Evaluation of 8-hydroxydeoxyguanosine, a typical oxidative DNA damage, in human leukocytes

Takeuchi, Torn; Nakajima, Masahiro; Ohta, Yukinobu; Mure, Kanae; Takeshita, Tatsuya; Morimoto, Kanehisa

doi:10.1093/carcin/15.8.1519

Cited by 76 publications

(30 citation statements)

References 18 publications

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“…In these detectors, the working electrodes are primarily carbon electrodes; silver chloride electrodes are usually used as reference electrodes. 37,[39][40][41][42][43][44][45][46][47][48] Only the mobile phases with the ability to conduct electric current can be used with HPLC-ECD.…”

Section: Direct Methodsmentioning

confidence: 99%

Analytics of oxidative stress markers in the early diagnosis of oxygen DNA damage

Dąbrowska¹,

Wiczkowski²

2017

Adv Clin Exp Med

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Under homeostatic conditions, an equilibrium state between amounts of free radicals formed and their scavenging is observed. Free radicals are destructive only when present in excess. Pathological changes within cells and tissues can result from a persistent excess of free radicals. Living organisms are increasingly exposed to oxidative stress, resulting in oxidative DNA modifications. One such modification is 8-hydroxy-2'-deoxyguanosine (8-OHdG). It is considered a biomarker of oxidative stress and oxidative DNA damage. It has been found both in physiological fluids and in cells. This paper presents methods found in the literature for determining 8-OHdG expression in various kinds of biological material -blood, urine or liver homogenates. Methods for determining the biomarker expression have been grouped into direct and indirect methods, and the various levels of 8-hydroxy-2'-deoxyguanosine that can be determined by the different techniques are presented. The basic pros and cons of the various techniques are also discussed.

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Section: Direct Methodsmentioning

confidence: 99%

Analytics of oxidative stress markers in the early diagnosis of oxygen DNA damage

Dąbrowska¹,

Wiczkowski²

2017

Adv Clin Exp Med

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“…8-OHgua and 7,8-dihydro-8-oxo-guanosine are abundant in urine, but they may originate in cellular RNA as well as in DNA, and they may be derived from nucleic acids in the diet (10 ..l with smoking (13,14). Therapeutic doses of ionizing radiation led to severalfold increases in 8-OHdG in leukocytes (15); however, others have reported no correlation of 8-OHdG in DNA with smoking or with age (16).…”

Section: Introductionmentioning

confidence: 99%

Oxidative Damage to DNA: Do We Have a Reliable Biomarker?

Collins¹,

Dušinská²,

Gedik³

et al. 1996

Environmental Health Perspectives

156

151

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“…The reaction was stopped by the addition of 1ml cold ethanol, and the DNA product was precipitated at Ϫ80°C for 1 h. The precipitated DNA was then dissolved in 0.3 M sodium acetate, and again precipitated with ethanol, rinsed in cold ethanol for 2 times, and dried. The amount of 8-OHdG was determined according to Takeuchi et al 15) Briefly, DNA was heat-denatured and then digested sequentially with nuclease P1 and alkaline phosphatase. Quantities of 8-OHdG and deoxyguanosine (dG) were determined by high performance liquid chromatography with electrochemical detection and UV absorp-tion.…”

Section: Synthesis Of Dihydropyrazine Derivativesmentioning

confidence: 99%

Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

Kashige

Takeuchi

Matsumoto

et al. 2005

Biological & Pharmaceutical Bulletin

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Dihydropyrazines (DHPs) revealed the single strand-cleavage 1) of covalently closed circular DNA of plasmid pBR322, especially in the presence of Cu 2ϩ . This cleavage was determined to be initiated by the radical species generated from DHPs. Therefore, further investigation of the reactivity of DHPs continues our laboratory.DNA damage is closely related to the variety of biological phenomena such as mutagenesis, carcinogenesis, aging and radiation effects. A better understanding of the mechanism of the biological effects caused by DHPs is of great interest, because DHPs were derived from sugar and universally existed in human body, and consequently might caused various internal injuries in vivo.2) The chemical reactivity 3,4) and the biological effects 2,5) of DHPs have been elucidated. The electron spin resonance (ESR) spectrum of DHPs showed the cogeneration of four radical species 6) such as · OH, · OOH, · CHR 2 and · CR 3 at the same time. Furthermore, the specificity 7) of the nucleotide sequence DNA strand cleavage sites produced by DHPs has been identified. However, whether oxygen radica'ls or carbon-centered radicals (C-radicals) primarily involved in the DNA strand-cleavage remains unclear. There are a number of reports of DNA strand-cleavage due to reactive oxygen species, including hydroxyl radical, however, there have been few reports related to C-radical. [8][9][10][11] The DNA damage caused by the copper-peroxide complex provides a unique example. 12) Herein, we use the results of DNA strand-cleavage activity and the ESR spectra with a spin-trapping agent to propose that the radical species that attacked the DNA strand formed 8-hydroxydeoxyguanosine (8-OHdG). MATERIALS AND METHODS Synthesis of Dihydropyrazine DerivativesThe dihydropyrazine derivatives ( Fig. 1) employed were all synthesized by condensation of diketones and diamines. 2,3-Dihydro-5,6-dimethylpyrazine (DHP-1), 2,3-dihydro-2,5,6-trimethylpyrazine (DHP-2) and 3-hydro-2,2,5,6-tetramethylpyrazine (DHP-3) were synthesized by the method of Yamaguchi et al. 1) 2,3,-Dihydro-5-methy, 6-phenylpyrazine 13) (DHP-4) was also synthesized by similar method.Assay of DNA Strand-Breaking Activity The method of assaying the DNA strand-cleavage activity of DHPs, using a covalently closed circular duplex DNA of plasmid pBR32 (ccc-DNA) was previously described. 14)ESR Spectroscopy of Dihydropyrazines The ESR spectra were recorded on a JES-FA200 spectrometer (JEOL Co., Tokyo) using a Mn 2ϩ marker as an external standard, and an ES-LC12 flat cell (JEOL Co., Tokyo). The spectra were measured in a 50 mM Tris-HCl buffer (pH 7.1) using 5,5-dimethy-1-pyrroline N-oxide (DMPO) as a spin trapping agent, according to previous paper. 6) The relative values were summarized in Table 3. The instrumental condition were: field center 335.9 mT, scan width Ϯ5 mT, modulation frequency 100 kHz, modulation width 0.14 mT, time constant 0.3 s, amplitude 7ϫ100, microwave power 10 mW, and microwave frequency 9.427 GHz. The spectra were recorded at 30 min after mixing.The Measureme...

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Evaluation of 8-hydroxydeoxyguanosine, a typical oxidative DNA damage, in human leukocytes

Cited by 76 publications

References 18 publications

Analytics of oxidative stress markers in the early diagnosis of oxygen DNA damage

Analytics of oxidative stress markers in the early diagnosis of oxygen DNA damage

Oxidative Damage to DNA: Do We Have a Reliable Biomarker?

Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

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