Development of a Method for Producing and Purifying N2-Carboxyethyl-2'-Deoxyguanosine for Molecular-Biological Research

2015 ◽  
Vol 1085 ◽  
pp. 436-440
Author(s):  
Sergey Krivoshchekov ◽  
Aleksandr O. Bogdanov ◽  
Ludmila M. Ogorodova ◽  
Natalya B. Dementeva ◽  
Irina V. Saltykova ◽  
...  
Keyword(s):  
Advanced Glycation End Products ◽  
Biological Research ◽  
Advanced Glycation ◽  
Reverse Phase Hplc ◽  
Biochemical Processes ◽  
Glycation End Products ◽  
End Products ◽  
The Cost

Methylglyoxal (MG) – is a high reactive α- oxoaldehyde, which can be synthesized through various biochemical processes in vivo. MG is capable of interacting with nucleophilic groups of proteins, lipids and nucleic acids which leads to their glycation. Covalent compounds that formed in this process were named advanced glycation end products. Advanced glycation end products play an important role in the pathogenesis of diabetes, chronic inflammation, cancer and Alzheimer's disease. One of the most stable compounds, which is formed by reacting of methylglyoxal with the nitrogenous bases of the DNA molecule is N2- carboxyethyl -2' - deoxyguanosine (CEdG). This compound can be used as a marker for monitoring of various diseases, investigation of the role of glycation end products in their pathogenesis, as well as to search for the therapeutic targets. To perform our molecular-biological investigations by mass spectrometry, we synthesized N2- carboxyethyl -2' - deoxyguanosine, and have proposed a new method for purifying compounds by reverse - phase HPLC. As a result, we achieved the optimal purity of the sample, while substantially reducing the cost of the purification procedure.

scholarly journals Examining the Role of Dietary Advanced Gycation End Products in Prostate Cancer Using Molecular Imaging Techniques (OR04-08-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Christian Konopka ◽  
Annaliese Paton ◽  
Aleksandra Skokowska ◽  
Joe Rowles ◽  
John Erdman ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Serum Albumin ◽  
Advanced Glycation End Products ◽  
Growth Media ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products ◽  
The Impact

Abstract Objectives The Receptor for Advanced Glycation End Products (RAGE) and its ligands have been shown to be both over expressed and critical to prostate cancer (PCa) development. Importantly, the overexpression of both RAGE and its ligands is associated with poor PCa patient survival, suggesting its promise as a molecular target. Additionally, one of the largest sources of ligands for RAGE, advanced glycation end products (AGEs), come from one's diet and their concentrations are directly related to disease. We hypothesized that dietary AGEs (dAGEs) significantly contribute to the progression of PCa through interactions with RAGE. In this study we explore the use of a novel imaging strategy targeted at RAGE in combination with conventional imaging and histological techniques to assess the role of dAGEs on RAGE expression and PCa progression in murine xenografts. Methods To examine the impact of AGEs on PCa cell function, experiments were performed in two PCa cell lines. Cells were grown in growth media enriched with carboxymethyl-lysine-modified human serum albumin (CML) (the most common AGE) or a control protein, bovine serum albumin (BSA). Western blot, confocal microscopy, clonogenic assays, and proliferations assays were performed. To study the effects of an enhanced consumption of dAGEs on PCa growth and progression in vivo, NU/J mice were fed a modified Ain-93 G diet, which was either CML or BSA enriched. PCa tumors were then initiated. Their growth was monitored, their perfusion measured using Speckle Contrast Imaging, and their metabolic rate and RAGE content quantified using 18FDG and a novel RAGE-targeted tracer using PET-CT. Finally, the tissues were excised for histological analysis. Results CML significantly enhanced in vitro expression of both RAGE and proliferation marker KI-67. Cell doubling time was also significantly quickened, (1.5 vs 2.4 days) in the CML vs control. In vivo data demonstrated significant differences in tumor growth (CML group up to 2-fold increase) and successful tumor implantation rate (30% vs 60%). Perfusion, metabolism, and RAGE imaging demonstrated unique patterns which varied over the course of PCa progression. Conclusions These studies indicate that dAGEs may play a significant role in the progression of PCa. The data suggests that RAGE and its ligands are promising targets for further therapeutic investigations. Funding Sources University of Illinois at Chicago Cancer Center Pilot Grant. Supporting Tables, Images and/or Graphs

Glycation in food and metabolic transit of dietary AGEs (advanced glycation end-products): studies on the urinary excretion of pyrraline

2003 ◽  
Vol 31 (6) ◽  
pp. 1383-1385 ◽  
Author(s):  
A. Foerster ◽  
T. Henle
Keyword(s):  
Urinary Excretion ◽  
Advanced Glycation End Products ◽  
Physiological Role ◽  
Dietary Control ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products ◽  
First Time

Pyrraline [∊-(2´-formyl-5´-hydroxymethyl-pyrrolyl)-l-norleucin] belongs to the group of AGEs (advanced glycation end-products) formed in the final stage of the Maillard reaction in foods and in vivo. As it is generally accepted that AGEs are pathophysiologically relevant in aging and in diseases such as diabetes and uraemia, physiological consequences resulting from the ingestion of dietary AGEs are discussed, but balance studies for well defined AGEs are still lacking. The aim of our study was to investigate the influence of nutrition on the urinary excretion of pyrraline. After the first day without dietary restrictions, seven healthy volunteers were asked, starting on the morning of day 2, to ingest a diet virtually free of Maillard compounds (i.e. no cooked or roasted foods, no bakery products, no coffee, etc.). Dietary control was stopped on the morning of day 5. We collected 24 h urine samples for these 5 days, which were analysed for free pyrraline by reverse-phase HPLC with UV detection at 297 nm. We found that urinary excretion of free pyrraline was directly affected by the composition of the diet, decreasing from 4.8±1.1 mg/day on day 1 to levels of 1.6, 0.4 and 0.3 mg/day on days 2, 3 and 4 respectively, followed by a significant increase to 3.2±1.4 mg/day on the 5th day. The results of this work prove, for the first time, that urinary excretion of pyrraline is strongly dependent on its dietary intake. Thus the influence of nutrition should be taken into consideration in studies directed to the physiological role of glycation compounds.

scholarly journals ROLE OF TOLL-LIKE RECEPTORS 2 AND 4, AND THE RECEPTOR FOR ADVANCED GLYCATION END PRODUCTS IN HIGH-MOBILITY GROUP BOX 1-INDUCED INFLAMMATION IN VIVO

Shock ◽  
2009 ◽  
Vol 31 (3) ◽  
pp. 280-284 ◽  
Author(s):  
Marieke A.D. van Zoelen ◽  
Huan Yang ◽  
Sandrine Florquin ◽  
Joost C.M. Meijers ◽  
Shizuo Akira ◽  
...  
Keyword(s):  
Advanced Glycation End Products ◽  
High Mobility ◽  
High Mobility Group ◽  
Toll Like Receptors ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products

scholarly journals Influence of Dopamine on Fluorescent Advanced Glycation End Products Formation Using Drosophila melanogaster

Biomolecules ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 453
Author(s):  
Ana Filošević Vujnović ◽  
Katarina Jović ◽  
Emanuel Pištan ◽  
Rozi Andretić Waldowski
Keyword(s):  
Drosophila Melanogaster ◽  
Advanced Glycation End Products ◽  
Model Organism ◽  
Covalent Modification ◽  
Advanced Glycation ◽  
Biomarkers Of Aging ◽  
Glycation End Products ◽  
End Products

Non-enzymatic glycation and covalent modification of proteins leads to Advanced Glycation End products (AGEs). AGEs are biomarkers of aging and neurodegenerative disease, and can be induced by impaired neuronal signaling. The objective of this study was to investigate if manipulation of dopamine (DA) in vitro using the model protein, bovine serum albumin (BSA), and in vivo using the model organism Drosophila melanogaster, influences fluorescent AGEs (fAGEs) formation as an indicator of dopamine-induced oxidation events. DA inhibited fAGEs-BSA synthesis in vitro, suggesting an anti-oxidative effect, which was not observed when flies were fed DA. Feeding flies cocaine and methamphetamine led to increased fAGEs formation. Mutants lacking the dopaminergic transporter or the D1-type showed further elevation of fAGEs accumulation, indicating that the long-term perturbation in DA function leads to higher production of fAGEs. To confirm that DA has oxidative properties in vivo, we fed flies antioxidant quercetin (QUE) together with methamphetamine. QUE significantly decreased methamphetamine-induced fAGEs formation suggesting that the perturbation of DA function in vivo leads to increased oxidation. These findings present arguments for the use of fAGEs as a biomarker of DA-associated neurodegenerative changes and for assessment of antioxidant interventions such as QUE treatment.

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