• 제목/요약/키워드: CM활성화

검색결과 345건 처리시간 0.022초

칡소 귀세포를 이용한 핵이식란의 배양방법이 배반포 발달율과 수태율에 미치는 영향

  • 윤종택;이호준;최은주
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.65-65
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    • 2001
  • 본 연구는 칡소 귀세포를 공여핵으로 이용한 체세포 복제송아지 생산에 있어서 배양방법이 배발생 및 배반포 발달율에 미치는 영향과 체세포 복제란의 이식후 수태율에 미치는 영향을조사하여 복제송아지의 생산 효율을 제고하고자 실시하였다. 실험에 공시된 공여핵은 칡소 의 귀세포를 회수하여 10%FBS가 첨가된 DMEM배지에서 3-4일 동안 배양하여 monolayar Confluent 형성 후 0.25% trypsin을 처리하여 준비하였으며 공여세포는 적어도 passage가 5회 이상의 세포만을 사용하였다. 복제수정란의 생산은 18-20시간 동안 체외성숙 된 난자의 핵을 제거하고 공여핵을 주입하여 2.2kv/cm, 10$\mu\textrm{s}$의 전압으로 2회 자극함으로 융합하였으며, 융합된 난자는 5$\mu\textrm{g}$/$m\ell$ ionomycin에서 4분간, 1.9mM 6-dimethyl aminopurine에서 4시간동안 배양하여 활성화처리를 하였다. 핵이식수정란의 배양은 39$^{\circ}C$, 5%$CO_2$ incubator에서 처리구 I은 CRlaa에서 4일간 배양 후 CR2aa배지에서 배양, 처리구II는 CRlaa에 4일간 배양후 CR2aa배지에 cumulus cell과 공배양, 처리구III은 CR2aa 배지에 camulus cell과 함께 배양하였다. 수정란이식은 발정발현 7일째에 비외과적 방법으로 젖소 미경산우에 이식하였으며 이식란수는 2~4개의 핵이식된 수정란을 이식하였다. 임신진단은 45~60일 사이에 직장검사 및 초음파 진단기를 이용하여 실시하였다. 배양방법에 따른 배발생율은 처리구 I에서 92.2 %(83/90)으로 처리구II와 III의 62.4%(63/101)와 77.8%(144/185)에 비하여 높게 나타났으나 배반포 발달율은 처리구II와III에서 65.1%(41/63)와 50.0%(72/144)로 처리구 I의 30.1%(25/83)보다 높게 나타났다. 각 처리구에 따른 수정란 이식후 수태율은 처리구II와 III에서 공히 20%의 수태율을 나타낸 반면 처리구 I에서는 수태가 되지 않았다. 따라서 체세포 복제수정란의 생산에 있어서 배반포 발달율과 수태율을 높이기 위해서는 단순배양보다 공배양이 더 효과적인 것으로 사료되지만 이런 결과가 복제송아지 생산효율에 있어서도 효과적일지는 향후 더 많은 연구가 있어야 할 것으로 사료된다.

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소 체세포 핵이식에서 체세포 계대에 따른 염색체 이상

  • 최은주;이호준;김병정;민관식;윤종택
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 국제심포지엄
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    • pp.77-77
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    • 2002
  • 본 연구는 체세포 및 체세포 핵이식란의 염색체 이상에 체세포의 계대의 영향성을 조사하고자 하였다. 한우 성축의 귀 조직으로 얻어진 세포를 공여세포를 체외성숙 후 제핵된 난자에 핵이식을 실시하였으며, 1.9kv/cm, 20$\mu\textrm{s}$/2times의 전기자극으로 융합후 5$\mu\textrm{g}$/ml의 ionomycin에서 4min, 1.9mM 6-DMAP에서 4h동안 배양함으로써 활성화를 유도하였다. 핵이식란은 CRlaa에서 4일간 배양 후 8 -cell단계에서 중기상의 유도를 위하여 상기 배양액 1ml당 0.05$\mu\textrm{g}$ colcemid에서 6-8시간 더 배양하였다. 이후, 6% Fetal bovine serum이 함유된 1% sodium citrate용액에 20분간 저장처리 후, methanol 5 : aceticacid 1 : distilled water 4로 1차, methanol 3: aceticacid 1 로 조성된 2차, methanol 4 : acetic acid 3 : distilled water 1의 3차고정액으로 1분간 재 침지시켰다. 고정 처리가 완료된 slide는 4% Giemsa용액으로 염색한 후 광학현미경 하에서 핵형 양상을 검경하였다. 체세포의 5계대에서는 684개의 spreads를 검경한 결과 염색체 수는 72%가 정상으로 60개이었고, 24%가 60개 이하였으며 4%가 60개 이상을 보였다. 10계대도 5계대와 비슷하여 71%가 정상, 26%가 60개 이하, 3%가 60개 이상이었고, 15계대에서는 55%가 정상이었고, 30%가 60개이하, 15%가 60개 이상을 보였다. 10계대 까지는 mixoploid의 비율의 변화가 없었으나 15계대에서 현저하게 늘어남을 볼 수 있었다. 또한 체외수정란과 핵이식란의 비교에서는 체외수정란은 250개의 spreads를 검경한 결과 염색체 수는 95.6%가 정상으로 60개이었고, 2.0%가 60개 이하, 2.4%가 60개 이상이었으나, 핵이식란은 204개를 검경하여 88%가 정상이었고, 4.9%가 60개이하, 7.1%가 60개 이상을 보임으로써, 핵이식란이 체외수정란에 비하여 염색체 이상의 비율이 높았다. 따라서 계대에 따라 체세포의 염색체이상의 비율이 상대적으로 증가하고, 체세포 핵이식에 따른 염색체 이상이 생길 수 있음을 알 수 있었다. (이 논문은 농림부 연구비에 의해 수행되었음)

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액상환원침전법에 의한 저온활성화소결용 복합W분말의 제조방법 및 소결특성에 관한 연구 (A Study on the Manufacture of Composite W Powder for Low Sintering Temperature by Liquid Reduction Precipitation Method)

  • 김창욱;이철;정인;윤성렬
    • 한국표면공학회지
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    • 제28권4호
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    • pp.207-218
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    • 1995
  • Tungsten(W) metal has excellent properties in heat-resistance, corrison-resistance and impact-resistance but W-Metal is hard to sinter because higher than $2,000^{\circ}C$ is required to sinter W-powder. Con-sequently, a deposit technique of Nikel Phosphorus(NiP) on W-powber by the liquid reduction precipitation method was performed. Sintering temperature of the resulting W-NiP composite was lowered around to $1,000^{\circ}C$, and the mechanical properties of the sintered body was studied. The most suitable conditions for NiP thin film deposit on W-Powder by the liquid reduction precipitation method, which are composition, concentration, pH and temperature of the liquid reduction solution, were considered. The activated sintering was carried out in a reducing condition furnace. Components and properties of the sintered body were investigated by the density and the hardness measurements, X- ray diffraction analysis, and microscopic photographs of the surface. Quantity of NiP thin film on W-powder could be varied by the change of the liquid reduction solution composition. The sintering temperature of W-NiP composite powder is lowered to $950^{\circ}C$ from $2,000^{\circ}C$ and the hardness is increased (ca. 720 Hv). Large shrinkage could be observed since density was increased from 5.5 to 11.0 g/$cm^2$ which 86.2% of theoretical density. W metal and $Ni_3P$ crystal were detected through X-ray diffraction on the sintered body. Perfectly activated sintering was observed by microscopic photographs.

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HSLA-100강 및 HY-100강의 응력제거처리 균열에 관한 연구 (A Study on the Stress Relief Cracking of HSLA-100 and HY-100 steels)

  • 박태원;심인옥;김영우;강정윤
    • Journal of Welding and Joining
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    • 제14권3호
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    • pp.48-57
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    • 1996
  • A study was made to examine the characteristics of base metal and stress relief cracking(SRC) of heat affected zone(HAZ) for HY-100 and Cu-bearing HSLA-100 steels. The Gleeble thermal/mechanical simulator was used to simulate the SRC/HAZ. The details of mechanical properties of base plate and SRC tested specimens were studied by impact test, optical microscopy and scanning electron microscopy. The specimens were aged at $650^{\circ}C$ for HSLA-100 steel and at $660^{\circ}C$ for HY-100 steel and thermal cycled from $1350^{\circ}C$ to $25^{\circ}C$ with a cooling time of $\Delta$t_${800^{circ}C/500^{circ}C}$=21sec. corresponds to the heat input of 30kJ/cm. The thermal cycled specimens were stressed to a predetermined level of 248~600MPa and then reheated to the stress relief temperatures of $570~620^{\circ}C$. The time to failure$(t_f)$ at a given stress level was used as a measure of SRC susceptibility. The strength, elongation and impact toughness of base plate were greater in HSLA-100 steel than in HY-100 steel. The time to failure was decreased with increasing temperature and/or stress. HSLA-100 steel was more susceptible to stress relief cracking than HY-100 steel under same conditions. It is thought to be resulted from the precipitation of $\varepsilon$-Cu phase by dynamic self diffusion of solute atoms. By the precipitation of $\varepsilon$-Cu phase, the differential strengthening of grain interior relative to grain boundary may be greater in the Cu-bearing HSLA-100 steel than in HY-100 steel. Therefore, greater strain concentration at grain boundary of HSLA-100 steel results in the increased SRC susceptibility. The activation energies for SRC of HSLA-100 steel are 103.9kcal/mal for 387MPa and 87.6kcal/mol for 437MPa and that of HY-100 steel is 129.2kcal/mol for 437MPa.

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배추 Polygalacturonase의 열안정성 (Thermostability of Polygalacturonase from Chinese Cabbage)

  • 정태규;문태화;박관화
    • 한국식품과학회지
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    • 제25권5호
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    • pp.576-581
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    • 1993
  • 김치 조직의 연화에 관여하는 효소인 polygalacturonase(PG)를 배추에서 추출하여 황산암모늄 분획, 이온교환크로마토그래피 및 FPLC를 이용하여 D-PG, C-1, C-2 PG 세 분획으로 분리 정제하여 특성을 조사하였다. 분리된 세 분획의 활성 최적 온도는 $65^{\circ}C$, 최적 pH가 5.2였으며 pH$4.5{\sim}8.0$ 범위에서 안정하였다. NaCl에 의한 영향은 0.3M NaCl에서 최대의 활성을 보였으나 0.6M 이상에서는 저해를 받았으며 $CaCl_2$의 경우 $0{\sim}0.5mM$ 농도에서는 활성이 크게 영향을 받지 않았으나 0.8mM에서는 저해를 받았다. 열불활성화 특성은 isozyme간에 큰 차이가 없었으며 1차 반응을 따랐다. 이 효소 isozyme의 z값은 $8.4{\sim}9.3^{\circ}C,\;80^{\circ}C$에세 D값은 $102{\sim}126$초였다. 이 호소의 불활성화 값을 이용하여 retort pouch 김치 살균공정에 적용하고 역가의 잔존 가능성에 대하여 고찰하였다.

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증착 조건에 따른 ITO 전극의 X-ray 변환물질에서의 특성 평가

  • 노성진;신정욱;이영규;송용근;이지윤;박성광;남상희
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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    • pp.366-366
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    • 2012
  • 현재 사용되고 있는 투명전극재료 중에 ITO(Indium Tin Oxide)가 가장 투명하면서 전기도 잘 통하고 생산성도 좋다. 투명전극은 비저항이 $1{\times}10^{-3}{\Omega}/cm$이하, 면 저항이 $10^3{\Omega}/sq$이하로 전기전도성이 우수하고 380에서 780 nm의 가시광선 영역에서의 투과율이 80%이상이라는 두 가지 성질을 만족시키는 박막이다. 본 연구에서 X-ray Film을 제작하기 위하여 상용화된 ITO Glass 전극 기판에 X-ray가 조사되면 직접 전자 전공 쌍(electron-hole pair)을 발생시켜 전기적 신호를 발생하는 광도전체 물질(Photoconductor)인 PbO, $PbI_2$, $HgI_2$를 스크린 프린팅(Screen Printing)법을 이용하여 각각 제작하였다. 상부 전극으로 마그네틱 스퍼터링(Magnetic Sputtering) 진공 증착 장치를 사용하여 전류, 전압, 아르곤 및 산소 유입량등을 조절하면서 상부 전극을 증착하였다. 이때 타켓으로 $In_2O_3;SnO_2$ (조성비:90:10wt%)를 사용 하였고, base pressure는 $9{\times}10^{-7}torr$, deposition pressure는 $3{\times}10^5torr$를 고정하였다. 또한 전류와 전압은 각각 0.4A, 800V로 유지하고, $O_2$:0.3 ppm, Ar의 경우 4.9 ppm에서 70 ppm까지 올려 플라즈마를 활성화 시킨 후 90초 동안 ITO를 증착하였다. 본 실험에 제작된 박막으로 X-ray을 조사하여 검출기로써 특성 평가를 실시하였으며, 실험결과 X-선 투과와 전도성 등 두가지 특성이 동시에 만족 될 만한 성능을 가질 수 있음을 확인 할 수 있었다.

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Ionomycin과 6-Dimethylaminopurine(6-DMAP)처리로 활성화된 토끼 난자의 단위발생 (Parthenogenetic Development of Rabbit Oocytes Activated by Ionomycin Plus 6-Dimethylaminopurine Treatments)

  • 윤희준;이효종;최상용;박충생
    • 한국가축번식학회지
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    • 제22권3호
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    • pp.221-227
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    • 1998
  • This study was carried out to determine the effects of ionomycin and 6-dimethylaminopurine (6-DMAP) treatments on parthenogenesis of rabbit oocytes. The oocytes were randomly assigned to the activation treatments with either ionomycin plus 6-DMAP or electric stimulation. The oocytes were colected from the oviducts of superovulated rabbits at 13~14 hours and 19~20 hours post hCG injection and were activated with 5$\mu$M ionomycin for 5 min and 2 hours incubation in 2mM 6-DMAP. The other oocytes were stimulated by three pulses of 3.6kV/cm for 60 $\mu$sec each 30 min apart, starting 19 hours post in hCG in 0.28M mannitol solution with 100$\mu$M Ca2+ and Mg2+. The results obtained were summarized as follows: 1. Following treatment of the oocytes with ionomycin plus 6-DMAP, the cleavage rate and in vitro developmental rate to blastocyst were significantly(P<0.01) higher in the oocytes collected bet ween 19~20 hours than between 13~14 hours after hCG injection. 2. When the oocytes were treated with ionomycin plus 6-DMAP, 85(98.8%) of 86 treated oocytes extruded the second polar bodies, with the entire chromatin complements outside ooplasm. However when the oocytes were restored during subsequent incubation in the drug-free medium, the cytoplasts regain their full capacity for parthenogenetic activation and nuclear remodelling. 3. The cleavage rate and the in vitro developmental rate to blastocyst were not significantly different in the oocytes activated by ionomycin plus 6-DMAP treatment(91.2 and 45.6%) or electrical stimulation(89.6 and 34.3%).

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옥상정원 활성화에 유용한 토질영양제에 대한 연구 (Study on Several Soil Conditioners for Roof Gardening)

  • 김기은
    • 한국환경복원기술학회지
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    • 제18권4호
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    • pp.63-71
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    • 2015
  • Recently the roof gardening has become very popular and even on the apartment-veranda and on the roofs in high buildings many kinds of vegetables and berries are cultivated. And the demand of the nutritional supplement for the effective plant growth is also increasing. The general urban conditions are to be adapted for plant growth. A different hygroscopic and temperature-conditions after regions, sunshine and wind have strong influences on the plant growth and usually it is not optimal enough. It is because why a nutritional condition in soil for plant growth so important and essential. The usual compound-soil or -fertilizer cannot offer enough quantities of nutrients for plant growth and additional soil conditioner becomes more necessary. There are many kinds of soil conditioners like hydrogel in the market and we studied on Geohumus, Montigel and Geko, which are widely used in Europe and other countries. Water absorption and microbial immobilization with effective microorganisms were tested and compared. The EM solution was identified as bacteria, fungi and azotobacter etc. and they were immobilized at the soil conditioners at first. And the cultivated and immobilized at the soil conditioners EM-solution was added to the plant soil. 1 g of the soil conditioners absorbed ca. 20 g of water. The plant grew 10 cm more, got 3 times more branches and 2 times more fruits in the soil with soil conditioners immobilized with microorganisms. With water addition the plant with both conditions in the soil could stay fresh in comparison to without soil conditioners.

유체 압력에 의한 흰쥐 심실근세포 pH의 감소 및 $Na^+-H^+$ 교환체의 활성화 (Decrease of Intracellular pH and Activation of $Na^+-H^+$ Exchanger by Fluid Pressure in Rat Ventricular Myocytes)

  • 김준철;우선희
    • 약학회지
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    • 제55권3호
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    • pp.247-250
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    • 2011
  • An increase in ventricular pressure can alter cardiac excitation and contraction. Recent report has demonstrated that fluid pressure (FP) suppresses L-type $Ca^{2+}$ current with acceleration of the current inactivation in ventricular myocytes. Since the L-type $Ca^{2+}$ channels known to be regulated by intracellular pH ($pH_i$), this study was designed to explore whether pressurized fluid flow affects pHi in isolated rat ventricular myocytes. A flow of pressurized (~16 dyne/$cm^2$) fluid, identical to that bathing the myocytes, was applied onto single myocytes, and intracellular $H^+$ concentration was monitored using confocal $H^+$ imaging. FP significantly decreased $pH_i$ by $0.07{\pm}0.01$ pH units (n=16, P<0.01). Intracellular acidosis enhances the activity of $Na^+-H^+$ exchanger (NHE). Therefore, we examined if the NHE activity is increased by FP using the NHE inhibitor, HOE642. Although HOE642 did not alter $pH_i$ in control conditions, it decreased $pH_i$ in cells pre-exposed to FP, suggesting enhancement of NHE activity by FP. In addition, FP-induced intracellular acidosis was larger in cells pre-treated with HOE642 than in cells under the control conditions. These results suggest that FP induces intracellular acidosis and that NHE may contribute to extrude $H^+$ during the FP-induced acidosis in rat ventricular myocytes.

유도결합 플라즈마 화학기상증착법에 의해 활성화된 탄소원자를 이용한 Ni/SiO2/Si 기판에서 그래핀 성장 (Graphene Formation on Ni/SiO2/Si Substrate Using Carbon Atoms Activated by Inductively-Coupled Plasma Chemical Vapor Deposition)

  • 람반낭;김의태
    • 한국재료학회지
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    • 제23권1호
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    • pp.47-52
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    • 2013
  • Graphene has been synthesized on 100- and 300-nm-thick Ni/$SiO_2$/Si substrates with $CH_4$ gas (1 SCCM) diluted in mixed gases of 10% $H_2$ and 90% Ar (99 SCCM) at $900^{\circ}C$ by using inductively-coupled plasma chemical vapor deposition (ICP-CVD). The film morphology of 100-nm-thick Ni changed to islands on $SiO_2$/Si substrate after heat treatment at $900^{\circ}C$ for 2 min because of grain growth, whereas 300-nm-thick Ni still maintained a film morphology. Interestingly, suspended graphene was formed among Ni islands on 100-nm-thick Ni/$SiO_2$/Si substrate for the very short growth of 1 sec. In addition, the size of the graphene domains was much larger than that of Ni grains of 300-nm-thick Ni/$SiO_2$/Si substrate. These results suggest that graphene growth is strongly governed by the direct formation of graphene on the Ni surface due to reactive carbon radicals highly activated by ICP, rather than to well-known carbon precipitation from carbon-containing Ni. The D peak intensity of the Raman spectrum of graphene on 300-nm-thick Ni/$SiO_2$/Si was negligible, suggesting that high-quality graphene was formed. The 2D to G peak intensity ratio and the full-width at half maximum of the 2D peak were approximately 2.6 and $47cm^{-1}$, respectively. The several-layer graphene showed a low sheet resistance value of $718{\Omega}/sq$ and a high light transmittance of 87% at 550 nm.