• 환경위생공학
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• 제13권3호
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• pp.37-42
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• 1998

2-Bromopropane has been implicated to be the reason for the mass intoxication of workers at an electronic company in Korea. 2-Bromopropane deposition and pattern of DNA replication in mouse fetuses were analyzed after intravenous injection of 2-bromopropane. Injections were administered to pregnant ICR mice in order to cytogenetically evaluate transplacental 2-bromopropane. The results are summarized as follows; 1. A dose-dependent effect on DNA replication was observed equally in the lung, liver and kideneys of fetuses has been exposed to 2-bromopropane transplacentally as reductions of the labeling index. 2. Deposition of transplacentally administred 2-bromopropane in the fetus was lower than placenta.

• Reproductive and Developmental Biology
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• 제33권4호
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• pp.237-242
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• 2009

Pregnancy is a unique event in which a fetus develops in the uterus despite being genetically and immunologically different from the mother, and the underlying mechanisms remain poorly understood. To analyze the differential gene expression profiles in nonpregnant and 7 days post coitus (dpc) pregnant uterus of mice, we performed a global proteomic study by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The uterine proteins were separated using 2-DE, Approximately 1,000 spots were detected on staining with Coomassie brilliant blue. An image analysis using Melanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between pregnant and nonpregnant uterus. Twenty-one spots were identified as differentially expressed proteins, of which 10 were up-regulated proteins such as alpha-fetoprotein, chloride intracellular channel 1, transgelin, heat-shock protein beta-1, and carbonic anhydrase II, while 11 were down-regulated proteins such as X-box binding protein, glutathione S-transferase omega 1, olfactory receptor Olfr204, and metalloproteinase-disintegrin domain containing protein TECADAM. Most of the identified proteins appeared to be related with catabolism, cell growth, metabolism, regulation, cell protection, protein repair, or protection. Our results uncovered key proteins of mouse uterus involved in pregnancy.

• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.101-105
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• 1999

N-Methylpurine-DNA glycosylase (MPG)는 DNA에서 N-methylpurine과 다른 손상된 purine기를 제거하는 DNA 회복 효소이다. 생쥐의 임신 8일 후의 태아조직부터 생후 400일까지의 조직들로부터 RT-PCR 방법으로 MPG mRNA 발현을 조사하였다. MPG mRNA는 태아 형성기동안 많은 양이 발현되었으며, 특히 15일에서 가장 높게 발현되었다. 태반에서의 MPG mRNA는 8 p.c. (post coitum)부터 18 p.c. 까지 계속적으로 감소하였다. 태아 형성기의 뇌와 간 조직에서 높은 mRNA 발현을 보였으나, 400일 되는 성체에서는 현저히 감소하였다. 부정소, 정낭, 정소, 정관, 난소, 난관 그리고 자궁 등의 생식기관에서의 MPG mRNA는 비교적 높게 발현되었고, 그들 중 부정소에서의 발현 정도가 가장 높았다. 이러한 결과들로 보아 MPG 유전자는 태아 발생단계 및 조직 특이적으로 발현됨을 알 수 있었다.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.93-98
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• 2009

In this study, bovine Dnmt1 cDNA was sequenced and detected Dnmt1 mRNA level in bovine tissues by northern blot, methylation pattern of genome by southern blot, specific localization of Dnmt1 in mouse and bovine preimplantation embryos by immunocytostaining and Dnmt1 protein level in ovary and testis by western blot. Bovine Dnmt1 cDNA sequence showed more homology with that of human than mouse and rat. The RNA level of Dnmt1 was 10 times higher expression in placenta than other tissues. This indicates that placenta was hypermethylated compared to others organs. The genomic DNA could not be cut by a specific restriction enzyme (HpaII) in placenta, lung and liver of bovine. It suggests that Dnmt1 in some somatic cells was already methylated. Dnmt1, which has the antibody epitope 1316~1616, was distributed in nucleus and cytoplasm including the stage of pronuclear stage and maturation of oocyte and gradually weaken to blastocyst stage compare to negative. In addition, Dnmt1 was strongly expressed in tetraploid embryo and cloned 8-cell than IVF 8-cell. An aberrant pattern of DNA methylation in cloned embryo may be abnormal development of fetus, embryonic lethality and placenta dysfunction. The somatic specific band (190kDa) was appeared in ovary and testis, but oocyte specific band (175kDa) was not. Further investigations are necessary to understand the complex links between the methyltransferases and the transcriptional activity of genes in the cloned bovine tissues.

• 대한수의학회지
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• 제34권4호
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• pp.821-831
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• 1994

Phenytoin(PHT), a commonly prescribed anticonvulsant, has been known as a teratogen in experimental animals and human. However, PHT has strain-specific teratogenic effects for mice and human. Dimethyl sulfoxids(DMSO) has been known to antagonize the teratogenic effects of secalonic acid D, a toxic mold metabolite that has similar teratogenic effects to PHT. Therefore, this study was performed to examine the embryopathic effects of PHT in terms of treatment period and the antiteratogenic effect of DMSO in ICR mice. PHT(75mg/kg, BW) was administered intrapetitoneally on day 10, 10-11 and 10-12 of gestation with or without DMSO(2ml/kg, BW), and the fetal malformation was observed on day 18. Major malformation of fetuses treated with PHT on day 10, 10-11 and 10-12 of gestation was cleft palate, and the percentages of fetus with cleft palate were 14.5%, 31.7% and 51.7%, respectively. Also, there was a significant decrease of cleft palate from 51.7% in PHT alone group to 30.8% in PHT plus DMSO group. Our findings suggest that cleft palate is one of major malformation by PHT treatment in ICR mouse and DMSO has strong antiteratogenic effect.

• Asian-Australasian Journal of Animal Sciences
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• 제17권12호
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• pp.1641-1646
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• 2004

The objective of this study was to generate transgenic mice expressing human resistin gene by using the tetraploidembryonic stem (ES) cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR, cloned into $pCR^{(R)}$ 2.1 $TOPO^{(R)}$ vector and constructed in pCMV-Tag4C vector. Mammalian expression plasmid containing human resistin was transfected into D3-GL ES cells by Lipofectamine 2,000, and then after 10-12 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec (fusion rate: 2,114/2,256, 93.5%) and cultured up to the blastocyst stage (development rate: 1,862/2,114, 94.6%). The selected 15-20 ES cells were injected into tetraploid blastocysts, and then transferred into the uteri of E 2.5 d pseudopregnant recipient mice. To investigate the gestation progress, two E 19.5 mused fetuses were recovered by Cesarean section of which one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, our findings demonstrate that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mice for the rapid analysis of gene function in vivo.

• 한국가축번식학회지
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• 제22권2호
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• pp.171-176
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• 1998

본 연구는 체외에서 배양된 생쥐 완전탈출 배반포기배를 동결보존액 EFS35를 이용하여 초자화 동결하였을 때 체내발달의 적합성 여부를 조사하기 위해 실시하였다. 공시된 완전탈출 배반포기배($\theta$ 130$\mu\textrm{m}$)는 체내에서 생산된 전핵기 수정란을 5일 동안 체외배양하여 얻었으며, 10% ethylene glycol(EG)에 5분 노출한 후 EFS35(35% EG, 18% Ficoll, 0.3 M sucrose)에 30초 동안 노출하거나, 초자화 동결하였다. 융해 후, 재팽창이 이루어진 완전탈출 배반포기배는 가임신 3일된 대리모의 한쪽 또는 양쪽 자궁각(4~6개/자궁각)에 이식하였다. 대리모의 임신율과 착상율은 임신 15일재 외과적 해부로 판정하였다. 그 결과를 요약하면 다음과 같다. 융해 30분 후, 완전탈출 배반포기배의 체외생존율은 노출군(65.5%)과 동결군(54.5%)간에 유의한 차이는 없었다. 또한, 체내발달율을 조사하였던 바, 착상율에 있어서 동결군(41.0%)과 대조군(58.5%)간에 유의한 차는 없었지만, 정상산자율에서는 동결군(24.0%)의 결과가 대조군(58.3%)보다 유의하게 낮게 나타났다(p<0.05). 이러한 결과는 EFS35를 이용한 완전탈출 배반포기배의 초자화 동결은 정상산자율은 감소하였지만, 완전탈출 배반포기배의 이용 효율성을 넓히는데 이용될 수 있다는 것을 알 수 있었다.

• Animal cells and systems
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• 제1권2호
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• pp.337-340
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• 1997

The purpose of this study is to evaluate whether water soluble chitosan, a natural nontoxic chelator, can reduce fetal contamination of radiostrontium in pregnant mice. Various forms of water soluble chitosans (10% or 1% powder, or 1% solution) were given to pregnant mice before or after contamination of 0.005 uCi/B.W(g) Sr-85. Transplacental transfer of Sr-85 to fetus was $6.8{\pm}2.7%$ of injected dose, when Sr-85 was administered at the 20th day of pregnancy. Fetal radioactivity was significantly reduced when mother mice were treated with water soluble chitosan before contamination of Sr-85. Water soluble chitosans of 10% or 1% powder, or 1% solution significantly reduced fetal retention of Sr-85 to $2.3{\pm}0.7%$, $2.7{\pm}0.8%$, and $2.0{\pm}0.9%$, respectively. However, fetal contamination was not reduced, when water soluble chitosans of 10% or 1% powder, or 1% solution were administered after maternal contamination of Sr-85. From these data we can conclude that water soluble chitosan could reduce fetal contamination of radiostrontium in pregnant mice, when given before the pregnant mice were exposed to radiostrontium.

• IMMUNE NETWORK
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• 제2권2호
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• pp.79-85
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• 2002

Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

• 대한임상검사과학회지
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• 제47권1호
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• pp.51-58
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• 2015

Ethanol has long been implicated in triggering apoptotic neurodegeneration. Alcohol also may indirectly harm the fetus by imparing the mother's physiology. We examined the effects of ethanol on immature brain of mice. Three-weeks-old female ICR strain mice daily intraperitoneally injected with ethanol at the concentration of 4 and 20% in saline for 0, 6, and 24 hours and 1 and 4 weeks. The mice were weighted and sacrificed, and the brains were ectomized for the present histological, immunohistochemical and TUNEL assays. Based on the histologic hematoxylin and eosin stain, immunohistochemical expression of glutamate receptor protein and neuronal cell adhesion molecule (NCAM) were evaluated. The cerebral cortex of the ethanol-treated group showed few typical symptoms of apoptosis such as chromosome condensation and disintegration of the cell bodies. TUNEL staining revealed DNA fragmentation in the 6 and 24 hours. This results demonstrated that acute ethanol administration causes neuronal cell death. I found that either glutamate receptor inhibition or activation could induce cerebellar degeneration as ethanol effect. Neuronal death also can be induced by excess activity of certain neurotransmitter, including glutamate. Neurons must establish cell-to-cell contact during growth and development in order to survive, migrate to their final destination, and develop appropriate connections with neighboring cell. Purkinje cell in cerebellar are especially vulnerable to the cell death and degeneration. After ethanol treatment in cerebellar, NCAM had decreased by 4 weeks. This result suggest that apoptosis seems to be involved in the slow elimination of neuron and cerebellar degeneration.