• The Korean Journal of Zoology
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• v.27 no.3
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• pp.151-164
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• 1984

In order to investigate the mechanism of myoblast fusion during muscle differentiation in culture, the effect of muscle-conditioned medium on the fusion was studied and possible release from cultured myoblast cells of proteins which may be responsible for the promotion of myoblast fusion was analyzed. The muscle-conditioned medium showed a marked fusion-promoting activity in a dose-dependent fashion. THis fusion-promoting activity of the muscle-conditioned medium appeared to be due to the accumulation of at least two proteins which were released from the myoblast into the culture medium. These released proteins were analyzed by electrophoresis and autoradiography and found to have molecular weights of 45,000 and 65,000.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.4
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• pp.291-299
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• 2000

Morphine or butorphanol was continuously infused into cerebroventricle (i.c.v.) with the rate of $26\;nmol/{\mu}l/h$ for 3 days, and the withdrawal from opioid was rendered 7 hrs after the stopping of infusion. The expression of physical dependence produced by these opioids was evaluated by measuring the naloxone-precipitated withdrawal signs. The withdrawal signs produced in animals dependent on butorphanol (kappa opioid receptor agonist) were similar to those of morphine (mu opioid receptor agonist). Besides the behavioral modifications, opioid withdrawal affected G protein expression in the central nervous system. The G-protein ${\alpha}-subunit$ has been implicated in opioid tolerance and withdrawal. The effects of continuous infusion of morphine or butorphanol on the modulation of G protein ${\alpha}-subunit$ mRNA were investigated by using in situ hybridization study. In situ hybridization showed that the levels of $G\;{\alpha}s$ and $G\;{\alpha}i$ were changed during opioid withdrawal. Specifically, the level of $G\;{\alpha}s$ mRNA was decreased in the cortex and cerebellar granule layer during the morphine and butorphanol withdrawal. The level of $G\;{\alpha}i$ mRNA was decreased in the dentate gyrus and cerebellar granule layer during the morphine withdrawal. However, the level of $G\;{\alpha}i$ mRNA was significantly elevated during the butorphanol withdrawal. These results suggest that region-specific changes of G protein ${\alpha}-subunit$ mRNA were involved in the withdrawal from morphine and butorphanol.

• Proceedings of the Korea Information Processing Society Conference
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• 2007.05a
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• pp.679-682
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• 2007

살아있는(in-vivo)실험체에서 여러 차례 획득된 영상의 관심영역 특성을 측정, 분석하기 위한 영상처리기법의 정확성은 동물을 희생시켜(in-vitro) 촬영한 영상과의 정량적 비교분석을 통해 검증할 수 있다. 하지만 육안검사에 의존한 기존 분석방법은 객관성이 떨어지는 단점이 있다. 따라서 본 논문에서는 in-vivo영상인 PET 영상과, in-vitro영상인 Autoradiography 영상에서 관심영역 특성을 객관적, 정량적으로 비교하는 방법을 제안한다. 종양을 심은 누드마우스에 방사성 동위원소를 표지하여 획득한 이 두 영상에서 종양 조직 성장 지표가 되는 체적과 조직의 활성도를 나타내는 방사능섭취량(SUV)을 각각 측정하고 이를 비교하였다. 또한 두 영상획득의 시간차에 의해 방사성동위원소가 붕괴되어 영상 전체의 명암도가 감소하게 되므로 시간에 반비례하게 변하는 방사성동위원소의 양을 고려하여 영상명암도를 보정하였다.

• Journal of Plant Biology
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• v.38 no.2
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Biomolecules & Therapeutics
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• v.1 no.1
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• pp.58-64
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• 1993

DNA addicts induced by putative chemical related to carcinogenesis were detected and determined by $^{32}$P-Postlabelling assay after exposure of 4 compounds comprising two auto dyes (amaranth, new coccine) and two flavonoid compounds (rutin, quercetin) to ICR mouse. DNA was isolated from mouse liver and digested enzymatically to deoxyribonucleoside 3'-monophosphate. The postincubation of DNA digests with nuclease Pl before $^{32}$P-labelling enhanced the technique's sensitivity. Nuclease Pl cleaves deoxyribonucleoside 3'-mono-phosphates of normal nucleotides to deoxyrihonucleosides which do not serve as substrates for polynucleotide kinase, while most of addicts were found to be totally or partially resistant to the 3'-dephosphorylating action of nuclease Pl. The adducted deoxyribonucleoside 3'-monophosphate was converted to 5'-$^{32}$P-labelled deoxynucleoside 3',5'-bisphosphate by T4 polynucleotide kinase. The nucleotides were separated by anion-exchange thin layer chromatography(TLC) on polyethyleneimine cellulose by 4-dimensional or 2-dimensional TLC then detected by autoradiography. The results show that DNA addicts were detected in liver DNA of ICR mouse after administration of amaranth and quercetin by 2-dimensional and/or 4-dimensional TLC.

• BMB Reports
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• v.39 no.6
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• pp.717-721
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• 2006

In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

• Proceedings of the KWS Conference
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• 2009.11a
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• pp.60-60
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• 2009

본 연구는 보론이 첨가된 저합금강 용접 열영향부에서의 보론 편석 거동 연구를 위해 보론이 10ppm 첨가된 저합금강을 이용하여 다양한 용접 입열량 및 외부 응력에 따른 용접부 CGHAZ의 보론 편석거동을 분석하였다. 이를 위해 Gleeble 시스템을 이용하여 다양한 입열량에 따른 CGHAZ를 열 및 열-응력 사이클을 통하여 재현하였다. 재현된 시편의 미세조직은 OM을 통하여 분석하고, 보론의 편석거동을 SIMS와 PTA 분석법을 통하여 분석하였다. 그 결과 입열량에 따른 보론의 편석 거동은 최초 입열량이 증가함에 따라 보론의 편석이 증가하다가 다시 감소하였는데 이는 비평형 편석 후 고온에서 유지시간이 길어짐에 따라 back diffusion 발생에 따른 영향으로 판단된다. 또한 외부 응력에 의한 보론 편석 거동 분석 결과, 용접 열 사이클 중 작용하는 외부 응력에 의해 결정립계 편석 감소하였는데 이는 외부 응력에 의한 오스테나이트 결정립 크기 감소에 따른 결정립계 증가의 영향으로 판단된다.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.135-135
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• 1993

신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2＋}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

• The Korean Journal of Nuclear Medicine
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• v.30 no.4
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• pp.532-540
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• 1996

목적 : 종양세포의 증식을 평가하기 위해 radiofluorinated ethyluracil (FEU)과 deoxyadenosine analogue(FAD)를 합성하여 종양의 영상화를 시도하였다. 대상 및 방법 : 5-(2-Fluoroethyl)uracil ([$^{18}F$]FEU)은 2, 4-dimethoxy-5-(2-hydroxyethyl) pyrimidine을 $K^{18}F$와 처리한 후 HBr로 가수분해하여 얻었으며 Fluorodeoxyadenosine은 adenosine의 triacetylated analogue를 $K^{18}F$와 처리하여 얻었다. 생물학적 조직분포는 유방암 세포(13762 NF, 100,000 cells per rat, im)를 쥐에 접종한 후 0.5, 1, 2 및 4시간에 주요장기를 적출하여 %ID/g을 측정하고 자가방사영상은 방사성의약품 투여 45분 후에 얻었다. PET 영상은 VX-2 종양을 접종한 가토를 이용하여 얻었다. In vitro cell proliferation assay는 사람의 말초단핵구를 이용하였다. 결 과 : In vitro assay상 ([$^{18}F$]FEU는 세포증식시 DNA/RNA에 결합함을 시사하였다. ([$^{18}F$]FAD와 ([$^{18}F$]FEU의 종양/비종양 방사능 섭취비는 시간경과에 따라 증가하였으며 ([$^{18}F$]FAD와 ([$^{18}F$]FEU를 이용한 자가방사영상과 ([$^{18}F$]FEU를 이용한 PET 영상에서 종양을 잘 관찰할 수 있었다. 결 론 : ([$^{18}F$]FAD 및 ([$^{18}F$]FEU를 이용하여 종양세포의 증식을 PET 영상에서 평가할 수 있으리라 사료된다.

• Journal of Microbiology
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• v.37 no.1
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• pp.21-26
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• 1999

Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and 45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).