• Analytical Science and Technology
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• v.36 no.1
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• pp.12-21
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• 2023

Protein glycation is vital to aging and disease. However, glycated proteins are low-abundant in plasma, rendering them difficult to identify using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Many studies have analyzed glycated peptides with high reproducibility. Here, glycated peptides in human serum were analyzed by LC-MS/MS using data-dependent acquisition (DDA) and parallel reaction monitoring (PRM). Boronic acid (BA) enrichment of in vitro glycated human serum peptides was performed. BA enrichment identified the most glycated peptides, and the glycated peptides of the more diversified proteins, excluding albumin, were analyzed. In PRM, glycated albumin PSMs were the most common, and this method exhibited the best reproducibility. The results of this study could help compare methods for identifying glycation-related biomarkers.

• Journal of Sensor Science and Technology
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• v.25 no.6
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• pp.435-439
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• 2016

We report a novel detection technology for glycated hemoglobin (HbA1c) that is measured primarily to identify the three-month average plasma glucose concentration. In enzymatic measuring of glycated hemoglobin, the generated hydrogen peroxide was then used as a reducing agent of gold (III) for the synthesis of gold (0). Gold nanoparticles obtained from this novel approach were measured by optical and piezoelectric methods. In optical method, we have developed polymer based film-type sensor cartridge filled with all the reagents for glycated hemoglobin analysis and the cartridge worked very well having the detection limit of 0.53% of glycated hemoglobin. On the other hand, quartz crystal microbalance (QCM) sensors also have been developed to determine the abilities of surface modified QCM sensors at various levels of the concentration of glycated hemoglobin to bind gold nanoparticles and limit of detection was 0.90%. Finally, despite of relatively lower sensitivities of QCM sensor and film-type optical sensor than well-plate based optical detection, these two sensors were available to measure the glycated hemoglobin level for diabetes patients and normal person.

• Journal of Korean society of Dental Hygiene
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• v.20 no.1
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• pp.19-27
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• 2020

Objectives: This study aimed to analyze the association between self-assessed periodontal symptoms and glycated hemoglobin levels in patients with type 2 diabetes. Methods: This cross-sectional study involved 156 patients with type 2 diabetes who were aged 50 years or older. Structured questionnaires were used to investigate the self-assessed periodontal symptoms of the patients. The glycated hemoglobin test was performed to evaluate their long-term blood glycemic control. Chi-square test and logistic multiple regression were performed to analyze the factors associated with glycated hemoglobin levels. Results: Compared with patients aged 65 years and above, more patients aged 64 years and below showed poor glycemic control (p=0.020). Further, compared with patients without self-perceived gingival bleeding and halitosis, more patients with these two conditions showed poor glycemic control (p<0.05). Compared with the group of patients without any periodontal symptoms, the group of patients that had at least one periodontal symptom had a higher proportion of patients with poor glycemic control (p<0.001). In the logistic regression model, gingival bleeding and halitosis were the factors most associated with hyperglycemia (p<0.05). Conclusions: The results of our study suggest that gingival bleeding and halitosis can predict hyperglycemia in patients with type 2 diabetes.

• BMB Reports
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• v.28 no.3
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• pp.249-253
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• 1995

The nonenzymatic glycation of copper, zinc-superoxide dismutase (Cu,Zn-SOD) led to inactivation and fragmentation of the enzyme. The glycated Cu,zn-SOD was isolated by boronate affinity chromatography. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA and the generation of strand breaks in pBhiescript plasmid DNA by a metal-catalyzed oxidation (MCO) system composed of $Fe^{3+}$, $O_2$, and glutathione (GSH) as an electron donor was enhanced more effectively by the glycated CU,Zn-SOD than by the nonglycated enzyme. The capacity of glycated Cu,Zn-SOD to enhance damage to DNA was inhibited by diethylenetriaminepentaacetic acid (DETAPAC), azide, mannitol, and catalase. These results indicated that incubation of glycated CU,Zn-SOD with GSH-MCO may result in a release of $Cu^{2+}$ from the enzyme. The released $Cu^{2+}$ then likely participated in a Fenton-type reaction to produce hydroxyl radicals, which may cause the enhancement of DNA damage.

• Analytical Science and Technology
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• v.35 no.1
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• pp.8-14
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• 2022

Many studies have shown that advanced glycation end-products (AGEs) and glycation adducts are significantly linked to aging and disease. Particularly, the level of glycation in human milk is important because the AGE intake is closely related to AGE levels in infants. In this study, we used human milk samples obtained from four primiparae and four multiparae. We isolated proteins using acetone and trichloroacetic acid (TCA) precipitation. A total of 67 glycated proteins and 122 glycated peptides was quantified; among them, 19 glycated peptides were differentially expressed. We confirmed that the degree of glycation differed according to fertility. The study provides a foundation for using proteomics to evaluate the mother's milk quality and link between maternal health and human milk quality.

• Molecules and Cells
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• v.19 no.1
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• pp.60-66
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• 2005

Low density lipoproteins (LDL) play important roles in the pathogenesis of atherosclerosis. Diabetes is associated with accelerated atherosclerosis leading to cardiovascular disease in diabetic patients. Although LDL stimulates the proliferation of arterial smooth muscle cells (SMC), the mechanisms are not fully understood. We examined the effects of native LDL and glycated LDL on the extracellular signal-regulated kinase (ERK) pathway. Addition of native and glycated LDL to rat aorta SMCs (RASMCs) stimulated ERK phosphorylation. ERK phosphorylation was not affected by exposure to the $Ca^{2+}$ chelator BAPTA-AM but inhibition of protein kinase C (PKC) with GF109203X, inhibition of Src kinase with PP1 ($5{\mu}M$) and inhibition of phospholipase C (PLC) with U73122/U73343 ($5{\mu}M$) all reduced ERK phosphorylation in response to glycated LDL. In addition, pretreatment of the RASMCs with a cell-permeable mitogen-activated protein kinase kinase (MEK) inhibitor (PD98059, $5{\mu}M$) markedly decreased ERK phosphorylation in response to native and glycated LDL. These findings indicate that ERK phosphorylation in response to glycated LDL involves the activation of PKC, PLC, and MEK, but is independent of intracellular $Ca^{2+}$.

• Bulletin of the Korean Chemical Society
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• v.31 no.7
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• pp.2103-2106
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• 2010

• Journal of Ginseng Research
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• v.26 no.4
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• pp.173-177
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• 2002

We examined effects of red ginseng on the formation of glycated protein in vivo and in vitro. The mixtures (1 : 1 : 1, v/v/v) with glucose (1.5 g/dl, hemoglobin (10 g/d) and red ginseng extract (0.5 g/dl) in 0.067 M phosphate saline buffer were incubated for 5 days in shaking water bath (37$\^{C}$, 70 RPM). Male rats were divided into three groups with one health and two diabetes, consisting of 20 heads in each group. Diabetic rats, induced by streptozotocin injection, were treated with or without red ginseng extract (100 mg/kg/day) for 3 months. The concentration of blood glucose and the rate of glycated hemoglobin were determined by commercial kits. The rate of glycated hemoglobin was significantly decreased by the addition of ginseng extract in comparison with non-addition group in vitro (12.17$\pm$ 1.01% vs 15.9$\pm$ 1.95%, meansd, p<0.01). Even though the levels of blood glucose in rats were not significantly different from each other, the rate of glycated hemoglobin in ginseng treated diabetic rats was $\pm$ se significantly lower than non-treated diabetic rats after 3 months (15.1$\pm$ 2.06% vs 20.1 $\pm$ 2.9%, mean$\pm$ sd, p<0.05). Additionally, the body weight was increased, drinking water volume was decreased non-significantly by the treatment of ginseng extract. These results suggest that ginseng can also inhibit the formation of glycated protein by other mechanisms which are not related with hyoglyemic effect of ginseng.

• Mass Spectrometry Letters
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• v.5 no.2
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• pp.52-56
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• 2014

Glycated hemoglobin (HbA1c) is used as an index of mean glycemia over prolonged periods. This study describes an optimization of enzyme digestion conditions for quantification of non-glycated hemoglobin (HbA0) and HbA1c as diagnostic markers of diabetes mellitus. Both HbA0 and HbA1c were quantitatively determined followed by enzyme digestion using isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) with synthesized N-terminal hexapeptides as standards and synthesized isotope labeled hexapeptides as internal standards. Prior to quantification, each peptide was additionally quantified by amino acid composition analysis using ID-LC-MS/MS via acid hydrolysis. Each parameter was considered strictly as a means to improve digestion efficiency and repeatability. Digestion of hemoglobin was optimized when using 100 mM ammonium acetate (pH 4.2) and a Glu-C-to-HbA1c ratio of 1:50 at $37^{\circ}C$ for 20 h. Quantification was satisfactorily reproducible with a 2.6% relative standard deviation. These conditions were recommended for a primary reference method of HbA1c quantification and for the certification of HbA1c reference material.

• The Korean Journal of Pharmacology
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• v.31 no.1 s.57
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• pp.135-140
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• 1995

Glycation occurs by covalent binding between the carbonyl group of monosaccharides and the epsilon amino group of amino acid. It can alter the physiological function of proteins and causes the development of diabetic complications. In this study, the influence of glycation on protein binding of warfarin and dansylsarcosine was studied by equilibrium dialysis which was performed for 3 hours at $37^{\circ}C$ in the water bath. The high glycated albumin which contained $50{\pm}16%$ of glycated albumin bound less than natural albumin which contained $8.5{\pm}5.28%$ of glycated albumin, if drugs concentration were more than the albumin concentration. But only warfarin binding showed a significant difference of 6% (P<0.05) when the molar concentration ratio of warfarin per albumin was 3. In consideration of low therapeutic concentrations, low glycated albumin concentrations in the body, and rapid elimination of excessive free drugs, these small increaes of free warfarin concentrations by glycation of albumin are not considered as risk. factors for drug intoxication for diabetics, if renal functions are intact.