• Preventive Nutrition and Food Science
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• 제17권2호
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• pp.116-121
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• 2012

Water extract from Pinus densiflora (WPD) was investigated for its antioxidant activity and its ability to provide protection from DNA damage. A series of antioxidant assays, including a 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging assay, a reducing power assay, a metal-chelating assay, a superoxide radical scavenging assay, and a nitrite scavenging ability, as well as a DNA damage protection assay were performed. Total phenolic content was found to be 211.32 mg Tan/g WPD. The extract scavenged 50% DPPH free radical at a concentration of 21.35 ${\mu}g/mL$. At that same concentration, the reducing power ability of WPD was higher than that of ${\alpha}$-tocopherol. The extract chelated 68.9% ferrous ion at the concentration of 4 mg/mL. WPD showed better nitrite scavenging effect at the lower pH. Meanwhile, WPD exhibited a strong capability for DNA damage protection at 1 mg/mL concentration. Taken together, these data suggest water extract from Pinus densiflora could be used as a suitable natural antioxidant.

• The Plant Pathology Journal
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• 제33권2호
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• pp.184-192
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• 2017

Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

• Natural Product Sciences
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• 제23권1호
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• pp.46-52
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• 2017

The aim of this study was to evaluate the anti-Helicobacter pylori activity of fractions and major aglycon compounds (baicalein, chrysin, oroxylin A, wogonin) of Scutellariae Radix. Minimum inhibitory concentration (MIC) measurement, DPPH radical-scavenging assay, DNA protection assay, and urease inhibition analysis were performed. The ethyl acetate (EtOAc) fraction showed the potent anti-Helicobacter activity, and therefore, compounds in the EtOAc fraction were subjected to further assay. The MICs of chrysin, oroxylin A, and wogonin against Helicobacter pylori 26695 were 6.25, 12.5 and $25{\mu}g/mL$, respectively. Baicalein exhibited the most effective DPPH radical-scavenging activity. DNA protection using Fenton reaction, chrysin, oroxylin A, and wogonin showed effective DNA protective effect. This result was also confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Regarding Jack bean urease (0.5 mg/mL, 50 unit/mg) inhibition, 20 mM ofbaicalein and chrysin inhibited urease activity by 88.2% and 72.5%, respectively.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6535-6539
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• 2015

This study was designed to assess, whether a new chemotherapeutic microtubule inhibitor, Epothilone B (EpoB, Patupilone), can induce DNA damage in normal ovarian cells (MM14.Ov), and to evaluate if such damage could be repaired. The changes were compared with the effect of paclitaxel (PTX) commonly employed in the clinic. The alkaline comet assay technique and TUNEL assay were used. The kinetics of DNA damage formation and the level of apoptotic cells were determined after treatment with IC50 concentrations of EpoB and PTX. It was observed that PTX generated significantly higher apoptotic and genotoxic changes than EpoB. The peak was observed after 48 h of treatment when the DNA damage had a maximal level. The DNA damage induced by both tested drugs was almost completely repaired. As EpoB in normal cells causes less damage to DNA it might be a promising anticancer drug with potential for the treatment of ovarian tumors.

• Journal of Radiation Protection and Research
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• 제24권2호
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• pp.93-99
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• 1999

Alkaline single cell gel electrophoresis (SCGE) assay는 일명 혜성분석이라고 부르며 in vivo 와 in vitro 에서 많은 화학적, 생물학적인 인자에 의한 DNA 손상을 감지하는데 유용한 기법으로 각각의 세포에서 DNA 단일 가닥 절단과 알칼리에 약한 장소를 평가하는 새로운 방법으로 인정되고 있다. 단세포 겔 전기영동법 (SCGE)을 사용하여 복숭아씨 추출물이 방사선에 의하여 사람 림프구 DNA에 나타나는 손상을 보호하는 지 여부를 평가하였다. 복숭아씨 추출물로 10 분간 전처리한 림프구를 0, 0.1, 0.3, 0.5, 1.0, 2.0 Gy 의 방사선으로 조사하였고 방사선만을 조사한 림프구 실험군과 비교평가하였다. 혜성분석에서 DNA 가닥 절단에 대한 표식인 tail moment의 증가는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었으며 각각의 농도별로 복숭아씨 추출물이 처리된 림프구의 DNA 손상은 현저히 감소하였다. 단세포 겔 전기영동법을 통한 평가결과 복숭아씨 추출물은 방사선에 의한 림프구 DNA 손상에 대한 탁월한 방어효과를 나타내었다.

• The Plant Pathology Journal
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• 제35권6호
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• pp.654-661
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• 2019

The soybean cyst nematode, Heterodera glycines, is a major plant-parasitic nematode that has caused important economic losses to Korea's soybean production. Four species of cyst nematodes, H. schachtii, H. glycines, H. trifolii, and H. sojae, all belong to schachtii group are coexist in field soil in Korea. The rapid identification of the nematode is crucial for preventing crop damage and in decision making for controlling this nematode. This study aimed to develop a species-specific primer set for quantitative PCR (qPCR) assay of H. glycines. The specific primer set (HGF1 and HGR1) for H. glycines was designed based on the cytochrome c oxidase subunit I (COI) sequence of mitochondrial DNA. After optimization, it is possible to identify the H. glycines using a qPCR assay with DNA extracted from a single cyst and single second-stage juvenile (J2). The specificity was confirmed by the absence of SYBR fluorescent signals of three other Heterodera species. A serial dilution of DNA extracted from a single cyst was obtained for the sensitivity test. The result showed that the standard curve of the test had a highly significant linearity between DNA concentration and Ct value (R² = 0.996, slope = -3.49) and that the detection limit concentration of DNA of the primer set was 10 pg of DNA per reaction. Our findings suggested that H. glycines could be distinguished from H. sojae and other Heterodera species when a qPCR assay is used with a specific primer set.

• Journal of Nutrition and Health
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• 제37권6호
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• pp.440-447
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• 2004

In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100％ orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50％ in vitamin C, 32％ in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55％ protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60％ with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.

• 한국환경과학회지
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• 제23권3호
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• pp.483-492
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• 2014

DNA damage such as genotoxicity was identified with comet assay, which blood cell of a marine parrot fish (Oplegnathus fasciatus) was exposed to an acidified seawater, lowered pH gradient making of $CO_2$ gas. The gradient of pH were 8.22, 8.03, 7.81, 7.55 with control as HBSS solution with pH 7.4. DNA tail moment of fish blood cell was $0.548{\pm}0.071$ exposed seawater of pH 8.22 condition, on the other hand, DNA tail moment $1.601{\pm}0.197$ exposed acidified seawater of pH 7.55 lowest condition. The approximate difference with level of DNA damage was 2.9 times between highest and lowest of pH. DNA damage with decreasing pH was significantly increased with DNA tail moment on blood cell of marine fish (ANOVA, p < 0.001). Ocean acidification, especially inducing the leakage of sequestered $CO_2$ in geological structure is a consequence from the burning of fossil fuels, and long term effects on marine habitats and organisms are not fully investigated. The physiological effects on adult fish species are even less known. This result shown that the potential of dissolved $CO_2$ in seawater was revealed to induce the toxic effect on genotoxicity such as DNA breakage.

• Preventive Nutrition and Food Science
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• 제14권4호
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• pp.277-282
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• 2009

The antioxidant potency of methanolic extract of Duchesnea indica (MDI; Indian strawberry) was investigated by employing various established in vitro systems, such as total phenolic content, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, reducing power assay, metal chelating assay, superoxide radical scavenging activity and protective ability of DNA damage and protein oxidation. MDI inhibited metal chelating by 75.57% at 2 mg/mL, scavenged 50% DPPH free radical at 29.13 ${\mu}$g/mL, and eliminated approximately 46.21% superoxide radical at the concentration of 1 mg/mL. In addition, MDI showed strong ability on reducing power, DNA damage protection and protein oxidation protection. Overall, results suggested that MDI might be beneficial as a potent antioxidant and effectively employed as an ingredient in food applications.

• Journal of Nutrition and Health
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• 제49권5호
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• pp.277-287
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• 2016

본 연구는 제5기 2차년도 국민건강영양조사 결과를 활용하여 한식 식품군의 총 페놀 함량, in vitro 항산화활성 및 인체세포를 이용한 ex vivo DNA 손상 감소효과를 비교하고, 각 지표간의 상관성을 분석하며, 한식의 총 식사 항산화능에 대한 각 식품군의 기여도를 알아보기 위해 수행되었다. 제5기 2차년도 국민건강영양조사 결과를 바탕으로 한식의 식물성 식품을 10가지 식품군 (곡류, 과일류, 채소류, 견과류, 김치류, 해조류, 감자류, 버섯류, 두류, 오일류)으로 분류한 후 각 식품군별로 총 섭취량의 1% 이상 섭취한 식품 84종을 한식의 식물성 식품으로 최종 선정하였다. 각 식품군의 총 페놀함량을 측정하였고, DPPH radical scavenging assay, TEAC assay, $ORAC_{ROO{\cdot}}$ assay를 사용하여 in vitro 항산화능을 측정하였다. 한식의 식품군별 항산화능 (dietary antioxidant capacity, DAC)은 in vitro 항산화활성 평균값과 각 식품군의 1일 섭취량을 고려하여 계산하였고 한식 TDAC는 각 식품군의 DAC로의 합으로 구하였으며, TDAC에 대한 각 식품군 항산화능의 기여도를 평가하였다. 인체 임파구에서의 ex vivo DNA 손상 정도는 comet assay를 사용하여 평가하였다. 한식 식품군의 총 페놀함량은 버섯류, 과일류, 채소류, 해조류, 김치류 등의 순으로 높았으며, 3가지 in vitro 실험법을 평균한 식품군의 항산화활성 순위는 버섯류, 해조류, 채소류, 김치류, 과일류 등의 순이었다. 각 식품군의 항산화활성에 식품섭취량을 고려하여 계산한 한식의 TDAC에 대한 식품군의 항산화능 기여도는 곡류가 33.4%로 가장 높았으며, 과일류 (23.9%), 채소류 (12.7%), 김치류 (11.2%) 등의 순으로 나타났다. 인체 임파구에서 ex vivo DNA 손상 보호효과는 버섯류에서 가장 높았으며, 그 다음 채소류, 과일류, 해조류, 김치류의 순으로 나타났다. 각 식품군의 페놀함량과 in vitro 항산화 활성, 그리고 ex vivo DNA 보호효과의 순위가 비슷하게 나타났으며 각 지표간의 상관성은 매우 높았다. 한식 식품군 중 버섯류, 과일류, 채소류, 해조류에서 총 페놀함량과 항산화 활성, DNA 손상 보호효과가 높게 나타났다. 각 식품군의 총 페놀함량과 in vitro 항산화 활성, ex vivo DNA 보호효과 지표 간의 상관성은 매우 높았다. 한식의 TDAC에 대한 식품군별 항산화능 기여도는 곡류가 가장 높았고, 그 다음이 과일류, 채소류, 김치류의 순이었다. 이러한 결과는 앞으로 한식의 우수성을 항산화 측면에서 밝히는데 매우 중요한 기초자료로 활용될 수 있을 것이다.