• Journal of Plant Biotechnology
• /
• v.30 no.1
• /
• pp.97-102
• /
• 2003

Lettuce (Lactuca sativa) was transformed with Agrobacterium tumefacience LBA4404 containing human granulocyte macrophage colony stimulating factor (hGM-CSF) gene to produce in cell suspension cultures. Cell suspension culture was established using callus from transgenic lettuce plant. Integration of hGM-CSF gene into plant chromosome was confirmed through genomic PCR and Southern blot analysis. In addition, Northern blot analysis indicated the expression of the introduced hGM-CSF gene in transformed lettuce. The recombinant hGM-CSF was expressed in transgenic cell cultures derived from transgenic plants as a yield of about 149.0 $\mu\textrm{g}$/L in culture filtrate, which was determined by ELISA. These results demonstrated that transformed lettuce cell suspension cultures could be used as a production system of therapeutic proteins such as hGM-CSF.

• 한국전산유체공학회:학술대회논문집
• /
• 2000.05a
• /
• pp.21-26
• /
• 2000

Pressure-flow control characteristics of a commercially available cerebrospinal flow(CSF) control shunt valve was tested using fluid-solid interaction analysis. Pre-stress of the valve diaphragm(membrane) was computed for proper valve opening. The results were ir good agreements with the valve specification listed in the commercially available CSF control valve. The results of the study can be effectively used to design variety of CSF control shunt valves.

• Journal of computational fluids engineering
• /
• v.6 no.1
• /
• pp.40-46
• /
• 2001

Pressure-flow control characteristics of a commercially available cerebrospinal flow(CSF) control shunt valve was studied using flow/structure interaction analyses. Pre-stress of the valve diaphragm(membrane) was accounted for the simulation of an actual valve. The present results were in good agreement with the valve specification listed in the commercially available CSF control valve. The flow/structure interaction analysis of the present study can be effectively used to design a variety of CSF control shunt valves.

• Journal of Life Science
• /
• v.20 no.11
• /
• pp.1577-1581
• /
• 2010

Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1993.04a
• /
• pp.148-148
• /
• 1993

본 실험에서는 유전자 재조합으로 제조한 [$^{125}$ I]-labeled human GM-CSF을 사용하여 GM-CSF의 HL-60 (promyelocytic leukemic cell)의 표면에 존재하는 GM-CSF의 수용체의 특성을 알고 GM-CSF의 수용체에 어떤 parameter로 결합하는지를 밝히고 나아가 현재 사용되고 있는 유전자 재조합으로 제조된 Prokine(Sargramostim)과 Lucky에서 제조된 GM-CSF (LDB-005)의 수용체에 대한 결합율을 측정, 비교하고자 하였다. 본 실험의 결과를 보면 유전자 재조립으로 제조된 Human[$^{125}$ I] GM-CSF가 HL-60 cell에 대하여 선택적으로 결합하고 표면수용체에 saturable하게 결합함을 알 수 있었으며 scatchard analysis 결과한 종류의 GM-CSF의 K3값은 2.03$\times$$10^{9}$/M로($IC_{50}$/=~493pM) 세포당 결합부위의 수는 75개 정도로 J. DiPersio et al과 Linda S. Park et al.의 보고와 비슷한 결과를 얻었다.

• Proceedings of the Botanical Society of Korea Conference
• /
• 1985.08b
• /
• pp.23-33
• /
• 1985

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• Asia pacific journal of information systems
• /
• v.1 no.1
• /
• pp.31-47
• /
• 1991

The purpose of this study is to assess and develop effective MIS planning method by applying the CSF method. With the comparative analysis of the Rockart's basic concept of CSF and the examples of the corporations which employed CSF methodology, a new CSF method more suitable to Korean business environment is proposed and applied to another company for evaluation. The new CSF methodology consists of three steps. The first step is to preexamine the business conditions. The second step contains an Introductory Workshop, interview with the managers, survey questionnaire, Focusing Workshop, and decision of the priorities for system development. The third step is a real development of an information system out of the first and second steps. As a result of this study, it is found that revised CSF method can induce top management to participate more in the activities of information system development and that it can make top management to focus on the critical area. The fulfillment of the information requirement out of the enhanced MIS planning will lead to more successful management activities.

• Asia pacific journal of information systems
• /
• v.12 no.4
• /
• pp.37-53
• /
• 2002

The three main purposes of this paper are to (1) identify critical success factors(CSF's) for electronic commerce(EC), (2) investigate the explanatory power of these CSF's on firm performance, and (3) compare differences in evaluating CSF's and explaining impact of CSF's on performance between Korean and U.S.A. EC managers. Through a literature review and interviews with managers in EC firms, a list of 16 CSF's consisting of 111 items was compiled. In the second stage, questionnaires were administered to managers of EC companies in Seoul, Korea and Texas, U.S.A. Survey results show that CSF's have very significant explanatory power for firm performance in both Korean and U.S.A. While security, privacy, technical expertise, information about goods/services, and variety of goods/services are the most explanatory CSF's in Korea, evaluation of EC operations, technical expertise, and ease of use show most explanatory power in U.S.A. This analysis confirms the fact that customers use EC if they feel comfortable about navigating EC for information about a variety of goods/services without technical difficulty and in a secure and private way.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.3
• /
• pp.287-292
• /
• 2000

Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

• Korean Journal of Veterinary Research
• /
• v.43 no.4
• /
• pp.607-613
• /
• 2003

Dendritic cells (DCs) play an essential role in a variety of immune reactions involving $CD4^+$ T cells and have been used to enhance tumor-specific immune responses. Immunosuppression in patients with cancer includes the downregulation of function and number of DCs. Although DCs have been studied, the apoptosis of Des induced by anticancer drugs for chemotherapy remains largely uncharacterized. This study demonstrated that GM-CSF protects DCs from 5-fluorouracil (5-FU) or mitomycin C-induced apoptosis. After 6 - 10 days culture, DCs were characterized by specific surface marker, CD11c and MHC class II. MTT assay revealed that GM-CSF significantly enhanced the viability of DCs treated with 5-FU or mitomycin C. The percentage of dead cells of DCs was determined by cell size using FACScan and GM-CSF was clearly effective. However, GM-CSF did not increase the expression of MHC class II on viable DCs gated, suggesting that GM-CSF may differentially regulate critical factors involved in the function of DCs. For the quantitative analysis of apoptosis, annexin V-FITC staining was performed. 5-FU induced the apoptosis of DCs and GM-CSF significantly protects DCs from 5-FU-induced apoptosis. Taken together, the results in this study that GM-CSF has an anti-apoptosis effect on DCs may provide patients with cancer with clinical benefits to overcome the immunosuppression induced by the decrease of number and functional insufficiency of DCs.