating interleukin-6 (r = -0.46, p < 0.01), tumour necrosis factor-α (r = -0.44, p < 0.05), high sensitivity C-reactive protein (r = -0.42, p < 0.05) and soluble receptor for advanced glycation end-products (r = -0.45, p < 0.01) concentrations. The administration of greater AGE load in nutritional formula potentially increases the amount of insulin required to maintain blood glucose within a normal range during critical illness. There was an inverse relationship between exogenous insulin and plasma inflammatory markers.

Maillard Reaction Products in Gluten-Free Bread Made from Raw and Roasted Buckwheat Flour.

The formation of Maillard reaction products (MRPs) in gluten-free bread made from roasted and raw buckwheat flour was examined. The levels of phenolic compounds such as flavonoids (catechin, naringenin, quercetin, rutin, and others) and phenolic acids (like 4-hydroxybenzoic, caffeic, dihydroxybenzoic, ferulic, gallic, syringic, vanillic, and p-coumaric) were measured using reversed-phase ultra-high performance liquid chromatography-electrospray ionization mass spectrometry (RP–UHPLC–ESI-MS). Early and advanced Maillard reaction products were analyzed using HPLC, whereas spectrofluorimetric analysis was used to determine the levels of fluorescent intermediate compounds (FIC). The total levels of phenolic compounds were higher in the case of buckwheat bread prepared from roasted buckwheat flour (156 and 140 µg/g of crumb and crust, respectively). Rutin, gallic acid, and catechin were the most abundant phenolic compounds detected in roasted buckwheat bread. The roasting process resulted in significantly lower radical scavenging capacities (ABTS) of the total phenolics and flavonoids in the buckwheat bread. Taking into consideration these Maillard reaction products, we observed a significant increase in FIC level in roasted buckwheat crumb and crust (at about 40%, and 38%, respectively). At the same time, the Nε-(carboxymethyl)lysine (CML) level did not change in roasted or raw buckwheat bread crumb, though in roasted buckwheat crust the concentration of CML increased by about 21%.

Influence of Dietary Metformin on the Growth Performance and Plasma Concentrations of Amino Acids and Advanced Glycation End Products in Two Types of Chickens.

Glycation is a non-enzymatic reaction inducing the bonding of glucose to amino acids and proteins. Glycated amino acids are not useful for protein synthesis, suggesting that glycation reduces the utilization of amino acids. Metformin (MF) is well known as a therapeutic drug for type II diabetes that inhibits glycation. It is possible that treatment with MF raises the utilization of amino acids by the inhibition of glycation, thereby improving the growth performance of chickens. In the present study, therefore, we investigated the influence of dietary MF on the growth performance, and plasma concentrations of free amino acids and Nε-(Carboxymethyl)lysine (CML), which is an advanced glycation end product, in layer (Experiment 1) and broiler (Experiment 2) chickens. From 7 d of age, chicks were allowed free access to one of the experimental diets containing MF at 3 supplementation levels (0, 150, and 300 mg/kg diet) for 14 days. Body weight and feed intake were measured every week. At the end of the experiments, blood and breast muscle (M. pectoralis major) were collected for further analysis. Dietary MF did not affect weight gain, feed intake, or feed efficiency in both layer and broiler chickens. Dietary MF at the level of 150 mg/kg diet increased breast muscle weight in both layer and broiler chickens. Dietary MF increased plasma concentrations of branched chain amino acids and decreased concentrations of CML in layer chickens, although it did not affect plasma concentrations of glucose. The present study suggested that dietary MF might have the potency to increase breast muscle weight of layer chickens with an increment in plasma concentrations of branched-chain amino acids.

In depth investigation of collagen non-enzymatic glycation by Raman spectroscopy

Non-enzymatic glycation is a post-translational modification of long-lived matrix proteins such as type I collagen. It occurs during aging and leads to the formation of advanced glycation end-products (AGEs). AGE accumulation is associated with severe complications in chronic and age-related diseases. The assessment of modifications induced by this (patho)physiological process represents an interest in biology and medicine for a better patient care. The objective of our work was to position the interest of Raman spectroscopy in the quantification of collagen glycation. Two types of in vitro glycation were used by incubating collagen samples, at different durations, with ribose or glyoxylic acid; these reducing agents acting on the chemical specificity of the glycation reaction. Glycation efficiency was evaluated by the liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) quantification of carboxymethyllysine (CML) and pentosidine, which are among the most studied AGEs. Raman data were processed by PCA coupled to validity indices and Lasso regression as multivariate analysis tools. Regression models were constructed by considering the LC-MS/MS results as reference values. A marked variability was observed within the Raman datasets making difficult the identification of spectral differences between control and ribose-treated collagen samples. By taking advantage of the chemical specificity of the glyoxylic acid treatment leading to CML formation, on one hand, and the feature selection included in the Lasso algorithm, on the other hand, Raman markers associated with glycation were identified. The assigned vibrations corresponded to modifications of side chains of collagen. In addition, a threshold of CML concentration was determined as quantitative indicator of the applicability of Raman spectroscopy for potential patient follow-up purposes. Although lacking in sensitivity to directly detect AGEs in collagen, Raman spectroscopy allows to highlight the molecular modifications of collagen induced by glycation.

Dietary protein intake and circulating advanced glycation end product/receptor for advanced glycation end product concentrations in the Health, Aging, and Body Composition Study

Advanced glycation end products (AGEs) promote adverse health effects and may contribute to the multi-system functional decline observed in aging. Diet is a major source of AGEs, and foods high in protein may increase circulating AGE concentrations. However, epidemiological evidence that high-protein diets increase AGEs is lacking. We examined whether dietary protein intake was associated with serum concentrations of the major AGE carboxymethyl-lysine (CML) and the soluble receptor for AGEs (sRAGE) in 2439 participants from the Health, Aging, and Body Composition study (mean age, 73.6±2.9 y; 52% female; 37% black). CML and sRAGE were measured by ELISA, and the CML/sRAGE ratio was calculated. Protein intake was estimated using an interviewer-administered FFQ and categorized based on current recommendations for older adults: <0.8 g/kg/d (n=1077), 0.8 to <1.2 g/kg/d (n=922), and ≥1.2 g/kg/d (n=440). Associations between protein intake and AGE-RAGE biomarkers were examined using linear regression models adjusted for demographics, height, lifestyle behaviors, prevalent disease, cognitive function, inflammation, and other dietary factors. CML concentrations were higher in individuals with higher total protein intake (adjusted least squares mean±SE: <0.8 g/kg/d, 829±17 ng/ml; 0.8 to <1.2 g/kg/d, 860±15 ng/ml; ≥1.2 g/kg/d, 919±23 ng/ml; P for trend=0.001), as were sRAGE concentrations (<0.8 g/kg/d, 1412±34 pg/ml; 0.8 to <1.2 g/kg/d, 1479±31 pg/ml; ≥1.2 g/kg/d, 1574±47 pg/ml; P for trend<0.0001). Every 0.1 g/kg/d increment in total protein intake was associated with a 13.3±3.0 ng/ml increment in CML and a 22.1±6.0 pg/ml increment in sRAGE (P<0.0001 for both). Higher CML and sRAGE concentrations were also associated with higher intakes of both animal and vegetable protein (all P values ≤0.01). There were no significant associations with the CML/sRAGE ratio. Higher dietary protein intake was associated with higher CML and sRAGE concentrations in older adults; however, the CML/sRAGE ratio remained similar across groups.

Free advanced glycation end product distribution in blood components and the effect of genetic polymorphisms

One hypothesis regarding the cause of diabetic complications is that advanced glycation end products (AGEs) bind to the AGE receptor and induce changes in gene expression. However, what AGEs exist in vivo and how individual AGEs are produced and impact body metabolic process to cause diabetes complications are not understood. We developed a new precise method to measure AGEs using LC-MS/MS with a new column and measured 7 free AGEs, including N(6)-carboxymethyllysine (CML), N(6)-(1-carboxyethyl)-l-lysine (CEL) and N5-(5-hydro-5-methyl-4-imidazolon-2-yl)L-ornithine (MG-H1), in human blood components. Blood was obtained from 9 people, and free AGEs were measured in individual blood components with LC-MS/MS before and after a meal. Free CML and CEL were abundant in erythrocytes, with 92% of free CML and 85% of free CEL localized in erythrocytes. In contrast, 60% of free MG-H1 was distributed in the serum. After the meal, free serum MG-H1 increased, but CML and CEL did not. CML and CEL are mainly distributed in erythrocytes and were not affected by the meal, indicating that they are produced in vivo. However, the main source of MG-H1 is the meal. The effect of genetic polymorphisms on AGEs was also investigated. Low activity type aldehyde dehydrogenase 2 (ALDH2) increased the CML concentration in the blood. This is the first observation that shows that the metabolic process of CML and CEL is different from that of MG-H1 and the effect of ALDH2 SNPs on CML.

Carboxymethyllysine and carboxyethyllysine in multiple sclerosis patients.

Advanced glycation end-products (AGE) are involved in the pathogenesis of many diseases, including neurodegenerative diseases such as multiple sclerosis (MS). The aim of the study was to evaluate the intensity of the protein glycation process in patients with multiple sclerosis and its possible involvement in disease activity. The study group (n = 45) consisted of patients suffering from MS, and the control group (n = 31) consisted of healthy adults. Concentrations of selected glycation markers such as carboxymethyllysine (CML) and carboxyethyllysine (CEL) in sera of patients with MS and healthy volunteers were determined by enzyme-linked immunosorbent assay (ELISA). Serum CML and CEL concentrations in patients with MS were higher than in healthy volunteers but only for CML the difference was statistically significant. CML concentrations positively correlated with CEL concentrations only in the healthy persons. In MS patients the serum CML and CEL concentrations did not differ significantly depending on the duration of the disease and depending on the EDSS (Expanded Disability Status Scale) score. Multiple sclerosis is accompanied by an intensification of protein glycation processes, especially within the pathways leading to the formation of carboxymethyllysine. The duration of the disease and the degree of motor impairment do not appear to affect the progression of the glycation processes. However, the disease process associated with multiple sclerosis may affect the relationship between CML and CEL concentrations.

PSIII-23 Nutrient and Maillard reaction product concentrations of commercially available pet foods and treats

Abstract Exposing ingredients to high temperatures may result in chemical reactions between amino acids and sugars, leading to the formation of Maillard reaction products (MRPs). These products may result in altered palatability, protein quality, and digestibility. This experiment quantified key MRPs in commercial pet foods, estimated daily intake of MRPs of pet animals, and correlated MRPs with macronutrient concentrations present in the foods. The analysis included 53 pet foods and treats commercially available in the United States: 11 dry dog foods, 29 wet dog foods, 2 dry dog treats, 3 wet dog treats, 2 dry cat foods, and 6 wet cat foods. Diets and treats were analyzed for dry matter, organic matter, crude protein (CP), acid-hydrolyzed fat, total dietary fiber, and gross energy using standard techniques. MRPs were analyzed using high-performance liquid chromatography and gas chromatography-mass spectrometry. All data were analyzed as a completely randomized design using the GLM procedure of SAS 9.4. On average, wet foods had higher (P &amp;lt; 0.001) fructoselysine (FRUC) and carboxymethyllysine (CML) than dry foods. Dry dog treats had the highest (P &amp;lt; 0.001) FRUC (29.64 µg/g) and CML (0.47 mg/g) concentrations of all products tested. Dry cat foods had the lowest (P &amp;lt; 0.001) CML (0.10 mg/g) concentrations of all commercial segments. Wet dog treats had the highest (P &amp;lt; 0.001) 5-hydroxymethyl-2-furfural (HMF) (10.92 µg/g) concentrations of all segments and wet cat foods had the lowest (P &amp;lt; 0.001) HMF (0.22 µg/g) content. Dogs and cats fed wet diets would be more likely to ingest higher amounts of CML and FRUC compared to animals fed dry diets. Dogs consuming dry treats would be more likely to ingest higher amounts of CML than wet treats. CP was negatively correlated to HMF (P = 0.0425; r = -0.27973), while TSF was positively correlated to CML (P = 0.0417; r = 0.28080).

N ε-(carboxymethyl)lysine formation from the Maillard reaction of casein and different reducing sugars.

Dietary advanced glycation end products (AGEs) are involved in the pathogenesis of diabetic complications, atherosclerosis, and kidney disease. Formation of N ε-(carboxymethyl)lysine (CML), a well-known AGEs, was evaluated from the reaction of casein from bovine milk with different reducing sugars (glucose, tagatose, and xylose) at various sugar concentrations and heating temperatures (75 and 120°C) used in food processing to determine the best sweetener to be used in dairy products. The concentration of CML was measured using an enzyme-linked immunosorbent assay. Additionally, SDS-PAGE was carried out to observe the changes in the molecular weight of casein. The results reveal that tagatose leads to a lower CML concentration at 75°C than glucose or xylose, whereas no significant differences are observed at 120°C. We conclude that it would be more appropriate to use tagatose rather than glucose or xylose as a sweetener, considering the AGEs contents in heat-treated dairy products.

Circulating Advanced Glycation Endproducts and Long-Term Risk of Cardiovascular Mortality in Kidney Transplant Recipients.

In kidney transplant recipients, elevated circulating advanced glycation endproducts (AGEs) are the result of increased formation and decreased kidney clearance. AGEs trigger several intracellular mechanisms that ultimately yield excess cardiovascular disease. We hypothesized that, in stable kidney transplant recipients, circulating AGEs are associated with long-term risk of cardiovascular mortality, and that such a relationship is mediated by inflammatory, oxidative stress, and endothelial dysfunction biomarkers. Prospective cohort study of stable kidney transplant recipients recruited between 2001 and 2003 in a university setting. We performed multivariable-adjusted Cox regression analyses to assess the association of AGEs (i.e., Nε-[Carboxymethyl]lysine (CML) and Nε-[Carboxyethyl]lysine (CEL), measured by tandem mass spectrometry) with cardiovascular mortality. Mediation analyses were performed according to Preacher and Hayes's procedure. We included 555 kidney transplant recipients (age 51±12 years, 56% men). During a median follow-up of 6.9 years, 122 kidney transplant recipients died (52% deaths were due to cardiovascular causes). CML and CEL concentrations were directly associated with cardiovascular mortality (respectively, hazard ratio, 1.55; 95% confidence interval, 1.24 to 1.95; P<0.001; and hazard ratio, 1.53; 95% confidence interval 1.18 to 1.98; P=0.002), independent of age, diabetes, smoking status, body mass index, eGFR and proteinuria. Further adjustments, including cardiovascular history, did not materially change these findings. In mediation analyses, free thiol groups and soluble vascular cell adhesion molecule-1 consistently explained approximately 35% of the association of CML and CEL with cardiovascular mortality. In stable kidney transplant recipients, circulating levels of AGEs are independently associated with long-term risk of cardiovascular mortality. This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2019_09_17_CJN00540119.mp3.

High-throughput quantification of carboxymethyl lysine in serum and plasma using high-resolution accurate mass Orbitrap mass spectrometry.

BackgroundCarboxymethyl lysine is an advanced glycation end product of interest as a potential biomarker of cardiovascular and other diseases. Available methods involve ELISA, with potential interference, or isotope dilution mass spectrometry (IDMS), with low-throughput sample preparation.MethodsA high-throughput sample preparation method based on 96-well plates was developed. Protein-bound carboxymethyl lysine and lysine were quantified by IDMS using reversed phase chromatography coupled to a high-resolution accurate mass Orbitrap Exactive mass spectrometer. The carboxymethyl lysine concentration (normalized to lysine concentration) was measured in 1714 plasma samples from the British Regional Heart Study (BRHS).ResultsFor carboxymethyl lysine, the lower limit of quantification (LLOQ) was estimated at 0.16 μM and the assay was linear between 0.25 and 10 μM. For lysine, the LLOQ was estimated at 3.79 mM, and the assay was linear between 2.5 and 100 mM. The intra-assay coefficient of variation was 17.2% for carboxymethyl lysine, 9.3% for lysine and 10.5% for normalized carboxymethyl lysine. The inter-assay coefficient of variation was 18.1% for carboxymethyl lysine, 14.8 for lysine and 16.2% for normalized carboxymethyl lysine. The median and inter-quartile range of all study samples in each batch were monitored. A mean carboxymethyl lysine concentration of 2.7 μM (IQR 2.0–3.2 μM, range 0.2–17.4 μM) and a mean normalized carboxymethyl lysine concentration of 69 μM/M lysine (IQR 54–76 μM/M, range 19–453 μM/M) were measured in the BRHS.ConclusionThis high-throughput sample preparation method makes it possible to analyse large cohorts required to determine the potential of carboxymethyl lysine as a biomarker.

Reduction of Nε-(carboxymethyl) lysine by (−)-epicatechin and (−)-epigallocatechin gallate: The involvement of a possible trapping mechanism by catechin quinones

(−)-Epicatechin (EC) and (−)-epigallocatechin gallate (EGCG) (0.001–5%) were found to reduce Nε-(carboxymethyl) lysine (CML) concentrations by up to 0.80 ± 0.11 mM and 0.85 ± 0.07 mM, respectively, as quantified by HPLC-MS during heating at 80 °C and 100 °C in a glucose-lysine model system at pH 5, 7 and 8, which mimic different food processing conditions. With the addition of 1 mM EC, the free CML concentration in coconut milk was reduced to 5.7 ± 0.1 μg/mL (∼2.8 × 10−2 mM), which was significantly lower than that in unheated coconut milk (6.1 ± 0.2 μg/mL (∼3.0 × 10−2 mM), p < 0.05) indicating that the free CML may be eliminated by the addition of EC in coconut milk. Products of EC/EGCG quinones reacted with CML by a Michael addition or Schiff base addition in model systems were tentatively identified by UPLC-ESI-MS/MS.