• 대한예방의학회:학술대회논문집
• /
• 대한예방의학회 1994년도 교수 연수회(환경)
• /
• pp.431-438
• /
• 1994

As currently conducted, standard rodent bioassays do not provide sufficient information to assess carcinogenic risk to humans at doses thousands of times below the maximum tolerated dose. Recent analyses indicate that measures of carcinogenic potency from these tests are restricted to a narrow range about the maximum tolerated dose and that information on shape of the dose-response is limited in experiments with only two doses and a control. Extrapolation from high to low doses should be based on an understanding of the mechanisms of carcinogenesis. We have postulated that administration of the maximum tolerated dose can increase mitogenesis which, in turn. increases rates of mutagenesis and, thus, carcinogenesis. The animal data are consistent with this mechanism, because about half of all chemicals tested are indeed rodent carcinogens, and about 40% of the positives are not detectably mutagenic. Thus, at low doses where cell killing does not occur, the hazards to humans of rodent carcinogens may be much lower than commonly assumed. In contrast, for high-dose exposures in the workplace, assessment of hazard requires comparatively little extrapolation. Nevertheless. permitted workplace exposures are sometimes close to the tumorigenic dose-rate in animal tests. Regulatory policy to prevent human cancer has primarily addressed synthetic chemicals, yet similar proportions of natural chemicals and synthetic chemicals test positive in rodent studies as expected from an understanding of toxicological defenses, and the vast proportion of human exposures are to natural chemicals. Thus, human exposures to rodent carcinogens are common. The natural chemicals are the control to evaluate regulatory strategies, and the possible hazards from synthetic chemicals should be compared to the possible hazards from natural chemicals. Qualitative extrapolation of the carcinogenic response between species has been investigated by comparing two closely related species: rats and mice. Overall predictive values provide moderate confidence in interspecies extrapolation; however, knowing that a chemical is positive at any site in one species gives only about a 50% chance that it will be positive at the same site in the other species.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제19권1호
• /
• pp.199-204
• /
• 2009

The newly cloned Bacillus thuringiensis cry1-5 gene showed high activity to both Plutella xylostella and Spodoptera exigua, while cry1Ac only showed high activity against P. xylostella but low to S. exigua. Through the alignment of amino acid sequences between Cry1Ac and Cry1-5, we found 12 different residues in domain I (6 residues) and domain II (6 residues). In this study, the modified cry1Ac gene, which is constructed according to a crop-preferring codon usage, was used as a template to construct mutant B. thuringiensis cry1Ac genes based on cry1-5 gene through multi site-directed mutagenesis. Total 63 various mutant cry genes were obtained at 12 positions randomly. Among them, ten mutant cry genes, whose domain I was totally converted and domain II was randomly, were selected to express in baculovirus expression system as a polyhedrin fusion form. The recombinant proteins were 95 kDa in size and were stably activated as 65 kDa by trypsin. The expressed mutant Cry proteins were applied to bioassays against P. xylostella and S. exigua. All mutants showed high insecticidal activity both to P. xylostella and S. exigua similar to cry1-5. These results suggest that these mutant cry genes might be expected of desirable cry genes for introduction to transgenic crops.

• BMB Reports
• /
• 제36권3호
• /
• pp.294-298
• /
• 2003

The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

• 한국환경성돌연변이발암원학회지
• /
• 제23권1호
• /
• pp.30-34
• /
• 2003

Recent industrial society has human widely exposed to PAHs (polynuclear aromatic hydrocarbons) that are comming from the incomplete combustion of organic material as wider spread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. Our laboratory have been studied the effect of PAHs in the mouse liver hepa 1 cells. In this study, we examined the mouse liver hepa-l cells as a new bioassay system to evaluate bioactivity of PAHs. We have selected 13 PAHs to examine bioassay using cyp1a1-luciferase reporter gene expression system where cyp1a1 1.6 Kb 5flanking region DNA was cloned in front of luciferase reporter gene and this plasmid was transfected into hepa 1 cells transiently. This cells then used for the study to observe the effect of PAHs. We demonstrated that PAHs induced the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase (EROD) activities in a concentration-dependant manner. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD. Acenaphthene, anthracene, fluorine, naphthalene, pyrene, phenanthrene, carbazole were weak responders to cyp1a1 promoter activity stimulation and EROD induction in hepa 1 cells and these chemicals seemed to respond less to EROD than cyp1a1 promoter activity. Benz(a)anthracene, benzo(b)fluoranthene, benzo(k)fluoranthene, chrysene, and dibenzo(a,h)anthracene showed strong response to cyp1a1 promoter activity stimulation and also EROD induction in hepa 1cells. Results of dose response study suggested that four strong responding PAHs, such as benzo(a)anthracene benzo(k)fluoranthene, chrysene, and dibenzo(a, h)anthracene might be mediated through arylhydrocarbon receptor system in hepa1 cells.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제4권1호
• /
• pp.43-49
• /
• 2002

We have cloned cDNAs encoding toxin from the spider, Araneus ventricosus, and constructed a recombinant baculovirus expressing the insecticidal toxin. The cDNAs encoding toxin were cloned from the cDNA library of A. ventricosus. Sequence analysis of the cDNAs encoding the toxin of A. ventricosus revealed that the 240 bp cDNA for AvTox-1 and 192 bp cDNA for AvTox-2 have an open reading frame of 80 and 64 amino acid residues, respectively. The deduced protein sequence of the toxin genes of AvTox-1 and AvTox-2 was aligned to that of the snack Anemonia sulcata and scorpion Centruroides limpidus limpidus, respectively. Northern blot analysis indicated that AvTox-2 toxin gene showed a fat body-spe-cific expression pattern at the transcriptional level. Furthermore, we have explored the possibility of improving baculovirus by incorporating the A. vontricosus toxin gene into Bombyx mori nuclear polyhedrosis virus genome under the control of polyhedrin promoter, The AvTox-2 toxin gene was expressed as approximately 5.8 kDa band in the recombinant baculovirus-injected silkworm larvae. Bioassays with the recombinant virus expressing AvTox-2 on 5th instar silkworm larvae demonstrated a decrease in the time to kill $(LT_{50} days)$ compared to wild-type BmNPV-Kl $(LT_{50} 6.72 days)$ in the injection of 10 viruses. These results indicate that A. ventricosus toxin is a novel member of the spider toxin family, suggesting that the toxin gene can be used in recombinant baculoviruses to reduce insect feeding damage and increase the speed of insect kill.

• Toxicological Research
• /
• 제17권
• /
• pp.227-236
• /
• 2001

The IARC Monographs Programme on the Evaluation of Carcinogenic Risks to Humans has reviewed, summarized and evaluated 869 environmental agents and exposures as oj June 2000. This large collection includes all relevant published epidemiological data on cancer in exposed humans and results of bioassays for carcinogenicity in experimental animals. Since 1986. cancer data have been systematically supplemented by summaries of other toxicological data that are relevant to assessments of carcinogenic hazard. These include summaries qf genetic and related effects of chemicals. which have been prepared as Genetic Activity Profiles (GAP) by the U.S. EPA in collaboration with IARC. As the Mono-graphs have proved increasingly valuable and influential worldwide. they have evolved into an encyclopedia on environmental carcinogenic risks to humans. However. the Monographs have historically been prepared only as printed books with limited distribution. and the Monographs Programme has needed to adjust to expectations oj wider availability. Since 1998 the evaluations and summaries have been globally accessible by Internet from IARC (http://www.iarc.fr) and the GAP profiles by Internet from EPA (http://www.epa.gov/gapdb/). with the two web sites linked. Improved EPN/ARC GAP database and software. GAP2000. now link GAP profiles directly to the appropriate IARC web pages for summaries of evaluations of a given compound and its overall IARC classification. During the year 2000. by means of optical character recognition (OCR) technology the entire series of IARC Monographs is being converted to an electronic version. The first edition is now available commercially in CD-ROM format and will soon become available on-line at .

• Journal of Microbiology and Biotechnology
• /
• 제19권8호
• /
• pp.749-759
• /
• 2009

A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

• 농약과학회지
• /
• 제20권4호
• /
• pp.355-360
• /
• 2016

화랑곡나방과 거짓쌀도둑거저리 유충 및 성충에 대한 Artemisia vulgaris, Achillea millefolium, Anethum graveolens, Cimicifuga heracleifolia, Citrus paradisi, Chrysanthemum indicum, Eucalyptus dives, Neroli bigard, Nigella sativa, Ruta graveolens 식물체 정유의 살충활성을 비교하였다. 그 결과 화랑곡나방 및 거짓쌀도둑거저리 유충에서는 N. bigard 및 A. graveolens 정유가 각각 139.11 및 $52.87{\mu}g\;cm^{-3}$의 접촉독성으로 가장 우수한 살충활성을 나타냈으며, 화랑곡나방 및 거짓쌀도둑거저리 성충의 경우 A. millefolium 및 A. graveolens 정유에서 각각 0.12 및 $93.14{\mu}g\;cm^{-3}$으로 훈증독성이 가장 우수한 살충활성을 나타냈다. 이러한 결과를 바탕으로 N. bigard, A. graveolens A. millefolium 및 A. graveolens 정유가 화랑곡나방과 거짓쌀도둑거저리 유충 및 성충을 방제하기 위한 친환경 방제 법으로 적용 가능성을 보여주었다.

• 한국수산과학회지
• /
• 제12권2호
• /
• pp.113-117
• /
• 1979

백합에 대한 수은, 구리 및 카드뮴의 독성정도를 구명하기 위하여 $1978.6.28\~7.15$간 정수식으로 실험을 실시하였다. 실험결과 수은 및 구리 용액에는 농도가 커질수록 점액을 분비하는 개체가 많아졌으나 카드뮴용액과 정상해수에 투입한 백합은 점액을 방출하지 않았다. 1mg/l 농도의 수조에서 15시간후에 분비한 점액은 수은에서는 $42.9\%$, 구리에서는 $14.3\%$ 나타냄으로서 수은이 구리보다 혐기도가 더 컸다. 농도와 사망률과의 관계에서 96시간후에 반치사농도(96hr-LC50)는 수은 0.67mg/l, 구리 7.04mg/l, 카드뮴 7.10mg/l, 로서 독성의 순위는 수은-구리-카드뮴이었다. 또한 각수조에서 $50\%$의 폐사를 일으키는데 소요한 시간(Lethal Tim 50)은 일반적으로 농도가 클수록 L. T.50값이 작아짐으로서 일정 농도에서는 노출시간이 반응을 유발시키는 자극이 되고 있으며 실제 중금속의 40mg/l에서 L.T.50값은 수은 55.8시간, 구리 104.8시간, 카드뮴 111.9시간으로서 동일농도에서는 수은-구리-카드뮴순으로 사망이 일어 났음을 밝혔다.

• Journal of Microbiology and Biotechnology
• /
• 제20권2호
• /
• pp.281-286
• /
• 2010

A Pseudomonas fluorescens strain was isolated and found to show antagonistic activity against phytopathogenic fungi and to possess a gene responsible for production of antibiotic 2,4-diacetylphloroglucinol. For the extension of biocontrol range, a gene for an Androetonus australis Hector insect toxin 1 (AaHIT1), one of the most known toxic insect-selective peptides, was designed and synthesized according to the preferred codon usage of Pseudomonas fluorescens, cloned, and transformed into the strain by pSUP106 vector, a broad-host-range plasmid. Bioassays indicated that the engineered strain was able to produce AaHIT1 with insecticidal activity, and at the same time retain the activity against plant pathogen. The experiments for nonplanted soil and rhizosphere colonization showed that, similar to the population of the wild-type strain, that of the engineered strain remained relatively constant in the first 10 days, and the subsequent 50 days, suggesting that AaHIT1 expression in the bacterial cell does not substantially impair its long-term colonization. It is first reported that a Pseudomonas fluorescens strain expressing an active scorpion neurotoxin has dual activity against phytopathogenic fungi and insects, making at attractive for agronomic applications.