• Title/Summary/Keyword: C1-2 fusion

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Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.293-302
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    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

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Study on Isothermal Crystallization Characteristics of PLA Film by Adding APP as a Nucleation Agent (APP 핵제를 첨가한 PLA 필름의 등온결정화 특성에 관한 연구)

  • Kim, Gyu-Sun;Kim, Moon-Sun;Kim, Byung-Woo
    • Korean Chemical Engineering Research
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    • v.50 no.3
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    • pp.582-587
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    • 2012
  • In this paper, it was studied on the crystallization characteristics of PLA film by adding ammonium phosphate (APP) as a nucleation agent. Crystallinity and crystallite size of PLA film were determined by Scherrer equation. Crystallization rate constant of PLA film was calculated through Avrami equation. Film samples in the study were prepared by two steps. PLA films were prepared by adding 1, 5, and 10 wt%, respectively, at first and was secondly annealed at 130, 140, and $150^{\circ}C$. Crystallinity of pure PLA film was average 4.6% and those of PLA film with adding 1, 5, and 10 wt% APP were 12.2, 47.7, and 50.0%, respectively. Crystallite size of PLA film was average 28.0 nm and those of PLA film with adding 1, 5, and 10 wt% APP were 26.8, 24.0, and 19.0 nm, respectively. Crystallization rate constants of PLA film with 1 wt% APP were 2.12, 3.86, and 0.27 by annealing at 130, 140, and $150^{\circ}C$, respectively, where was higher than pure PLA film and those with adding 5 and 10 wt% APP, respectively.

Interspecific Variation in the Protoplast Formation of the Genus Cellulomonas (Cellulomonas속 종간의 원형질체 형성조건의 차이에 대하여)

  • Lee, Eun-Ju;Bae, Moo
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.154-160
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    • 1986
  • In order to develope interspecific fusion of the genus Cellulomonas capable of assimilation cellulose, the optimun conditions for the protoplast formation was investigated to examine the susceptibility of cell wall, between different species of the same genus using scanning electron microscope. The variation in the susceptibilities of Cellulomonas sp. CS 1-1 and C. flavigena to lysozyme treatment were considerably remarkable, although they belong to the same genus. The rate of protoplast formation of CS1-1 was 99.9% being treated with lysozyme $(100{\mu}g/ml)$ for 30 minute and that of C. flavigena was about 80% being treated at the concentration of $600{\mu}g/ml$ of lysozyme for 6 hours. The susceptibility of cell wall to the lysozyme treatment on protoplast formation of the strain, CS1-1 seems not to be depend on the cultural periods of cells. On the contrary, that of C. flavigena was considerably depend on the periods. Cells of C. flavigena at mid exponential phase could be more efficiently converted to protoplast cells than those at late exponential phase be done. The rate of the protoplast formation was 95%, when cells of C. flavigena at mid exponential phase were treated with lysozyme $600{\mu}g/ml$ for 6 hours and observed by SEM. In the evalution of protoplast formation of the CS1-1 results of counting method in plate after osmotic shock treatment were similar to the results of the direct observation method by means of SEM. But in the case of C. flavigena the latter method was much more reliable than the former, because the differences between the number of spheroplasts and protoplasts were not able to figure out on conuting the number of protoplast after osmotic shock tretment.

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Subarachnoid Space Reconstruction for Treatment of Posttraumatic Syringomyelia - A Case Report - (외상성 척수공동증의 치료를 위한 지주막하강 재건술 - 증례보고 -)

  • Chung, Dai Jin;Kim, Sung Min;Kim, Hun;Shim, Young Bo;Park, Yong Kee;Choi, Sun Kil
    • Journal of Korean Neurosurgical Society
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    • v.29 no.2
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    • pp.255-260
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    • 2000
  • The authors report a posttraumatic syringomyelia in a 30-year-old man who has complained pain, weakness of upper arm and dissociation sensory loss since 2 months before. He was underwent by decompressive laminectomy from T12 to L1, reduction of encroached bony fragments, transpedicular screw fixation from T12 to L2 and posterolateral bony fusion due to burst fracture of L1 at other hospital 3 years ago. Preoperative spinal MRI was highly suggestive of wide-spread, multiseptated syringomyelia from C3 to thoracolumbar junction. We performed wide decompressive laminectomy from T10 to L2 and subarachnoid space reconstrucion composed of microdissection of meningeal fibrosis widely, iatrogenic meningocele formation with lefting the dura mater opened for treatment of spinal-spinal pressure dissociation. Clinical manifestations and radiological findings of the patient were improved after the operation. This technique was thought to be superior to shunting procedures in cases of wide-spread, multiseptated post-traumatic syringomyelia.

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IRRADIATION EMBRITTLEMENT OF CLADDING AND HAZ OF RPV STEEL

  • Lee J.S.;Kim I.S.;Jang C.H.;Kimura A.
    • Nuclear Engineering and Technology
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    • v.38 no.5
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    • pp.405-410
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    • 2006
  • Microstructural features and their related mechanical property changes in the 309L cladding and the heat affected zone (HAZ) of SA508 cl.3 steel were investigated through the use of TEM, tensile and small punch (SP) tests. The specimens were irradiated at 563 K up to the neutron fluences of $5.79{\times}10^{19}n/cm^2$ (>1MeV). The microstructure of the clad was mainly composed of a fcc ${\gamma}-phase$, a low percentage of bcc ${\delta}-ferrite$, and a brittle ${\sigma}-phase$. Along the weld fusion line there formed a heavy carbide precipitation with a width of $20{\sim}40{\mu}m$, showing preferential cracking during plastic deformation. The yield stress and ductile-to-brittle transition temperature (DBTT) of the irradiated clads increased. The origin of the hardening and the shift of the DBTT are discussed in terms of the irradiation-produced defect clusters of a fine size and brittle ${\sigma}-phase$.

Characterization of Microbial Fuel Cells Enriched Using Cr(VI)-Containing Sludge

  • Ryu, Eun-Yeon;Kim, Mi-A;Lee, Sang-Joon
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.187-191
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    • 2011
  • Microbial fuel cells (MFCs) were successfully enriched using sludge contaminated with Cr(VI) and their characteristics were investigated. After enrichment, the charge of the final 10 peaks was 0.51 C ${\pm}$ 1.16%, and the anodic electrode was found to be covered with a biofilm. The enriched MFCs removed 93% of 5 mg/l Cr(VI) and 61% of 25 mg/l Cr(VI). 16S rDNA DGGE profiles from the anodic electrode indicated that ${\beta}$-Proteobacteria, Actinobacteria, and Acinetobacter sp. dominated. This study is the first to report that electrochemically active and Cr(VI)-reducing bacteria could be enriched in the anode compartment of MFCs using Cr(VI)-containing sludge and demonstrates the Cr(VI) removal capability of such MFCs.

The Effects of Visual Training on Binocular Anomalies (양안시 기능 이상의 시훈련(Visual Training)의 효과)

  • Kim, In-Suk;Hong, Dong-Gyun;Kim, Jung-Hee
    • Journal of Korean Ophthalmic Optics Society
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    • v.10 no.1
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    • pp.53-61
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    • 2005
  • In this study, we measured Near Point of Convergence(N.P.C) tests, Phoria tests using Von Grafe method and relative convergence tests on 138 men and 162 women, so a total of 300 subjects aged between 8~45 to examine the improvement of the fusion vergence through visual training and obtained as follows. 1. According to the results, the near point of convergence of 57 (19%) subjects were shorter than 7cm, and 243 (81%) were 7cm or longer, having a problem in convergence. After visual training, the number of subjects have the value shorter than 7 cm increased from 57 to 111 (37%), and the number of those have the value 7cm or longer decreased significantly form 243 to 189 (63%). 2. The results of the measure of lateral Phoria at far distance by Von Grafe method showed orthophoria 18 (6%), exophoria 198 (66%), esophoria 84 (28%). After phoria test, we examined the N.R.C and P.R.C test. The results showed that the hope finger was improved after V.T using B.l, B.O card. 3. The results of the measure of lateral Phoria at near distance by Von Grafe method showed orthophoria 6 (2%), exophoria 222 (74%), esophoria 72 (24%). After phoria test, we examine the N.R.C and P.R.C test. The results showed that the hope finger was improved after V.T using B.I, B.O card.

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Delivery of Ti Plasmid into Nicotiana sanderae Protoplasts via Liposomes (Liposome을 이용한 Ti Plasmid의 꽃담배 원형질체내 도입)

  • Lim, Myung-Ho;Jeong, Jae-Dong;Kim, In-Soo
    • Applied Biological Chemistry
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    • v.37 no.5
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    • pp.343-348
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    • 1994
  • Ti plasmid of A. tumefaciens was labeled with $^3H-thymidine$, purified and encapsulated into phosphatidylserine (PS) and PS-cholesterol (Chol; 1 : 1 molar ratio) liposomes by lyophilization-rehydration method. PS was supplemented with 1 mole percent octadecyl rhodamine B for fluorometric measurement of PS. Liposomes entrapping $^3H-Ti plasmid$ were fused with Nicotiana sanderae protoplasts by treating with 5 mM $CaCl_2$ and 10% PEG. The fusion was evidenced by fluorescence microscopic technique. The amounts of Ti plasmid and PS associated with protoplasts were assayed by the radioactivity of $^3H-Ti plasmid$ and by the fluorescence of rhodamine B. About 7.9% of the PS liposome and 7.2% of PS-Chol liposome were fused with protoplasts. During the fusion process, about 30% of the liposomal contents of PS-Chol liposome was leaked, in contrast to about 60% leakage of its contents in PS liposome. Accounting the number of liposomes fused with protoplasts together with the encapsulation efficiency and the leakage of liposomal contents, it was calculated that ca. 1,700 Ti plasmid was transfered into one protoplast by the present method. This result may indicates that the present method transfers enough Ti plasmid into plant protoplast to elicit genetic transformation of plants.

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Post Annealing Effects on Iron Oxide Nanoparticles Synthesized by Novel Hydrothermal Process

  • Kim, Ki-Chul;Kim, Young-Sung
    • Journal of Magnetics
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    • v.15 no.4
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    • pp.179-184
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    • 2010
  • We have investigated the effects of post annealing on iron oxide nanoparticles synthesized by the novel hydrothermal synthesis method with the $FeSO_4{\cdot}7H_2O$. To investigate the post annealing effect, the as-synthesized iron oxide nanoparticles were annealed at different temperatures in a vacuum chamber. The morphological, structural and magnetic properties of the iron oxide nanoparticles were investigated with high resolution X-ray powder diffraction (XRD), high resolution transmission electron microscopy (HRTEM), Mossbauer spectroscopy, and vibrating sample magnetometer analysis. According to the XRD and HRTEM analysis results, as-synthesized iron oxide nanoparticles were only magnetite ($Fe_3O_4$) phase with face-centered cubic structure but post annealed iron oxide nanoparticles at $700^{\circ}C$ were mainly magnetite phase with trivial maghemite ($\gamma-Fe_2O_3$) phase which was induced in the post annealing treatment. The crystallinity of the iron oxide nanoparticles is enhanced by the post annealing treatment. The particle size of the as-synthesized iron oxide nanoparticles was about 5 nm and the particle shape was almost spherical. But the particle size of the post annealed iron oxide nanoparticles at $700^{\circ}C$ was around 25 nm and the particle shape was spherical and irregular. The as-synthesized iron oxide nanoparticles showed superparamagnetic behavior, but post annealed iron oxide nanoparticles at $700^{\circ}C$ did not show superparamagnetic behavior due to the increase of particle size by post annealing treatment. The saturation of magnetization of the as-synthesized nanoparticles, post annealed nanoparticles at $500^{\circ}C$, and post annealed nanoparticles at $700^{\circ}C$ was found to be 3.7 emu/g, 6.1 emu/g, and 7.5 emu/g, respectively. The much smaller saturation magnetization value than one of bulk magnetite can be attributed to spin disorder and/or spin canting, spin pinning at the nanoparticle surface.

Point Mutations in the Split PLC-γ1 PH Domain Modulate Phosphoinositide Binding

  • Kim, Sung-Kuk;Wee, Sung-Mo;Chang, Jong-Soo;Kwon, Taeg-Kyu;Min, Do-Sik;Lee, Young-Han;Suh, Pann-Ghill
    • BMB Reports
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    • v.37 no.6
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    • pp.720-725
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    • 2004
  • A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-${\gamma}1$ has two putative PH domains, an $NH_2$-terminal (PH1) and a split PH domain ($nPH_2$ and $cPH_2$). We previously reported that the split PH domain of PLC-${\gamma}1$ binds to phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)$P_2$) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)$P_2$, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-1) region of the PLC-${\gamma}1$ $nPH_2$ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-${\gamma}1$ nPH2 domain mutants, P500A and H503A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-${\gamma}1$ molecules showed reduced PI(4,5)$P_2$ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both $PH_1$ and $nPH_2$ domains are responsible for membrane-targeted translocation of PLC-${\gamma}1$ upon serum stimulation. Together, our data reveal that the amino acid residues $Pro^{500}$ and $His^{503}$ are critical for binding of PLC-${\gamma}1$ to one of its substrates, PI(4,5)$P_2$ in the membrane.