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Factors Affecting the Productivity of Germ-line Chimeras from Jl Embryonic Stem Cells (Jl 배아주세포를 이용한 효율적인 생식선 이행 카이미라의 생산)

  • Kim, S.U.;Koo, B.S.;Jeong, S.;Lee, T.H.;Yu, S.L.;Nam, Y.I.;Kim, J.L.;Hyun, B.H.;Shin, H.S.;Lee, K.K.;Sang, B.C.;Yu, D.Y.
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.71-77
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    • 2001
  • This experiment was designed to improve the production efficiency of germ-line chimeric mice from phospholipase C (PLC)-$\beta$3 or peroxiredoxin (Prx) E -targeted ($\Delta$) ES cells by the investigating the manipulation conditions and characteristics of Jl ES cells. Four targeting clones were isolated to investigate the karyotypical and morphological stability prior to injection. All clones ($\Delta$PLC$\beta$-3 C3, $\Delta$Prx II C3, C10 and I5) showed more than 80% euploidism, however, most of $\Delta$PLC$\beta$-3 C3 clones were extensively differentiated compared to the other clones. Nine of 13 $\Delta$Prx II chimeras appeared to have at least 80% chimerism, whereas $\Delta$PLC$\beta$-3 C3 chimeras had 20% chimerism at most. Therefore, the morphological stability of ES cells under stable euploidism might mainly affect the production rate of high-coat chimeric mice. To increase the collection rate of injectable blastocysts (IBs), 5 to 10 week -aged C57BL/6J female mice were sacrificed at 3.5 days post-coitum. Ten week-aged mice were the most optimal IB donors by showing the highest collection rate (2.94/mouse) of injectable blastocysts without increase of non-injectable embryos (0.29/mouse). Foster mothers might be another factor because ICR x C57BL/6J F1 foster mother showed more increased productivity in litter size (2.8 vs. 5.6) and chimera (0 vs. 35.3%) than those of ICR foster mothers. In conclusion, the efficient production of germ-line chimeras mainly depends on the maintenance of ES cell morphology during targeting procedure, and the establishment of manipulation conditions might be a key point to maximize it.

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Comparison of Viability on In Vivo and In Vitro-derived Bovine Embryos in Korean Native Cattle (Hanwoo) (한우 체내.외 유래 소 수정란의 생존성 비교)

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Son, Jun-Kyu;Ko, Yeoung-Kyu;Lee, Poong-Yeon;Cho, In-Cheol;Han, Sang-Hyun;Ko, Moon-Suck;Kim, Hyun-Jong
    • Journal of Embryo Transfer
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    • v.26 no.3
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    • pp.195-199
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    • 2011
  • The objective of this study was to evaluate the efficiency of the conventional slow freezing and vitrification methods for cryopreserving in vivo and in vitro-produced bovine embryos. Morphology of post-thawed embryos was evaluated and normal embryos were used for successive culture for 72 h. In experiment I, In embryo viability, There was no significant differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos(89.6% vs. 81.5%). whereas hatched-BL and total cell number rates was significantly higher (p<0.05) for in vivo-derived embryos (76.9%, 136${\pm}$3.6 vs. 43.4%, 107${\pm}$3.8). In experiment II, There was no significant differences in blastocyst re-expansion and Expansion-BL rates were found between in slow freezing and vitrification methods (91.3% vs. 85.7% and 71.4% vs. 75.0%, respectively). in conclusion, These results suggested that the field application for bovine embryo transfer is in part supported by improvements of technologies in embryo conventional slow freezing and vitrification cryopreservation.

Study on Metagonimus yokogawai(Katsurada, 1912) in Korea VII. Susceptibility of Various Strains of Mice to Metagonimus Infection and Effect of Prednisolone (요꼬가의흡충에 관한 연구 Vll. 마우스 Strain별 감자성 및 Prednisolone의 영향)

  • 채종일;서병설이순형
    • Parasites, Hosts and Diseases
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    • v.22 no.2
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    • pp.153-160
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    • 1984
  • An experimental study was undertaken to observe the difference in susceptibility of mouse strains to Metagonimus yokogawai infection by estimating it from worm recovery rate and dimension of worms. It was also studied the effects of prednisolone injection on the chronological pattern of worm recovery in ICR mice. The metacercariae were obtained from sweetfish and 300 in each number were given to 5 strains (CBH, A, DBA, $C^{57}BL$ and KK) of mice, and after 7 days period, the worms were collected from their intestine. Prednisolone at the dose of 10 mg/kg was injected to ICR mice every other day from 7 days prior to infection until sacrificed at 6 hours to 35th post-infection day. ICR mice infected with M. yokogawai but untreated were used for controls. The success rate in infection of mice ranged 25.0-83. 3% by strains, the worm recovery rate 1. 2-18. 9%, and the average size of worms O. 554-0. 683 mm long and 0.214-0.244 mm wide. The higher rates and larger size of worms were observed in KK and $C^{57}BL$ strains than others and the difference was statistically significant. In ICR mice for control, the worm recovery rate until 1 day after infection was relatively high (38-66%) but it became much lower (less than 0.7%) during 1-35 days. However, prednisolone injection brought about persistently high recovery rates (16-80%) until 21 days. It was concluded that the susceptibility to M. yokogawai infection is different by strains of mice but it can be elevated by prednisolone injection probably due to suppression of Immune respon3e3 in ICR mice.

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T Regulatory Cell Responses to Immunization with a Soluble Egg Antigen in $Schistosoma$ $mansoni$-Infected Mice

  • El-Ahwany, Eman;Bauiomy, Ibrahim Rabia;Nagy, Faten;Zalat, Rabab;Mahmoud, Ola;Zada, Suher
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.29-35
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    • 2012
  • The aim of the study is to characterize the phenotypes of $CD4^+$ $CD25^+$ T regulatory cells within the liver granulomas and association with both Foxp-3 gene expression and splenic cytokines. Naive C57BL/6 mice were intravenously injected with multiple doses of the soluble egg antigen (SEA) 7 days before cercarial infection. The immunized and infected control groups were sacrificed 8 and 16 weeks post-infection (PI). Histopathology, parasitological parameters, splenic phenotypes for T regulatory cells, the FOXP-3 expression in hepatic granuloma using real-time PCR, and the associated splenic cytokines were studied. Histopathological examination of the liver revealed remarkable increase in degenerated ova within hepatic granuloma which decreased in diameter at weeks 8 and 16 PI ($P$<0.01). The percentage of T regulatory cells ($CD4^+$ $CD25^+$) increased significantly ($P$<0.01) in the immunized group compared to the infected control at weeks 8 and 16 PI. The FOXP-3 expression in hepatic granulomas increased from 10 at week 8 to 30 fold at week 16 PI in the infected control group. However, its expression in the immunized group showed an increase from 30 at week 8 to 70 fold at week 16 PI. The splenic cytokine levels of pro-inflammatory cytokines, IFN-${\gamma}$, IL-4, and TNF-${\alpha}$, showed significant decreases ($P$<0.05) compared to the infected control group. In conclusion, the magnitude and phenotype of the egg-induced effects on T helper responses were found to be controlled by a parallel response within the T regulatory population which provides protection in worm parasite-induced immunopathology.

Relationships between Radiation-induced Prostaglandin E2 and Natural Killer Cell Activity in Mice (방사선조사(放射線照査)에 의한 Prostaglandin E2 및 자연살해세포(自然殺害細胞) 활성도(活性度)의 변화(變化))

  • Kim, Sung-ho
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.185-189
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    • 1987
  • The number of splenic lymphocyte, serum prostaglandin $E_2$ level and natural killer cell activity were assayed after single whole body irradiation of a sublethal dose of $^{60}Co-{\gamma}$ ray to C57BL/6J mice. With a view to knowing the relationships between radiation induced prostaglandin $E_2$ level and the normal natural killer cell activity after natural killer cell-target cell conjugation, The change of normal natural killer cell activity were measured by administration of prostaglandin $E_2$ containing serum from irradiated mice. The results were summarized as follows; 1. The total number of splenic lymphocyte was significantly decreased by irradiation and the number was not affected by indometacin, prostaglandin synthesis inhibitor, treatment. 2. Serum prostaglandin $E_2$ level was increased in irradiated mice, but indometacin treated mice group showed low level of prostaglandin $E_2$. 3. In the case of irradiated mice, natural killer cell activity was not shown any difference between irradiated group and indometacin combined group. But when natural killer cell-target cell conjugations were exposed to the serum of each group during cytotoxic activity assay, whereas the normal natural killer cell activity was significantly decreased by treatment of serum from irradiated mice, the activity was not changed by treatment of indometacin pretreated mice serum. This result indicated that the prostaglandin $E_2$ induced by the radiation inhibited the post-target binding cytolytic process of natural killer activity.

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A Clinical Study of Bee Venom Acupuncture Therapy on Shoulder Pain Patients in Stroke Sequelae (중풍후견증(中風後遣症)으로 인한 견비통(肩譬痛) 환자(患者)의 봉약침치료(蜂藥鍼治療)에 대한 임상적(臨床的) 고찰(考察))

  • Lee, Dae-Yong;Yeom, Seung-Chul;Kim, Do-Ho;Kim, Dae-Joong;Lee, Geon-Mok
    • Journal of Acupuncture Research
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    • v.23 no.4
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    • pp.69-80
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    • 2006
  • Objectives : The occurrence of shoulder pain after attack of stroke varies from 15% to about 80% in patients. Hemiplegic shoulder pain has been shown to affect stroke outcome in a negative way that it interferes with recovery after a stroke. These following processes have been all postulated as causes of a shoulder pain: glenohumeral subluxation, spasticity, impingement, soft tissue trauma, glenohumeral capsulitis, shoulder hand syndrome. And stroke patients may suffer from pain caused by stroke itself(central post-stroke pain). The aim of this study is to investigate the effectiveness of Bee venom therapy for shoulder pain in stroke patients. Methods : To evaluate the effectiveness of Bee Venom Acupuncture Therapy, 40 patients were allocated into control and treatment group. They were monitored for 3 weeks and followed up with VAS score(with the interval of Initial(YAS1)), 1 week later(VAS2), 2 weeks later(VAS3), 3 weeks later(VAS4), Motor Grade and Passive ROM. Results : VAS score decrease in treatment group compared to control group. Bee Venom Acupuncture Therapy seems to decrease hemiplegic shoulder pain and this effect was statistically significant after 3 weeks. Therefore this therapy could be recommended for the treatment of patients with shoulder pain after stroke and further extensive clinical studies are expected. Conclusion : We suggest that GDS oral administration and electro-acupuncture at $BL_{52}$ & $GB_{39}$ are available for prevention and curing about the postmenopausal osteoporosis.

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Dependence of Mouse Embryonic Development in vitro on the Exposed Period to Oviductal Environment (난관체류시간에 따른 생쥐초기배의 체외발생능력)

  • Song, H.B.;Seo, B.B.;Kim, K.S.;Park, S.E.;Lee, S.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.117-123
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    • 1992
  • Development in vitro of 2-cell mouse embryos was examined after appropriate exposure to oviductal milieu to demonstrate biological activity present in the oviducts. ICR and ($C57Bl/6{\times}Balb/c$) $F_1$ hybrid mice were superovulated and mated for the recovery of early embryos. Embryos were recoverd at every 2h intervals from 32h post-hCG(hph) to 56 hph. The proportions of developmental stages were determined in the recovered embryos. Development in vitro of 2-cell embryos was more rapid in $F_1$ hybrid than in ICR, showing high proportions of 4-cell embryo and blastocyst at 120 hph. 100% of blastocyst development was obtained at 38hph in $F_1$ hybrid and at 50 hph in ICR when 2-cell embryos were cultured upto 120hph in vitro. Moreover, in vitro culture of oviducts containing 2-cell embryos in ICR mice for 12h from 34hph to 46hph increased developmental capacity of ICR mouse embryo in vitro. The results indicate that oviductal environment contains substances having mitogenic activity and overcoming early cell block in vitro. The mitogenic activity is effective in vitro as well as in vivo.

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High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo;Hong, Min-Seon;Shin, Hang-Chel;Lee, Jeewon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.301-305
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    • 2001
  • The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

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Promotion of cAMP Responsive Element-Binding Protein Activity Ameliorates Radiation-Induced Suppression of Hippocampal Neurogenesis in Adult Mice

  • Kim, Joong-Sun;Yang, Mi-Young;Cho, Jae-Ho;Kim, Sung-Ho;Kim, Jong-Choon;Shin, Tae-Kyun;Moon, Chang-Jong
    • Toxicological Research
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    • v.26 no.3
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    • pp.177-183
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    • 2010
  • This study was performed to examine whether elevated activity of cAMP responsive element-binding protein (CREB) attenuates the detrimental effects of acute gamma ($\gamma$)-irradiation on hippocampal neurogenesis and related functions. C57BL/6 male mice were treated with rolipram (1.25 mg/kg, i.p., twice a day for 5 consecutive days) to activate the cAMP/CREB pathway against cranial irradiation (2 Gy), and were euthanized at 24 h post-irradiation. Exposure to $\gamma$-rays decreased both CREB phosphorylation and immunohistochemical markers for neurogenesis, including Ki-67 and doublecortin (DCX), in the hippocampal dentate gyrus (DG). However, the rolipram treatment protected from $\gamma$-irradiation-induced decreases of CREB phosphorylation, and Ki-67 and DCX immunoreactivity in the hippocampal DG. In an object recognition memory test, mice trained 24 h after acute $\gamma$-irradiation (2 Gy) showed significant memory impairment, which was attenuated by rolipram treatment. The results suggest that activation of CREB signaling ameliorates the detrimental effects of acute $\gamma$-irradiation on hippocampal neurogenesis and related functions in adult mice.

Neuroprotective effect of lithium after pilocarpine-induced status epilepticus in mice

  • Hong, Namgue;Choi, Yun-Sik;Kim, Seong Yun;Kim, Hee Jung
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.1
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    • pp.125-131
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    • 2017
  • Status epilepticus is the most common serious neurological condition triggered by abnormal electrical activity, leading to severe and widespread cell loss in the brain. Lithium has been one of the main drugs used for the treatment of bipolar disorder for decades, and its anticonvulsant and neuroprotective properties have been described in several neurological disease models. However, the therapeutic mechanisms underlying lithium's actions remain poorly understood. The muscarinic receptor agonist pilocarpine is used to induce status epilepticus, which is followed by hippocampal damage. The present study was designed to investigate the effects of lithium post-treatment on seizure susceptibility and hippocampal neuropathological changes following pilocarpine-induced status epilepticus. Status epilepticus was induced by administration of pilocarpine hydrochloride (320 mg/kg, i.p.) in C57BL/6 mice at 8 weeks of age. Lithium (80 mg/kg, i.p.) was administered 15 minutes after the pilocarpine injection. After the lithium injection, status epilepticus onset time and mortality were recorded. Lithium significantly delayed the onset time of status epilepticus and reduced mortality compared to the vehicle-treated group. Moreover, lithium effectively blocked pilocarpine-induced neuronal death in the hippocampus as estimated by cresyl violet and Fluoro-Jade B staining. However, lithium did not reduce glial activation following pilocarpine-induced status epilepticus. These results suggest that lithium has a neuroprotective effect and would be useful in the treatment of neurological disorders, in particular status epilepticus.