• Molecules and Cells
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• v.37 no.1
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• pp.43-50
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• 2014

Jumonji domain-containing proteins (JMJD) catalyze the oxidative demethylation of a methylated lysine residue of histones by using $O_2$, ${\alpha}$-ketoglutarate, vitamin C, and Fe(II). Several JMJDs are induced by hypoxic stress to compensate their presumed reduction in catalytic activity under hypoxia. In this study, we showed that an H3K27me3 specific histone demethylase, JMJD3 was induced by hypoxia-inducible factor (HIF)-$1{\alpha}/{\beta}$ under hypoxia and that treatment with Clioquinol, a HIF-$1{\alpha}$ activator, increased JMJD3 expression even under normoxia. Chromatin immunoprecipitation (ChIP) analyses showed that both HIF-$1{\alpha}$ and its dimerization partner HIF-$1{\beta}$/Arnt occupied the first intron region of the mouse JMJD3 gene, whereas the HIF-$1{\alpha}/{\beta}$ heterodimer bound to the upstream region of the human JMJD3, indicating that human and mouse JMJD3 have hypoxia-responsive regulatory regions in different locations. This study shows that both mouse and human JMJD3 are induced by HIF-1.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.269-280
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• 2003

Objective: We reported the overcoming effect of $Ni^{2+}$ on the in vitro 2-cell block of mouse embryos. In this study, we aim to investigate whether $Ni^{2+}$ should induce intracellular $Ca^{2+}$ transient in the mouse embryos. Materials and Methods: Embryos were collected at post hCG 32hr from the oviduct of the ICR mouse and cultured in M2 medium omitted phenol red. Intracellular $Ca^{2+}$ was checked by using a confocal laser scanning microscope and fluo-3AM by using various intracellular $Ca^{2+}$ antagonists. Results: In 1mM $Ni^{2+}$ treated medium which contained $Ca^{2+}$(1.71mM), 75.7% of the embryos showed $[Ca^{2+}]i$ transient about 200 sec later. In the $Ca^{2+}$-free medium, 69.8% of the embryos showed $[Ca^{2+}]i$ transient. In U73122, phospholipaseC(PLC) inhibitor (5uM, 10min) pretreated group, 33.3% of the embryos showed $[Ca^{2+}]i$ transient. Heparine, inositol 1, 4, 5-triphosphate receptor(IP3R) antagonist preinjected embryos showed no response with 1mM $Ni^{2+}$. In danthrolene treatment, ryanodine receptor(RyR)-antagonist, 43% embryos showed $[Ca^{2+}]i$ transient but they showed delayed response about 340sec in the presence of $Ca^{2+}$. Conclusions: Summing up the above results, $Ni^{2+}$ seems to induce $Ca^{2+}$-release from the $Ca^{2+}$-store even in the $Ca^{2+}$-free medium. IP3 receptors of the mouse 2-cell embryos might have an essential role for the intracellular $Ca^{2+}$ increase by $Ni^{2+}$.

• Journal of Embryo Transfer
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• v.15 no.3
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• pp.247-253
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• 2000

This study was undertaken to optimize enucleation and reconstitution methods for the production of cloned mice by somatic cell nuclear transfer Outbred ICR mouse oocytes at the metapahse- II stage were retrieved from female mice superovulated by PMSG and hCG. In Experiment 1, oocytes were enucleated in medium supplemented with cytochalasin B (CCB) of 3 levels (0, 7.5 or 15 $\mu\textrm{g}$/mL), and higher rate of encleation was obtained at 7.5 and 15 $\mu\textrm{g}$/mL than at $\mu\textrm{g}$/mL. In Experiment 2, oocytes enucleated in 7.5 $\mu\textrm{g}$/mL CCB-containing medium were reconstituted with different types of somatic cell by following methods; 1) cumulus cells by direct cell injection, 2) cumulus cells by electric fusion (1.25 kV/cm, 2 pulses for each 70 $mutextrm{s}$) or 3) STO cells by the electrofusion. Electrofusion of STO cells with enucleated oocytes yielded the greatest (P<0.05) rate of reconstitution without lysis (76%) than any other combinations. Although significant decrease in the rate of somatic cell introduction was found, the electrofusion of cumulus cells yielded better rate of reconstitution than direct injection (0 vs. 18%). In Experiment 3, the duration of electric stimulation for the fusion was changed to either 50 $mutextrm{s}$ or 90 $mutextrm{s}$, but no significant improvement of reconstitution efficacy was obtained. In conclusion, this study showed that ICR mouse oocytes could be used for the production of reconstituted oocytes and a fusion method of 1.25 KV/cm with 2 pulses using 570 cell was the optimal.

• Korean Journal of Veterinary Research
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• v.54 no.3
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• pp.171-177
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• 2014

Garlic and mugwort have long been used in traditional medicine to prevent various diseases. Several in vitro studies have reported protective efficacies of garlic and mugwort in cases of gastric cancer. In the present study, we investigated the cancer preventive effects of garlic and mugwort mixture extract (GME) in a Helicobacter (H.) pylori-associated gastric carcinogenesis mouse model. To induce gastric cancer, C57BL/6 mice were treated with N-methyl-N-nitrosourea and H. pylori. Various concentrations of GME (0, 100, 500, and 1,000 ppm) were then fed to the mice for 38 weeks, after which the tumor tissues were examined for histopathology, mucin histochemistry and ${\beta}$-catenin. The incidence of gastric tumors was significantly lower in the highest dose GME-treated mice (46.7%) than control mice (85.7%) (p < 0.05). The multiplicity and size of tumors were also significantly reduced by GME feeding in a dose-dependent manner (p < 0.01). Furthermore, GME suppressed the H. pylori-associated chronic inflammation measured by histologic grading of H. pylori density, chronic gastritis, glandular atrophy and intestinal metaplasia in non-tumorous gastric mucosae. Our data suggest that GME suppresses gastric tumorigenesis via suppression of H. pylori-associated chronic inflammation.

• Archives of Pharmacal Research
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• v.22 no.2
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• pp.130-136
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• 1999

Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either $interferon-{\gamma} (IFN-\gamma)$ or lipopolysaccaride (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-\gamma (100 U/ml) or LPS ($1{\mu}g/ml$) , and remained at palteau for at least 24 h. Cycloheximide ($10{\mu}g/ml$) had no effect on the $IFN-\gamma-$ or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by $IFN-{\gamma}$. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-\gamma and LS induce the mGBP3 expression through distinct signal transduction pathways.

• Journal of Embryo Transfer
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• v.3 no.1
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• pp.48-52
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• 1988

우수정란의 이식전 성판별이 관한 연구를 수행하기 위하여 GTH와 PGF$_2$$\alpha$투여에 대한 난소반응과 회수난자의 발유단계별 동결융해후 생존성을 조사하였으며, 이식전 수정라느이 성판별을 위하여 H-Y항체 처리후 정상발육 난자의 염색체를 분석하여 다음과 같은 결과를 얻었다. 웅성 비장세포(male, spleen cells)를 면역원으로 mouse와 rat에 투여, 항혈청의 항체를 확인한 결과 mouse에서는 C57 BL계통과 rat에서는 DonRyu 계통이 항체생산능력이 우수하였다. 공란우 87두에 hormone(2500IU PMSG, 25mg PGF$_2$alpha)처리하여 평균 57.8%의 채란유과 두당 4.9개의 난자가 회수되었으며, 전체회수란자(427개)중 moula(162개)와 blastocyst(190개)의 정상발육란자는 82.4%였다. 동결융해후 회수된 난자 (312개)중, 형태적으로 정상인 난자(241개)의 비율은 77.2% 발육단계별 성적은 blastocyst(83.4%)가 morula(71.0%)보다 우수하였다. 항체와 보체(Guinea pig serum)로 처리된 82개의 morula중 15개(18.3%)가 blastocyst로 발육되어 이중 5개(33.3%)가 성이 판별되었으며, 모두 xx형 성염색체를 갖는 자성수정란으로 판명되었다.

• Journal of Preventive Medicine and Public Health
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• v.21 no.1 s.23
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• pp.163-171
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• 1988

The paralytic shellfish poisoning was occurred among 25 laborers who worked at breaking-up of ships in Pusan for 5 days from March 29 to April 2 of 1956. For the purpose of accurately defining the paralytic shellfish poison(PSP) , the authors carried out mouse bioassay and chemical analysis. The results were summarized as follows: 1. The mean amount of Paralytic shellfish toxin was $1,207.8{\mu}g$ Per 100gm meat, and the mean death time of mouse was 5 minutes 16 second. 2. The properties of the PSP were mainly gonyautoxin group by chemical analysis(TLC, IR, $^{1}H-NMR$).

• Journal of Sasang Constitutional Medicine
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• v.11 no.2
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• pp.283-299
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• 1999

1. Purpose : Systemic injection of kainic acid in experimental animals induces the limbic seizure and structural damages in hippocampus and amygdala which resembles the changes in human temporal lobe epilepsy. The author performed this study to investigate the neuroprotective effects of Yuldahansotang, on the neurotoxicity induced by kainic acid in the hippocampus in rats. 2. Method : Kainic acid was administered intraperitoneally. And feeding with Yuldahansotang for 3 weeks after kainic acid administration. Seizure were induced in male mice (kainate 10-40 mg/kg i.p) and animals were sacrified at various time-points after injection. The experimental animals were sacrificed at 1, 2, 3day and 1, 3weeks while Yuldahansotang administrations. Seizure were graded using a behavioral scale developed in our laboratory. c-fos belong to immediate early genes(IEGs) known to have rapid and brief responses. And neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. 3. Results & Conclusion : Seizure severity paralled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variable in CA1, thalamus, amygdala within 24 h, is maximal within 72 h, and is large gene by 7 days after administration of kainate. And we can't see the expression of DNA fragmentation and c-fos in the mice what feeded by Yuldahansotang after 7 days from kainic acid administration. It is consequently suggested that Yuldahansotang may attenuate the kainic acid-induced neuronal degeneration and increase the immunoreactivity of hippocampus in mouse.

• Journal of Life Science
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• v.23 no.10
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• pp.1223-1229
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• 2013

Ursolic acid (UA) a bio-active ingredient found in a variety of fruits and vegetables, and it has potent antioxidant activity. However, the role of UA in mouse embryonic stem (ES) cells is poorly understood. This study investigated the functional role of UA in regulating the development of mouse ES cells under hypoxia. Hypoxia did not exert a significant effect on the undifferentiated state of mouse ES cells. However, it induced reactive oxygen species (ROS) generation and increased the level of lactate dehydrogenase (LDH) production at 48 h of hypoxic exposure. Conversely, oxidative stress induced by hypoxia was significantly inhibited by UA ($30{\mu}M$) pretreatment. Hypoxia significantly decreased cell survival and the level of [$^3H$] thymidine incorporation, both of which recovered following pretreatment of UA. In addition, UA decreased the apoptotic effect of hypoxia by attenuating caspase-3 cleavage or by recovering cellular inhibition of the apoptotic protein (cIAP)-2 and Bcl-2 expression. We further found that UA decreased senescence-associated beta-galactosidase activity. We suggest that UA is a natural antioxidant and one of the functional modulators of hypoxia-induced survival, apoptosis, proliferation, and aging in mouse ES cells.

• Journal of Microbiology and Biotechnology
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• v.12 no.3
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• pp.526-529
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• 2002

Lysis of erythrocytes isolated from human, rabbit, and mouse blood samples was investigated with the culture supernatant of Xenorhabdus nematophilus in a primary form. Prior to use, the culture supernatant of the bacteria was concentrated and the concentrate was dialyzed against Tris-HCl buffer (10 mM, pH 8.1) by ultrafiltration using PM-5 membrane with a molecular weight cut-off of 5,000. At $30^{\circ}C$, the supernatant exhibited no lytic activity towards three types of erythrocytes. However, at $4^{\circ}C$, the supernatant showed selective lytic activity towards rabbit erythrocytes within 90 min. yet did not lyze human or mouse erythrocytes. Microscopic examination clearly revealed that most of the rabbit erythrocytes had been fumed into ghost forms.