The overall objective of this lecture will be to present an overview of the evolution of the science of immunotoxicology by highlighting a few key milestones, namely a solid mechanistic foundation and recent applications in a regulatory context. Neither time nor space will allow a discussion of the full extent of the evolution of immunotoxicology from a historical perspective.

GLP regulations were initially “promulgated to address assuring the validity of data in the wake of investigations by EPA and FDA during the mid -1970's which revealed that some studies submitted to the agencies had not been conducted in accordance with acceptable laboratory practices.” [1] In the early 1970s, results of an investigation by the FDA in about 40 laboratories revealed many cases of badly managed studies, poor training of personnel and some cases of deliberate fraud. The general findings were that there were poorly trained study directors and study personnel, poorly designed protocols, protocols not followed, procedures not conducted as described, raw data badly collected, data not correctly identified, data without traceability, data not verified and approved by responsible persons, lack of standardised procedures, poor animal husbandry, inadequate characterisation of test items and test systems, inadequate resources, equipment poorly calibrated or otherwise qualified, reports not sufficiently verified, not an accurate account of the actual study, not a proper reflection of raw data and inadequate archiving of data. These problems are not just past history, since they resurface time and time again, even in quite recent times as the experience of GLP inspectors shows [1]. The GLPs specify minimum practices and procedures in order to ensure the quality and integrity of data submitted in accordance with a regulatory requirement

인류는 1,500 만 종류에 달하는 화학물질을 합성 혹은 분리ㆍ동정 해왔다. 그리고 현재 약 10 만 종류 정도의 화학물질이 상업적으로 이용되고 있다. 이들 화학물질 중에 유기염소화합물, 공업용화합물, 농약류, 유기취소(bromine)화합물, 중금속 및 유기금속, 식물 및 합성에스트로겐 등을 포함한 수십 종의 화합물이 내분비계장애물질 (Endocrine Disruptors, EDs)로서 생각, 혹은 의심되고 있다. 그러나, 대부분의 다른 화합물에 대해서는 그것들이 내분비계장애 작용을 가지고 있는지에 대한 충분한 지견이 얻어지지 않고 있어 이들 화학물질의 내분비계장애 작용을 평가하기 위해서는 신속 또는 간편한 검출계의 확립은 매우 중요하다.

Crop Protection Products (CPPs) are a useful weapon in our fight against the pests, diseases and weeds that harm or destroy our food. To be effective CPPs need to exhibit some persistence. The consequence of this is that residues of the original material or its metabolites may remain on food and thus there is a potential for the intake of such residues. The issue of CPP residues in food is an emotional subject and is rarely debated rationally or supported by scientific facts. Media headlines like “Poison in our food” or “These days enough Pesticides (CPPs) are sprayed onto vegetables to turn you into one” are testimony to the high level of emotion that surrounds the subject of CPP residues in food. Recent surveys of consumers have indicated that more than 80 percent view CPP residues in food as a “serious hazard”. This significantly exceeds concerns over drugs, hormones in meat, nitrates in food, irradiated foods, food additives, or artificial colours.

수돗물의 염소소독 과정에서 부식질의 초기 분해생성물인 MX(Mutagen X)는 수돗물의 전체 돌연변이유발성의 최고 60%까지 차지하는 것으로 알려져 있다. 그러나 존재하는 농도는 수 ng/L로 정량분석을 위해서는 resin을 사용하여 10 ~ 20L의 물을 농축하여야 한다. 이 연구에서 MX의 농축에 사용된 resin은 XAD-4와 XAD-7HP의 1:1 mixture이었으며, 사용 전에 Soxhlet extractor를 사용하여 ethyl acetate와 methanol로 각각 12시간씩 세척하였다. 15L의 물 시료는 37%의 염산으로 pH 2에 맞춘 후 직경 2cm의 유리관에 15cm의 높이로 채워진 resin을 40mL/min의 유속으로 통과시켰다. 유리관에 잔류하는 물은 질소가스로 대부분 불어 제거한 후 ethyl acetate 200mL를 가해 1mL/min의 유속으로 흘려주어 MX를 용출시킨 후 회전증발기로 5mL의 부피까지 농축시키고 10% H$_2$SO$_4$ in MeOH 0.1mL를 가하여 6$0^{\circ}C$에서 methyl ester화하였다. 과포화된 $Na_2$SO$_4$ 수용액 0.3ml를 가한 후 MTBE 0.3mL로 추출하고 GC/ECD로 분석하였다. MX의 회수율은 60% 이상이었으며 일부수돗물 시료에 대해 분석하였다.

Benzoyl peroxide is a High Production Volume Chemical, which is produced about 1,375 tons/year in Korea as of 2001 survey. The substance is mainly used as initiators in polymerization, catalysts in the plastics industry, bleaching agents for flour and medication for acne vulgaris. The substance is one of seven chemicals of which human health and environmental risks are being assessed by National Institute of Environmental Research (NIER) under the frame of OECD SIDS Program. In this study, Quantitative Structure-Activity Relationships (QSAR) is used for getting adequate information on the physical-chemical property and the environmental fate of this chemical. For the assessment of benzoyl peroxide, models such as MPBPWIN for vapor pressure, KOWWIN for octanol/water partition coefficient, HENRYWIN for Henry's Law constant, AOPWIN for photolysis and BCFWN for bioconcentration factor (BCF) were used. These (Q)SAR model programmes were worked by using the SHILES (Simplified Molecular Input Line Entry System) notations. The physical-chemical properties and the environmental fate of benzoyl peroxide were estimated as followed : vapor pressure =0.00929 Pa, Log Kow = 3.43, Henry's Law constant = 0.00000354 atm-㎥/mole at 25 $^{\circ}C$, the half-life of photodegradation = 3 days, bioconcentration factor (BCF) = 92

Ambient urban particulate matters contain various transition metals. When the particulate matters are inhaled into the lung, not all but some part of metals from the particles might be mobilized to participate in a reaction that can damage various biomolecules, such as DNA and proteins. The dust particle size as well as organic acids may influence the metal mobilization. Thus, the mobilization of the metal from SRM1648 (NIST, USA) and urban particulate matters (PM2.5 and PM10) was measured in the presence of artificial or biological chelator with or without reductant. The degree of the mobilization was higher with the artificial or biological chelator than the control with saline. In some cases, a reductant increased the mobilization as much as about 5 times the control without the reductant. Especially, the mobilization of Fe was greatly influenced by the presence of reductants. In general, the degree of the mobilization of the transition metal was higher with PM2.5 than with PM10. Therefore, it is expected that, considering the previously known toxicities of the transition metals, the PM2.5 is more damaging to various biomolecules than PM10. The results also suggest that not the total amount but the mobilizable fraction of the metal in the dust particles should be considered with regard to the toxicity of the urban particulate matters.

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.

본 연구에서는 한강을 세 구간으로 구분하여 구간별 대표지점(의암호, 팔당호, 및 동호대교 정점)의 하상퇴적물을 grap으로 각각 채취한 후, Tandem Bioassays 을 적용하여 환경독성 수준을 평가하고 정점간 상호 비교하였다. Tandem Bioassays 이란 유해물질 오염에 의한 다양한 생태계 반응을 표현할 수 있는 최소한의 상호 보완적 환경독성 평가 체계를 의미한다. 본 연구에서 적용한 Tandem Bioassays 은 발광성미생물을 사용하는 Microtox 시험, 수서 물벼룩인 daphnia와 ceriodaphnia 시험, 그리고 개구리 배아를 이용하는 기형유발능 시험(최기형성 시험, FETAX)등으로 구성하였다.

최근 들어, 일반적으로 알려진 중금속 외에 As 및 Se의 생래독성(예를 들면, 어류 및 양서류의 번식독성)이 많은 연구자나 환경관리자들의 관심대상으로 부각되고있다. 상수원 보호구역내 위치하는 하천이나 호소 인근에 As이나 Se의 오염원이 분포되어 있는 경우, 수서 생태계뿐만 아니라 인체의 건강에 큰 위협이 될 수 있기 때문이다. 그러므로, 본 연구에서는 하천수나 하상 퇴적토의 잠재적 오염 물질인수 종의 중금속에 대한 급성독성과, As 및 Se의 급성, 그리고 번식 독성을 수서곤충을 이용하여 측정하고 이를 상호 비교하였다. 본 실험에 적용된 생물은 생활사가 짧고 배양이 용이하며, 번식력이 높은 물벼룩 (Daphnia magna)과 소형물벼룩 (Ceriodaphia dubia)이었다.

양서류를 이용한 내분비계장애물질의 환경독성시험기법을 확립하기 위해 Xenopus laevis의 변태 과정에 thyroid계 물질을 처리하여 이들의 효과에 의한 내분비계 장애물질의 판정 기법을 확립하고자 하였다. 올챙이의 발달 단계에 의한 내분비계 장애물질에 대한 민감도를 고려하여, embryo 단계에서 tail resorption 과정의 전단계인 stage 11로부터 30일간(방법1), stage 49로부터 28일간(방법2)와 tail resorption 과정 및 이후 stage 57로부터 17 내지 21일간(방법3)으로 구분하여 시험물질을 노출시켰으며 대조군과 비교하여, 시험물질에 노출된 올챙이의 기형적 변태 정도는 developmental stage 변화 속도, head-tail length 차이, 및 body weight 변화 등을 endpoints로 하였다.

호소 수질에 영향을 미치는 대표적 인자로 외부 유입물질과 퇴적물을 들 수 있다. 따라서, 호소 수질 관리를 위해서는 호소로 유입되는 점ㆍ비점 오염원 관리와 함께, 퇴적층 관리도 수반되어야만 한다. 특히, 호소수가 주요 식수원으로 사용되는 경우에는 엄격한 퇴적토질 평가가 이루어져야 할 것이다. 본 연구에서는 소양호 퇴적층에 대한 환경지질학적 조사 연구의 일환으로 다양한 bioassays 기법(the battery of bioassays)을 적용하여 퇴적토의 오염상태를 평가하였다.

최근, 무분별한 농약사용으로 인해 조리 후에도 잔류하는 농약성분의 독성이 문제시 되고 있어 인체 내로의 섭취 전에 독성 유무와 그 정도를 확인할 수 있는 방법 개발을 위한 연구가 활발히 진행되고 있다. 본 연구에서는 채소류, 곡류, 과일류 등에 빈번히 사용되는 농약성분의 독성 확인과 함께 이들 식품 내에 잔류하는 농약성분의 검출을 위한 방법을 연구하였으며, 이들의 독성 유무를 유전자 재조합된 5종의 발광성 미생물을 이용하여 확인하였다. 또한 이들 농약류가 식품 내에 잔류하여 발생하는 독성을 분석하기 위해 식품내 잔류하는 농약 성분의 추출에 필요한 효율적인 추출 용매를 제작하였으며, 농약성분의 화학적 성질에 따라 추출 방법을 두 단계로 분리하여 추출률을 향상시켰다. 뿐만 아니라, 식품류의 특성에 적합하게 식품별로 특정한 추출방법을 연구하였으며, 추출된 농약 성분의 독성 및 안정성 역시 발광성 박테리아를 이용하여 확인하여, 전체적인 추출방법의 효율을 진단하였다.

염소소독 결과 수돗물 중에 존재하는 Dichloroacetic acid(DCAA)와 Trichloroacetic acid(TCAA)는 동물실험 결과 발암성 등 악영향을 일으키는 것으로 알려져 있다. 이러한 화합물들은 수돗물을 사용할 때 열을 가하면 그 농도가 변하게 된다. 이 연구에서는 설문조사를 통해 개인이 하루 동안에 섭취하는 수돗물의 양과 섭취패턴을 조사하고, 각 가정마다 수돗물의 농도를 측정하여 DCAA 및 TCAA에 대한 경구섭취 노출량을 평가하고자 실시하였다. 춘천에 위치하고 있는 60가정을 2002년 1월부터 2월 사이와 7월부터 8월 사이에 방문하여 수돗물을 채취하고, 주부를 대상으로 24시간 동안의 물 사용에 대한 설문을 실시하였으며, 아침 첫 소변을 회수하였다. 수돗물 및 소변 중 DCAA 및 TCAA에 대한 분석은 methyl ester 형태의 유도체를 만든 후 GC/PDECD로 분석하였다. 수돗물을 가열하면 DCAA의 농도는 증가하고, TCAA의 농도는 감소하였고, DCAA 및 TCAA의 경구 섭취량은 수돗물 중의 농도에 섭취부피와 가열시간에 따른 농도변화율을 곱하여 산출하였다. 실험 대상자들의 평균 수돗물 섭취부피는 1.7$\ell$이었고, 평균 가열시간은 10~15분이었다. 추정한 DCAA의 섭취량은 찬 수돗물만을 마시는 것을 가정한 것에 비해 182% 높았고, TCAA에 대해서는 30% 낮았다 이와 같은 결과는 우리나라 사람들의 식생활이 국이나 찌개 및 보리차 등과 같이 가열한 음식이나 음용수를 많이 섭취하기 때문이다. 그러므로 음용수 중 DCAA 및 TCAA의 인체 섭취 노출평가를 하기 위해서는 섭취한 부피뿐만 아니라 가열에 따른 농도 변화도 동시에 고려하여 야 한다.배 센타의 재고 완충역할을 수행하게 한다. 이에 따라 안전 재고 및 역 배송을 최소화 할 수 있을 뿐 아니라 미 출고로 인한 손실을 최소화하며 고객의 서비스를 일정하게 유지시킬 수 있을 것으로 기대된다.증진, 페기물 발생 억제 분야에 상당한 성공을 거두었으며, 지금도 지속적으로 유지 관리 및 미비한 부분(CAER, 등)의 향상에 노력하고 있다. Dow Korea의 수입 판매 부분에 주로 관련된 Product Stewardship 및 Distribution Code는 여러 사업부 및 여러 지역(미국, 유럽, 아시아 등)에 위치한 담당자가 관련된 까닭에 가장 복잡한 부분이어서 많은 인력과 시간을 소비하면서 노력하고 있으나 아직까지 소정의 목표에 도달하지 못했으며, 2000년 말에 모든 실행지침이 실행되도록 목표, 실행 계획 및 조직을 수정하여 노력하고 있다.lumn density at distant position from MD5 is larger than that in the (:entral region. We have deduced that this hot-core has a mass of 10sR1 which i:s about an order of magnitude larger those obtained by previous studies.previous studies.업순서들의 상관관계를 고려하여 보다 개선된 해를 구하기 위한 연구가 요구된다. 또한, 준비작업비용을 발생시키는 작업장의 작업순서결정에 대해서도 연구를 행하여, 보완작업비용과 준비비용을 고려한 GMMAL 작업순서문제를 해결하기 위한 연구가 수행되어야 할 것이다.로 이루어 져야 할 것이다.태를 보다 효율적으로 증진시킬 수 있는 대안이 마련되어져야 한다고 사료된다.$\

국내에서 사용중인 농약에 대한 농민의 농약 노출량 및 양상을 조사하고자 내분비계 장애 물질로 추정되는 Mancozeb의 살포 시 작업자의 피부 노출량 및 호흡 노출량 측정을 감귤농장에서 실시하였다. 피부노출 측정은 Patch법(50$\textrm{cm}^2$)을 사용하였고 손 노출은 순면장갑을 사용하였다. 호흡노출은 XAD-2 resin 흡착제와 Personal air monitor(2L/min)를 이용, 측정하였다. Mancozeb 분석은 CS$_2$법을 이용하였고 patch, resin, 장갑, 마스크, 소변에서 Mancozeb의 회수율 및 포장회수율 모두 90%이상이었다. 감귤 재배시 Mancozeb 사용에 따른 피부노출량은 여자가 약 1800mg/hr, 남자는 약 1300mg/hr 이었고 주로 배, 등, 상박 그리고 허벅지에 노출량이 많았다. 호흡노출은 관찰되지 않았고 소변에서도 Mancozeb는 관찰되지 않았다. 위해성 평가를 위해 PDE, AQE 및 MOS를 구하였으며 MOS는 Severn의 공식을 이용하였다. MOS는 여자가 0.37, 남자가 0.58이었다.

에스트로젠을 처리한 송사리의 간으로부터 choriogenin vitellogenin estrogen receptor의 발현량을 전사수준에서 Real-time을 사용하여 정량.비교하였다. 시험어종으로는 부화 후 5개월 이상된 성숙한 수컷 송사리(Oryzias latipes)를(체중 약 250mg/마리)를 사용하여 17$\beta$-estradiol(25ppt, 50ppt, 100ppt)에 24시간 노출시켰다. Fluorescence dye는 choriogenin vitellogenin estrogen receptor의 경우 FAM (6-carboxyfluorescein)을 사용하였으며, $\beta$-actin의 경우는 VIC를 사용하였다. 프로브에 사용하는 quencher dye는 TAMRA(6-carboxy-N',N',N',N'-tetramethyl rhodamine)을 사용하였다. Internal control로 사용된 $\beta$-actin은 17$\beta$-estradiol의 농도에 상관 없이 0~10pM 범위에서 일정하게 발현됨을 보여주었다. vitellogenin choriogenin L 및 choriogenin H는 17$\beta$-estradiol의 농도에 의존하여 발현이 증가되는 용량-반응양상(Dose-dependent)을 나타내었다. 반면, estrogen receptor는 모든 처리군에서 $10^{-2}$pM 정도로 발혐됨에 따라 본 시험농도의 17$\beta$-estradiol에 의해서는 거의 유도발현이 되지 않음을 보여주었다. choriogenin L, choriogenin H, vitellogenin I 및 estrogen receptor 발현감수성을 비교한 결과, 25ppt 및 50ppt의 17$\beta$-esoadiol 농도에서는 ChgL > ChgH > VTG I >ER의 순으로 감수성이 높았으며, 100ppt 노출에서는 ChgL > VTG I > Chg H > ER의 순으로 감수성이 높게 나타났다. 결론적으로 choriogenin이 에스트로젠물질에 의한 가장 민감한 생체지표임을 알 수 있었다.

Hypertension is considered to be caused by a complicated combination of genetic and environmental factors. Atrial natriuretic peptide (ANP) has been to suppress renin activity and inhibit the synthesis and release of aldosterone. Therefore, Abnormalities of this peptide caused by genetic variation may be influence the blood pressure. The aim of present study was to examine the relationship between hypertension and Sca I RFLP of ANP gene in Korean population. The genotype distribution of this RFLP was significantly different between normotensives and hypertensives (P<0.05). However, this genetic marker was not significantly associated with any anthropometric parameters or plasma lipid concentrations in our study group. Therefore, our result suggest that Sca I RFLP of ANP gene may be useful as genetic marker in the ethiology of hypertension in Korean population, independent of any cardiovascular risk. factors studied.

The response of environmental pollutants can be detected bioanalytically focusing on the source and matrices of concern. Cell culture bioassays are rapid and inexpensive, and thus have great potential for determination of environmental pollution. We have examined the estrogenic and dioxin-like activites of industrial wastewater using E-screen assay and EROD microbioassay. Influent and effluent wastewater were collected from four different industrial wastewater treatment plants, such as cosmetics, paints, textile producing and metal coating plant, and extracted using solid-phase extraction with Oasis＠HLB plus cartridge. Pollutants adsorbed to the cartridge were eluted with MTBE. MCF-7 cells were treated with extracts showed various estrogenic potential. The textile wastewater showed strong estrogenic activity and the others showed weak estrogenic activity, No effect was observed in the wastewater from paints producing plant. All extracts showed CYPIA inducing effects, indicating these samples contain dioxin-like chemicals. Bioanalytical results of effluents compared with influents could give us information about the incomplete wastewater treatment and biological potency caused by pollutants. [Supported by a Grant from the Korea Science and Engineering Foundation]

Methylmercury (MeHg), one of the heavy metal compound, can cause severe damage to the central nervous system in humans. Many reports have contributed MeHg poisoning to contaminated foods and release into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established. To find genes differentially expressed by MeHg in neuronal cell, we peformed forward and reverse suppression subtractive hybridization (SSH) method on mRNA derived from neuroblastoma cell line, SH-SY5Y treated with solvent (DMSO) and 6.25 uM (IC$\sub$50/) MeHg. Differentially expressed CDNA clones were sequenced and the mRNAs were re-examined on Northern blots. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences has provided an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as common environmental pollutants.

To establish a test protocol for the rodent 20-day thyroid/pubertal assay, flutamide and diethylstilbestrol (DES) were administered to intact male Sprague-Dawley rats from postnatal day 33 for 20 days. Flutamide (1, 5, and 25 mg/kg/day) or DES (10, 20, and 40 ug/kg/day) was given once daily by oral gavage to immature male rats. Prepuce separation was significantly delayed in flutamide group and in DES group. One day after the last dose, the rats were killed and pituitary, thyroid, and reproductive organs were removed and weighed. Flutamide treatment resulted in a significant reduction in the weights of epididymides, ventral prostate, seminal vesicles plus coagulating glands and fluid (SVCGF), levator ani. bulbocarvenus muscles (LABC), Cowper's glands, and glans penis. The weight of adrenal glands decreased at % mg/kg/day, while testes and any other organ weights were unaffected. No microscopic changes were observed in the thyroid glands. Serum levels of testosterone wert significantly increased in the flutamide-treated groups and serum levels of estradiol were also increased. A significant reduction in the weights of testes, epididymides, ventral prostate, SVCGF, LABC, Cowpers glands, and glans penis of DES treated group. Serum testosterone and LH decreased significantly in DES group. Decrease of estradiol was observed, but not significant. These results indicate that flutamide and DES delay puberty in the male rat and its mode of action appears to be via altered secretion of steroids, which subsequently affect the development of the reproductive tract. (Supported by the grant from NITR/Korea FDA for Endocrine Disrupter Research.)

The mobilizable amount of transition metals is a fraction of the total amount of the metal from urban particulate matter. Although the fraction is small, some metals (Fe, Cu) are the major participants in a reaction that generates reactive oxygen species (ROS), which can damage various biomolecules. Damaging effects of the metals can be measured by the single strand breakage (SSB) of X174 RFI DNA or the carbonyl formation of protein. In another study, we have shown that more metals are mobilized by PM2.5 than by PM10 in general. DNA SSB of >20% for PM2.5 and >15% for PM10 was observed in the presence of chelator (EDTA or citrate)/reductant (ascorbate), compared to the control (<3%) only with the chelator. The carbonyl formation by both PMs was very similar in the presence of the chelator, regardless of the kind of proteins. Compared to the control in the absence of chelator/reductant, 3.3 times and 4.9 times more carbonyl formation for PM2.5 and PM10, respectively, was obtained with BSA in the presence of chelator/reductant, showing that PM10 induced 33% more damage than PM2.5. However, 4.8 times and 1.9 times more carbonyl formation for PM2.5 and PM10, respectively, was observed with lysozyme in the presence of chelator/reductant, showing that PM2.5 induced 250% more damage than PM10. Although different proteins showed different sensitivities toward ROS, all these results indicate that the degrees of the oxidation of or damage to the biomolecules by the mobilized metals were higher with PM2.5 than with PM10. Therefore, it is expected that more metals mobilized from PM2.5 than from PM10, more damage to the biomolecules by PM2.5 than by PM10. We suggest that when the toxicity of the dust particle is considered, the particle size as well as the mobilizable fraction of the metal should be considered in place of the total amounts.

College of Pharmacy, Ewha womans University, Seoul, 120-750, Korea 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is the most potent halogenated aromatic hydrocarbon congener that induces expression of several genes including CYP1A1. Exposure to TCDD results in many toxic actions such as carcinogenesis, hepatotoxicity, immune suppression, and reproductive and developmental toxicity. Dramatic differences in dioxin toxicity have been observed between the sexes of some animal species, suggesting hormonal modulation of dioxin action. Many studies have been reported and propose several mechanisms of anti-estrogenic effects of TCDD. In contrast, the effect of estrogen on the regulation of CYP1A1 are not clear at present. There are several reports showing conflicting results. It seems that induction/inhibition of CYP1A1 may be dependent on cell-type and concentration. The purpose of this study was to investigate the regulation of TCDD-induced CYP1A1 gene expression by estradiol and its metabolites. We examined whether estradiol and its metabolites altered TCDD-mediated induction of CYP1A1 enzyme activity. 17 ${\beta}$ estradiol and 16 ${\alpha}$ estriol at non cytotoxic concentrations caused a significant concentration dependent decline of TCDD-induced EROD activity To determine whether reduced EROD activity reflected altered CYP1A1 mRNA expression, we measured CYP1A1 mRNA level by RT-PCR. And to examine whether estradiol and its metabolites have effects on TCDD-induced CYP1A1 gene expression at the transcription level, we also peformed transient transfection with an AhR responsive reporter plasmid containing the 5' flanking region of the human CYP1A1 gene to examine whether estradiol and its metabolites have effects on TCDD-induced CYP1A1 gene expression at the transcription level.

Recent industrial society has human widely exposed to PAHs that are comming from the incomplete combustion of organic material as widerspread environmetal contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. PAHs in the mammalian cells affect CYP1A1 gene expression as well as other phase II drug metabolizing enzymes as UDPGT, NMOR etc. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYP1A1 in human breast cancer. Our labolatory have been studied the effect of PAHs in the human breast cancer cell lind MCF7. In this study, we examined the ZR-75-1 human breast cancer cells as a new system to evaluate bioactivity of PAHs. ZR-75-1 human breast cancer cell line has been estabilished from the breast cnacer patient, has estrogen receptors and progesteron receptors. We have been able to estbilish long term culture system of this cells then used for the study to observe the effect of PAHs. We demonstrate that PAHs induced the transcription of an aryl hydrocarbon-responsive reporter vector containing the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase(EROD) activity of CYP1A1 enzyme in a concentration-dependant manner. RT-PCR analysises indicated that PAHs significantly up-regulate the constitutive level of CYP1A1 mRNA. Apparently, ZR-75-1 cells have Aryl hydrocarbon recetors, therefore it would be good experimental tool to study the cross-talk between PAHs and steroid actions.

Previously, we reported a novel polymeric micellar solubilizer, Aceporol 330, that showed relatively low toxic effects when it was compared with that of Cremophor EL which is currently being used for paclitaxel. In this study, we have developed a new micellar solubilizer, Aceporol 460, that has 3-4 times higher loding capacity for paclitaxel than Aceporol 330. The single-dose and the repeated-dose toxicity of Aceporol 460 were evaluated in ICR mice. For single dose toxicity test, male and female mice were randomly assigned to one of five study groups to receive, and injected intravenously with dosages of 0, 3, 4mL Cremophor EL/kgbody weight, and 3, 4mL Aceporol 460/kg body weight, respectively. In both male and female mice, LD50 for Aceporol 460 can not he determined even at the maximal administrable dosage, 4mL/kg due to the high viscosity of chemical and there was no significant change in body weight, hematological and serum biochemical analysis, organ weight, and histopathological examination compared with that of Cremophor EL. For the repeated dose toxicity test, male and female mice were given the dosage of 0, 1.6mL Cremophor EL/kgbody weight/day, and 1.6mL Aceporol 460/kg body weight/day for 2 weeks. Results of repeated dose toxicity tests for 2 weeks suggested that Aceporol 460 treated group show no significant toxicological findings with body weight, hematological and serum biochemical analysis, organ weight, urinalysis, and ophthalmoscopic and histopathological examination compared with that of Cremophor EL. These results indicate that Aceporol 460 have higher paclitaxeL-loading capacity than Aceporol 330 and less toxic effects than Cremophor EL in male and female mice.

In order to understand the mechanism of action of TCDD, we have examined the effect of COX-inhibitors on cyp1a1 activity. We observed the effect of COX-inhibitor on EROD activity in C57BL/6 mouse in vovo. And we also evaluated the effect of COX-inhibitors on cyp1a1 mRNA, mouse cyp1a1 promoter activity and EROD activity in Hepa cell. When Aspirin were pretreated with 3MC in vivo, the EROD activity that was stimulated by 3MC was inhibited. And Pretreatment of Aspirin, Celecoxib, Nimesulide and other several Cox-inhibitors in vitro, inhibited the TCDD stimulated EROD activity and Luciferase acitivity. In case of cyp1a1 mRNA level, Nimesulide and SB100 were able to decrease cyp1a1 mRNA that was stimulated by TCDD, but other tested COX-inhibitors were not decrease. We don't know this different result exactly. For the action of Cox-inhibitors on the Cyp1a1, it seems to be important to do pretreatment of these chemicals as apposed to TCDD. In this study, thus, we have suggested that COX-inhibitors such as aspirin, celecoxib, Nimesulide and other several Cox-inhibitors decrease the TCDD induced Cyp1a1.