This study was investigated to test whether the zygote recognized the topoisomerase II beta (TOP2B) mediated DNA fragmentation in epididymal spermatozoa or the nuclease degradation in vas deferens spermatozoa by testing for the presence of gammaH2AX (γH2AX). The γH2AX is phosphorylation of histone protein H2AX on serine 139 occurs at sites flanking DNA double-stranded breaks (DSBs). The presence of γH2AX in the pronuclei of mouse zygotes which were injected with DNA broke epididymal spermatozoa was tested by immunohistochemistry at 5 and 9 h post fertilization, respectively. Paternal pronuclei that arose from epididymal spermatozoa treated with divalent cations did not stain for γH2AX at 5 h. On the other hand, in embryos injected with vas deferences spermatozoa that had been treated with divalent cations, γH2AX was only present in paternal pronuclei, and not the maternal pronuclei at 5 h. Interestingly, both pronuclei stained positively for γH2AX for all treatments and controls at 9 h after sperm injection. In conclusion, the embryos recognize DNA that is damaged by nuclease, but not by TOP2B because H2AX in phosphorylated in paternal pronuclei resulting from spermatozoa treated with fragmented DNA from vas deferens spermatozoa treated with divalent cations, but not from epididymal spermatozoa treated the same way.

The effects of extracts from Lentinula edodes (L. edodes) and Aquilariae agallocha (A. agallocha) on the DNA damage response in ultraviolet A (UVA)-exposed HaCaT cells and on the allergic contact dermatitis caused by 2,4-dinitro-chlorobezene (DNCB) were investigated. When UVA-exposed cells were incubated for 24 hours in medium containing L. edodes or A. agallocha extract, the level of 8-OHdG and CPD decreased in a concentration-dependent manner. The combined treatment with both extracts potentiated the decrease in UVA-induced 8-OHdG and CPD levels as compared with those following treatment with a single extract. In addition, the two extracts showed preventive effects against the UVA-induced reduction in collagen levels. Furthermore, the blood levels of IgE, IL-6, and histamine decreased more significantly upon combined treatment with L. edodes and A. agallocha extracts as compared with those following treatment with single extracts in DNCB-induced allergic contact dermatitis in the ICR mouse. The results of the present study suggest that the components with in the extracts of L. edodes and A. agallocha can help to prevent of UVA-induced genomic instability via a decrease in DNA damage, and to decrease the DNCB-induced allergic dermatitis via modulation of relevant proteins including IgE and IL-6. Further study is needed to clarify the purified components related to the preventative effects of the two extracts against UVA- or DNCB-induced genomic damage.

본 연구는 식용 버섯의 조리방법에 따른 항산화 생리활성의 평가를 위해 수행되었으며, 산화적 스트레스에 의한 DNA 손상 감소 효과를 통해 조리방법을 달리한 버섯 추출물의 유전독성학적 방호효과를 살펴보았다. Human lymphocyte에 조리방법을 달리한 3가지 버섯(느타리, 팽이, 표고)의 추출물을 처리하고, hydrogen peroxide(H2O2)로 산화적 손상을 준 후, DNA 감소 효과를 Comet assay로 평가한 결과, 모든 시료군에서 산화적 손상에 의한 DNA 손상 감소 효과를 나타냈다. 3가지 버섯 모두 비조리군이 조리군보다 높은 효과를 나타냈는데, 이는 조리과정에 의한 페놀성 화합물의 감소로 인한 것으로 보이며, 조리군 중에서 볶기와 전이 비교적 낮은 DNA 손상 감소 효과를 나타낸 것은 조리 시 첨가되었던 대두유의 가열 산화에 의한 것으로 사료된다. 결론적으로, 조리된 버섯은 생버섯에 비해 산화적 스트레스에 의한 DNA 손상 감소효과가 낮으나, 양성 대조군과 비교하였을 때 손상을 유의적으로 감소시킨 것으로 나타났다. 또한, 본 연구에서 사용한 네 가지 조리법(굽기, 데치기, 볶기, 전) 중 DNA 손상 감소에 효과적인 조리법은 대두유를 사용하지 않은 굽기와 데치기인 것으로 판단된다.

수은이 DNA 수복에 미치는 영향을 알아보기 위해 E. fetida를 염화수은(II)과 이온화 방사선에 순차적으로 노출시킨 후, 단세포 겔 전기영동 기법을 이용하여 DNA의 손상 수준과 방사선 조사 후 시간 경과에 따른 수복 양상을 관찰하였다. 염화수은(II)의 농도를 40 mg kg-1으로 하여 48시간 동안 in vivo 노출 시험을 수행한 뒤 20Gy의 감마선을 조사한 결과, 시간이 지날수록 대체로 DNA 손상의 수준이 감소했다. 이온화

E. fetida를 방사선과 수은에 각각 노출시킨 후, 체강세포를 추출하고 단세포 겔 전기영동 기법을 이용하여 DNA의 손상정도와 시간의 경과에 따른 수복 양상을 평가해 보았다. 그 결과, 방사선 조사 후의 시간이 경과할수록 대체로 DNA 손상정도가 감소했으며, 12시간 내에 모든 실험군의 DNA가 완전히 수복되었다. 정확한 수복 완료 시간을 알아보기 위해 OTM 값을 대조군과 비교해 보면 2.5와 5Gy는 방사선 조사 후 약 2시간, 10과 20 Gy

각종 유전독성학적 물질로 인한 생물체내의 영향을 평가해보기 위해 E. fetida를 대상으로 본 연구를 수행하였다. 염화수은에 대한 DNA 손상을 알아보는 실험에서는 노출 시간에 상관없이 노출 농도에 비례한 유전자의 손상이 나타났다. 방사선이 지렁이의 DNA 손상에 미치는 영향을 알아본 실험에서도 역시 방사선 총 선량의 증가에 따라 DNA 손상이 증가하는 경향을 보였다. 염화수은에 48시간 동안 노출시키고 방사선을 조사한 지렁이의 세포를 comet as

UV는 방사선이나, 엑스선과 함께 DNA 손상을 일으켜 돌연변이를 일으키는 돌연변이원으로 잘 알려져 있다. 이러한 돌연변이원인 UV나 방사선을 이용해 해충을 방제하려는 노력들이 많이 이루어지고 있는데, 본 연구에서는 UV-A의 해충방제 이용가능성을 조사하기 위해, 벼의 엽육을 전체적으로 식해해서 피 해를 주는 혹명나방의 번데기 단계에 장파형 자외선인 UV-A(360nm)을 실험 실내에서 1시간, 6시간동안 조사하여 이들의 우화율, 생존율, 산란수, 성비 등 을 조사한 결과, 우화율에 있어 30%이상의 감소를 나타내었으며, 무처리구에 비해 성비가 6.4:3.6, 5.4:4.6으로 수컷이 적게 발생하며 성비에 있어 불균형을 나타내는 것을 확인하였다. 또한 자외선 조사 후 정상적으로 우화한 성충이라 하더라도, 생존율이 낮게 나타났으며, 암컷 한 마리당 산란 수도 낮게 나타나 자외선이 혹명나방의 생태학적 특성에 부정적인 영향을 미치는 것으로 확인 되었다. 또한 자외선 처리에 의한 DNA손상정도를 확인하기 위해 Comet assay 를 실시한 결과, 처리후 생존하여 우화한 성충의 일부 개체에서 Comet tail이 확인되며 DNA가 자외선에 의해 손상된 것을 확인할 수 있었다. 따라서 혹명 나방 방제시 논주변이나 포장주변에 자외선 등을 설치하여 해충의 우화율이 나 성비불균형, 핵산의 손상을 유도함으로 다음세대의 해충개체군 크기를 감 소시킬 수 있는 측면에서의 해충방제를 기대할 수 있을 것으로 여겨진다.

세포에 미치는 염화수은(II)과 이온화 방사선의 영향과 수은 처리 전 후 방사선 조사 시 그 상호 작용에 관해 알아보고자 본 연구를 수행하였다. 염화수은(II)의 독성정도를 알아보기 위하여 사람의 자궁암 세포에 농도별로 염화수은(II)을 처리하였다. 세포의 생존율은 3가지 농도(1,0. 1,0. 0.01 μM)모두에서 유의하게 감소하였으며 이미 0.1 μM에서 약 73%의 생존율이 감소하는 것으로 나타났다. 염화수은(II)과 방

Cell response to genotoxic agents is complex and involves the participation of different classes of genes including cell cycle control, DNA repair and apoptosis. In this report, we presented a approach to characterize the cellular functions associated wit

양강추출물(Alpinia officinarum, 70% ethanol extract)과 함유 주성분인 galangin은 프리라디칼소거작용과 지질과산화억제활성을 나타내었으며, H_2O_2 또는 KO_2유도 세포독성에 대해서도 억제적으로 작용하여 세포보호효과를 나타내었다. DNA single strand breakage와 같은 산화적 스트레스에 대해서도 보호작용을 하고 있다. 또한, 마우스 소핵시험에 의하여 adriamycin과 같은 superoxide유발물질에 의한 염색체 수준에서의 손상에 대해서도 억제효과를 나타내었다. 양강추출물은 galangin을 유효성분으로하여 산소프리라디칼들에 의한 산화적 손상에 억제적으로 작용하는 기전으로 산화적 스트레스에 대한 항노화, 암예방제로서의 응용가능성이 높은 추출물로서 향후 기능성 식품으로서의 응용가능성이 높을 것으로 판단되었다.

병해충을 막기 위해 농업용 살충제가 광범위하게 사용되고 있다. 농약사용에 따른 생물학적 위해가 우려되며 농약이 또 다른 환경유해요인과 인체에 상승적으로 작용할 경우 농업재해로 이어질 가능성이 있다. 다양한 인자에 의한 DNA손상을 감지하는데 유용한 단세포 겔 전기영동법을 이용하여 살충제와 방사선에 의한 사람 림프구 DNA손상을 평가하였다. 각기 다른 농도로 살충제를 10분간 전처리한 림프구와 정상 림프구에 0-2.0 Gy의 방사선으로 조사한 다음 DNA

Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatac agent. This study was devised to develop a chemopreventive agent from panax ginseng to be able to suppress the genotoxicity and oxidative damage by reactive oxygen species, which are involved with cancer or aging. Ginseng petroleum ether extract (GPE) and one of its fraction, P2, showed an antioxidative effect on the lipid peroxidation of ethyl linoleate with Fenton's reagents and free radical scavenging effect to 1,1-diphenyl-2-picryl hydrazil (DPPH) radical generation. They also showed the suppressive effect of H₂O₂ or KO₂ induced DNA damage by single cell gel electrophoresis (SCGE). Results from our study indicate that GPE and P2 are capable of protecting lipid peroxidation, and oxidative DNA damage. Therefore, GPE and P2 may be useful chemopreventive agents which are involved with cancer and aging.

This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at 50 μg/mL had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.

Background: Valeriana fauriei (Valerianaceae) has been used to as a traditional medicine to treat a variety of symptoms, including headache, insomnia, hypertension, and menstrual irregularity. However, the present study investigates the species' antioxidant activity and its inhibition of oxidative DNA damage, which have yet to be studied.Methods and Results:The antioxidant activity was assessed using radical scavenging assays with 1,1-diphenyl-2-picryl hydrazyl (DPPH) and, 2, 2'-azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) and a reducing power assay. The total phenol content was also analyzed, and phenolic compounds were detected using HPLC/UV, whereas the inhibitory effect of Valeriana fauriei on oxidative DNA damage was measured using φ-174 RF I plasmid DNA cleavage assay. The DPPH and ABTS radical scavenging activity were 75.17 ± 3.55% and 95.83 ± 0.63%, repectively, and the reducing power was 93.14 ± 1.74 at 200 μg/ml. The total phenol content was 10.24 ± 0.04 ㎎/g, whereas chlorogenic acid, catechin, caffeic acid and epicatechin were identified using HPLC/UV, and the φ-174 RF I plasmid DNA cleavage assay indicated that V. fauriei provided protection against oxidative damage.Conclusions:The results of the present study suggest that V. fauriei has powerful antioxidant activity that can provide protective effects against the oxidative DNA damage caused by free radicals. The species, therefore, provides a valuable resource for the development of natural pharmaceutical to treat aging, cancer, and degenerative diseases.

Background : Lactuca sativa (LS) is a member of lactuca genus and the Asteraceae family. To its usual purpose as an edible leafy vegetable, lettuce has had a number of uses in ancient times as a medicinal herb. Depending on the variety, lettuce is an excellent source vitamin K and vitanin A. Methods and Results : The objective of this study is to find out the antioxidant and oxidative DNA damage prevention capacity of LS for both water and ethanol extract. The total phenolic and flavonoid contents have been estimated in this study. The extracts have been tested to assess the 1, 1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2ʹ -azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and nitrite scavenging activity. We have also evaluated the reducing power activity of LS. LS extracts showed a good radical scavenging activity on DPPH and ABTS free radical as concentration dependent manner. The IC50 values on DPPH radical scavenging activity were 0.75 mg/mL and 0.54 mg/mL for water and ethanol extract respectively. ABTS radical scavenging activity ranges from 38-89 % for water extract and 33-96% for ethanol extract. Although ethanolic extract showed a higher radical scavenging activity as compared to the water extracts. All the extracts exhibited reducing power activity, dose dependently. Moreover, when the DNA was treated with the extracts, supercoiled DNA was restored in a concentration-dependent manner. Conclusion : According to the results, we suggest that LS contains considerable amount of phenolic and flavonoid content and could be used as a source of natural antioxidant substances and oxidative DNA damage preventer.

Antioxidant activity and inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Cone of Red Pine (Pinus densiflora) were investigated to find utilization of Cone, by-product of Red Pine, thrown out after berry shatter, as a new natural plant resource. Cone from P. densiflora was extracted with methanol (MeOH) and separated to petroleum ether, ethyl acetate and water fraction. Among them, ethyl acetate fraction was used. The antioxidant activity was conducted by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay, Fe 2+ chelating assay and reducing power assay. The inhibitory effect on oxidative DNA damage was determined by DNA cleavage assay using φX-174 RF I plasmid. The results of DPPH and ABTS radical scavenging activity at 200 ㎍/㎖ of extracts were 86.50% and 95.80% respectively, which were similar figures compared with L-ascorbic acid as control. Fe 2+ chelating activity was 77.96% and reducing power was 0.77 at 200 ㎍/㎖. Total phenolic component was 27.29±0.3 ㎎/g and Vitamin C content was 1.84±0.1 ㎎/g. Also ethyl acetate fraction from Cone has inhibitory effect, using φX-174 RF I plasmid on DNA cleavage assay. In conclusion, Cone, by-product of P. densiflora, showed high antioxidant activity and inhibitory effect on oxidative DNA damage. Therefore this study suggests Cone, useless by-product, can be developed as a new natural plant resource with lots of utilization such as an effective antioxidant, natural medicine, food, cosmetics and so on.

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 CO2 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 ± 0.071 exposed seawater of pH 8.22 condition, on the other hand, DNA tail moment 1.601 ± 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 CO2 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 CO2 in seawater was revealed to induce the toxic effect on genotoxicity such as DNA breakage.

Dieldrin, one of the organochlorine pesticides (OCPs), induced the damages in neuroblastoma cells and DNA damages in lymphocytes. The ethanol extracts of A. sessiliflorus leaves were examined for the suppressive effects on the dieldrin-induced cell damages. Moreover, the extract was used to test whether it might inhibit the oxidative DNA damage of lymphocytes using Comet assay. The cell and DNA damage by dieldrin were suppressed in vitro upon treating A. sessiliflorus extract. This result suggests that A. sessiliflorus extract might be useful to reduce dieldrin toxicity.