본 연구는 분열형 효모 Schizosaccharomyces pombe에서 여러 가지 DNA 절제회복 및 유전자 발현에 관여하는 SNF2/SW12유전자의 기능을 연구하기 위하여 이에 관련되는 유전자를 분리하고 그 특성을 연구하였다. SNF2 motif 의 conserved sequence를 primer로 하여 중합효소 연쇄반응(PCR) 방법으로 480 bp 크기의 DNA fragment를 분리하여, 이를 probe로 하여 효모에서 hrp2＋ 유전자를 분리하였다. 분리한 hrp2＋ 유전자의 sequence homology를 비교한 결과 3개의 SNF2 motif를 포함하고 있었다. hrp2＋유전자의 전사체 크기는 4.7 kb임을 Northern hybridization으로 확인하였다. hrp2＋유전자의 전사 개시 부위를 알기 위하여 primer extension분석을 한 결과, 첫 번째 ATG에서 약47 base pair 위쪽에 위치함을 확인하였다. 또한 특성 연구를 위하여 Northern hybridization으로 hrp2＋ 유전자의 UV와 MMS에 대한 유도성을 조사한 결과 자외선에 대해서만 유전자의 발현이 유도되었다. 이 결과 분리한 hrp2＋는 UV-inducible 유전자임을 확인하였다.

Although ionizing radiation (IR) has been used to treat the various human cancers, IR is cytotoxic not only to cancer cells but to the adjacent normal tissue. Since normal tissue complications are the limiting factor of cancer radiotherapy, one of the major concerns of IR therapy is to maximize the cancer cell killing and to minimize the toxic side effects on the adjacent normal tissue. As an attempt to develop a method to monitor the degree of radiation exposure to normal tissues during radiotherapy, we investigated the transcriptional responses of human peripheral blood lymphocytes (PBL) following IR using cDNA microarray chip containing 1,221 (1.2 K) known genes. Since conventional radiotherapy is delivered at about 24 h intervals at 180 to 300 cGy/day, we analyzed the transcriptional responses ex-vivo irradiated human PBL at 200 cGy for 24 h-period. We observed and report on 1) a group of genes transiently induced early after IR at 2 h, 2) of genes induced after IR at 6 h, 3) of genes induced after IR at 24 h and on 4) a group of genes whose expression patters were not changed after IR. Since Biological consequences of IR involve generation of various reactive oxygen species (ROS) and thus oxidative stress induced by the ROS is known to damage normal tissues during radiotherapy, we further tested the temporal expression profiles of genes involved in ROS modulation by RT-PCR. Specific changes of 6 antioxidant genes were identified in irradiated PBL among 9 genes tested. Our results suggest the potential of monitoring post-radiotherapy changes in temporal expression profiles of a specific set of genes as a measure of radiation effects on normal tissues. This type of approach should yield more useful information when validated in in vivo irradiated PBL from the cancer patients.

The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, to regulate and to evaluate the chemical hazard will be important to environment and human health. The clastogenicity of 18 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 4-Chloro-3,5-dimethyl phenol (CAS No. 88-04-0) induced chromosomal aberrations with significance at the concentration of 15.7 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. Phenoxybenzene (CAS No. 101-84-8) which is one of the most cytotoxic chemical among 18 chemicals tested revealed no clastogenicity in the range of 0.11-0.43 $\mu\textrm{g}$/$m\ell$ both in the presence and absence of metabolic activation system. From the results of chromosomal aberration assay with 18 synthetic chemicals in Chinese hamster lung cells in vitro, 4-chloro-3,5-dimethyl phenol (CAS No. 88-04-0) revealed weak positive clastogenic results in this study.

The role of the kidney in initiating hypertension has been much debated. The SA gene is expressed in the kidney and is association with hypertension in man and in experimental animal models. Also, increased plasma concentrations of homocysteine have been found in patients with coronary artery disease (CAD) and hypertension. The genetic variation of methlene tetrahydrofolate reductase (MTHFR) gene is related to its enzyme activity and to the plasma homocysteine concentration. In view of the effect of SA and MTHFR as risk factor for cardiovascular diseases, we investigated the Pst I RFLP of the SA gene and C667T mutation of the MTHFR gene in the Korean patients with hypertension. There were no significant differences in the allele and genotype frequencies of these polymorphisms between normotensive and hypertensive subjects. Therefore, our results do not support a possible role of these genes on hypertension in Korean population.

Phthalate analogues are a plasticizer and solvent used in industry and were reported to be a potential carcinogen classified in the category of suspected endocrine disruptors. Most common human exposure to these compounds may occur with contaminated food. They may migrate into food from plastic wrap or may enter food from general environmental contamination. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. To determine whether seven phthalate analogues i.e. diallyl phthalate, diisodecyl phthalate, di-n-nonyl phthalate, butyl benzyl phthalate, di-n-octyl phthalate, di-tridecyl phthalate, and dibutyl phthalate, can induce DNA strand breakage that is one of the various factors related to the mechanism of carcinogenicity, the comet assay which has been widely used for the detection and measurement of DNA strand breaks, was conducted in L5178Y mouse lymphoma cells. From these results, seven phthalates revealed dose-dependent decrease of cell viability, however, no remarkable cytotoxicity was observed even at high concentration of 100 $\mu\textrm{g}$/$m\ell$ phthalates. And also, the results showed that the induction of DNA strand breaks by seven phthalates was not significantly different from the control in this study.

Resveratrol is known to have potent cancer chemopreventive activity against tumorigenesis caused by 7,12-dimetylbenz［$\alpha$］anthracene(DMBA) which is known to be oxidized to reactive products by cytochrome P450 1B1 (CYP1B1). The effects of resveratrol on the activity of recombinant human P450 1 family enzymes, expressed in Escherichia coli membranes with human NADPH-P450 reductase, were determined by measuring alkoxyresorufin O-dealkylation activity, e.g., ethoxyresorufin O-deethylation (EROD) CYP1A1, methoxyresorufin O-demethylation (MROD), CYP1A2, benzyloxyresorufin-O-debenzylation (BROD), CTP1B1. Resveratrol inhibited CYP1B1 and CYP1A1 activities in a dose-dependent manner with $IC_{50}$/ values of 59 and 10$\mu$M for EROD activity and 1.8 and 30$\mu$M for BROD activity, respectively. Resveratrol had only weak inhibitory effect on CYP1A2 activity ($IC_{50}$/ values of 0.44 mM for EROD and ＞2 mM for MROD). Furthermore, resveratrol did not affect NADPH-P450 reductase activity significantly. Resveratrol inhibited the CYP1B1-dependent EROD activity with a $K_{i}$ of 28 $\mu$M in a non-competitive type manner. these results suggest that resveratrol-derived inhibited of CYP1B1 and CYP1A1 activities may contribute to the suppression of DMBA inducible tumorigenesis observed in extrahepatic tissues.s.

Endocrine disruptors (EDs) are the chemicals that affect endocrine systems through activation or inhibition of steroid hormone response. It is necessary to have a good system to evaluate rapidly and accurately endocrine-disrupting activities of suspected chemicals and their degradation products. The key targets of EDs are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. We constructed a co-expression system of Gal4p DNA binding domain (DBD)- ligand binding domain of human estrogen receptor $\alpha$ or $\beta$, and Gal4p transactivation domain (TAD)-co-activator AIB-1, SRC-1 or TIF-2 in Saccharomyces cerevisiae with a chromosome-integrated lacZ reporter gene under the control of CYC1 promoter and Gal4p binding site (GAL4 upstream activating sequence, GAL4$_{UAS}$). Expression of this reporter gene was dependent on the presence of estrogen or EDs in the culture medium. We found that the two-hybrid system with combination of the hER$\beta$ LBD and co-activator SRC-1 was most effective in the xenoestrogen-dependent induction of reporter activity. The extent of transcriptional activation by those chemicals correlated with their estrogenic activities measured by other assay systems, indicating that this assay system is efficient and reliable for measuring estrogenic activity. The data in this research demonstrated that the yeast detection system using steroid hormone receptor and co-activator is a useful tool for identifying chemicals that interact with steroid receptors.s.

Hypertension is a multifactorial disease. Both genetic and environmental factors have been implicated in its etiology. Since the impairment of nitric oxide (NOS) production plays an important role in the pathogenesis of hypertension, endothelial nitric oxide synthase (ecNOS) gene is supposed to be a candidate gene of hypertension. Our study group investigated the 27 bp insertion/deletion (Ins/Del) polymorphism of ecNOS gene in 99 Korean normotensives and 98 hypertensives, respectively. There was no significant association with any cardiovascular risk factors as well as hypertension in Koreans. The Ins/Del polymorphism of the ecNOS gene indicated the similar allele distribution among ethnic groups studied. Further studies using larger sample size and subject information is required to describe the general picture of the association between the ecNOS gene polymorphic loci and hypertension

Chlorpyrifos is an organophosphate insecticide and one of the most commonly and widely used insecticide. However, a little known about the dermal risk of chlorpyrifos on human being. Therefore, this study was conducted for the dermal risk assessment after exposure to chlorpyrifos in Korean farmers. First, skin irritation by chlorpyrifos (10 mg/$\textrm{cm}^2$, 50 mg/$\textrm{cm}^2$, 100 mg/$\textrm{cm}^2$, 250 mg/$\textrm{cm}^2$ in acetone) was determined in rabbits for 5 days considering the usage of chlorpyrifos short term highly exposure. The index of skin irritation by chlorpyrifos was increased in each dose and length of exposure dependent manners. Next, using benchmark dose (BMD$_{5}$) approach, the dose-response relationship was assessed to calculate the reference dose (RfD). The value of RfD was 2.84 $\mu\textrm{g}$/kg/day from 142.16 $\mu\textrm{g}$/kg/day BMD5 value divided uncertainty factor 50. Finally, we assessed human dermal risk of chlorpyrifos with exposure level and RfD. Skin absorbed levels were assumed with several exposure scenarios encounting the circumstances of exposure that application method, protection equipment and cloth, exposure time and exposure frequency during chlorpyrifos spraying. By the comparison of skin absorbed dose with the reference dose, it was identified that risk values (risk index) to skin chlorpyrifos exposure were 0.958 from the point of above results and it was recommended that the occurrence of hazard effect (skin irritation toxicity) of chlorpyrifos would not be expected. Risk index was smaller than 1 in the case of spraying vehicle mounted application, 1hour exposure time and wearing protective cloth exposure. Whereas, risk index was above 1 in the case of hand-held application, 2hour exposure time and wearing common cloth. Comparing two kinds of application method, total risk index of the hand held application (1.67) was higher than vehicle mounted (0.27). Therefore, chlorpyrifos skin exposure was mainly affected by application equipment and applied form. The results of risk assessment on the human dermal toxicity of chlorpyrifos should be required to control in keeping safety rules, skin surface area available for contact, spraying time ,and spraying frequency.y.

It has been known that Ganoderma lucidum and Lentinus edodes have anticancer activity and immune enhancing activity. These two mushrooms were grown in liquid culture and harvested. From these mycelia, DNA was isolated and EtBr-CsCl density gradient ultracentrifugation was performed to purify it further. Then mitochondrial DNA was isolated by bisbenzimide-CsCl density ultracentrifugaton. Mitochondrial DNA of Ganoderma lucidum was digested by restriction enzymes, EcoR I, Hind Ⅲ, and Pst I, then electrophoresed. It showed 12, 22, 4 fragments. Mitochondrial DNA of Lentinus edodes was digested by EcoR I. Electric pattern showed 6 fragments. 4 fragments had appeared by Pst 1 digested mitochondrial DNA. Hind ill couldn't digest mitochondrial DNA of Lentinus edodes. Mitochondrial DNA of fusants was isolated to compare to those of parents. The results showed that fusant P₂S₄has new, recombined mitochondrial DNA. But P₂S₄had the same DNA that Ganoderma lucidum had.

In irradiated human lymphocytes, translocation of chromosome has been more frequently observed than dicentric chromosome. Differences in the misrepair process leading to translocation and dicentric chromosomes may explain the above observations. In order to find out whether dicentric and translocation are originated from different mechanism, the frequencies of radiation induced translocation and dicentric in lymphocytes were examined following treatment of irradiated lymphocytes with two DNA repair inhibitors, 3AB for inhibition of poly(ADP-ribose) synthesis and Ara C for inhibition of DNA-polymerase $\alpha$. Ara C potentiated the frequencies of radiation induced dicentric and translocation. 3AB also potentiated the frequencies of radiation induced dicentric, but not translocation. These results suggest the potential differences in the mechanisms in the formation of translocation and dicentric chromosomes.