imidazolone and pentosidine

We report a comprehensive density functional theory (DFT) study of the mechanism of pentosidine formation. This work is a continuation of our earlier studies in which we proposed pathways for formation of glucosepane (J. Mol. Model. 2011, pp 1-15, DOI 10.1007/s00894-011-1161-x), GODIC (glyoxal-derived imidazolium cross-link), and MODIC (methyl glyoxal-derived imidazolium cross-link; J. Phys. Chem. 2011, 115, pp 13542-13555). Here we show that formation of pentosidine via reaction of α-oxoaldehydes with lysine and arginine in aqueous solution is possible thermodynamically and kinetically, in good agreement with the available experimental evidence. Five pathways, A-E, were characterized, as in our previous GODIC and MODIC work. In pathways A and B, a Schiff base is first formed from lysine and methyl glyoxal (MGO), and this is followed by addition of arginine and glyoxal (GO). By contrast, in pathways C, D, and E, addition of arginine to MGO occurs first, resulting in the formation of imidazolone, which then reacts with lysine and GO to give pentosidine. Our calculations show that the reaction process is highly exergonic and that the three pathways A, C, and E are competitive. These results serve to underline the potentially important role that α-oxoaldehydes play as precursors in pentosidine formation in the complex field of glycation.

Topics: Arginine; Glycosylation; Glyoxal; Imidazoles; Kinetics; Lysine; Molecular Structure; Pyruvaldehyde; Quantum Theory; Schiff Bases; Thermodynamics; Water

Advanced glycation end products (AGEs) are known to be deposited in the target organ of ageing. In addition, the deposition of AGEs accelerate the process of ageing. We investigated the immunohistochemical localization of AGEs in pinguecula, one of the ocular changes related with ageing process.. Surgical specimens of conjunctiva with or without pinguecula were prepared from nine patients, respectively. Immunohistochemical localization of AGEs was investigated using monoclonal antibodies to N(epsilon)-(carboxymethyl)lysine, pentosidine, imidazolone, and pyrraline.. Moderate to strong immunoreactivities to AGEs were detected in the subepithelial amorphous deposits of all the surgical specimens with pinguecula. In contrast, no or weak immunoreactivities to AGEs were detected in the surgical specimens without pinguecula.. Pinguecula is an aggregation of AGEs-modified proteins. The presence of pinguecula would be an index of local irradiation of ultraviolet rays and decreased antioxidant activities.

Topics: Adult; Aged; Aging; Antibodies, Monoclonal; Arginine; Conjunctival Diseases; Female; Glycation End Products, Advanced; Humans; Imidazoles; Immunoenzyme Techniques; Lysine; Male; Middle Aged; Norleucine; Pyrroles

Hemodialysis (HD) patients are frequently in a state of increased oxidative stress, and hyperglycemia appears to be a major factor. We recently found that oxidized human serum albumin (HSA) is a reliable marker of oxidative stress in HD patients. However, the issue of whether oxidized HSA is associated with the progression of oxidative stress in HD patients with or without diabetes is not clear. In the present study, we examined the effect of a qualitative modification of HSA in HD patients with or without diabetes. Blood samples from 10 HD patients with diabetes, 7 HD patients without diabetes, and 10 healthy age-matched controls were examined. The increase in plasma protein carbonyl content and advanced glycation endproducts (AGEs) in HD patients was largely due to an increase in the levels of oxidized HSA. Furthermore, these increases were greatest in HD patients with diabetes. Purified HSA from HD patients (non-DM-HSA) was carbonylated and AGE-modified. The amount of modified HSA was the highest in HD patients with diabetes (DM-HSA). Carboxy methyl lysine (CML)-modified HSA triggered a neutrophil respiratory burst, and this activity was closely correlated with the increase in the CML/HSA ratio. These findings indicate that uremia plays an important role in the progression of oxidative stress in HD patients via an increase in CML-modified HSA. They also indicate that diabetic complications further exacerbate the progression of oxidative stress by further increasing the amount of these modified HSA molecules.

Topics: Adult; Aged; Aged, 80 and over; Arginine; Blood Proteins; Case-Control Studies; Diabetes Mellitus; Female; Glycation End Products, Advanced; Humans; Imidazoles; Kidney Failure, Chronic; Lysine; Male; Middle Aged; Neutrophils; Norleucine; Oxidative Stress; Protein Carbonylation; Pyrroles; Renal Dialysis; Respiratory Burst; Serum Albumin

The levels of plasma 3-deoxyglucosone (3-DG) increase under hyperglycemic conditions and are associated with the pathogenesis of diabetic complications because of the high reactivity of 3-DG with proteins to form advanced glycation end products (AGE). To investigate potential markers for 3-DG-mediated protein modification in vitro and in vivo, we compared the yield of several 3-DG-derived AGE structures by immunochemical analysis and HPLC and measured their localization in human atherosclerotic lesions. When BSA was incubated with 3-DG at 37 degrees C for up to 4 wk, the amounts of N(epsilon)-(carboxymethyl)lysine (CML) and 3-DG-imidazolone steeply increased with incubation time, whereas the levels of pyrraline and pentosidine increased slightly by day 28. In contrast, significant amounts of pyrraline and pentosidine were also observed when BSA was incubated with 3-DG at 60 degrees C to enhance AGE-formation. In atherosclerotic lesions, CML and 3-DG-imidazolone were found intracellularly in the cytoplasm of most foam cells and extracellularly in the atheromatous core. A weak-positive immunoreaction with pyrraline was found in the extracellular matrix and a few foam cells in aortic intima with atherosclerotic lesions. Our results provide the first evidence that CML and 3-DG-imidazolone are major AGE structures in 3-DG-modified proteins, and that 3-DG-imidazolone provides a better marker for protein modification by 3-DG than pyrraline.

Topics: Adult; Aged; Antibodies, Monoclonal; Aorta; Arginine; Arteriosclerosis; Binding, Competitive; Chromatography; Chromatography, High Pressure Liquid; Cytoplasm; Deoxyglucose; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Foam Cells; Glycation End Products, Advanced; Humans; Imidazoles; Immunochemistry; Immunohistochemistry; Lysine; Male; Middle Aged; Models, Biological; Models, Chemical; Norleucine; Pyrroles; Temperature; Time Factors

Chronic renal failure (CRF) is characterized by enhanced formation and accumulation of advanced glycation end products (AGEs), which are involved in the pathogenesis of vascular damage. Their role as risk factors for cardiovascular complications is still unknown. This study aims to investigate whether elevated serum levels of the AGEs pentosidine, N(epsilon)-carboxymethyllysine (CML), and the 3-deoxyglucosone-derived imidazolone involve a greater risk for cardiovascular events (CVEs) and left ventricular hypertrophy (LVH).. Patients with CRF (n = 99), on maintenance hemodialysis (HD) therapy (n = 84), and renal transplant recipients (RTRs; n = 50) were included. Pentosidine was measured by high-performance liquid chromatography, and CML and imidazolone, by enzyme-linked immunosorbent assays. Statistical analyses were performed using Mann-Whitney U test, logistic regression analysis, and Cox proportional hazards model.. At baseline in all investigated groups, patients with a history of CVEs or LVH showed greater mean serum AGE levels. By retrospective data analysis, significant odds ratios for increases in CML and imidazolone levels were calculated for LVH in HD patients, as well as for increases in CML levels for CVEs in RTRs, respectively. By prospective data analysis, serum AGE levels could not be evaluated as independent risk factors for CVEs in all investigated groups.. From these preliminary results, serum AGE levels could not be identified as independent risk factors for CVEs or LVH in patients with CRF. Prospective studies are needed to answer this question.

Topics: Adult; Aged; Aged, 80 and over; Arginine; Cardiovascular Diseases; Diabetes Mellitus; Female; Glycation End Products, Advanced; Humans; Hypertrophy, Left Ventricular; Imidazoles; Kidney Failure, Chronic; Kidney Transplantation; Lysine; Male; Middle Aged; Renal Dialysis; Retrospective Studies; Risk Factors

To investigate whether the advanced glycation end-products (AGEs) pentosidine, N( epsilon )-(carboxymethyl)lysine (CML), and imidazolone are present in the aqueous of cataract patients and how AGE levels correlate to cataract type or the diabetic condition of the patient.. Departments of Ophthalmology and Internal Medicine, University of Jena, Jena, Germany.. Aqueous and serum samples from 77 cataractous patients (33 nondiabetics, 44 diabetics; 14 with dense posterior, 63 with nuclear cataracts) were investigated. The mean age of the patients was 69 years +/- 14 (SD). The aqueous protein concentration was examined using a laser flare-cell meter. In the samples, pentosidine was measured by high-performance liquid chromatography and CML using a competitive enzyme-linked immunosorbent assay. Western blot analysis was used to detect imidazolone, pentosidine, and CML in the aqueous.. The aqueous samples contained CML, pentosidine, or imidazolone. These AGEs occurred mainly bound to albumin. Significant correlations existed between serum pentosidine and aqueous CML and flare levels as well as between serum and aqueous CML. Patients with nuclear cataract had insignificantly higher pentosidine and CML levels than patients with posterior cataract, whereas the flare was significantly higher. No significant differences were found between the aqueous AGE levels in nondiabetic and diabetic patients.. The aqueous of cataractous eyes contained the AGEs CML, pentosidine, and imidazolone. All 3 AGEs occurred mainly albumin-bound, providing evidence they may originate from the blood. Further investigation is needed to determine the relevance of aqueous AGEs in cataractogenesis.

Topics: Aged; Albumins; Aqueous Humor; Arginine; Blotting, Western; Cataract; Chromatography, High Pressure Liquid; Electrophoresis, Polyacrylamide Gel; Female; Glycation End Products, Advanced; Humans; Imidazoles; Lysine; Male; Middle Aged; Protein Binding

To investigate the occurrence of advanced glycation end products (AGEs) formed oxidatively (pentosidine and N(epsilon)-carboxymethyl lysine [CML]) or nonoxidatively (imidazolone) in human lenses and the relation of AGEs to lens coloration, cataract type, and patients' diabetic state.. Departments of Ophthalmology and Internal Medicine III, University of Jena, Jena, Germany.. Pentosidine, CML, and imidazolone concentrations were measured in the water-soluble protein fraction of 44 cataractous lenses (from 24 nondiabetic and 20 diabetic donors) and 6 noncataractous control lenses.. Pentosidine, CML, and imidazolone were higher in cataractous lenses than in control lenses (pentosidine, 3.7 pmol/mg +/- 5.3 (SD) and 1.9 +/- 1.7 pmol/mg, respectively; CML, 3.0 +/- 2.2 nmol/mg and 1.3 +/- 0.7 nmol/mg, respectively; imidazoline, 80.4 +/- 93.3 AU/mg and 19.6 +/- 18.5 AU/mg, respectively). Among the cataractous lenses, the highest AGE concentrations were found in mature cataracts, with a statistically significant increase in CML. The AGE content increased relative to the intensity of brown coloration of the lens; the brown coloration also indicated the highest rise of imidazolone compared to pentosidine and CML. Lenses from diabetic donors had generally similar pentosidine values and elevated CML and imidazolone levels compared to lenses from nondiabetic donors. The pentosidine, CML, and imidazolone levels in the lenses correlated significantly with one another but not with patient age.. Advanced glycation end products formed oxidatively and nonoxidatively occurred to a higher degree in cataractous lenses than in noncataractous lenses. The strong relationship between the lenses' AGE content, color/opacity, and the state of the cataract may indicate that advanced glycation plays a pivotal role in cataract formation.

Topics: Aged; Antibodies, Monoclonal; Arginine; Cataract; Diabetes Mellitus; Glycation End Products, Advanced; Humans; Imidazoles; Immunoenzyme Techniques; Lens, Crystalline; Lysine; Middle Aged; Oxidation-Reduction

To determine the concentrations of methylglyoxal-derived advanced glycation end-products (AGEs), the hydroimidazolones MG-H1 and -H2, in soluble human lens proteins and compare them with the concentrations of other methylglyoxal-derived AGEs and pentosidine.. Lens protein samples were hydrolyzed enzymatically. AGEs were assayed without derivatization by HPLC with tandem mass spectrometry; the fluorescent AGEs argpyrimidine and pentosidine were assayed by fluorometric detection. MG-H1 and -H2 were resolved and assayed by fluorometric detection after derivatization with 6-aminoquinolyl-N-hydroxysuccimidylcarbamate (AQC).. The methylglyoxal-derived hydroimidazolones MG-H1 and -H2 were detected and quantified in human lens proteins. AGE concentrations (mean +/- SEM) were: MG-H1 4609 +/- 411 pmol/mg protein, MG-H2 3085 +/- 328 pmol/mg protein, argpyrimidine 205 +/- 19 pmol/mg protein, and pentosidine 0.693 +/- 0.104 pmol/mg protein. The concentration of MG-H1 in human lens protein correlated positively with donor age (correlation coefficient = 0.28, P < 0.05), the concentration of MG-H2 (correlation coefficient = 0.78, P < 0.001) and argpyrimidine (correlation coefficient = 0.42, P < 0.01). The concentrations of AGEs were increased in cataractous lenses in comparison with noncataractous lenses: the increases were MG-H1 85%, MG-H2 122%, argpyrimidine 255%, and pentosidine 183% (P < 0.001). Multiple logistic regression analysis showed a significant link of cataract to donor age (regression coefficient beta = 0.094, P = 0.026) and argpyrimidine (beta = 0.022, P = 0.002).. Methylglyoxal hydroimidazolones are quantitatively major AGEs of human lens proteins. These substantial modifications of lens proteins may stimulate further glycation, oxidation, and protein aggregation leading to the formation of cataract.

Topics: Aged; Arginine; Cataract; Chromatography, High Pressure Liquid; Crystallins; Female; Fluorometry; Glycation End Products, Advanced; Humans; Hydrolysis; Imidazoles; Lens, Crystalline; Lysine; Male; Mass Spectrometry; Middle Aged; Pyruvaldehyde

Previous studies from our laboratory demonstrated that N(epsilon)-(carboxymethyl)lysine (CML), one of the major advanced glycation end products (AGE), was accumulated in human pyramidal neurons in the hippocampus in an age-dependent manner. This suggests a potential link between AGE-accumulation and the aging process in neurons. The purpose of the present study was to examine whether this notion could be extended to other AGE structures, such as imidazolone and pentosidine. This was done using 19 human brains that were not affected by dementia. The immunohistochemical survey on distribution in brain tissues of imidazolone and pentosidine was carried out with monoclonal antibodies specific for imidazolone and pentosidine. A parallel control experiment was carried out with anti-CML antibody. The results showed that pentosidine and imidazolone were localized in neurons in different areas of human brain tissue, especially in neurons of CA4 in the hippocampus. The characteristic distribution of pentosidine and imidazolone is very similar to that of CML. Furthermore, when the accumulation of these AGE structures was compared with the age of individual brains it was found that accumulation of imidazolone, pentosidine and CML in the CA4 region increased with age. These findings taken together support the notion that the accumulation of AGE structures in the CA4 region might be closely related to the aging process in neurons.

Topics: Adult; Aged; Aged, 80 and over; Aging; Arginine; Female; Glycation End Products, Advanced; Humans; Imidazoles; Immunohistochemistry; Lysine; Male; Middle Aged; Pyramidal Cells

To assess a role for oxidative stress in the pathogenesis of amyotrophic lateral sclerosis (ALS), we analyzed the immunohistochemical localization of 8-hydroxy2'-deoxyguanosine (OHdG) as a nucleic acid oxidation product, acrolein-protein adduct and 4-hydroxy-2-nonenal (HNE)-protein adduct as lipid peroxidation products, Nepsiloncarboxymethyl-lysine (CML) as a lipid peroxidation or protein glycoxidation product, pentosidine as a protein glycoxidation product, and imidazolone and pyrraline as nonoxidative protein glycation products in the spinal cord of three familial ALS patients with superoxide dismutase(SOD 1) A4V mutation, six sporadic ALS patients, and six age-matched control individuals. The spinal cord sections of the control cases did not show any distinct immunoreactivities for these examined products. In the familial ALS cases, intense immunoreactivities for pyrraline and CML were confined to the characteristic Lewy body-like hyaline inclusions, and imidazolone immunoreactivity was located in the cytoplasm of the residual motor neurons. No significant immunoreactivities for other examined products were detected in the familial ALS spinal cords. In the sporadic ALS cases, intense immunoreactivities for pentosidine, CML and HNE-protein adduct were seen in the cytoplasm of the degenerated motor neurons, and OHdG immunoreactivity was located in the cell nuclei of the residual neurons and glial cells. The present results indicate that oxidative reactions are involved in the disease processes of sporadic ALS, while there is no evidence for increased oxidative damage except for CML deposition in the familial ALS spinal cords. Furthermore, it is likely that the accumulation of pyrraline and imidazolone supports a nonoxidative mechanism in SOD1-related motor neuron degeneration.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Acrolein; Adult; Aged; Aldehydes; Amyotrophic Lateral Sclerosis; Arginine; Deoxyguanosine; Glycosylation; Humans; Imidazoles; Lipid Peroxidation; Lysine; Male; Middle Aged; Motor Neurons; Oxidative Stress; Spinal Cord; Superoxide Dismutase