[표제지 등]
제출문
요약문
SUMMARY
목차
제1장 OECD GLP 운영을 위한 체계확립 78
제1절 서론 78
제2절 OECD GLP 체제분석 78
제3절 단기독성시험에서의 OECD GLP 도입 및 SOP 확립 100
제4절 독성시험수행의 전산처리에 대한 OECD GLP 적용 및 SOP 확립 157
제2장 소동물을 이용한 장기투여 독성시험법의 확립 및 보완 166
제1절 랫트에서의 미정맥내 연속투여법의 확립 166
1. 서론 166
2. 미정맥내 카테터 장착법의 개발[원문불량;p.96] 167
3. Infusion pump를 이용한 K-378의 미정맥내 연속 또는 반복투여가 암컷 랫트에 미치는 영향 차이 174
4. Infusion pump를 이용한 K-566의 미정맥내 연속 또는 반복투여가 수컷 랫트에 미치는 영향 차이 195
5. Infusion pump를 이용한 K-799의 암컷 랫트를 이용한 미정맥내 연속투여 급성독성시험 218
6. 고찰 및 결론 230
7. 참고문헌 232
제2절 독성동태 이론을 이용한 시험물질 용량설정기술의 확립 233
1. 서론 233
2. 재료 및 방법 234
3. 결과[원문불량;p.170] 235
4. 고찰 및 결론 250
5. 참고문헌 252
제3절 병리조직학적 변화를 검색하기 위한 다양한 특수염색방법의 확립 253
1. 서론 253
2. 재료 및 방법 256
3. 결과 260
4. 고찰 및 결론 267
5. Legends for Figures 276
6. 참고문헌 302
제3장 소동물을 이용한 특수독성 시험법의 확립 및 보완 311
제1절 신경독성 시험방법의 확립 311
1. 서론 311
2. Measurement of Kainate-induced increase in GFAP(Glial Fibrillary Acidic Protein) by Sandwich Elisa assay 312
3. Measurement of memory behavior using Morris water maze and Radial 8-arm maze 325
제2절 면역독성시험방법의 확립 및 보완 337
1. 서론 337
2. 재료 및 방법 341
3. 결과 351
4. 고찰 및 결론 404
5. 참고문헌 411
제3절 유전자전이동물(Transgenic animal)을 이용한 유전독성시험법의 확립 419
1. 서론 419
2. 재료 및 방법 424
3. 결과 428
4. 고찰 및 결론 431
5. 참고문헌 432
제4절 In Vitro 최기형성 시험법의 확립 444
1. 서론 444
2. 재료 및 방법 445
3. 결과[원문불량;p.373~376] 448
4. 고찰 및 결론 456
5. 참고문헌 461
제4장 중동물을 이용한 일반 및 특수독성 시험법의 확립 및 보완 464
제1절 중동물을 이용한 독성시험에서의 시험물질 용량설정기술의 적용 464
1. 서론 464
2. 재료 및 방법 465
3. 결과 467
4. 고찰 및 결론 478
5. 참고문헌 480
제2절 광독성 및 광감작성시험방법의 확립 및 SOP 작성 481
1. 서론 481
2. 재료 및 방법 481
3. 결과 483
4. 고찰 및 결론 484
5. 참고문헌 484
제3절 중동물을 이용한 항원성시험방법의 확립 및 보완 485
1. 서론 485
2. 재료 및 방법 486
3. 결과 490
4. 고찰 및 결론 500
5. 참고문헌 502
제5장 환경독성시험법 및 GLP체계 확립 503
제1절 서론 503
제2절 생태독성시험법 연구 505
1. 송사리를 이용한 생육초기독성시험법 확립 505
2. 어류의 혈액화학, 혈액생화학적 및 조직학적 지표를 이용한 독성시험법 연구 517
제3절 환경내 대사 및 동태시험법 연구 541
1. 토양대사 연구 541
2. 식물대사 연구 563
3. 수중광분해 연구 585
제4절 환경독성시험 GLP 체제확립 601
1. 환경독성팀의 GLP 조직 및 운영체계 확립 601
2. SOP 분류, SOP 번호 및 관리 602
제6장 실험동물 육종기술 확립 612
제1절 서론 612
제2절 재료 및 방법 613
1. 표준화된 양질의 SPF 실험동물 대량 육종기술 확립 (II) 613
2. Nude mouse의 육종기술 확립 615
3. 새로운 질환모델 동물의 확립과 연구 620
제3절 결과 624
1. 표준화된 양질의 SPF 실험동물 육종기술 확립 (II) 624
2. Nude mouse의 육종기술 확립 630
3. 새로운 질환모델 동물의 확립과 연구 647
제4절 고찰 및 결론 649
1. 표준화된 양질의 SPF 실험동물 육종기술 확립 (II) 649
2. Nude mouse의 육종기술 확립 649
3. 새로운 질환모델 동물의 확립과 연구 653
제5절 참고문헌 657
제7장 국제공동 위탁과제 663
제1절 서론 663
제2절 위탁과제 내용 664
제3절 결론 677
제8장 위탁과제 678
1. 새로운 면역독성 평가방법의 개발[원문불량;p.618~619,623~625] 679
2. 발암성 물질에 대한 분자유전학적 독성평가방법의 확립[원문불량;p.677~679,704] 707
3. 분산계를 이용한 난용성약물의 가용화기술확립 793
[title page etc.]
Contents
Chapter 1. Establishment of the system for OECD GLP accreditation 78
Section 1. Introduction 78
Section 2. Analysis of OECD GLP operation 78
Section 3. Study of OECD GLP system and establishment of SOPs for the short-term toxicity testing 100
Section 4. Study of OECD GLP system and establishment of SOPs for the operating computer-system on toxicity tests 157
Chapter 2. Establishment and improvement of repeated dose toxicity testing methods using rodent 166
Section 1. Establishment of continuous infusion method in the rat tail vein 166
1. Introduction 166
2. Development of catheterization in the rat tail vein[원문불량;p.96] 167
3. A 14-day continuous dosing toxicity study of K-378 in the rats 174
4. A 14-day continuous dosing toxicity study of K-566 in the male rats 195
5. A 24-hour continuous dosing acute toxicity study of K-799 in the female rats 218
6. Discussion and Conclusions 230
7. References 232
Section 2. Toxicokinetic application for dose-range finding in the long-term toxicity studies 233
1. Introduction 233
2. Materials and Methods 234
3. Results[원문불량;p.170] 235
4. Discussion and Conclusions 250
5. References 252
Section 3. Establishment of special staining methods in histopathology 253
1. Introduction 253
2. Materials and Methods 256
3. Results 260
4. Discussion and Conclusions 267
6. References 302
Chapter 3. Establishment of special toxicity testing methods using rodent 311
Section 1. Establishment of neurotoxicity testing methods 311
1. Introduction 311
Section 2. Development of methods for studying chemical-induced immunotoxicity in rodents 337
1. Introduction 337
2. Materials and Methods 341
3. Results 351
4. Discussion and Conclusions 404
5. References 411
Section 3. Establishment of in vivo mutagenicity test methods using transgenic animals 419
1. Introduction 419
2. Materials and Methods 424
3. Results/(Restuls) 428
4. Discussion and Conclusions 431
5. References 432
Section 4. Establishment of in vitro teratogenicity method 444
1. Introduction 444
2. Materials and Methods 445
3. Results[원문불량;p.373~376] 448
4. Discussion and Conclusions 456
5. References 461
Chapter 4. Establishment of general and special toxicity testing methods using non-rodent 464
Section 1. Application of dose-range finding technique in the toxicity studies using non-rodents 464
1. Introduction 464
2. Materials and Methods 465
3. Results 467
4. Discussion and Conclusions 478
5. References 480
Section 2. Establishment of phototoxicity and photosensitization test and document of SOP 481
1. Introduction 481
2. Materials and Methods 481
3. Results 483
4. Discussion and Conclusions 484
5. References 484
Section 3. Development of methods for antigenicity tests in dogs 485
1. Introduction 485
2. Materials and Methods 486
3. Results/(Restuls) 490
4. Discussion and Conclusions 500
5. References 502
Chapter 5. Establishment of environmental toxicological testing methods and GLP system 503
Section 1. Introduction 503
Section 2. Study on ecological toxicological testing methods 505
1. Establishment of early-life stage toxicity testing methods using killifish 505
2. Study on toxicological testing methods using blood and histopathological parameters 517
Section 3. Study on metabolism and fate testing methods in environment 541
1. Study on soil metabolism 541
2. Study on plant metabolism 563
3. Study on aquatic photolysis 585
Section 4. Establishment of GLP system in environmental toxicological testing 601
1. GLP organization and operating system of environmental toxicology team 601
2. Classification, numbers and management of SOP 602
Chapter 6. Establishment of breeding methods and technologies of laboratory animals 612
Section 1. Introduction 612
Section 2. Materials and Methods 613
1. The establishment of mass production of high quality laboratory animals 613
2. The establishment of breeding methodology for nude mice 615
3. Research on a new animal model for human disease 620
Section 3. Results 624
1. The establishment of mass production of high quality laboratory animals 624
2. The establishment of breeding methodology for nude mice 630
3. Research on a new animal model for human disease 647
Section 4. Discussion and Conclusions 649
1. The establishment of mass production of high quality laboratory animals 649
2. The establishment of breeding methodology for nude mice 649
3. Research on a new animal model for human disease 653
Section 5. References 657
Chapter 7. Tasks for collaborative research between SRI and KRICT 663
Section 1. Introduction 663
Section 2. Content 664
Section 3. Conclusions 677
Chapter 8. Final reports for tasks of subcontract 678
1. Development of immunotoxicity test methods[원문불량;p.618~620,623~626] 679
2. Establishment of toxicological evaluation system of carcinogenic materials based on molecular genetic methodology[원문불량;p.677~679,704] 707
3. Solubilization of poorly water soluble drugs using disperse systems 793
Table 1-3-1. Observation of clinical sign on the GLP regulation 102
Table 2-1-1. Mortality of female rats in the 14-day study of K-378 179
Table 2-1-2. Clinical findings of female rats in the 14-day study of K-378 179
Table 2-1-3. Mean body weights of female rats in the 14-day study of K-378 181
Table 2-1-4. Mean food consumption of female rats in the 14-day study of K-378 181
Table 2-1-5. Mean water consumption of female rats in the 14-day study of K-378 182
Table 2-1-6. Hematological values of female rats in the 14-day study of K-378 183
Table 2-1-7. Differential leucocyte count of female rats in the 14-day study of K-378 184
Table 2-1-8. Serum biochemical values of female rats in the 14-day study of K-378 185
Table 2-1-9. Gross findings of female rats in the 14-day study of K-378 187
Table 2-1-10. Absolute organ weights of female rats in the 14-day study of K-378 188
Table 2-1-11. Relative organ weights of female rats in the 14-day study of K-378 190
Table 2-1-12. Histopathological findings of female rats in the 14-day study of K-378 192
Table 2-1-13. Mortality of male rats in the 14-day study of K-566 200
Table 2-1-14. Clinical findings of male rats in the 14-day study of K-566 200
Table 2-1-15. Body weights of male rats in the 14-day study of K-566 203
Table 2-1-16. Food consumption of male rats in the 14-day study of K-566 203
Table 2-1-17. Water consumption of male rats in the 14-day study of K-566 204
Table 2-1-18. Urinalysis of male rats in the 14-day study of K-566 205
Table 2-1-19. Hematological values of male rats in the 14-day study of K-566 207
Table 2-1-20. Differential leucocyte count male rats in the 14-day study of K-566 208
Table 2-1-21. Serum biochemical values of male rats in the 14-day study of K-566 209
Table 2-1-22. Gross findings of male rats in the 14-day study of K-566 210
Table 2-1-23. Absolute organ weights of male rats in the 14-day study of K-566 211
Table 2-1-24. Relative organ weights of male rats in the 14-day study of K-566 213
Table 2-1-25. Histopathological findings of male rats in the 14-day study of K-566 215
Table 2-1-26. Mortality of female rats in the acute toxicity study of K-799 222
Table 2-1-27. Clinical findings of female rats in the acute toxicity study of K-799 223
Table 2-1-28. Incidence of clinical signs of female rats in the acute toxicity study of K-799 225
Table 2-1-29. Body weights of female rats in the acute toxicity study of K-799 226
Table 2-1-30. Gross findings of female rats in the acute toxicity study of K-799 228
Table 2-2-1. Findings at cesarean section in dams treated with LGC-40863 during the organogenesis period 236
Table 2-2-2. Pharmacokinetics of LGC-40863 after an intravenous bolus administration of 100 mg/kg to rats 237
Table 3-1-1. GFAP level in specific regions of rat brain as determined by Sandwich ELISA 319
Table 3-1-2. Mean response latencies on 4 retention trials of 3 weeks old male rats 334
Table 3-1-3. Mean response latencies on 4 retention trials of 3 weeks old male rats after first trial session 334
Table 3-1-4. Mean response distances on 4 retention trials of 3 weeks old male rats 334
Table 3-1-5. Mean response distances on 4 retention trials of 3 weeks old male rats after first trial session 335
Table 3-1-6. Mean response latencies on 4 retention trials of 3 weeks old male rats 335
Table 3-1-7. Mean response distances on 4 retention trials of 3 weeks old male rats 336
Table 3-2-1. Testing panels for detecting drug-induced immunotoxicity: a guideline from the Ministry of Health and Welfare, Korea. 338
Table 3-2-2. Effects of Ethyl Carbamate, Vinyl Carbamate, Ethyl N-Hydroxycarbamate, and Methyl Carbamate on the Antibody Response to Lipopolysaccharide in Splenocyte Cultures Isolated From female Balb/C Mice 360
Table 3-2-3. Effects of ethyl carbamate, methyl carbamate, and ethyl N-hydroxycarbamate on lymphoproliferative responses in splenocyte cultures from Balb/C mice 361
Table 3-2-4. Summary of histopathology in spleen and thymus 370
Table 3-2-5. Effects of β-ionone on liver microsomal monooxygenases in Sprague Dawley rats. 383
Table 3-2-6. Effects of α- and β-ionone on ethoxyresorufin O-deethylase in male Sprague Dawley rats. 393
Table 3-2-7. Effects of α- and β-ionone on methoxyresorufin O-demethylase in male Sprague Dawley rats. 395
Table 3-2-8. Effects of α- and β-ionone on pentoxyresorufin O-depentylase in male Sprague Dawley rats. 396
Table 3-2-9. Effects of α- and β-ionone on benzyloxyresorufin O-debenzylase in male Sprague Dawley rats. 397
Table 3-2-10. Effects of pretreatment with β-ionone on thioacetamide induced changes of serum clinical parameters in male BALB/c mice 402
Table 3-3-1. Induction of mutant plaques in livers of male transgenic mice 428
Table 3-3-2. Induction of mutant plaques in kidneys of male transgenic mice 429
Table 3-3-3. Induction of mutant plaques in spleen of male transgenic mice 430
Table 3-3-4. Induction of mutant plaques in livers of male transgenic rats 430
Table 3-4-1. Embryonic growth parameters and morphogenesis of rat whole-embryos grown from GD 9.5 for 48hrs in vivo and in vitro. 454
Table 3-4-2. Effects of in vitro exposure to PHT on embryonic growth and differentiation of GD 11.5 rat embryos explanted on GD 9.5 and cultured 48hrs. 454
Table 3-4-3. Morphological defects induced by in vitro exposure to PHT in GD 11.5 rat embryos explanted on GD 9.5 and cultured 48hrs. 455
Table 4-3-1. Sensitization of mice for heterologous passive cutaneous anaphylaxis and indirect hemagglutination test. 491
Table 4-3-2. Sensitization of guinea pigs for active systemic anaphylaxis and homologous passive cutaneous anaphylaxis. 492
Table 4-3-3. Active systemic anaphylaxis in guinea pigs. 493
Table 4-3-4. Four-hours passive cutaneous anaphylaxis test in guinea pigs with sera from sensitized guinea pigs. 494
Table 4-3-5. 24-hours heterologous passive cutaneous anaphylaxis test in rats with sera from sensitized mice. 495
Table 4-3-6. Indirect hemagglutination test with sera isolated from sensitized mice. 496
Table 5-2-1. Test conditions 507
Table 5-2-2. Hatching success of the embryo exposed to the various exposure conditions 509
Table 5-2-3. Days to end of hatching of the embryo exposed to the various exposure conditions 511
Table 5-2-4. Post-hatching success of the embryo exposed to the various exposure conditions 512
Table 5-2-5. Times to reach the developmental stage of embryo exposed to different water temperature 513
Table 5-2-6. Water quality used in this test 519
Table 5-2-7. Haematological results of korean catfish(Silurus asotus) following to expose at 1.78 mg/l of cadmium 524
Table 5-2-8. Haematological results of korean catfish(Silurus asotus) following to expose at 0.35 mg/l of cadmium 525
Table 5-2-9. Blood chemistry values of korean catfish, Silurus asotus, exposed to 1.78 mg/l of cadmium 528
Table 5-2-10. Blood chemistry values of korean catfish, Silurus asotus, exposed to 0.35 mg/l of cadmium 530
Table 5-2-11. Histopathological changes of liver, skin, gill, brain, kidney and intestine of korean catfish, Silurus asotus, exposed to 0.35 mg/l and 1.78 mg/l of cadmium 533
Table 5-2-12. Cadmium concentrations in the tissue and water during the test 534
Table 5-3-1. Physicochemical properties of the test soil. 545
Table 5-3-2. Total 14C material balance from a 14C-alachlor treated soil(이미지참조) 553
Table 5-3-3. Recovery of radiochemicals from plant 572
Table 5-3-4. TRR, and radioactivity of extract and residue 574
Table 5-3-5. Relative % of absorbed radioactivity 578
Table 5-3-6. Flupyrazofos recovery 591
Table 5-3-7. Rate constant and half-life of Flupyrazofos 596
Table 5-3-8. Photo bleaching of SHW 597
Table 5-4-1. Kinds of SOP regarding environmental toxicity testing, classification code 603
Table 6-2-1. The list of microorganisms for microbiological monitoring 613
Table 6-2-2. Chemical composition of diet 617
Table 6-2-3. Formula and composition of diet 618
Table 6-3-1. The results of microbiological monitoring of animals in SPF condition 625
Table 6-3-2. Results of microbiology monitoring of animal rooms 626
Table 6-3-3. Results of microbiology monitoring of animal rooms (falling microbes) 626
Table 6-3-4. Results of microbiology monitoring of animal rooms(Smear test) 626
Table/Talbe 6-3-5. Microbiology test of diet 627
Table/Talbe 6-3-6. Environmental control of animal rooms 627
Table 6-3-7. Reproductive performance of rate and mice 628
Table 6-3-8. The number of animals distributed 628
Table 6-3-9. The list of institutes using KRICT animals and the strains 629
Table 6-3-10. Changes of body weight in nude and BALB/c Mice with age. 630
Table 6-3-11. Changes of organ weight in Nude and BALB/c male mice with age. (Unit : g) 632
Table 6-3-12. Changes of organ weight in Nuce and BALB/c female mouse with age. (Unit : g) 633
Table 6-3-13. Abbreviations, units and analysis methods of the hematology. 634
Table 6-3-14. Changes of blood hematology in Nude and BALB/c male mice with age. 635
Table 6-3-15. Changes of blood hematology in nude and BALB/c female mice with age. 636
Table 6-3-16. WBC differential counts in nude and BALB/c male mice with age. (unit : percent Mean ± S.D) 637
Table 6-3-17. WBC differential counts in nude female mice with age (unit : percent Mean ± S.D) 638
Table 6-3-18. Abbreviations, units and analysis methods of the biochemistry. 639
Table 6-3-19. Changes of biochemistry in nude and BALB/c male mice with age. 640
Table 6-3-20. Changes of biochemistry in Nude and BALB/c female mice with age. 641
Table 6-3-21. Urinalysis in Nude and BALB/c male mice with ages. 643
Table 6-3-22. Urinalysis in Nude and BALB/c female mice with ages. 644
Table 6-3-23. Comparison of spleen B & T-cell in nude and BALB/c mice(FACs anlaysis). 645
Table 6-3-24. Reproductive performance in BALB/c, BALB/c nu/+, and Nude mice. 646
Fig. 2-1-1. The setting of needle catheter, syringe and tubings 173
Fig. 2-1-2. A rat in a polycarbonate cage connected to the infusion assembly[원문불량;p.96] 173
Fig. 2-2-1. Linearity for the standard K-585 spiked in the plasma by HPLC analysis. 240
Fig. 2-2-2. Stability of K-585 under the various conditions. Each point represent the mean±SE of the three different determination. 241
Fig. 2-2-3. Retention and peak response characteristics of K-585 on the HPLC chromatogram at various pHs of buffer or at various ACN fractions in the mobile phase. Each point represents the mean±SE of the different three determination. 243
Fig. 2-2-4. Plasma time vs concentration curves of K-585 after a single oral administration to the rats at various doses. 244
Fig. 2-2-5. Nonlinear toxicokinetics for the AUC0-120min(이미지참조) of K-585 after the oral at various doses in the rats. 245
Fig. 2-2-6. Nonlinear kinetics for the CLapp-1(이미지참조) of K-585 after the oral at various doses in the rats. 246
Fig. 2-2-7. Steady-state kinetics for initial repeated dosing and serum concentration curves of K-585 after the oral 3 month-repeated administration of K-580 to the rats. Each point represents the mean±SE of three rats.[원문불량;p.170] 247
Fig. 2-2-8. Time course of LGC-40863 plasma concentration after a single intravenous dose of 100 mg/kg to female rats. Dotted curve represents the calculated curve obtained by fitting the two-compatrtment open model. Each value and vertical bar represent the mean of three rats. 248
Fig. 2-2-9. Cumulative excretion of total radioactivity in the urine and feces after a single oral administration of LGC-40863 at doses of 500 and 2000 mg/kg spiked with (14C)LGC-40863 (200 Ci/kg) to female rats. 249
Fig. 2-3-1. Scheme of rat spermatogenesis. 281
Fig. 2-3-2. Photomicroscopic finding of the testis shows severe atrophy comparing with normal(N). 42 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Scale=3cm. 282
Fig. 2-3-3. Photomicroscopic finding of the epididymis shows severe atrophy comparing with normal(N), 42 day after 3.5g/kg/day gavage of 2-broopropane for 3 consecutive days. Bar=1cm. 282
Fig. 2-3-4. Normal seminiferous tubule in stage I. 15St; step 15 spermatid, S; Sertoli cell, Sc; spermatocyte, Sg; spermstogonia. H-E. Bar=10㎛ 283
Fig. 2-3-5. a,b; seminiferous tubules in stage I have degenerating sprmatogonia(↑). 1 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 283
Fig. 2-3-6. Normal seminiferous tubule in stage VII. P;pachytene spermatocyte, PL;preleptotene spermatocyte. H-E. Stain. Bar=10㎛ 284
Fig. 2-3-7. Seminiferous tubule in stage VII shows depletion of preleptotene spermatocytes. P; pachytene spermaticyte, St; step 7 spermatid. 5 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive day. PAS. Bar=10㎛ 284
Fig. 2-3-8. Normal seminiferous tubule in stage X. P; pachytene spermatocyte, L; leptotene spermatocyte. H-E. Bar=10㎛ 285
Fig. 2-3-9. a; seminiferous tubules in various stages show depletion of spermatogonia and early spermatocytes, and spermatid retention in stage IX. Arrow; residual body. Bar=20㎛. b; seminiferous tubule in stage X shows spermatid retention. P; pachytene spermatocyte, arrow heads; step 19 spermatids. 7 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 285
Fig. 2-3-10. Seminiferous tubule in stage VII shows complete delpletion of spermatocytes. 14 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 286
Fig. 2-3-11. Seminiferous tubule shows delpletion of spermatocytes and round spermatid and regenerating spermatogonia. 28 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 286
Fig. 2-3-12. Epididymal tubules show oligospermia or aspermia. 28 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 287
Fig. 2-3-13. Seminiferous tubules show complete delpletion(*(이미지참조)) or regeneration(↑) of germ cells. 42 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=20㎛ 287
Fig. 2-3-14. Seminiferous tubules show complete depletion of germ cells and Leydig cell hyperplasia. 70 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=20㎛ 288
Fig. 2-3-15. Seminiferous tubule shows regeneration of germ cells and degenerating multinucleated giant cells(↑). 70 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 288
Fig. 2-3-16. Epididymal tubules show oligospermia and exfolated germ cells. 70 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=10㎛ 289
Fig. 2-3-17. Atrophy and vacuolation of spermatogonia by hypertonic fixative without detachment of spermatocytes, and spermatid from the Sertoli cells. 1 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Toluidine blue, Bar=10㎛ 289
Fig. 2-3-18. Electron micrograph of normal seminiferous tubule in stage IV. Spermatogonia(Sg), step 4 spermatid(St) ad Sertoli cells(Se) in the stag IV. Bar=2㎛ 290
Fig. 2-3-19. a,b,c; electron micrographs of degenerating spermatogonia show extensive heterochromatin, chromatin margination, or pyknosis. 1 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Bar=2㎛ 290
Fig. 2-3-20. Electron micrograph of interstitium shows normal appearance. 1 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. L; Leydig cell Bar=4.66㎛ 291
Fig. 2-3-21. Electron micrograph of interstitium shows normal appearance. 70 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. L; Leydig cell Bar=3.18㎛ 291
Fig. 2-3-22. Electron micrograph of severe atrophic seminiferous epithelium. Note the presence of an infolded Sertoli cell(S) nucleus, convoluted basement mmbrane(↑). M; myeloid cell. 70 days 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Bar=4.66㎛ 291
Fig. 2-3-23. Lanthanum particle is deposited between the Sertoli cell and spermatogonia(Sg), Sc; Spermatocyte, M; myeloid cell. 1 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Bar=3.18㎛ 291
Fig. 2-3-24. Normal seminiferous tubules in varoius stages. Spermatogonia(arrow head) spermatocytes(↑) show positive reaction for PCNA antibody. Immunohisochemical stain for PCNA antibody. Bar=20㎛ 292
Fig. 2-3-25. Seminiferous tubules in stages VI-VII show depletion of PCNA positive cells. 5 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=20㎛ 292
Fig. 2-3-26. Seminiferous tubules in stages XIII and III have PCNA positive cells. 7 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=20㎛ 293
Fig. 2-3-27. Seminiferous tubules in stages I-III have PCNA positive cells. 14 day after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=20㎛ 293
Fig. 2-3-28. Seminiferous tubules show increased PCNA positive cells. 28 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=20㎛ 294
Fig. 2-3-29. Seminiferous tubules show markedly increased PCNA positive cells. 42 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=20㎛ 294
Fig. 2-3-30. a; numerous interstitial cells show PCNA antibody positive reaction(↑), Bar=20㎛. b,c; high magnification of PCNA positive cells. 70 days after 3.5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Immunohistochemical stain for PCNA antibody. Bar=10㎛ 295
Fig. 2-3-31. Relative proportions of cells with haploid, diploid and tetraplid states of DNA ploidy in the testicular cell suspensions of the Sprague-Dawley rats treated with 3.5g/kg/day gavage of 2-bromopropane for once daily 3 consecutive days. a. control. b. 3 day after the completion of administration, showing proportions of decreased diploid cells. c. 5 days after, showing proportions of decreased diploid and tetraploid cells. d. 7 days after, showing proportions of decreased diploid and tetraploid cells. e. 14 days after, showing proportions of decreased diploid and tetraploid cells. f. 28 days after, showing proportions of markdly decreased diploid and tetraploid cells. g. 42 days after, showing proportions of markdly increased diploid and tetraploid cells. h. 70 days after, showing proportions of decreased diploid and tetraploid cells comparing with g. 296
Fig. 2-3-32. Seminiferous tubules in various stages show depletion of spermatogonia and early spermatocytes or severe exfoliation and degeneration of germ cell, and cytoplasmic vacuolation of Sertoli cells. 1 day after 5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=20㎛ 297
Fig. 2-3-33. Seminiferous tubules show severe degeneration and exfoliation of germ cell, and cytoplasmic vacuolation of Sertoli cells. Arrow; exfolated multinucleated giant cells. 7 days after 5g/kg/day gavage of 2-bromopropane for 3 consecutive days. H-E. Bar=20㎛ 297
Fig. 2-3-34. Electron micrograph of seminiferous tubule shows degenerating spermatocytes(↑) and Sertoli cell(S). Sertoli cells show irregular shape of nucleus and cytoplasmic vauolation. Spermatocyte shows a extensive heterochromatin 1 day after 5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Bar=2㎛ 298
Fig. 2-3-35. Electron micrograph of multinucleated giant cell formed by degenerating spermatids. 7 day after 5g/kg/day gavage of 2-bromopropane for 3 consecutive days. Bar=2㎛ 298
Fig. 2-3-36. Seminiferous tubules show complete delpletion of germ cells(*(이미지참조)) or degeneration of spermatids(↑), and interstitium shows Leydig cell hyperplasia with eosinophilic materials. 8 weeks after 0.5g/kg/day gavage of 2-bromopropane for 8 consecutive weeks. H-E. Bar=20㎛ 299
Fig. 2-3-37. Epididymal tubules show oligospermia and exfolated germ cells(↑) in lumen. 0.5g/kg/day gavage of 2-bromopropane for 8 consecutive weeks. H-E. Bar=10㎛ 299
Fig. 2-3-38. Relative proportions of cells with haploid, diploid and tetraplid states of DNA ploidy in the testicular cell suspensions of the Sprague-Dawley rats treated with 0.5g/kg/day of 2-bromopropane orally for 8 weeks. a. control b. 8 weeks after the completion of administration, showing proportions of decreased haploid cells. 300
Fig. 2-3-39. Seminiferous tubules show germ cell degeneration, exfoliation, and multinucleated giant cells. Intratesticular injection of 10㎕ 2-bromopropane. H-E. Bar=20㎛ 301
Fig. 2-3-40. Seminiferous tubule shows severe germ cell degeneration, vacuolation, and exfoliation. Intratesticular injection of 30㎕ 2-bromopropane. H-E. Bar=10㎛ 301
Fig. 3-1-1. Sample preparation flow chart for GFAP and second-tier assays 315
Fig. 3-1-2. GFAP level in specific regions of male rat brain after intravenous administration of kainic acid(1 mg/kg) determined by Sandwich ELISA 320
Fig. 3-1-3. GFAP level in specific regions of female rat brain after intravenous administration of kainic acid(1 mg/kg) determined by Sandwich ELISA 321
Fig. 3-1-4. GFAP level in specific regions of male rat brain after intravenous administration of kainic acid(0.1 mg/kg) determined by Sandwich ELISA 322
Fig. 3-1-5. GFAP level in specific regions of female rat brain after intravenous administration of kainic acid(0.1 mg/kg) determined by Sandwich ELISA 323
Fig. 3-1-6. Mean response latencies on 4 retention trials of 3 weeks old male rats in morris water maze test 328
Fig. 3-1-7. Mean response latencies on 4 retention trials of 3 weeks old male rats after first trial session in morris water maze test 329
Fig. 3-1-8. Mean response distances on 4 retention trials of 3 weeks old male rats in morris water maze test 330
Fig. 3-1-9. Mean response distances on 4 retention trials of 3 weeks old male rats after first trial session in morris water maze test 331
Fig. 3-1-10. Mean response latencies on 4 retention trials of 3 weeks old male rats in radial 8-arm maze test 332
Fig. 3-1-11. Mean response latencies on 4 retention trials of 3 weeks old male rats in radial 8-arm maze test 333
Fig. 3-2-1. Effects of 28-day exposure to 2-bromopropane on hematological parameters in male Sprague Dawley rats. 2-Bromopropane in com oil (10 ml/kg) was administered orally to rats for 28 consecutive days. Four days prior to the necropsy, individual animals(aniamls) were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells. Following the overnight fasting, the blood was collected to an EDTA vial for hematology. Each bar represents the mean ± SE of five animals. The asterisks indicate the value significantly different from the vehicle control at P<0.05(*) or P<0.01(**)(이미지참조) 353
Fig. 3-2-2. Effects of 28-day exposure to 2-bromopropane on biochemical parameters in male Sprague Dawley rats. 2-Bromopropane in com oil (10 ml/kg) was administered orally to rats for 28 consecutive days. Four days prior to the necropsy, individual animals/(aniamls) were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells. Following the overnight fasting, the blood was collected to prepare serum for clinical chemistry. Each bar represents the mean ± SE of five animals. The asterisks indicate the value significantly different from the vehicle control at P<0.05(*) or P<0.01(**)(이미지참조) 354
Fig. 3-2-3. Effects of 28-day exposure to 2-bromopropane on T-dependent antibody response in male Sprague Dawley rats. 2-Bromopropane in com oil (10 ml/kg) was administered orally to rats for 28 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). Each bar represents the mean ± SE of five animals(animals). The asterisks indicate the value significantly different from the vehicle control at P<0.05(*) or P<0.01(**)(이미지참조) 355
Fig. 3-2-4. Flow cytometric analyses of immune cells. To analyze macrophages. B-cells, T-cells and T-cell subsets (CD4 helper and CD8 suppressor T-cell), each of the respective cell typer was labeled with an appropriat monoclonal antibody conjugated to a fluorescent probe 356
Fig. 3-2-5. Effects of 28-day exposure to 2-bromopropane on splenic lymphocyte subpopulation in male Sprague Dawley rat. 2-Bromopropane in corn oil (10 ml/kg) was administered orally to rats for 28 consecutive days. Each bar represents the mean ± SE of five animals. 357
Fig. 3-2-6. Effects of 28-day exposure to 2-bromopropane on thymic lymphocyte subpopulation in male Sprague Dawley rats. 2-Bromopropane in corn oil (10 ml/kg) was administered orally to rats for 28 consecutive days. Each bar represents the mean ± SE of five animals. The asterisks indicate the value significantly different from the vehicle control at P<0.05(*) or P<0.01(**)(이미지참조) 358
Fig. 3-2-7. Effects of 7-day exposure to ethyl carbamate on splenic lymphocyte subpopulation in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Each bar represents the mean±SE of five animals. 363
Fig. 3-2-8. Effects of 7-day exposure to ethyl carbamate on splenic lymphocyte subpopulation in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). Each bar represents the mean ± SE of five animals. An asterisk indicates the value significantly different from the vehicle control at P<0.05 364
Fig. 3-2-9. Effects of 7-day exposure to ethyl carbamate on thymic lymphocyte subpopulation in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). Each bar represents the mean ± SE of five animals. An asterisk indicates the value significantly different from the vehicle control at P<0.05 365
Fig. 3-2-10. Effects of 7-day exposure to ethyl carbamate on histopathiogy of spleen in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). The spleen was stained with hematoxylin and eosin. 366
Fig. 3-2-11. Effects of 7-day exposure to ethyl carbamate on histopathlogy of thymus in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). The thymus was stained with hematoxylin and eosin. 368
Fig. 3-2-12. Effects of adrenalolectomy (ADX) in ethyl carbamate-induced changes of splenic lymphocyte subpopulation in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to either naive, sham operated, or adrenolectomized mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). Each bar represents the mean ± SE of five animals. The asterisks indicate the value significantly different from the individual naive controls at P<0.01. 372
Fig. 3-2-13. Effects of adrenalolectomy (ADX) in ethyl carbamate-induced changes of thymic lymphocyte subpopulation in female BALB/c mice. Ethyl carbamate in saline (10 ml/kg) was administered intraperitoneally to mice for 7 consecutive days. Four days prior to the necropsy, individual animals were immunized with 1 ml of 2.5% sheep red blood cells to enumerate the antibody-forming cells (AFCs). Each bar represents the mean ± SE of five animals. 373
Fig. 3-2-14. Effects of sera from ethyl carbamate-treated mice on LPS-induced lymphoproliferation of splenocyte cultures. The sera ware prepared from mice pretreated with 1 g/kg of ethyl carbamate and added to a normal splenocyte culture isolated from female BALB/c mice in the presence of 100 ㎍/ml of lipopolysaccharide. Each bar represents the mean ± SE of quadruplicate cultures. 374
Fig. 3-2-15. Optimization of LPS and ConA mitogenicity using the nonradioactive colorimetric lymphoproliferation assay. Various number of cells were cultured for either 48 or 72 hr in the presence of either 100 ㎍/ml of lipopolysaccharide or 1.0㎍/ml of concanavalin A. Twenty μl of proliferation assay reagent was added into each well for the given time. The absorbance at 490 nm was monitored in an ELISA reader. Each value represents the mean of duplicate wells. 376
Fig. 3-2-16. An example of LPS and ConA mitogenicity using the nonradioactive colorimetric lymphoproliferation assay. 2 × 105(이미지참조) cells were cultured with a test substance for 72 hr in the presence of either 100 ㎍/ml of lipopolysaccharide or 1.0 ㎍/ml of concanavalin A. Twenty μl of proliferation assay reagent was added into each well for 4 hr. The absorbance at 490 nm was monitored in an ELISA reader. Each value represents the mean ± SD of quadruplicate wells. 378
Fig. 3-2-17. An example of mixed lymphocyte response using the nonradioactive colorimetric lymphoproliferation assay. 2 × 105 responder cells from female BALB/c mice were co-cultured with 8 ×105 mitomycin C-inactivated stimulator cells from female DBA/2 mice in the presence of a test substance for 120 hr. Twenty μ1 of proliferation assay reagent was added into each well for 4 hr. The absorbance at 490 nm was monitored in an ELISA reader. Each value represents the mean ± SD of quadruplicate wells.(이미지참조) 379
Fig. 3-2-18. Effect of intravenous and subcutaneous immunization on IgM production in rats. Male Fischer 344 rats were immunized once per week for two times with keyhole limpet hemocyanin (KLH). Seven days after the last immunization, the sera were prepared and the KLH-specific IgM antibody was assayed by the ELISA method. Each bar represents the mean absorbance at 405 nm of two animals. 380
Fig. 3-2-19. Effect of intravenous and subcutaneous immunization on IgG production in rats. Male Fischer 344 rats were immunized once per week for two times with keyhole limpet hemocyanin (KLH). Seven days after the last immunization, the sera were prepared and the KLH-specific IgG antibody was assayed by the ELISA method. Each bar represents the mean absorbance at 405 nm of two animals. 382
Fig. 3-2-20. RT-PCR amplification of P450 1A2, 2B1, 2B2, 2C6 and NADPH-P450 reductase mRNAs. β-Ionone at 600 mg/kg were treated to rats subcutaneously for 6 hr and poly(A+(이미지참조)) mRNAs were prepared. A, P450 1A2; B, P450 2B1; C, P450 2B2; D, P450 2C6; E, NADPH-P450 reductase; F, glucose 3-phosphate dehydrogenase. Lane 1, mw marker showing 100-bp ladder; lane 2, untrated control; lane 3, β-Ionone-treated; lane 4, TCDD-treated at 10㎍/kg intraperitoneally once for 32 days; lane 5 in E, mw marker showing 2000-, 1500-, 1000-, 750-, 500-, 300-, 150-, and 50-bp ladders from top; lane 5 in F, 100-bp markers from 1000-bp. 384
Fig. 3-2-21. Induction of P450 2B by α-and β-ionone in liver microsomes of male Sprague Dawley rats. The rats were administered subcutaneously with either α- or β-ionone in corn oil 72 and 48 hr before sacrifice. The microsomal proteins (10 mg/well) were resolved on a 10% SDS-PAGE and transferred onto a nitrocellulose filter for the Western immunoblotting. The filter was incubated with the primary antibody against P450 2B proteins, followed by an incubation with alkaline phosphatase-conjugated goat anti-rabbit IgG. Then the filter was developed using the 1:1:10 mixture of 5-bromo-4-chloro-3-indolyl phosphate, nitroblue tetrazolium and 0.1 M Tris buffer. The upper band indicates p450 2B proteins and the lower band indicates a non-specific binding. 386
Fig. 3-2-22. An RT-PCR amplification of P450 2B1 mRNAs from rat livers treated with either α- or β-ionone. The rats were treated with ionones subcutaneously for 6 hr. M, molecular weight size markers showing 100-bp ladders; VH, corn oil-treated vehicle control. 387
Fig. 3-2-23. Time-course studies of P450 mRNA induction by β-ionone. A) P450 2B1; B) P450 1A2. The rats were treated with 600 mg/kg β-ionone subcutaneously for the designated time. M, molecular weight size markers showing 100-bp ladders. 388
Fig. 3-2-24. Effects of the routes of administration on P450 2B induction by β-ionone. β-Ionone was administered either subcutaneously or orally at 300 mg/kg 72 and 48 hr before sacrificing the animals. The liver microsomal proteins (10 mg/well) prepared from individual inducers-administered male Sprague Dawley rate were resolved on a 10% SDS-PAGE. Lane 1, untreated control; lane 2, 10 mg/kg TCDD-treated once for 3 days, ip; lane 3, 5 g/kg ethanol-treated daily for 3 days, po; lane 4, 80 mg/kg phenobarbital-treated daily for 3 days, ip; land 5, β-ionone treated subcutaneously; lane 6, β-ionone treated orally. 389
Fig. 3-2-25. Sex difference in the induction of P450 2B by α- and β-ionone. α- and β-ionone were administered orally at 100 mg/kg for 24 hr. The live microsomal proteins (10 mg/well) were resolved on a 10% SDS-PAGE and transferred onto a nitrocellulose filter for the Western immunoblotting. The lower band indicates a non-specific binding. α-I, α-ionone; β-I, β-ionone. 390
Fig. 3-2-26. Effects of α- and β-ionone on the expression of P450 1A proteins in male Sprague Dawley rats. Ionones were administered orally at 100 mg/kg for 24 hr. The liver microsomal proteins (10 mg/well) were resolved on a 10% SDS-PAGE and transferred onto a nitrocellulose filter for the Western immunoblotting. α-I, α-ionone; β-I, β-ionone. 391
Fig. 3-2-27. Nomenclature used for lobes of rat liver. A view of the dorsal surface of the liver. 392
Fig. 3-2-28. Western immunoblotting for P450 2B protein in rat liver S-9 fractions isolated from livers treated with α- and β-ionone. The proteins from S-9 fractions isolated from liver lobes were used. A. caudate; B, right posterior; C, right anterior; D, right median; E, left median; F, left lobe. 398
Fig. 3-2-29. Pretreatment of male BALB/c mice with β-ionone potentiates thioacetamide-induced elevation of SGPT and SGOT activities. Mice were pretreated with 600 mg/kg of β-ionone 72 and 48 hr prior to an administration with thioacetamide. Twenty four hr later, the serum was prepared. Each bar represents the mean activity ± SE of five animals. The asterisks indicate the values significantly different from each corn oil-treated control at P<0.05(*) or P<0.01(**).(이미지참조) 400
Fig. 3-2-30. Liver histopathology. 200X magnification. A, vehicle control; B, 100 mg/kg of thioacetamide, C, 200 mg/kg of thioacetamide; D, 600 mg/kg of β-ionone; E, β-ionone plus 100 mg/kg thioacetamide; F, β-ionone plus 200 mg/kg thioacetamide. Severe hepatic necrosis and congestion and mild infiltration of polymorphonuclear cells were obvisously observed in panel F. 401
Fig. 3-2-31. Effects of β-ionone on P450 2B1-selective benzyloxyresorufin O-debenzylase (BROD) and pentoxyresorufin O-depentylase (PROD) activities in liver S-9 fractions. Mice were pretreated with 600 mg/kg of β-ionone 72 and 48 hr prior to an administration of thioacetamide. Twenty four hr later, livers were removed to prepare the S-9 fractions. 403
Fig. 3-4-1. Photograph of a GD 9.5 rat decidua removed from uterus. Scale bar is 1mm.[원문불량;p.373] 450
Fig. 3-4-2. Photograph of a GD 9.5 rat conceptus. Reichert's membrane has been torn open. The embryo is in head-fold stage. Scale bar is 1mm.[원문불량;p.373] 450
Fig. 3-4-3. Photograph of a control conceptus, showing normal appearance. Scale bar is 1mm.[원문불량;p.374] 451
Fig. 3-4-4. Photograph of a control embryo, showing no abnormalities. Scale bar is 1mm.[원문불량;p.374] 451
Fig. 3-4-5. Photograph of a conceptus treated with 50 ㎍/㎖ PHT, showing abnormal rotation. Scale bar is 1mm.[원문불량;p.375] 452
Fig. 3-4-6. Photograph of a conceptus treated with 100 ㎍/㎖ PHT, showing altered yolk sac and embryonic circulation. Scale bar is 1mm.[원문불량;p.375] 452
Fig. 3-4-7. Photograph of a embryo treated with 100 ㎍/㎖ PHT, showing growth retardation, abnormal rotation, craniofacial hypoplasia, missing optic vesicle, enlarged cardiac tube, open caudal neuropore, limb bud hypoplasia, and blunted tail. Scale bar is 1mm.[원문불량;p.376] 453
Fig. 3-4-8. Photograph of a embryo treated with 100 ㎍/㎖ PHT, showing growth retardation, abnormal rotation, craniofacial hypoplasia, absence of the second branchial bar, limb bud hypoplasia, enlarged cardiac tube and pericardium, and blunted tail. Scale bar is 1mm.[원문불량;p.376] 453
Fig. 4-1-1. Linearity for the standard SA spiked in the plasma by HPLC analysis. 468
Fig. 4-1-2. Serum concentration curves of SA after the oral single administration of K-580 to the beagle dogs. Each point represents the mean ± SE of three dogs. 469
Fig. 4-1-3. Serum concentration curves of SA after the oral 3 month-repeated administration of K-580 to the beagle dogs. Each point represents the mean ± SE of three dogs. 470
Fig. 4-1-4. Steady-state kinetics of SA after the oral multiple administration of K-580 during a period of 5days in the beagle dogs. Each point represents the mean ± SE of three dogs. 471
Fig. 4-1-5. Toxicokinetic linearity for the AUC0-24hr(이미지참조) of SA in the oral single administration and repeated administration of K-580 at the three doses to the beagle dogs. Each value represents the mean ± SE of three dogs. 472
Fig. 4-1-6. Toxicokinetic linearity for the Cmax(이미지참조) of SA in the oral single and repeated administration of K-580 to the beagle dogs. Each value represents the mean ± SE of three dogs. 473
Fig. 4-1-7. Toxicokinetic linearity for the Tmax(이미지참조) of SA in the oral single and repeated administration of K-580 to the beagle dogs. Each value represents the mean ± SE of three dogs. 474
Fig. 4-1-8. Toxicokinetic linearity for the Kel(이미지참조) of SA in the oral single and repeated administration of K-580 to the beagle dogs. Each value represents the mean ± SE of three dogs. 475
Fig. 4-1-9. Toxicokinetic linearity for the t1/2(이미지참조) of SA in the oral single and repeated administration of K-580 to the beagle dogs. Each value represents the mean ± SE of three dogs. 476
Fig. 4-1-10. Toxicokinetic linearity for the AUC0-5day(이미지참조) of SA relating to steady-state condition after the oral multiple administration of K-580 at the beagle dogs. Each value represents the mean ± SE of three dogs. 477
Fig. 4-3-1. Detection of IgG in sera from male beagle dogs by ELISA. Male beagle dogs were treated intravenously with a test substance for 28 consecutive days. The sera were prepared from individual animals to analyze the test substance-specific antibodies by the ELISA. In a 96-well plate coated with the test substance, sera were added. Rabbit anti-dog IgG-alkaline phosphatase conjugate was used to detect to antibodies. Each value represents the absorbance at 405 nm of each animal. 498
Fig. 4-3-2. Detection of IgG in sera from female beagle dogs by ELISA. Female beagle dogs were treated intravenously with a test substance for 28 consecutive days. The sera were prepared from individual animals to analyze the test substance-specific antibodies by the ELISA. In a 96-well plate coated with the test substance, sera were added. Rabbit anti-dog IgG-alkaline phosphatase conjugate was used to detect to antibodies. Each value represents the absorbance at 405 nm of each animal. 499
Fig. 5-2-1. Continuous flow system used in this test. 508
Fig. 5-2-2. Experimental scheme for this study. 520
Fig. 5-3-1. Soil metabolism apparatus 547
Fig. 5-3-2. DAR Autoradiogram of 14C-alachlor(이미지참조) 550
Fig. 5-3-3. Chemical structure of alachlor, 2-chloro-2', 6'-diethylacetamide, 2, 6-diethylacetanilide 550
Fig. 5-3-4. Mass spectrum of 2-chloro-2', 6'-diethylacetamide 551
Fig. 5-3-5. 14CO₂evolution rate from 14C-alachlor treated soil.(이미지참조) 551
Fig. 5-3-6. Distribution of radiocarbon in acetone extract and soil 552
Fig. 5-3.7. DAR autoradiograms of alachlor and metabolites 554
Fig. 5-3-8. Degradation rate of 14C-alachlor(이미지참조) 555
Fig. 5-3-9. Formation of metabolites. 556
Fig. 5-3-10. DAR autoradiogram of 14C-Flupyrazofos(이미지참조) 568
Fig. 5-3-11. Chemical structure of 14C-Flupyrazofos, 14C-PTMHP and 14C-Flupyrazofos oxon(이미지참조) 568
Fig. 5-3-12. Time course of TRR, and radioactivity of extract and residue. 573
Fig. 5-3-13. Photodegradation rate of Flupyrazofos 574
Fig. 5-3-14. DAR autoradiogram of solvent extract 575
Fig. 5-3-15. Image autoradiogram of solvent extract 576
Fig. 5-3-16. Time course of Flupyrazofos and metabolites. 577
Fig. 5-3-17. Radioactivity absorption pattern for plant parts 578
Fig. 5-3-18. Image autoradiogram of seelings 579
Fig. 5-3-19. Chemical structure of Flupyrazofos 588
Fig. 5-3-20. Flupyrazofos photodegradation rate by light 592
Fig. 5-3-21. Flupyrazofos degradation rate under dark condition 593
Fig. 5-3-22. Photodegradation of Flupyrazofos under air saturated condition 594
Fig. 5-3-23. Photodegradation of Flupyrazofos under nitrogen saturated condition 595
Fig. 5-4-1. GLP organization of toxicology research center. 601
Fig. 5-4-2. cover page of environmental toxicology research team's SOP. 604