• 제목/요약/키워드: Glycated

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Identification of glycated peptides in human serum using LC-MS/MS: A comparison of data-dependent acquisition and Parallel reaction monitoring

  • Seonghyeon Cho;Yejin Jeon;Van-An Duong;Jong-Moon Park;Hookeun Lee
    • 분석과학
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    • 제36권1호
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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.

금 나노입자를 이용한 새로운 당화혈색소의 검출 기술 (Novel Detection Technology for Glycated Hemoglobin using Gold Nanoparticles)

  • 이수석
    • 센서학회지
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    • 제25권6호
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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.

제2형 당뇨병 환자에서 자가 인식한 치은 출혈 및 구취와 당화혈색소 간의 연관성 (Association between self-assessed gingival bleeding and halitosis, and glycated hemoglobin levels in patients with diabetes)

  • 최준선
    • 한국치위생학회지
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    • 제20권1호
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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.

Glycation of Copper, Zinc-Superoxide Dismutase and its Effect on the Thiol-Metal Catalyzed Oxidation Mediated DNA Damage

  • Park, Jeen-Woo;Lee, Soo-Min
    • BMB Reports
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    • 제28권3호
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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.

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An Analysis of the proteomics approach to the glycated peptides of human milk

  • Cho, Seonghyeon;Park, Jong-Moon;Lee, Hookeun;Song, Jun Hwan;Kang, Nam Mi
    • 분석과학
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    • 제35권1호
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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.

The Src/PLC/PKC/MEK/ERK Signaling Pathway Is Involved in Aortic Smooth Muscle Cell Proliferation induced by Glycated LDL

  • Cho, Hyun-Mi;Choi, Sung Hee;Hwang, Ki-Chul;Oh, Sue-Young;Kim, Ho-Gyung;Yoon, Deok-Hyo;Choi, Myung-Ae;Lim, So Yeon;Song, Heesang;Jang, Yangsoo;Kim, Tae Woong
    • Molecules and Cells
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    • 제19권1호
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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 $Ca^{2+}$.

당화단백질의 형성에 미치는 인삼의 효과 (Effects of Ginseng on the Formation of Glycated Protein)

  • 맹성호;전강웅;배진우
    • Journal of Ginseng Research
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    • 제26권4호
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    • pp.173-177
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    • 2002
  • 당화헤모글로빈의 형성을 포함한 당뇨병에 대한 인삼의 보호작용에 대하여 조사하고자 인산완충용액으로 포도당(1.5 g/dl), 헤모글로빈(10 g/dl) 및 홍삼액기스(0.5 g/dl)의 혼합용액(1 : 1 : 1, v/v/v)을 생산하여 시험관에 옮긴 후 37$^{\circ}C$에서 5일간 배양하였다. 또한 웅성 백서를 각 군당 20두씩 할당하여 건강한 군과 streptozotocin으로 유발된 당뇨군으로 나누고 이를 다시 인삼투여군과 그 대조군으로 나누어 홍삼액기스를 음용수에 용해하여 100 mg/kg/day의 용량을 3개월 동안 투여하였다. 혈당의 농도와 당화헤모글로빈의 비율은 시판되는 키트를 사용하여 측정하였다. 홍삼액기스를 첨가한 시험관의 당화헤모글로빈이 첨가하지않는 대조군에 비하여 유의하게 감소되었으며(12.17$\pm$1.01% : 15.9$\pm$ 1.95%, p<0.01) 홍삼액기스를 투여한 당뇨쥐의 혈당은 그 대조군에 비하여 유의한 차이가 없음에도 불구하고 3개월 후의 당화헤모글로빈의 비율이 유의하게 감소되었다(15.1$\pm$2.06% : 20.1$\pm$2.9%, p<0.05). 또한 3개월 후 홍삼액기스를 투여한 당뇨쥐는 그 대조군에 비하여 체중이 증가 되었으며 음수량이 감소 되었다. 이런 결과들로 보아 인삼은 당화단백질의 형성을 저혈당 작용이 아닌 다른 방법으로도 억제할 수 있을 것으로 생각한다.

Optimization of Enzyme Digestion Conditions for Quantification of Glycated Hemoglobin Using Isotope Dilution Liquid Chromatography-Tandem Mass Spectrometry

  • Jeong, Ji-Seon
    • Mass Spectrometry Letters
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    • 제5권2호
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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.

알부민 Gylcation이 약물의 단백질결합에 미치는 영향 (Influence of Albumin Glycation on the Protein Binding of Drugs)

  • 배진우
    • 대한약리학회지
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    • 제31권1호
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    • pp.135-140
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    • 1995
  • Glycation이란 단당류의 카보릴기와 아미노산의 입실론 아미노기가 공유결합에 의하여 형성되는 반응으로 이는 단백질의 생리적 기능을 변화시키며 아울러 당뇨합병증을 유발한다. 본 연구에서는 warfarin과 dansylsarcosine의 단백질결합에 미치는 glycation의 영향을 평형투석법을 이용하여 연구하였으며 평형투석은 섭씨 37도의 진탕수조에서 3시간 동안 실시하였다. 약물의 농도가 알부민의 농도보다 높을 경우, $50{\pm}16%$가 glycation된 알부민은 $8.5{\pm}5.28%$ glycation 알부민을 함유한 정상알부민에 비해 약물과의 결합도가 낮았으나 warfarin의 농도가 알부민의 3배가 될 경우에만 유의성이 인정되었고(P<0.05) 6%의 차이를 보였다. 본 실험에 나타난 glycation에 의한 유리약물의 미미한 상승효과는 glycated albumin 농도가 낮은 생체내의 여건과, 실제로 사용되는 약물의 적정 치료농도가 낮고 또한 과도한 유리약물은 신장을 통하여 신속히 배설되는 이유로 당뇨환자의 신기능 손상이 없는한 glycation에 의한 유리약물의 상승은 약물중독의 위험요소로 작용되지 않으리라 생각된다.

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