• Journal of environmental and Sanitary engineering
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• v.13 no.3
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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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• v.33 no.4
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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.

• Development and Reproduction
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• v.3 no.1
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• pp.101-105
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• 1999

N-Methylpurine-DNA glycosylase (MPG) removes N-methylpurine and other damaged purines in DNA. RT-PCR analysis revealed MPG mRNA expression at various tissues of fetal development from day 8 to day 18 fetus and day 400 mature adult. The MPG transcripts were abundant during fetal development in mice. In placenta, the MPG mRNA was continuously decreased from day 8 post coitum (p.c) to day 18 p.c. fetus. The high level of mRNA in fetal brain and liver was drastically declined in day 400 mature adult. The expression of MPG, originally characterized by its highest level of expression in the epididymis of adult mouse, was detected with high level in several other reproductive organ, including the ovary, oviduct, testis, vas deference, uterus, and seminal vesicles. These results demonstrate developmental stage- and tissue-specific variation of MPG gene expression.

• Reproductive and Developmental Biology
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• v.33 no.2
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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.

• Korean Journal of Veterinary Research
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• v.34 no.4
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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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• v.17 no.12
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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.

• Korean Journal of Animal Reproduction
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• v.22 no.2
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• pp.171-176
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• 1998

This study was carried out to confirm whether the in vivo developmental potential of mouse hatched blastocysts (HBs) can be obtained by vitrification method using the cryoprotectant EFS35. The HBs ($\theta$ 130$\mu\textrm{m}$) were cultured in vitro until day 5 from zygotes produced in vivo and were equilibrated in 10% ethylene glycol(EG) for 5 min, and then were exposed or vitrified in EFS35 (35% EG, 18% Ficoll and 0.3M sucrose). After 30 min thawing, re-expanding HBs were transferred into one or both uterine horns of pseudopregnant recipients on day 3 (4~6 embryos /horn). Pregnancy rates of recipients and implantation were assessed by autopsy on day 15 of gestation. The results obtained in these experiments were summarized as follows : After thaw-ing, in vitro survival of HBs was not significantly different between exposed (65.5%) and vitrified(54.5%) group. Also, when the in vivo development potential was examined, total implantation was not different between control (58.5% ) and vitrified (41.0%) group, although the live fetus formation of vitrified group (24.0%) was significantly lower than that of control (58.3%) group (p< 0.05). These results suggested that vitrification freezing method of mouse HBs using EFS35 can be used to make wide the utility of embryo transfer of the more embryos produced in vitro.

• Animal cells and systems
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• v.1 no.2
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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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• v.2 no.2
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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.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.1
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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.