• Toxicological Research
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• 제16권3호
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• pp.221-227
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• 2000

A 6-sulfooxymethylbenzo[a]pyrene (SMBP), the ultimate metabolite of methyl-substituted benzo[a]pyrene (BP), has been found to be carcinogenic in mice. These properties may be attributable to its strong reactivity with cellular macromolecules such as DNA. However, serum and its major constituent albumin attenuated significantly the cytotoxicity and mutagenicity of 5MBP in bacterial and mammalian cell systems. This inhibitory activity of serum against 5MBP-induced cytotoxicity and mutagenicity in Chinese hamster V79 cells appears to be caused by the reduced macromolecular adducts such as DNA and proteins, but serum failed to reduce 5MBP binding to naked calf thymus DNA. A number of proteins in the serum could act as nucleophiles that are able to intercept reactive chemicals through covalent binding. Albumin present in the plasma seems to be one of major components responsible for direct binding with 5MBp, thereby reducing its reactivity to genetic materials. We here determined which fraction is preferential for 5MBP binding through fractionation of 5MBP-treated serum with ammonium sulfate. The albumin-containing fraction had slightly more affinity for 5MBP than the immunoglobulin-containing fraction. Our results indicate that the covalent modification of plasma proteins may reduce 5MBP-induced damage.

• 셀메드
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• 제3권3호
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• pp.25.1-25.8
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• 2013

Homoeopathically prepared Condurango 30C is traditionally used in amelioration of certain types of cancer by homeopathic practitioners. In this study, ability of Condurango 30C in amelioration of the conventional benzo[a]pyrene (BaP)-induced lung cancer in rat has been tested. After one month of scheduled oral feeding of BaP, lung cancer is routinely developed after four months in rats. Tumorbearing rats were then treated with Condurango 30C for the next one ($5^{th}$), two ($6^{th}$) and three ($7^{th}$) months, respectively, and sacrificed. Efficacy of post-cancer treatment by Condurango 30C was evaluated against controls (placebo) by different study parameters like: body and lung weights, number and diameter of lung tumour nodules, lung architecture, DNA damage, anti-oxidant activity and reactive oxygen species (ROS) accumulation. Administration of this homeopathic remedy caused increase of body weight and decrease of lung weight, decrease in number and diameter of lung tumour nodules, particularly after one and two months of drug treatment. BaP intoxication significantly increased lipid peroxidase (LPO) with concomitant decrease in activities of different antioxidants, while Condurango 30C administration certainly reduce their levels than normal and cancerous groups, notably after one and two months' of drug treatment. Condurango 30C showed capability to induce ROS-mediated cell death evidenced from the study of ROS activities at different time-points. Further, the remedy possibly achieved its anticancer goal through mediation of DNA-nicks that possibly led cancer cells to the apoptotic pathway. Thus, Condurango 30C has anticancer potential in BaP-induced lung cancer of rats via tissue damage recovery and ROS-mediated programmed cell death.

• 한국식품과학회지
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• 제19권3호
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• pp.212-219
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• 1987

재래종 적색 자두 (Prunus salicina)에서 효소를 추출하여 4종류의 polyphenol화합물과 반응시켜 얻어진 갈변반응 생성물에 대하여 Bacillus subtilis H17과 M45를 이용한 rec-assay와 Salmonella typhimurium TA98과 TA100 두 균주를 이용한 Ames test, 그리고 calfcthymus DNA를 이용하는 DNA절단시험을 이용하여 돌연변이원성과 돌연변이 억제작용을 조사하였다. 포자 rec-assay 에서는 pyrogallol, hydroxyhydroquinone, 3,4-dihydroxytoluene, chlorogenic acid 의 갈변반응 생성물은 모두 DNA손상능력이 없었으며 8가지 금속이온 중 ${Zn}^{2+}$과 ${Ni}^{2+}$의 첨가로 고초균 DNA손상에 약한 영향을 나타내었다. DNA절단시험 결과 4종류 갈변반응 생성물 모두 DNA절단작용이 없었으며 금속이온의 영향에 있어서는 pyrogallol 갈변반응 생성물이 ${Cu}^{2+}$의 영향을 받아 ${Cu}^{2+}$의 농도가 증가함에 따라 강한 절단작용을 나타내었으며 3,4-dihydroxytoluene 과 hydroxyhydroquinone갈변반응 생성물은 금속이온의 영향을 전혀 받지 않았다. 또한 chlorogenic acid갈변반응 생성물은 DNA 절단을 억제하는 효과를 나타내었다. Ames test에서는 4가지 갈변반응 생성물 모두 변이원성은 없었으며 benzo$[{\alpha}]$pyrene을 사용한 변이원성 억제작용 실험결과 benzo$[{\alpha}]$pyrene의 활성을 강하게 억제하는 것으로 나타났다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1992년도 제1회 신약개발 연구발표회 초록집
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• pp.53-53
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• 1992

galangin은 1차 발암물질인 MNNG, EMS, 2차 발암물질인 B(a)P 및 DMBA에 의해 유도된 소핵생성에 대하여 억제효과를 나타내었으나 ADM에 대해서는 억제효과를 나타내지 않았다. 한편, S-9 mixture 존재하 benzo(a)pyrene에 의한 자매염색분체교환 빈도에 대해서도 억제효과를 나타내었다. 또한, radiomimetic agent인 BLM 유도 염색체 이상에 대해서도 억제효과를 나타내었다. 이같은 결과를 종합하면 galangin은 DNA alkylation이나 adduct를 형성하는 발암물질에 대하여 억제효과가 큰 것으로 보아 DNA alkylation 및 DNA adduct를 형성을 차단하거나 방향족 탄화수소의 대사 활성화를 방해하여 특정 발암물질들의 염색체 손상작용을 억제해주는 Anticlastogen으로서의 가능성을 보여주었다.

• 한국환경성돌연변이발암원학회지
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• 제21권2호
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• pp.89-94
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• 2001

Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

• 약학회지
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• 제52권5호
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• pp.376-383
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• 2008

Chungkookjang (CKJ) is a fermented soybean product and one of favorite traditional foods in Korea. In this study, the alcoholic extract from Korean fermented soybean (CKJ) and its one of major flavonoids, genistein were evaluated for their protective effect against B(a)P induced cytotoxicity and DNA damage in HepG2 cells. CKJ extract and genistein decreased B(a)P-induced cell cytotoxicity. CKJ extract inhibited DNA single strand breaks evaluated by single cell gel electrophoresis. From RT-PCR study, it was revealed that CKJ extract decrease DNA damage induced in HepG2 cells expressing CYP1A1 and 1A2 by B(a)P. The metabolizing activities of CYP1A1 and CYP1A2, as measured by the 7-alkoxy resorufin O-deethylation (AROD) assay, showed that CKJ extract and genistein inhibited CYP1A1 and CYP1A2 activities. Genistein may contribute to these biological effects of CKJ extract at least in part. All these results indicate that CKJ extract and genistein may be useful for protection against B(a)P-induced cytotoxicity and DNA damage. Therefore, the alcoholic extract of Korean fermented soybean (CKJ) is suggested to be promising functional food which can prevent the cellular genotoxicity of dietary and lifestyle related carcinogens.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2003년도 추계학술대회
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• pp.133-133
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• 2003

In recent years. attention has focused on the application of the alkaline single cell gel electrophoresis (SCGE or Comet) assay in environmental mutagenesis. To evaluate the suitability of the assay as a monitoring. technique, the DNA damages in liver cells and erythrocytes of carp (Cyprinus carpio) exposed to benzo[${\alpha}$]pyrene (B[${\alpha}$]P) were estimated comparatively with the in vivo Comet assay and the micronucleus test (MNT).(omitted)

• Molecular & Cellular Toxicology
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• 제2권3호
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• pp.170-175
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• 2006

The extensive isolation of genes responsive to stressful conditions from a soft coral Scleronephthya gracillimum was described. Soft coral colonies were exposed to thermal and chemical stressors to induce the expression of stress related genes. Differentially expressed genes by natural or anthropogenic stressors were identified by construction of standard and stress exposed-paired subtractive cDNA library. Thirty-two and thirty-seven kinds of candidate genes were identified from thermal or benzo[a]pyrene stress exposed group, respectively, which are associated with cell cycle, cell signaling, transcription, translation, protein metabolism, and other cellular functions. The expected function of each gene was described. The isolated and identified differentially expressed genes have a great potential to identify environmental stressors in global environmental changes and could act as molecular biomarkers for biological responses against environmental changes. Finally, it may open a new paradigm on soft coral health assessment.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.20-45
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• 2002

In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.

• Applied Biological Chemistry
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• 제31권1호
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• pp.38-45
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• 1988

10종류(種類)의 산채류가열즙(山菜類加熱汁)에 대한 돌연변이원성(突然變異原性)을 Spore rec-assay, Ames test 그리고 DNA 절단실험(切斷實驗)으로 검토(檢討)하였으며 가열즙(加熱汁)의 조제(調製)는 $100^{\circ}C$에서 20분간 가열하여 가열즙(加熱汁)을 얻었다. Spore rec-assay 결과 10종류(種類)의 시료(試料)모두 변이원성(變異原性)은 없었으며 금속이온의 영향에서 소리쟁이가열즙은 $Pb^{2+}$, 곰취와 참나물가열즙의 경우는 $Zn^{2+}$의 존재하에서 변이원성을 나타내었다. Ames 시험과 DNA 절단시험에서도 시료모두 변이원성은 없었으며 DNA 절단(切斷)시험에서는 곰취, 머위, 잔대, 개미취가열즙의 경우는 25mM $Cu^{2+}$ 공존하에서 절단활성(切斷活性)이 촉진되었다. 그러나 두릅, 원추리, 잔대, 참나물 그리고 참취가열즙(加熱汁)은 $Fe^{2+}$의 공존시 DNA 절단활성(切斷活性)을 억제(抑制)시키는 결과(結果)를 얻었다. Benzo${\alpha}$pyrene에 대한 돌연변이(突然變異) 억제작용(抑制作用)에서는 시료(試料) 모두 농도증가(濃度增加)에 따라 강(强)한 변이원(變異原) 억제작용(抑制作用)을 나타내었다.