• The Korean Journal of Nuclear Medicine Technology
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• v.13 no.3
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• pp.143-146
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• 2009

Purpose: We use the centrifugation of refrigeration state in separation of blood serum, Anti-ds-DNA, Vitamin $B_{12}$/Folate and GAD-Ab assay. However, Cortisol urine and 25-Hydroxyvitamin $D_3$ ($25OHD_3$) are conducted centrifuge at room temperature. This is troublesome that change centrifugation temperature into room temperature due to using of most assays at cold temperature. Therefore when using centrifuge after extraction of Cortisol urine and $25OHD_3$, we conducted researches on effect of the centrifugation temperature in assay results. Materials and Methods: In Cortisol urine, add dichloromethane 1.0 mL in urine $500\;{\mu}L$, mix for 15 minutes, and then centrifuge for 8 minutes at 2600rpm. In $25OHD_3$ add acetonitrile 0.5 mL in serum $200\;{\mu}L$, and then centrifuge for 8 minutes at 2600rpm. Those experiments were conducted centrifuge at room temperature and $4^{\circ}C$. And experiments conducted immediately after centrifugation at $4^{\circ}C$ and standing for 20 minutes after centrifugation $4^{\circ}C$. Results: In Cortisol urine, room temperature result in 1.93, 2.18, 2.43, 9.45, 14.2 (${\mu}g/dL$). Experiments of performing immediately after centrifuge at $4^{\circ}C$ result in 1.8, 2.0, 2.3, 8.1, 13.7 (${\mu}g/dL$). Experiments of performing after 20 minutes result in 2.1, 2.1, 2.7, 9.95, 14.35 (${\mu}g/dL$). On the other hand, the $25OHD_3$ tests conducted at room temperature result in 7.13, 26.6, 35.8, 48.2, 74.8 (ng/dL). Experiments were conducted immediately by pipetting after $4^{\circ}C$ centrifugation result in 7.53, 30.9, 40.3, 61.5, 89.1 (ng/dL) as results are higher than experiments at room temperature. The experiments that conducted centrifuge at $4^{\circ}C$ and then left at room temperature for 20 minutes result in 7.40, 32.4, 41.3, 51.6, 85.6 (ng/dL). Conclusions: Experiments were conducted by using centrifuge at $4^{\circ}C$ are higher or lower than room temperature. The differences between results of standing for 20 minutes after centrifuge at $4^{\circ}C$ and those of centrifuge at room temperature are less than conducting immediately. It is concerned that experiments conducted immediately after centrifuge at $4^{\circ}C$ are incorrect, because tubes become dim due to temperature differences between $4^{\circ}C$ and room temperature. Therefore, it is desirable to centrifuge at room temperature as manual and we should pipet promptly without stopping.

• Development and Reproduction
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• v.15 no.3
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• pp.205-213
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• 2011

Ethanol treatment during the brain growth spurt period has been known to induce the death of Purkinje cells. The underlying molecular mechanisms and the role of reactive oxygen species (ROS) in triggering ethanol-induced Purkinje cell death are, however, largely unresolved. We undertook TUNEL staining, western blotting assay and immunohistochemistry for the cleaved forms of caspase-3 and -9, with calbindin D28K double immunostaining to identify apoptotic Purkinje cells. The possibility of ROS-induced Purkinje cell death was immunohistochemically determined by using anti-8-hydroxy-2'deoxyguanosine (8-OHdG), a specific cellular marker for oxidative damage. The results show that Purkinje cell death of PD 5 rat cerebellum following ethanol administration is mediated by the activation of caspase-3 and -9. However, unexpectedly, TUNEL staining did not reveal any positive Purkinje cells while there were some TUNEL-positive cells in the internal and external granular layer. 8-OHdG was detected in the Purkinje cell layers at 8 h, peaked at 12-24 h, but not at 30 h post-ethanol treatment. No 8-0HdG immunoreactive cells were detected in the internal and external granular layer. The lobule specific 8-OHdG staining patterns following ethanol exposure are consistent with that of ethanol-induced Purkinje cell loss. Thus, we suggest that ethanol-induced Purkinje cell death may not occur by the classical apoptotic pathway and oxidative damage is involved in ethanol-induced Purkinje cell death in the developing cerebellum.

• Proceedings of the Korean Society of Toxicology Conference
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• 2003.10b
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• pp.187-187
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• 2003

This study evaluated the potential usefulness of urinary 1-hydroxypyrene glucuronide (1-OHPG) and 8-hydroxy-deoxyguanosine (8-OHdG) as biomarkers of the Asian Dust event. Urine samples were collected from 224 subjects (112 children and 112 their mothers) from Seoul (n=60), Inchon (n=104) and Pohang (n=60) in South Korea. (omitted)

• Biomolecules & Therapeutics
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• v.23 no.4
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• pp.374-378
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• 2015

The purpose of the present study was to investigate the effects of resveratrol supplementation on oxidative damage and lipid peroxidation induced by strenuous exercise in rats. The rats were randomly divided into five groups: a sedentary control group, an exercise control group, and three treatment exercise groups administered increasing doses of resveratrol (25, 50, and 100 mg/kg body weight). Resveratrol was administered by oral gavage once daily for four weeks. At the end of the four-week period, the rats performed a strenuous exercise on the treadmill, and the levels of lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA), 4-hydroxy-2-nonenal (4-HNE), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were measured. The results showed that resveratrol supplementation had protective effects against strenuous exercise-induced oxidative damage and lipid peroxidation by lowering the levels of LDH, CK, MDA, 4-HNE, and 8-OHdG in the serum or muscle of rats. These beneficial effects are probably owing to the inherent antioxidant activities of resveratrol.

• Toxicological Research
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• v.20 no.3
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• pp.187-193
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• 2004

This study aimed to determine the association of maternal oxidative stress and adverse pregnancy outcome with serum vitamin C concentration and a myeloperoxidase (MPO) genetic polymorphism during pregnancy. We investigated 450 pregnant women who visited Ewha Womans University Hospital for prenatal care during gestational weeks 24～28. During the second trimester, we measured serum vitamin C levels and urinary 8-hydroxyde-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) as an oxidative stress biomarker. We determined the presence of a maternal MPO polymorphism (G-to-A substitution at nucleotide 463) using a PCR-RFLP assay. We compared the level of oxidative stress and birth weight with the vitamin C concentration and the presence of the MPO polymorphism. The mean level of maternal oxidative stress tended to be higher and the birth weight lower for MPO type A/A than for types A/G and G/G. Vitamin C levels above the 75 percentiles were associated with reduced concentrations of urinary MDA and 8-OHdG but increased birth weight. Our data demonstrate that oxidative stress and neonatal birth weight are associated with the MPO genetic polymorphism, with the association modified by the maternal vita-min C levels.

• Environmental Mutagens and Carcinogens
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• v.25 no.3
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• pp.118-123
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• 2005

Nicotine has been implicated as a potential factor in the pathogenesis of human lung cancer, however its mechanism of action in the development of lung cancer remains largely unknown. To explore the role of nicotine in the development of lung cancer, we first investigated the effects of nicotine on the expression of tumor associated genes by treating Sprague-Dawley rats with nicotine (10 mg/kg) by gavage once daily for 10 days. We determined the expression of proteins and mRNAs of the ras, raf, myc, jun, fos oncogenes and p53, Rb tumor suppressor genes by Western and Northern blotting, respectively. We did not detect any changes on the levels of proteins and mRNAs of these tumor associated genes in the lung of Sprague-Dawley rats from 3 days to 12 weeks after the last treatment of nicotine, indicating that nicotine appears to have no effect on expression of these oncogenes and tumor suppressor genes at an early stage in multistage chemical carcinogenesis. In a second experiment, we investigated the possibility that 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) could be formed endogenously by treating with nicotine and sodium nitrite. We treated groups of Fischer 344 rats with nicotine ($60{\mu}mol/kg$) and sodium nitrite ($180{\mu}mol/kg$), nicotine, sodium nitrite and NNK (120 nmol/kg) alone by gavage once daily for 7 days, respectively and determined the 8-hydroxydeoxyguanosine (8-OHdG), as an indicator of NNK formation, in the lungs of rats 24 hours and 48 hours after the last treatment by HPLC/ECD method. We detect increased level of 8-OHdG in the lungs of rats treated with NNK, but in the case of nicotine plus sodium nitrite, nicotine and sodium nitrite alone we could not detected any changes of 8-OHdG, respectively.

• Journal of Periodontal and Implant Science
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• v.43 no.3
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• pp.121-129
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• 2013

Purpose: The present study has two aims; firstly, it attempts to verify the presence of oxidative stress by estimating the reactive oxygen species (ROS) levels in periodontal pockets ${\geq}5$ mm as compared to controls. The second aim is to evaluate the effect of lycopene as a locally delivered antioxidant gel on periodontal health and on the gingival crevicular fluid (GCF) levels of 8-hydroxydeoxyguanosine (8-OHdG), a marker of oxidative injury. Methods: Thirty-one subjects participated in this study. In the pretreatment phase, the ROS levels in pockets ${\geq}5$ mm were measured by flow cytometry. Three sites in each subject were randomly assigned into each of the following experimental groups: sham group, only scaling and root planing (SRP) was done; placebo group, local delivery of placebo gel after SRP; and lycopene group, local delivery of lycopene gel after SRP. Clinical parameters included recording site-specific measures of GCF 8-OHdG, plaque, gingivitis, probing depth, and clinical attachment level. Results: The gel, when delivered to the sites with oxidative stress, was effective in increasing clinical attachment and in reducing gingival inflammation, probing depth, and 8-OHdG levels as compared to the placebo and sham sites. Conclusions: From this trial conducted over a period of 6 months, it was found that locally delivered lycopene seems to be effective in reducing the measures of oxidative stress and periodontal disease.

• Journal of Nutrition and Health
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• v.55 no.3
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• pp.348-358
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• 2022

Purpose: Hyperglycemia accelerates the formation of advanced glycation end products (AGEs), a group of compounds formed via non-enzymatic glycation/glycoxidation. Type 2 diabetes mellitus (T2DM) is related to oxidative stress, resulting in some overgeneration of AGEs. The accumulation of AGEs in T2DM patients leads to increased inflammation, DNA damage, tissue damage, progression of diabetic microvascular disease, and nephropathy. Heme oxygenase-1 (HO-1) is an intracellular enzyme that catalyzes the oxidation of heme. Expression of HO-1 in the endothelium and in muscle monocytes/macrophages was upregulated upon exposure to reactive oxygen species or oxidized low-density lipoprotein. Cells activated by oxidative stress are reported to release HO-1 in the serum. In the current study, we discuss the oxidative status according to the level of AGEs and the association of HO-1 with AGEs or urinary DNA damage marker in type 2 diabetic Korean patients. Methods: This study enrolled 36 diabetic patients. Subjects were classified into two groups by serum AGEs level (Low AGEs group: < 0.85 ng/mL serum AGEs; High AGEs group: ≥ 0.85 ng/mL serum AGEs). Body composition was measured using bioelectrical impedance analysis. Blood and urinary parameters were measured using commercial kits. Results: No significant differences were observed in the general characteristics and body composition between the two groups. Serum HO-1 concentration was significantly higher in the High AGEs group than in the Low AGEs group. After adjustment of age and gender, a correlation was performed to assess the association between serum HO-1 and serum AGEs or urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG). Our results indicate that serum HO-1 is positively correlated with serum AGEs and urinary 8-OHdG. Conclusion: Taken together, our results indicate that in diabetes patients, a high level of HO-1 is associated with a high concentration of AGEs and 8-OHdG, probably reflecting a protective response against oxidative stress.

• Korean Security Journal
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• no.18
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• pp.21-37
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• 2009

The purpose of study were the effect of stress levels and duty type of guarder on 8-OHdG(hydroxydeoxyguanosin), Immunoglobin, T-cell, NK-cell and antioxidant enzyme. The subject of this study is people who have the guard's career over 2 years. A group of object sample consists of sixty guards carrying the guardbusiness. The group division and distribution of the number of persons a group divided into the guard group of international factor(N=23), private guard group(N=23),and the others(N=14) as a group following the guardbusiness. The group following a stress level divided into 3groups as their score of stress measure; low stress group(N=14), middle stress group(N=26),and high stress group(N=20). In the office type, the experiment sampling the blood, analysing result of following the office type and stress had a conclusion after measuring stress as the followings. The stress level in the office type of guard showed highly in the guard group of international factor(p<.05). Finally we knew that the guard have different stress levels in the office type, even the immune function and the activation of superoxide dismutase affected them in the stress level. Stress showing in the guard career can do them an injury so it is desirable for the guard themselves to participate in the activities settling their stresses and in the development of exercise programs.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.5
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• pp.1131-1136
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• 1999

The total contents of polyphenolics in potatoes measured by Folin Denis method were 42~76mg/100g in fresh weight. A major phenolic component contained in potato polyphenolics was identified as chlorogenic acid(3.6~15mg/100g in fresh weight). The antioxidative effects of potato polyphenolics and chlorogenic acid on the lipid peroxidation of liver microsome were studied in vivo and in vitro systems by measuring the formation of thiobarbituric acid reactive substances(TBARS) and the content of urine 8 hydroxy deoxyguanosine(OHdG). The TBARS contents of liver showed an increase in the cholesterol diet compared to those in the normal diet. This trend, however, was minimized when potato polyphenolics and chlorogenic acid were supplemented in the cholesterol diet. On the other hands, urinary 8 OHdG contents showed a marked increase with the supplementation of potato polyphenolics in the cholesterol diet. However, there was a trend of marked decrease by the supplementation of chlorogenic acid. In vitro study, potato poly phenolics and chlorogenic acid effectively inhibited the formation of TBARS in liver microsomal system in a dose dependent manners. These results suggest that potato polyphenolics exerts an antioxidative activity in cholesterol fed rat liver.