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A new purification method for the Fab and F(ab)2 fragment of 145-2C11, hamster anti-mouse CD3ε antibody

  • Kwack, Kyu-Bum
    • BMB Reports
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    • 제33권2호
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    • pp.188-192
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    • 2000
  • Recombinant protein G has been utilized in the purification of antibodies from various mammalian species based on the interaction of antibodies with protein G. The interaction between immunoglobulin and protein G may not be restricted to the Fc protion of antibodies, as many different $F(ab)_2$ or Fab fragments can also bind to protein G. I found both FAb $F(ab)_2$ of 145-2C11, a hamster anti-mouse $CD3{\varepsilon}$ antibody, bound to the protein G-sepharose. Interestingly, Fab and $F(ab)_2$ of 145-2C11 did not bind to the protein A-sepharose. The binding of Fab and $F(ab)_2$ of 145-2C11 to protein G provided a useful method to remove proteases, chopped fragments of the Fc region, and other contaminating proteins. The remaining intact antibody in the protease reaction mixture can be removed by using a protein A-sepharose, because the Fab and $F(ab)_2$ portions of 145-2C11 did not bind to protein A-sepharose. The specific binding of Fab and $F(ab)_2$ portions of 145-sC11 to a protein G-sepharose (though not to a protein A-sepharose) and binding of intact 145-2C11 to both protein A- and G-sepharose will be useful in developing an effective purification protocol for Fab and $F(ab)_2$ portions of 145-2C11.

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Temperature Dependence of the Vibration-Vibration Energy Transfer for HF(v = n) + $H_2$(v = 0) and DF(v = n) + $D_2$(v = 0)

  • Lee, Chang-Soon;Kim, Yoo-Hang
    • Bulletin of the Korean Chemical Society
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    • 제13권1호
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    • pp.11-17
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    • 1992
  • Vibration-to-vibration energy transfer probabilities for $HF(v=n)+H_2(v=0){\to}HF(v=n-1)+H_2(v=1)$ and $DF(v=n)+D_2(v=0){\to}DF(v=n-1)+D_2(v=1)$ including both the vibration-to-vibration and translation (V-V, T) and vibration-to-vibration and rotation (V-V, R) energy transfer paths have been calculated semiclassically using a simplified collision model and Morse-type intermolecular interaction potential. The calculated results are in reasonably good agreement with those obtained by experimental studies. They also show that the transition processes for $HF(v=1-3)+H_2(v=0){\to}HF(v=0-2)+H_2(v=1)$ and $DF(v=1,\;4)+D_2(v=0){\to}DF(v=0,\;3)+D_2(v=1)$ are strongly dependent on the V-V, T path at low temperature but occur predominantly via the V-V, R path with rising temperature. The vibration-to-vibration energy transfer for $HF(v=4)+H_2(v=0){\to}HF(v=3)+H_2(v=1)$ and $DF(v=2-3)+D_2(v=0){\to}DF(v=1-2)+D_2(v=1)$ occur predominantly via V-V, R path and V-V, T path through whole temperatures, respectively.

Endoplasmic Reticulum Ca2+ Store: Regulation of Ca2+ Release and Reuptake by Intracellular and Extracellular Ca2+ in Pancreatic Acinar Cells

  • Kang, Yun Kyung;Park, Myoung Kyu
    • Molecules and Cells
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    • 제19권2호
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    • pp.268-278
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    • 2005
  • We investigated the effect of cytosolic and extracellular $Ca^{2+}$ on $Ca^{2+}$ signals in pancreatic acinar cells by measuring $Ca^{2+}$ concentration in the cytosol($[Ca^{2+}]_c$) and in the lumen of the ER($[Ca^{2+}]_{Lu}$). To control buffers and dye in the cytosol, a patch-clamp microelectrode was employed. Acetylcholine released $Ca^{2+}$ mainly from the basolateral ER-rich part of the cell. The rate of $Ca^{2+}$ release from the ER was highly sensitive to the buffering of $[Ca^{2+}]_c$ whereas ER $Ca^{2+}$ refilling was enhanced by supplying free $Ca^{2+}$ to the cytosol with $[Ca^{2+}]_c$ clamped at resting levels with a patch pipette containing 10 mM BAPTA and 2 mM $Ca^{2+}$. Elevation of extracellular $Ca^{2+}$ to 10 mM from 1 mM raised resting $[Ca^{2+}]_c$ slightly and often generated $[Ca^{2+}]_c$ oscillations in single or clustered cells. Although pancreatic acinar cells are reported to have extracellular $Ca^{2+}$-sensing receptors linked to phospholipase C that mobilize $Ca^{2+}$ from the ER, exposure of cells to 10 mM $Ca^{2+}$ did not decrease $[Ca^{2+}]_{Lu}$ but rather raised it. From these findings we conclude that 1) ER $Ca^{2+}$ release is strictly regulated by feedback inhibition of $[Ca^{2+}]_c$, 2) ER $Ca^{2+}$ refilling is determined by the rate of $Ca^{2+}$ influx and occurs mainly in the tiny subplasmalemmal spaces, 3) extracellular $Ca^{2+}$-induced $[Ca^{2+}]_c$ oscillations appear to be triggered not by activation of extracellular $Ca^{2+}$-sensing receptors but by the ER sensitised by elevated $[Ca^{2+}]_c$ and $[Ca^{2+}]_{Lu}$.

결정화 유리에 관한 연구 저 Li$_2$O 유리에 관하여 (Studies on the Crystallizing Glass on Low Li$_2$ O Glass)

  • 박용완;이종근;고영신;김정은
    • 한국세라믹학회지
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    • 제13권1호
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    • pp.30-34
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    • 1976
  • In general the chemical composition of glass ceramics in Li2O-Al2O3-SiO2 system is similar to the composition of $\beta$-spodumene (Li2O-Al2O3-4SiO2). With the object to manufacture the glass ceramics which can be produced in the domestic pot the composition of glass was so settled at 1.0 Li2O.0.9Al2O3.6.0SiO2 in order to reduce the contents of Li2O, to prevent the corrosion of the pot and to decrease the cost of raw materials. 0.2 mole and 0.1 mole of the mixture of TiO2 and ZrO2 as nucleants were added to the basic composition of 1.0 Li2O-0.9Al2O3-6.0SiO2. Each sample was divided into two kinds with a TiO2/ZrO2 ratio of 2 to 1 and the other with a TiO2/ZrO2 ratio fo 1 to 1. Thermal expansion coefficient, the most important property of glass ceramics, was tested. The softening point and the melting point of the samples were observed by the use of a heating microscope. The results obtained were as follows. The manufacturing of glass ceramics seems to be possible in the industrial plant using the domestic pot. 1) The composition of the glass which can be melted in the domestic pot process was near 1.0 Li2O.0.9Al2O3.6.0SiO2. 2) The temperature range of crystal creation and crystal growth was between 850-94$0^{\circ}C$, and 5 hours holding the samples at the temperature range was enough to crystallize them. The major crystal was $\beta$-spdumene and there existed petalite partialy. 3) The thermal expansion coefficient fo the crystallized glass was negative. 4) The deforming point of the crystallized glass was 1435$^{\circ}C$.

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P2P(peer-to-peer) 모델의 e비즈니스 적용 가능성에 관한 연구 (A Study on the Possibility of Applying Peer-to-Peer(P2P) Model to e-Business)

  • 문형남
    • 통상정보연구
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    • 제6권3호
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    • pp.63-79
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    • 2004
  • Peer-to-peer(P2P) has potential strength and possible factors which enable it to develop toward a new business by supplementing the limitation of the existing central server-type network. In various business sectors, various attempts have already been made to apply P2P. The computing power which was possible to be formed through peer-to-peer networking will become a powerful tools for expanding toward new business area depending the utilization methods by sector. Although the technology-centric P2P concept is a major interest now, its development direction will head toward service-centric P2P concept for satisfying desires of end-users(individual peers) for reliable contents and improving their purchasing convenience. But the solution requisites should be solved for the efficient application of P2P to business such as the establishment of and supplement for the industrial standard and legal system for P2P, the enhancement of peers recognition of P2P. Presently e-commerce using P2P is being implemented by a few enterprises locally by means of B2C. But if many enterprises enter this type of e-commerce, and the standardization of peer programmes, goods and documents is made, it will be able to be extended to B2B. Also it seems that it can be utilized in international transactions as well as in domestic transactions. So there is much room for the application of P2P to be extended further. Also there has been presented the necessity for related industries to support their work with theories and for academic studies to be made actively to set the direction of its development. It is hoped that multi-angled and in-depth studies on the utilization of P2P and various attempts to application of it to business be made on the basis of this study. For the development of P2P, continuous study and attempts are needed. The field of P2Ps application is very large and it will become a powerful tool, depending upon its application methods.

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Activation of JNK and c-Jun Is Involved in Glucose Oxidase-Mediated Cell Death of Human Lymphoma Cells

  • Son, Young-Ok;Jang, Yong-Suk;Shi, Xianglin;Lee, Jeong-Chae
    • Molecules and Cells
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    • 제28권6호
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    • pp.545-551
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    • 2009
  • Mitogen-activated protein kinases (MAPK) affect the activation of activator protein-1 (AP-1), which plays an important role in regulating a range of cellular processes. However, the roles of these signaling factors on hydrogen peroxide ($H_2O_2$)-induced cell death are unclear. This study examined the effects of $H_2O_2$ on the activation of MAPK and AP-1 by exposing the cells to $H_2O_2$ generated by either glucose oxidase or a bolus addition. Exposing BJAB or Jurkat cells to $H_2O_2$ affected the activities of MAPK differently according to the method of $H_2O_2$ exposure. $H_2O_2$ increased the AP-1-DNA binding activity in these cells, where continuously generated $H_2O_2$ led to an increase in mainly the c-Fos, FosB and c-Jun proteins. The c-Jun-$NH_2$-terminal kinase (JNK)-mediated activation of c-Jun was shown to be related to the $H_2O_2$-induced cell death. However, the suppression of $H_2O_2$-induced oxidative stress by either JNK inhibitor or c-Jun specific antisense transfection was temporary in the cells exposed to glucose oxidase but not to a bolus $H_2O_2$. This was associated with the disruption of death signaling according to the severe and prolonged depletion of reduced glutathione. Overall, these results suggest that $H_2O_2$ may decide differently the mode of cell death by affecting the intracellular redox state of thiol-containing antioxidants, and this depends more closely on the duration exposed to $H_2O_2$ than the concentration of this agent.

Ca$^{2+}$ 농도에 따른 생쥐 초기배의 발생 (The Development of Early Mouse Embryos Depend on $Ca^{2+}$ Concentration)

  • 양정숙;배인하
    • Clinical and Experimental Reproductive Medicine
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    • 제27권3호
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    • pp.275-282
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    • 2000
  • Objective: This study was to determine the effect of different concentration of calcium III medium on the preimplantational development of zygotes and early 2 cell embryos. Methods: Female mice of ICR strain ($5{\sim}8$ weeks old) were superovulated and mated with fertile males. Zygotes or early 2-cell embryos were collected by flushing the oviducts $31{\sim}32$ hours after hCG injection. The embryos were cultured in various concentrations of $Ca^{2+}$ in medium or with EDTA, EGTA and $Ni^{2+}$. Result and Conclusion: Treatment of high concentraion of $Ca^{2+}$ (3.42 mM $(2X){\sim}17.l$ mM (10X)) in medium didn't develop well compared to the control. Low concentrations of $Ca^{2+}$ (0.214mM $(1/8X){\sim}0.855$ mM (1/2X) were deterimental to development beyond 2-cell stage. EDTA, $Ca^{2+}$ chelating agent was treated with ranged concentrations of EDTA (0.014 $mM{\sim}0.107$ mM) to medium contaning 1.71 mM $Ca^{2+}$ showed beneficial effect to development to blastocyst compared to the control. EGTA, extracellular $Ca^{2+}$ chelator, was treated with ranged concentrations of EGTA ($0.014{\sim}0.107$ mM) to the medium contaning 1.71 mM $Ca^{2+}$. There is no significant difference with the control. $Ni^{2+}$ (50 ${\mu}M$), T-type $Ca^{2+}$-channel blocker was treated to medium contaning low concentration of $Ca^{2+}$. It overcame 2-cell block significantly. Rate of degenerated embryos decreased and developmental rate to morula and blastocyst increased more than low $Ca^{2+}$ concentration alone. Further studies are needed for the overcoming effect of 2-cell block by $Ni^{2+}$.

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P2P 개요 및 P2P-based Collaborative Machine의 Scheme (Scheme of P2P and P2P-based Collaborative Machine)

  • 김동훈;송준엽
    • 한국공작기계학회:학술대회논문집
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    • 한국공작기계학회 2005년도 춘계학술대회 논문집
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    • pp.477-482
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    • 2005
  • Recently, client computing trend has been changed from server oriented information application to network based P2P(Peer to Peer) services. The conventional client/server method has the merit of accessing abundant information, on the other side P2P has the merit of synchronized community support and information exchange. P2P has four meaning of point to point, peer to peer, person to person and path to profitability. In manufacturing system field, the second meaning is interested. P2P is classified to three type such as conventional client/server, hybrid P2P and pure P2P. The third is really peer to peer concept. The related technologies with P2P are P2P searching, XML, cooperation, IPv6, computing sharing and P2P communication. This paper describes the scheme of P2P and related contents. And through the P2P based technology, a P2P-based collaborative machine and a vertical portal machine are introduced in this paper. The scheme of the machines mentioned above is suggested for cooperation in manufacturing system and u-Manufacturing.

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생쥐배 분할구의 in vitro 발달에 관한 연구 (Studies on in vitro Development of Blastomeres Separated from Mouse Embryos)

  • 정덕수;이상진;정길생
    • 한국가축번식학회지
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    • 제12권3호
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    • pp.132-140
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    • 1988
  • These experiments were carried out to examine the development capacity of mouse blastomers separated from 2 to 8-cell stage mouse embryos. The female ICR and C3H mice were subjected to supervolution by intraperitoneal injection of PMSG and HCG and then mated with males of the same strain. Embryos were flushed from oviducts and uteri on a proper time after injection of HCG. After removal of zona pellucida with 0.5% pronase, each embryos were separated into 1/2, 1/4, 2/4, 1/8, 2/8 and 4/8 embryos by pipetting or a fine glass needle in Ca2+$.$Mg-2+ free Hoppe& Pitts medium containing 0.02% EDTA. Splitted embryos were cultured in Hoppe & Pitts medium for 48h to 72h. The embryos developed to blastocyst were transferred to recipients on 2 or 3 days of pseudopregnancy. On the other hand, a monozygotic pairs of 1/2 embryos developed to blastocyst after 48h in vitro culture were transferred to recipients on 2 days of pseudopregnancy or pregnancy. The results obtained were summarized as follows. 1. Success rates of separation of blastomeres from 2-, 4- and 8-cell embryos were 91.7%, 68.5-92.4% and 60.8-90.6%, respectively. 2. Development rates of various type of blastomeres to blastocyst after 72h in vitro culture were ranged 64.7-87.1%. 3. Blastocysts obtained after 48h in vitro culture were transferred to recipients on 2 or 3 days of pseudopregnancy. The production rates of live fetuses after transfer on 2 days, only 1/2, 2/4 and 4/8 embryos, were 13.2%, 13.5% and 17.2%, respectively and those of embryos transferred on 3 days were 11.8%, 9.6% and 11.5%, respectively. However, the production rates of live fetuses 1/2 embryos following 72h in vitro culture and transfer to recipients on 2 or 3 days of pseudopregnancy were 7.7% and 12.5%, respectively. 4. From 29 and 31 pairs of 1/2 embryos transferred to recipients on 2 days of pseudopregnancy or pregnancy, 4 sets of monozygotic twins were produced from only pregnant recipients.

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$SO_2$에 대한 참나무속 식물의 반응 (Responses of Quercus spp. to $SO_2$)

  • 이창석;배정오
    • 한국대기환경학회지
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    • 제7권3호
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    • pp.219-226
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    • 1991
  • Results from study on physiological responses to $SO_2$ of Quercus spp. selected for restoration of vegetation damaged by air pollution in the field study were as follows. Tolerance of Quercus aliena, Q. acutissima and Q. mongolica used in this study to $SO_2$ was higher in that order and tolerance of these plants to $SO_2$ was high comparatively among trees composing of major forest vegetation of Korea including natural forests and plantations. Stomatal resistance of these plants was increased after exposure to $SO_2$ and range of increase was different among species. From discrepancy between order of tolerance to $SO_2$ and range of increase in stomatal resistance, we were estimated that resistance mechanisms of Quercus spp. were different among species as mechanism originated in avoidance and resistance, respectively. Water potential of plant leaves reduced after exposure to $SO_2$, degree of reduction accorded with order of tolerance to $SO_2$. Reduction of water potential of plants after exposure to $SO_2$ was initiated before appearance of visible damage in plant leaves and water potential of plants exposed to $SO_2$ of low concentration, in which plants were not showed viaible damage was also reduced.

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