• KSBB Journal
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• v.16 no.4
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• pp.376-380
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• 2001

The human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was produced from cell suspension culture of transgenic tobacco which was transformed by using Agrobacterium harboring the hGM-CSF gene. To improve the production of hGM-CSF in batch culture system, the effects of initial sucrose concentration and inoculum size were investigated. The results show that the hGM-CSF production was not affected by small inoculum size in medium containing either low or high concentration of sucrose. However, the production of hGM-CSF was increased under increasing of the inoculum sizes and sucrose concentration. Under the combination of inoculum and sucrose concentration, the maximum hGM-CSF production of 720 $\mu$g/L was obtained at 90 g/L of initial sucrose concentration and 110 g/L of inoculum size.

• Korean Journal of Plant Tissue Culture
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• v.28 no.4
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• pp.179-184
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• 2001

We evaluated a possibility of the use of chloroplast number per guard cell pairs as a measure for ploidy level in the different ploidy levels of tobacco plant (Nicotiana tabacum L. cv. BY-4) . The guard-cell chloroplast numbers of leaves of haploid plant were a half of wild-type plant. Furthermore, the number of chloroplast per guard cell pairs of the leaves of doubled-haploid plant increased in two times compared with that of haploid plant. In addition, the chloroplast number was not changed in the F$_1$ progenies. The change of the chloroplast number by leaf stage was not observed. The results indicate that there is a strong relationship between ploidy level (2x and 4x) and chloroplast number per guard cell pairs. This relationship was also, observed in both in vitro and pot cultured plants. It was determined that the measurement of chloroplast number in guard cells of leaf epidermis is simple to use and less labour intensive, and hence can be considered a practical alternative to the chromosome counting methods or flow cytometry in the tobacco plant.

• Korean journal of applied entomology
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• v.56 no.1
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• pp.19-28
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• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.475-490
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• 2005

본 실험은 흡연이 치주인대세포의 기능에 미치는 영향을 알아보기 위해 담배 부산물 중 하나인 니코틴이 지주인대세포의 부착과 성장 및 세포의 가역성에 미치는 영향을 관찰하였다. 니코틴이 치주인대세포의 부착에 미치는 영향을 관찰하기 위하여 치주인대세포에 니코틴을 투여한 후 1, 3, 6, 12, 24시간째 trypan blue 염색 후 부착된 세포수의 측정과 H&E stain후 형태학적 관찰을 하였고 니코틴이 치주인대세포의 성장에 미치는 영향을 관찰하기 위하여 치주인대세포에 니코틴을 투여한 후 1, 4, 7, 11, 14일째 trypan blue 염색 후 증식된 세포수의 측정과 H&E stain후 형태학적 관찰을 하였다. 또 니코틴의 세포독성효과의 가역성 평가를 위하여 니코틴을 투여하고 1, 4, 7, 11일째 니코틴이 없는 새로운 배지로 갈아주어 2일 더 배양한 후 trypan blue로 염색하여 세포수를 측정, 비교하였고 H&E 염색 후 형태학적 관찰을 하였다. 실험결과 니코틴 농도가 증가함에 따라 치주인대세포의 부착율은 감소되었고 24시간 배양 후 2mg/ml, 0.5mg/ml, 0.1mg/ml니코틴 투여군에서 통계학적으로 유의성 있는 부착억제가 관찰되었고(P<0.01) 형태학적 관찰결과 2mg/ml, 0.5mg/ml 니코틴 투여군에서는 대조해서 관찰되는 세포질의 확장이 관찰되지 않았다. 니코틴이 치주인대세포의 증식에 미치는 효과를 살펴본 결과 2mg/ml, 0.5mg/ml, 0.1mg/ml 니코틴 투여군에서 증식억제가 관찰되었고(P<0.01) 0.005mg/ml이하의 니코틴 투여군에서는 아무런 변화도 관찰되지 않았다. 14일 배양 후 형태학적 관찰결과 0.1mg/ml 니코틴 투여군에서 세포질내 공포를 관찰할 수 있었고 2mg/ml, 0.5mg/ml 니코틴 투여군에서는 세포의 괴사가 나타났다. 니코틴의 세포독성효과의 가역성평가결과 2mg/ml이상의 니코틴은 세포에 비가역적인 손상을 일으키며 0.5mg/ml, 0.1mg/ml 니코틴에 의한 세포독성은 초기에는 가역적이나 장기간 세포에 노출시키면 비가역적인 손상을 야기하는 것으로 나타났다.(P<0.05) 본 실험결과 담배의 구성성분 중 니코틴은 농도 의존적으로 치주인대세포의 부착과 증식에 영향을 주었고 니코틴의 치주인대세포에 대한 독성효과는 농도가 증가할수록, 시간이 지닐수록 비가역적으로 나타났다. 따라서 흡연은 치주조직재생에 영향을 미칠 것으로 생각되며 흡연의 기간과 정도가 치주질환의 심도 및 치주치료 후 불량한 치유반응과 관계가 있을 것으로 사료된다.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.289-292
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• 2003

Effects of immobilization on the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) by Nicotiana tabacum cells were investigated using alginate and polyurethane foam as immobilization matrices. Encapsulation of the cells in alginate decreased protein production by 50% compared with that of suspension culture. Maximum hGM-CSF concentration was obtained by the cells immobilizaed in polyurethane foam. High hGM-CSF production could be possible when polyurethane foam was used because of high specific production and easy immobilization for cell recycling process with high cell density.

• Journal of Life Science
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• v.12 no.5
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• pp.577-583
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• 2002

The growth kinetics and the production of $\beta$-glucuronidase from transgenic tobacco's suspension culture was investigated in the flask culture and a 2.5 L bubble column reactor. The growth of bubble column reactor was similar to that of flask culture. However, in the bubble column reactor, the production of $\beta$ -glucuronidase reached 2850 U/mg (85-fold higher than that of flask culture). In both case, the production level of $\beta$ -glucuronidase was fluctuated, which was resulted from periodical degradation of the protein. Sucrose is important component in plant culture medium. Twice addition of sucrose in bubble column reactor could not improve cell growth, since other components in a medium were already depleted. However, the addition of sugar decreased cell size, which facilitated the operation of bioreactor. The production of $\beta$ -glucuronidase was continuously increased, however final concentration of $\beta$ -glucuronidase was similar to that without sucrose addition.

• Journal of Plant Biology
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• v.29 no.1
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• pp.67-75
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• 1986

An experimental system for the possibility of association between cultured plant cells and a cyanobacterium (N. muscorum) was investigated. The association was established in nitrogen-free medium, and nitrogen source was provided by N. muscorum through its own nitrogen fixation. N. muscorum was distributed on the outer part and intercellular spaces of the cultured plant cells. Polyamines were treated to the nitrogen-free medium to improve the association. Polyamines increased nitrogenase activity and protein amount in the association between N. muscorum and the cultured plant cells, as compared to the association without polyamines. Especially, spermine showed an increasing effect on the nitrogenase activity and the protein content in the association of tobacco and N. muscorum, while spermidine showed similar effect in case of soybean and N. muscorum.

• Tuberculosis and Respiratory Diseases
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• v.59 no.5
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• pp.530-535
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• 2005

Background and Aims : Cigarette smoking induces an inflammatory response in the airways, which may play a key role in the pathogenesis of chronic obstructive pulmonary disease. Interleukin-6 (IL-6) is one of the cytokines that plays an important role in inducing bronchial inflammation. The aim of this study was to determine if the level of the pro-inflammatory cytokine, Interleukin-6, is increased when the bronchial epithelial cells are exposed to a cigarette smoke extract (CSE) and an extract from stop smoking-aiding cigarettes, and examined the safety of these commercially available stop smoking-aiding cigarettes. Method : Bronchial epithelial cells were exposed to CSE from cigarette and stop smoking-aiding cigarettes for 24 hours. ELISA was used to measure the IL-6 levels in the supernatant from each condition. The IL-6 mRNA levels were measured by Taqman Real time RT-PCR. N-acetyl-L-cysteine(NAC) was added to each condition to determine if NAC can inhibit the release of IL-6 from the bronchial epithelial cells when they are exposed to CSE from cigarette and stop smoking-aiding cigarettes. Result : When bronchial epithelial cells were exposed to a CSE from cigarettes and stop smoking-aiding cigarettes, each type of CSE stimulated IL-6 production from the bronchial epithelial cells. The IL-6 mRNA level in the Bronchial epithelial cells was also elevated and NAC was found to inhibit the release of IL-6 from bronchial epithelial cells when they were exposed to the CSE from cigarettes and stop smoking-aiding cigarettes. Conclusion : Commercially available stop smoking-aiding cigarette can induce bronchial inflammation and can be harmful to smokers. Therefore, the safety of these cigarettes for smoking cessation should be evaluated.

• KSBB Journal
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• v.14 no.5
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• pp.534-538
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• 1999

For the optimization of operating conditions for plant cell suspension culture in bioreactors, effects of bioreactor types, various kinds of impellers, and aeration rates were examined using Nicotiana tabacum cells as a model system. Stirred tank bioreactor and airlift bioreactor were used for the comparison of bioreactor type. Growth rates in both bioreactors were lower than in shake flasks. In terms of final cell concentration, stirred tank bioreactor supported a little bit better growth compared to airlift bioreactor. Impeller type did not affect cell growth significantly, but it was apparent that cell size index decreased in the case of using hollowed paddle impeller. When the aeration rate was maintained at 0.3 vvm, cell growth was the best. At above 1.0 vvm, growth inhibition as well a browning was noticed. In addition, it was found that cell size index reduced proportionally to the increased of aeration rate.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.341-345
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• 2003

Effect of immobilization on the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) by Nicotiana tabacum cells was investigated using polyurethane foam as immobilization matrices. The cell activity and the hGM-CSF production were maintained for 16 days in spite of 3 times of media exchange. Under the same conditions of temperature and agitation rate, maximum concentrations of hGM-CSF in a 500-mL spinner flask and 100-mL Erleuneyer flasks were 17.3 ${\mu}g/L$ and 9.8 ${\mu}g/L$, respectively. Consequently high hGM-CSF production could be possible in spinner flask when the rate and amount of media exchange were optimized.