• 한국가축번식학회지
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• 제18권1호
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• pp.55-62
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• 1994

This study was carried out to test the effects of fluorescein diacetate(FDA 0${mu}ell$/ml, 0.5${mu}ell$/ml, 5${mu}ell$/ml, 10${mu}ell$/ml or 0${mu}ell$/ml, 5${mu}ell$/ml in PBS) treated before culture on the survival of vitrified mouse morulae in vitrification solution(20% glycerol＋glycerol＋10% ethylene glycol＋30% ficoll＋10% sucrose). The results were summarized as follows; 1. The survival rate of FDA-tested fresh mouse morulae after 24 hours culture was over 96%((P4.8) in the control or treatment groups with various levels of FDA. Because the rate of mouse morulae which developed to hatched blastocysts was higher with the various levels of FDA treatment(67%) than control(50%), it was considered that toxicity of FDA did not affect the survival of mouse morulae. 2. When mouse morulae were FDA-tested in FDA 0(control), 0.5, 5, and 10${mu}ell$/ml treatment after vitrification, the development rate to expanded blastocyst were 66, 82, 64 and 76%, and FDA scores were P4.2(84%), P4.7(94%), P4.2(84%) and P3.9(78%), respectively. There were no significant differences between control and FDA treatments, but there were significant difference between 0.5${mu}ell$/ml)94%) and 10${mu}ell$/ml(78%) treatment(P<0.01). 3. The survival rates of cultured mouse morulae according to FDA-scores(P0=non-fluorescence; P1~P5=according to their fluorescence) after vitrification were P5;92%, P4;67%, P3;42% and P2.P0;0%, respectively. 4. Implantation rates of morulae stage embryos cultured into early blastocysts and implanted into uterine hornes vitrification were 14 and 11% embryos treated control(0${mu}ell$/ml) and FDA 5${mu}ell$/ml and the normal fetus development was 2% embryos for both treatments. Results of this percent study indicated that toxicity of FDA does not affect not only the survival of fresh and vitrified mouse morulae but also the development rate and implantation of fetus after transfer as well. The development rates of mouse morulae with the FDA score of P5, P4 and P3 were 92, 67 and 42%, respectively, it was considered that FDA-test was fit for the judge of survival.

• 한국해양생명과학회지
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• 제3권2호
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• pp.74-80
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• 2018

수은은 다양한 산업활동에 사용되어 해양 환경 내에 지속적으로 유입되며 먹이사슬을 따라 축적되며 생물체 내로 유입 시 해양 생물의 성장, 발생, 생식, 대사 등에 악영향을 미칠 수 있다. 본 연구에서는 기수산 물벼룩 Diaphanosoma clelbensis에 대한 수은의 급성 독성과 산화적 스트레스 지표(총 glutathione 함량, GST, GR, GPx 활성)를 이용한 항산화 반응을 조사하였다. 24시간 수은을 노출시킨 결과 생존율이 농도 의존적으로 감소하는 양상을 보였으며, 24시간 LC₅₀ 값은 0.589 mg/l (95% C.I. 0.521~0.655 mg/l)로 나타났다. 수은(0.08과 0.4 mg/l)을 24시간 노출시킨 D. celebensis에서 총 glutathione 함량은 유의하게 감소하는 양상을 보였으며, GST, GR, GPx 활성은 유의하게 증가하는 양상을 보였다. 이러한 결과는 수은 노출에 의해 D. celebensis에서 산화적 스트레스가 유도되었음을 의미하며, 이들 산화적 스트레스 지표의 변화는 세포내에서 방어기전으로 작용하였음을 나타낸다. 본 연구는 D. celebensis에서 수은 독성의 분자메커니즘을 이해하는데 도움이 되며, 중금속 오염된 해양 생태계 모니터링과 해양 생물의 건강성 평가에서 이들 분자지표의 활용 가능성을 제시한다.

• Toxicological Research
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• 제24권2호
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• pp.137-150
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• 2008

Sexually mature male and female rats were orally intubated with the organophosphorus insecticide, Pestban at a daily dosage of 7.45 or 3.72 mg/kg bwt, equivalent to 1/20 and 1/40 $LD_{50}$, respectively. Male rats were exposed for 70 days, while the female rats were exposed for 14 days, premating, during mating and throughout the whole length of gestation and lactation periods till weaning. The results showed depressed acetylcholinesterase(AChE) activity in the brain of parents, fetuses and their placentae in a dose-dependent manner. The fertility was significantly reduced with increasing the dose in both treated groups, with more pronounced suppressive effects in the male treated group. The number of implantation sites and viable fetuses were significantly reduced in pregnant females of both treated groups. However, the number of resorptions, dead fetuses, and pre-and postimplantation losses were significantly increased. The incidence of resorptions was more pronounced in treated female compared to male group and was dose dependant. The behavioral responses as well as fetal survival and viability indices were altered in both treated groups during the lactation period. The incidence of these effects was more pronounced in the treated female group and occurred in a dose-related manner. The recorded morphological, visceral, and skeletal anomalies were significantly increased with increasing the dose in fetuses of both treated groups, with more pronounced effects on fetuses of treated females. In conclusion, the exposure of adult male and female rats to Pestban would cause adverse effects on fertility and reproduction.

• 한국가축번식학회지
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• 제22권2호
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• pp.177-186
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• 1998

While tubal pregnancy is frequently observed in human, it has been reported to rarely occur in other mammals. To investigate the reason of the absence of tubal pregnancy in other mammals, the ability of bovine tubal(oviductal) fluid to su, pp.rt the outgrowth of mouse embryos waw examined by using an in vitro model system wherein the trophoblast cells of hatched mouse blastocysts attach to and outgrow on tissue culture plates coated with FBS. When mouse blastocysts grwon in vitro from 2-cell embryos were cultrued in the dishes coated with FBS, human follicular fluid(hFF) and bovine follicular fluid(bFF), respectively, underwent outgrowth by spreading onto the plastic dishes during 48 hr. In contrast, none of the embryos cultured in the dishes coated with BSA or bovine obiductal fluid(bOF) did outgrow but remained as late blastocysts. Since addition of bOF at 5mg/ml or higher conc. to the culture medium resulted in degeneration of all embryos during 48 hr culture, 10mM conc. of glutathione(GSH) was added to the bOF-containing medium to circumvent the toxicity of bOF. In addition, bOF was heated $65^{\circ}C$ for 30 min(hbOF) to get rid of its precipitating properties and then added to the culture medium. When blastocysts were cultured in the presence of both hbOF and GSH 45.4% of embryos attached to the culture dishes. However, none of these embryos underwent outgrowth. Fially embryos were cultured in the presence of both hbOF and GSH but in the dishes coated with FBS. When they were examined after 48 hr, all of the blastocysts exhibited well-developed outgrwoth. Based upon these results, it is concluded that bovine oviductal fluid is capable of su, pp.rting the attchment of mouse blastocysts onto the culture plaste whereas it cannot promote the outgrwoth of mouse blastocysts in vitro, probably due to the lack of outgrwoth factor.

• 한국가축번식학회지
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• 제16권3호
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• pp.269-276
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• 1992

Studies were carried out to find the freezing media which gives no ice crystals in single(glycerol, ethylene glycol, dimethyl sulfoxide(DMSO)) and mixture solutions(glycerol＋propylene glycol, glycerol＋ethylene glycol) of permeable cryoprotectants in vitrification solution and to study effects of VS on the survival of vitrified mouse morulae. The results are summarized as follows: 1. In toxicity test of permeable cryoprotectants, 30% glycerol of single solution showed the highest FDA-score(4.1) in mouse morulae frozen compared among other single solutions. The FDA-score(4.1) of 30% glycerol was higher than 30% ethylene glycol(3.6) and DMSO(1.4( (P<0.05). 2. 20, 30 or 40% single solution of permeable cryoprotectants containing m-PBS with 10% sucrose and 20% BSA was not crystallized during cooling, but crystallized during warming. However, the 30% mixture solution of the two permeable cryoprotectants was not crystallized both during cooling and warming.3. When mouse morulae were frozen in 30% mixture solutions of two permeable cryoprotectants(glycerol and propylene glycol, glycerol and ethylene glycol), highest FDA-score(4.5) was obtained in a mixture solution of 20% glycerol and 10% ethylene glycol(20G10E) than other 30% mixture solutions(10G20E, 15G15E, 20G10P, 15G15P, 10G20P) and there was significant difference between 20G10E and 10G20E(P<0.05).

• The Plant Pathology Journal
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• 제39권3호
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• pp.290-302
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• 2023

Abamectin offers great protection against Bursaphelenchus xylophilus, a well-known devastating pathogen of pine tree stands. Trunk injection of nematicides is currently the most preferred method of control. This study aimed to evaluate the potency of the commonly used formulations of abamectin against B. xylophilus. Twenty-one formulations of abamectin were evaluated by comparing their sublethal toxicities and reproduction inhibition potentials against B. xylophilus. Nematodes were treated with diluted formulation concentrations in multi-well culture plates. And, populations preexposed to pre-determined concentrations of the formulations were inoculated onto Botrytis cinerea culture, and in pine twig cuttings. Potency was contrastingly different among formulations, with LC₉₅ of 0.00285 and 0.39462 mg/ml for the most, and the least potent formulation, respectively. Paralysis generally occurred at an application dose of 0.06 ㎍/ml or higher, and formulations with high sublethal toxicities caused significant paralysis levels at the tested doses, albeit the variations. Nematode reproduction was evident at lower doses of 0.00053-0.0006 ㎍/ml both on Botrytis cinerea and pine twigs, with significant variations among formulations. Thus, the study highlighted the inconsistencies in the potency of similar product formulations with the same active ingredient concentration against the target organism, and the need to analyze the potential antagonistic effects of the additives used in formulations.

• 농약과학회지
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• 제14권4호
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• pp.478-489
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• 2010

펜티오피라드에 대한 독성을 평가하고 일일섭취허용량을 설정하기 위하여 급성독성시험 등 32항목의 시험성적서를 검토하였다. 펜티오피라드는의 소변과 대변으로 24 시간이내에 대부분 배살되었다. 급성독성은 낮았으며, 안점막 자극성은 경도의 자극성을 보였고 피부자극성 및 피부감작성은 없었다. 랫드, 개 90일 및 1년 반복투여 경구독성시험에서 간에 영향이 나타났다. 랫드 2세대 번식독성시험에서 수정율, 임신율 등 번식에 영향을 나타내지 않았고, 랫드 및 토끼 기형독성시험에서 기형독성은 없었다. 복귀돌연변이시험 등 6종의 유전독성시험에서 유전독성에 대한 영향은 없었으며, 랫드 24개월 발암성시험에서 갑상선 여포세포선종, 마우스 18개월 발암성시험에서 간세포 선종 발생 빈도가 증가 되었으나 간장효소 유도 시험에서 고농도 반복투여에 의한 대사효소 유도로 발생하는 영향이므로 역치가 존재한다고 판단되어 펜티오피라드는 비발암성물질로 평가하였다, 따라서 펜티오피라드의 독성시험성적서 중 가장 낮은 최대부작용량은 개 1년 반복투여독성시험의 8.10 mg/kg bw/day으로 선정하였고 번식독성, 기형독성, 발암성 등에 대한 영향이 없으므로 안전계수를 100으로 정하여 펜티오피라드의 일일섭취허용량을 0.081 mg/kg/day로 설정하였다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.16-16
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• 2001

Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

• 환경생물
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• 제32권1호
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• pp.1-15
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• 2014

양서류는 육상과 수상생태계를 연결하는 먹이사슬의 연결자로 진화적 생태적 지이를 갖는다. 양서류의 배아와 유생은 모체와 독립되어 수환경 내에서 초기발생 및 성장하기 때문에 수환경에 존재하는 다양한 화학물질에 직접적으로 노출될 수 있다. Azole계열 항곰팡이제는 농업 및 의료용으로 널리 사용되는 화학물질로서 농지, 하수처리장 등으로 부터 수계로 유입된다. 최근, 양서류에서 이러한 azole계 물질에 의한 기형유발, 내분비계장애 효과가 증가하고 있다. 본 소고에서는 azole계 물질의 양서류 독성을 파악하고 azole계 물질의 안전한 이용을 위한 가이드라인을 제공하고자 azole계열에 속하는 imidazole, triazole, thiazole, oxazole, pyrazole 항곰팡이 물질이 양서류의 발생, 분화, 생식 등에 미치는 영향에 대해 최근까지의 연구결과를 정리하였다.

• 환경생물
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• 제22권2호
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• pp.300-307
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• 2004

The chloroacetanilide herbicide alachlor (2-chloro-2', 6'-diethyl-N-(methoxymethyl)-acetanilide) and organochlorine insecticide endosulfan (6, 7, 8, 9, 10, 10-hexachloro-l, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 3, 4-benzodioxathiepin-3-oxide)are the highly toxic agricultural chemicals. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. B. orientalis spawns in the farming regions at Spring when the massive application of agricultural chemicals occurs. These chemicals in farmland may threaten the reproduction of this frog. Therefore, we examined the embryotoxic and survival effects of alachlor and endosulfan at various concentrations in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with alachlor and endosulfan were decreased with concentration dependent manner. Also, developmental malformations appeared by alachlor and endosulfan in B. orientalis embryos. The malformations showed in order of frequency with bent trunk, tail dysplasia, bent tail, thick-set body and ventral blister in alachlor treated embryos. The exposure of endosulfan produced 7 types of severe external malformations with tail dysplasia, pectoral blister, bent trunk, bent tail, cephalic dysplasia, ventral blister, and thick-set body. Following exposure to alachlor and endosulfan the types of malformations were diverse, suggesting these chemicals target multiple events in embryonic and larval development in this species. These results suggested that alachlor and endosulfan were detrimental for survival and development of B. orientalis embryos.