• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• Journal of Chest Surgery
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• v.32 no.12
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• pp.1093-1099
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• 1999

배경 : 류마티스성 승모판막 폐쇄부전증과 퇴행성 승모판막 폐쇄부전증에서 승모판막 성형술의 결과와 비교하여 류마티스성 승모판막 폐쇄부전증에서도 승모팍막성형술이 적합한 치료방법이 될 수 있는지를 알아보았다. 대상 및 방법 : 95년 1월부터 98년 12월 까지 승모판막 성형술을 시행받은 184명의 환자중에서 류마티스성 승모판막 폐쇄부전증 49례(1군)의 퇴행성 승모판막 폐쇄부전증 78례(2군)를 대상으로 하였다. 평균연령은 1군이 36.3$\pm$14.6(16-74세) 2군은 52.5$\pm$13.4(14-77)세였다 총 추적 관찰기간은 1군이 72.2인년 2군이77.2인년이었다 두군에서 수술후와 수술후 6개월 1년 및 이후 1년 단위로 주기적인 심초음파를 시행하였고 이를 통계적 검정하였다. 결과 : 두 군간에 수술전 혈류역학적인 차이를 보이지 않았고 수술전 평균 승모판막 폐쇄 부전의 정도는 1군이 3.0$\pm$0.4, 2군이 3.9$\pm$0.3였으나 수술후 추적 관찰에서 각각 0.9$\pm$0.9와 0.8$\pm$0.7정도의 양호한 판막 성형술의 결과를 보였고 승모판막 면적의 변화나 승모판막에서의 평균압력차이 등 혈류역학적인 결과에도 차이를 보이고 있지 않았다 수술조기 사망과 후기 사망은 없었으며 재수술율은 1군이 인년대비 1.4% 2군이 인년대비 2.6%였고 색전발생율은 1군이 인년대비 2.8% 2군이 1.3%였다. 심내막염발생은 1군에서만 1례있었으며 상기 결과들에서 두군간에 의미있는 차이를 보이고 있지는 않았다 결론 : 향후 장기적인 추적 관찰이 필요하나 중기 성적에서 승모판막 성형술이 류마티스성 승모판막 폐쇄 부전증에서도 효과적인 치료방법임을 알수 있었다.다 출생후 폐포막의 FN의 활성은 출생후 5일 및 7일에 최고주에 달했다. 출생직후 1-2일경에 혈관의 조직내 FN의 활성이 양성을 나타내지만 3일이후 활성이감소되었다. 폐포대식세포내 FN의 활성은 출생후 증가되었다. 폐조직내 소기관지의 FN의 활성은 출생후 완만하게 상승되었다. 큰 폐포세포는 출생 1-3일에 일정량의 FN 반응이 세포질과 미세융모내에 관찰되었다. 결론 : 이상과 같은 결과로 흰쥐의 폐포의 분화과정이 계속되는 출생후 폐에서 FN의 분비는 7일이내에 성숙흰쥐의 폐포내 반응과 비슷한 반응으르 보이며 이때 폐의 실질조직은 분화가 거의 완료되었을 것으로 사료되었고 큰 폐포세포에서도 FN이 분비되는 것으로 결론지울수 있다.X>에서 $1,332.75{\mu}g/mL$으로 최 대값을 나타내었으며, 추출시간 4.24시간 및 시료에 대한 용매비 9.71 mL/g에서 가장 높게 나타났다. 추출온도가 높고, 추출시간이 증가할수록 총 polyphenol 함량이 증가하는 경향을 나타내었다. Gallic acid 함량은 $65.84^{\circ}C$에서 $30.51{\mu}g/mL$으로 최대값을 나타내 었으며, 추출시간 1.65시간 및 시료에 대한 용매비 17.17mL/g에서 가장 높은 추출율을 보였다. Gallic acid 함량에 대한 추출조건의 영향은 추출시간과 용매비에 영향을 받는 것으로 나타났으며, 설정된 범위 내에서 온도에 대한 영향은 거의 나타나지 않는 것으로 나타났다. 실험연구가 더 필요하리라 본다. 혈액학적 변화를 유도하고 환자의 연령, 혈소판 수, 대동맥 차단 시간, 체외 순환 시간, 술후 PT 및 aPTT와 같은 다인적 상황들이 술후 출혈에 영향을 미친다는 점들을 시사하고

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.171-178
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• 2003

This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM＋0.05％ FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3％ BSA or 10％ FBS at 37$^{\circ}C$, 90％ $N_2$, and 5％ $CO_2$in incubator for 6 days. The fusion rates were 51.6％(chamber) and 68.9％(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7％, 11.5％), respectively. The cleavage rates were significantly different between the chamber method(73.2％) and needle method(80.3％), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1％, 61.1％ and 18.4％, 26.3％ respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7％) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Proceedings of the Korean Society for Bio-Environment Control Conference
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• 1998.05a
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• pp.153-157
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• 1998

무등산 수박의 유묘 생산에 있어 적정 배양액 농도를 구명하기 위하여 일본원시균형배양액을 기준으로 하여 NO$_3$-N, K 및 Ca의 농도별 첨가량에 따른 유묘생장반응을 조사하였다. 1. 배양액내 질소농도을 증가시킬수록 초장, 엽면적, 엽수, 엽장, 엽폭이 증가하였으며 생체중과 건물중도 증가하였다. 2. 배양액내 칼륨의 농도를 증가시켰을 경우 수박 유묘의 초장은 200ppm까지는 약간 증가하지만 엽면적, 엽수, 엽장, 생체중 및 건물중은 200ppm이상으로 증가시키면 감소하는 경향이었다. 3. 칼슘처리의 경우 농도의 증가가 생장의 증가를 나타내지는 않았으며 원시균형 배양액의 표준이라 할 수 있는 80ppm처리구의 생장이 양호한 것으로 나타났다. 4. 배양액(일본원시균형배양액 1/2농도)에 N의 농도를 200ppm으로 증가시킬 경우 수박유묘의 엽병내 N, K 및 Mg의 함량이 증가하였지만 P 및 Ca의 증가나 감소의 경향은 보이지 않았다. 5. K의 농도를 100ppm으로 증가시킬 경우 수박유묘의 엽병내 N, K 및 Mg의 함량이 증가하였지만 P 및 Ca의 증가나 감소의 경향은 보이지 않았다. 5. K의 농도를 150ppm으로 증가시킬 경우 수박유묘의 엽병내 N, K 및 Mg의 함량이 증가하였지만 P 및 Ca의 증가나 감소의 경향은 보이지 않았다. 그러나 200ppm이상으로 증가시킬 경우 N과 Mg의 감소가 나타났다. 6. 양액내 Ca의 농도를 증가시킬수록 엽병내 N, K Ca 및 Mg의 농도가 증가하는 반면 P의 경우 거의 변화가 없었으며,N의 경우는 200ppm 이상의 농도에서 감소하는 경향을 나타내었다. Mg의 경우는 150ppm까지는 함량의 변화가 없었지만 200ppm 이상의 처리구에서 급격한 함량증가를 나타내었다.~21.2ml가 흘러나와 배지의 공극이 일정하지 않아 시간당 통과하는 양이 일정하지 않았다고 생각되었다. $\bigcirc$ 펄라이트 : 합섬A(비스코스+레이온)급액천의 유입은 소(1$\times$60cm)에서 21.8ml, 중(2$\times$60cm) 33.5ml, 대(3$\times$60cm) 43.4ml가 통과되었고 합섬(폴리에스텔)에서는 19.0~30.7ml로서 급액천의 규격에 따라 통과되는 차이가 있었다. 배지가 규격화되어 있어 급액천의 규격별로 일정하게 유입되었으며 급액천의 재질이 유입에 영향을 미친 것으로 사료되었다. (2) 급액관과 베드상과의 높이에 따른 유출양 : 급액과 베드상과의 낙차가 클수록 유출이 증가함을 알수 있었으나 합섬C(인견)실험구에서는 낙차가 유출에 영향을 미치지 않았다. (4) 급액된 양액의 EC 및 pH조사 : 급액된 양액의 EC 및 pH에 전혀 변화가 없어 재배 적응에 문제가 없을것으로 사료되었다.이가 가장 이상적인 것으로 생각된다.세포수에 대한 내부세포괴세포(ICM/total cells)가 20~40% 범주에 드는 비율은 처리구가 대조구보다 낮은 결과를 나타냈다. 결론적으로 돼지난포란을 이용하여 체외성숙을 유기할 때 효과적인 cysteamine의 농도는 50$\mu$M이 적당하며, 초기배발달을 유기할 때의 효과적인 cysteamine의 농도는 25~50$\mu$M인 것으로 판단된다.N)A(N)/N을 제시하였다(A(N)=N에 대한 A값). 위의 실험식을 사용하여 헝가리산 Zempleni 시료(15%$S_{XRD}$)의 기본입자분포로부터 %$S_{XRD}$를 계산한

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.235-243
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• 2002

This study was designed to examine the ability of the bovine (MII) oocytes cytoplasm to support several mitotic cell cycles under the direction of differentiated somatic cell nuclei of bovine, porcine, mouse and human. Bovine GV oocytes were matured in TCM-199 supplemented with 10% FBS. At 20h after IVM, recipient oocytes were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst and their 1st polar body (PB) and MII plate were removed by enucleation micropipette under UV filter. Ear skin samples were obtained by biopsy from an adult bovine, porcine, mouse and human and cultured in 10% FBS added DMEM. Individual fibroblast was anlaysed chromosome number to confirm the specificity of species. Nuclear transferred (NT) units were produced by electrofusion of enucleated bovine oocytes with individual fibroblast. The reconstructed embryos were activated in 5 $\mu$M ionomycin for 5 min followed by 1.9 mM 6-dimethylaminopurine (DMAP) in CR1aa for 3 h. And cleaved NT embryos were cultured in CR1aa medium containing 10% FBS on monolayer of bovine cumulus cell for 8 days. Also NT embryo of 4~8 cell stage was analysed chromosome number to confirm the origin of nuclear transferred somatic cell. The rates of fusion between bovine recipient oocytes and bovine, porcine, mouse and human somatic cells were 70.2%, 70.2%, 72.4% and 63.0%, respectively. Also, their cleavage rates were 60.6%, 63.7%, 54.1% and 62.7%, respectively, there were no differences among them. in vitro development rates into morula and blastocyst were 17.5% and 4.3% in NT embryos from bovine and human fibroblasts, respectively. But NT embryos from porcine and mouse fibroblasts were blocked at 16~32-cell stage. The chromosome number in NT embryos from individual fibroblast was the same as chromosome number of individual species. These results show that bovine MII oocytes cytoplasm has the ability to support several mitotic cell cycles directed by newly introduced nuclear DNA.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.175-180
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• 2005

This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for $16\~20$ h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated $0.4\;{\mu}L/mL$ demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs ($86.2\%$) treated with demecolcine protruded a chromosome mass and enucleated efficiently ($98.8\%$, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB ($18.2\%\;vs.\;4.6\%\cdot$, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes ($69.4\%\;and\;5.9\%$, respectively) were significantly lower than those of embryos reconstituted with MII oocytes ($96.7\%\;and\;23.9\%$, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.

• Journal of Chest Surgery
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• v.35 no.7
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• pp.542-547
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• 2002

Background: Postpneumonectomy empyema(PPE) is an infrequent but potentially life-threatening complication. To date, various surgical efforts have been made to manage this complication. We reviewed our 20-year surgical experience of PPE and long-term follow-up data. Material and Method: Total of 37 patients who were treated for PPE between fan, 1980 and Jun, 2000 were included. Various clinical factors such as micro-organism, operative method and timing, presence of bronchopleural fistula(BPF), underlying disease and fate of empyema cavity were retrospectively reviewed and analyzed. Result: Majority of patients(34) underwent Eloesser operation for effective drainage. There was only one operative mortality. The causative organisms were Staphylococcus species and Pseudomonas species in 46% BPF was found in 20 cases, among which spontaneous closures took place in 4 cases. The chest wall was closed in 40%(8/20) of patients with BPF, compared to 59%(10/17) without BPF. The closure rate was statistically better in patients without BPF(p=0.006). Even though the patients with benign disease showed higher closure rate(50%) than those with lung cancer (31%), the difference was not significant(p=0.25). Conclusion: Eloesser procedure was an effective method for initial drainage of PPE cavity with low operative mortality. Given the findings of low spontaneous closure rate of BPF, aggressive approach to close the BPF is mandatory to achieve the final goal of chest wall closure. It was found that majority of patients still left their chest cavity opened, even after controlling the active inflammation of the empyema cavity. More aggnessive approach for chest wall closure is recommended in all patents with benign disease and in selective patients with lung cancer if there is no evidence of recurrence at several years after the initial operation.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.65-70
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• 2006

This study investigated apoptosis and in vitro development of parthenogenetic preimplantation porcine embryos. In vitro matured oocytes for $42{\sim}44h$ were used. Apoptotic cell death was analyzed by using a terminal deoxynucleatidyl transferase mediated deoxyuridine 5-triphosphate nick-end tabling (TUNEL) assay. In experiment 1, oocytes were activated with two electric pulses (CH) of 1.2 kV/cm for $30{\mu}sec$ (E), E + 6-dimethylaminopurine (6-DMAP) or E + cycloheximide (CH) and cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$. In experiment 2, oocytes were activated by E and cultured in PZM-3 or NCSU-23 under a gas atmosphere of 20% $O_2$ ($5%\;CO_2$, in air) or 5% $O_2$ $(5%\;CO_2,\;5%\;O_2\;90%\;N_2)\;at\;38.5^{\circ}C$. Oocytes activated with E+6-DMAP or E+CH showed higher blastocyst rates (36.3% and 32.5%) compared to E alone (27.7%). The frequency of apoptosis according to treatments were 5.3%, 7.7% and 7.1% respectively. Oocytes activated with E alone showed lower (P<0.05) frequency of apoptosis compared to other groups. In experiment 2, parthenotes cultured in PZM-3 showed slightly higher blastocyte rates (28.2% and 29.7%) compared to NCSU-23 (22.6% and 24.4%) regardless of atmosphere. Blastocysts generated in PZM-3 showed lower (P<0.05) apoptosis rate under 20% $O_2$ (9.2% vs 16.9%), whereas those in NCSU-23 had slightly lower apoptosis rate under 5% $O_2$ (14.0% vs 18.4%). This result represents that activation method and culture condition could affect the frequency of apoptosis as well as in vitro developmental rate.

• Korean Journal of Food Science and Technology
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• v.13 no.4
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• pp.314-318
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• 1981

In order to improve the storage stability of spray and freeze dried red ginseng extract powder packed in a bottle, the water vapor permeability of Al-foil laminate paper used for cap closure and shelf life of those products on various storage temperatures and relative humidities were investigated. The thickness of the laminate paper was $93{\pm}3\:{\mu}m$ and its physical properties were equal to standard of ASTM (B-377-66) The absorption rate of the freeze dried powder was 2-6 times greater than that of the spray dried powder at $37^{\circ}C$. Therefore it was considered that the laminate could be used for cap closures for the spray dried powder but unsuitable for the freeze dried powder. The shelf life of the spray dried powder was longer than that of the freeze-dried powder at $37^{\circ}C$.