Proceedings of the Korean Society of Plant Biotechnology Conference
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2002.04b
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pp.49-58
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2002
Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21st century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.
Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21st century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (lpomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.
Lee, Ji Hyung;Amarsanaa, Khulan;Wu, Jinji;Jeon, Sang-Chan;Cui, Yanji;Jung, Sung-Cherl;Park, Deok-Bae;Kim, Se-Jae;Han, Sang-Heon;Kim, Hyun-Wook;Rhyu, Im Joo;Eun, Su-Yong
The Korean Journal of Physiology and Pharmacology
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v.22
no.3
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pp.311-319
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2018
Mitochondrial calcium overload is a crucial event in determining the fate of neuronal cell survival and death, implicated in pathogenesis of neurodegenerative diseases. One of the driving forces of calcium influx into mitochondria is mitochondria membrane potential (${\Delta}{\psi}_m$). Therefore, pharmacological manipulation of ${\Delta}{\psi}_m$ can be a promising strategy to prevent neuronal cell death against brain insults. Based on these issues, we investigated here whether nobiletin, a Citrus polymethoxylated flavone, prevents neurotoxic neuronal calcium overload and cell death via regulating basal ${\Delta}{\psi}_m$ against neuronal insult in primary cortical neurons and pure brain mitochondria isolated from rat cortices. Results demonstrated that nobiletin treatment significantly increased cell viability against glutamate toxicity ($100{\mu}M$, 20 min) in primary cortical neurons. Real-time imaging-based fluorometry data reveal that nobiletin evokes partial mitochondrial depolarization in these neurons. Nobiletin markedly attenuated mitochondrial calcium overload and reactive oxygen species (ROS) generation in glutamate ($100{\mu}M$)-stimulated cortical neurons and isolated pure mitochondria exposed to high concentration of $Ca^{2+}$ ($5{\mu}M$). Nobiletin-induced partial mitochondrial depolarization in intact neurons was confirmed in isolated brain mitochondria using a fluorescence microplate reader. Nobiletin effects on basal ${\Delta}{\psi}_m$ were completely abolished in $K^+-free$ medium on pure isolated mitochondria. Taken together, results demonstrate that $K^+$ influx into mitochondria is critically involved in partial mitochondrial depolarization-related neuroprotective effect of nobiletin. Nobiletin-induced mitochondrial $K^+$ influx is probably mediated, at least in part, by activation of mitochondrial $K^+$ channels. However, further detailed studies should be conducted to determine exact molecular targets of nobiletin in mitochondria.
Transactions of the Korean Society of Mechanical Engineers A
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v.36
no.5
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pp.475-479
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2012
We present a novel polymer fabrication process involving direct UV patterning of a hyperbranched polymer, AEO3000. Compared to PDMS, which is the most widely used polymer in bioMEMS devices, the present polymer has advantages with regard to electrode integration and fast fabrication. We designed a four-chip microelectrofluidic bench having three electrical pads and two fluidic I/O ports. We integrated a microfluidic mixer and a cell separator on the bench to characterize the interconnection performance and sample manipulation. Electrical and fluidic characterization of the microfluidic bench was performed. The measured electrical contact resistance was $0.75{\pm}0.44{\Omega}$, which is small enough for electrical applications, and the pressure drop was 8.3 kPa, which was 39.3% of the value in the tubing method. By performing yeast mixing and a separation test in the integrated module on the bench, we successfully showed that the interconnected chips could be used for bio-sample manipulation.
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.9
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pp.1270-1278
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2015
In vitro anticancer effects of black soybean doenjang on HT-29 human colon cancer cells were studied. SD (soybean doenjang prepared with nine-time baked bamboo salt) and BD (black soybean doenjang prepared with nine-time baked bamboo salt) were compared with CD (commercial doenjang). There were no significant differences between experimental groups in terms of pH, amino-type nitrogen, and ammonia-type nitrogen levels of the doenjang samples. BD showed the highest antioxidative effect, followed by SD and CD in that order. BD also showed the highest total polyphenol concentration of all samples. CD, SD, and BD extracts showed no toxic effects on normal RAW 264.7 cells at a concentration ranging from 0.1 to 0.5 mg/mL. BD exhibited anticancer effect on HT-29 cells by MTT assay. Also, BD manipulated mRNA expressions in certain factors; it suppressed pro-inflammatory cytokines such as $TNF-{\alpha}$, IL-6, and COX-2, promoted cell-cycle-related genes of p21, and p53, suppressed expression of cyclin D1, and suppressed anti-apoptotic Bcl-2; such manipulation by BD was the strongest, followed by SD and CD in order. From the results above, BD exhibited the highest anticancer effects by inhibiting growth of HT-29 cells, probably by regulating pro-inflammatory cytokines, cell cycling related genes, etc. These results might be due to using black soybeans containing high levels of polyphenol, including anthocyanins.
From the concepts of cellular pathology and of exfoliative cytology, as elucidated by Virchow and Papanicolaou respectively in the late 19th and early 20th century, have evolved the primary methods for the diagnosis of cancer today. From Papanicolaou's concept of exfoliative cytology developed fine needle aspiration biopsy in the early 1960's, this has become a major diagnostic procedure and has contributed to a significant reduction in open biopsies and, therefore, to medical cost-effectiveness immunobiochemical techniques provided us with a supplement to cancer diagnosis in the 1980's. The immunoperoxidase method, using monoclonal antibodies, is applied primarily as an ancillary measure to elucidate the nature of cancers The availability of specific monoclonal antibodies has greatly facilitated the identification of cell products or surface markers. For example, antibodies directed against intermediate filaments have proved to be of value in determining the histogenesis oi poorly differentiated neoplasms. Tumor markers may serve as biochemical indicators of the presence of a neoplasm. They can be detected In plasma and other body fluids. Their concentration can be applied as a diagnostic test, for monitoring the clinical course of known cancer, and as a screening measure to detect certain cancers in a population at risk. Flow cytometry is a useful tool for distinguishing several cell characteristics, such as the immunophenotype of leukemia-lymphoma cells, the DNA content of neoplastic cells, and cell proliferation rate. Molecular biologic techniques provided a giant step for the management of cancer patients encompassing diagnosis, prognostic evaluation, and therapy. Nucleic acid hybridization techniques are utilized as Southern, Northern, and dot blots and in situ hybridization. Molecular biology and its techniques may bring a blight new horizon for understanding cancer biology and in designing therapy on the basis of gene manipulation.
Takahashi, J.;Mwenya, B.;Santoso, B.;Sar, C.;Umetsu, K.;Kishimoto, T.;Nishizaki, K.;Kimura, K.;Hamamoto, O.
Asian-Australasian Journal of Animal Sciences
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v.18
no.8
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pp.1199-1208
/
2005
Abatement of greenhouse gas emitted from ruminants and promotion of biogas energy from animal effluent were comprehensively examined in each anaerobic fermentation reactor and animal experiments. Moreover, the energy conversion efficiency of biomass energy to power generation were evaluated with a gas engine generator or proton exchange membrane fuel cell (PEMFC). To mitigate safely rumen methanogenesis with nutritional manipulation the suppressing effects of some strains of lactic acid bacteria and yeast, bacteriocin, $\beta$1-4 galactooligosaccharide, plant extracts (Yucca schidigera and Quillaja saponarea), L-cysteine and/or nitrate on rumen methane emission were compared with antibiotics. For in vitro trials, cumulative methane production was evaluated using the continuous fermented gas qualification system inoculated with the strained rumen fluid from rumen fistulated Holstein cows. For in vivo, four sequential ventilated head cages equipped with a fully automated gas analyzing system were used to examine the manipulating effects of $\beta$1-4 galactooligosaccharide, lactic acid bacteria (Leuconostoc mesenteroides subsp. mesenteroides), yeast (Trichosporon serticeum), nisin and Yucca schidigera and/or nitrate on rumen methanogenesis. Furthermore, biogas energy recycled from animal effluent was evaluated with anaerobic bioreactors. Utilization of recycled energy as fuel for a co-generator and fuel cell was tested in the thermophilic biogas plant system. From the results of in vitro and in vivo trials, nitrate was shown to be a strong methane suppressor, although nitrate per se is hazardous. L-cysteine could remove this risk. $\beta$1-4 galactooligosaccharide, Candida kefyr, nisin, Yucca schidigera and Quillaja saponarea are thought to possibly control methanogenesis in the rumen. It is possible to simulate the available energy recycled through animal effluent from feed energy resources by making total energy balance sheets of the process from feed energy to recycled energy.
GM-CSF is a multipotent growth factor, which also plays an important role during the process of wound healing. rrhGM-CSF was specifically produced from rice cell culture in our laboratory (Hanson Biotech Co., Ltd, Daejeon). The rrhGMCSF contains more oligosaccharide side chains than any other types of GM-CSF. This work was taken to evaluate the influence on wound healing of rrhGM-CSF in male golden hamsters. Full thickness skin defects of 9 mm in diameter were made in the back of hamsters, and 100 ${\mu}L$ ointment containing rrhGM-CSF 50 ${\mu}g/mL$ was applied. Control groups were given ointment without rrhGM-CSF. The wound sizes were relatively reduced and skin was well regenerated in the experimental group compared with the control group. Structurally, reepithelialization and architecture of the skin following injury were well accomplished in the experimental group. And also, positive reaction of PCNA of the skin following injury was more prominent in rrhGM-CSF containing ointment treatment group. Since this type of GM-CSF has highly glycosylated side chains, the effectiveness might be retain longer and stable, regarding acceleration of wound healing in the animal model. The present study has important implications for further development of the therapeutic manipulation of wound healing using rrhGM-CSF.
Animation movie is a non-photorealistic animated art that consists of formative language forming a frame based on a story and cuts describing frames that form the cuts. Therefore, in expressing an image, artistic expression methods and devices for a formative space are should be provided in a frame while cuts have the images between frames faithfully. Short animation movie is produced by various image experiments with unique image expressions rather than narration for expressing subjective discourse of a writer. Therefore, image style that forms unique images and various image directions are important factors. This study compared the experimental image directions of and , both of which showed a production method of film manipulation. First, while uses pixilation that produces images obtained from live images through painting and many optical disclosure process on a cell mat, was made with diverse collage techniques such as tearing, cutting, pasting, and folding hundreds of scenes from action movies. Second, expresses non-causal relationship of characters by their repetitive behaviors and circulatory image structure through a fixed camera angle, resisting typical scene transition. On the other hand, has an advancing structure that progresses antagonistic relationship of characters through diverse camera angles and scene transition of unique images. Third, in terms of editing, uses a long-take short cut technique in which the whole image consists of one short cut, though it seems to be many scenes with the appearance of various characters. On the other hand, maximizes visual fun and commitment by image reconstruction with hundreds of various short cuts. That is, both works have common features of an experimental work that shows expansion of animated image expressions through film manipulation that is different form general animation productions. On top of that, delivers routine life of diverse human beings without clear narration through image of conceptualized spaces. expresses it in a new image space through image reconstruction with collage technique and speedy progress, setting a binary opposition structure.
Kim, Jong-Myung;Yu, Ji-Min;Bae, Yong-Chan;Jung, Jin-Sup
Journal of Life Science
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v.21
no.5
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pp.631-646
/
2011
Mesenchymal stem cells (MSC) are multipotent and can be isolated from diverse human tissues including bone marrow, fat, placenta, dental pulp, synovium, tonsil, and the thymus. They function as regulators of tissue homeostasis. Because of their various advantages such as plasticity, easy isolation and manipulation, chemotaxis to cancer, and immune regulatory function, MSCs have been considered to be a potent cell source for regenerative medicine, cancer treatment and other cell based therapy such as GVHD. However, relating to its supportive feature for surrounding cell and tissue, it has been frequently reported that MSCs accelerate tumor growth by modulating cancer microenvironment through promoting angiogenesis, secreting growth factors, and suppressing anti-tumorigenic immune reaction. Thus, clinical application of MSCs has been limited. To understand the underlying mechanism which modulates MSCs to function as tumor supportive cells, we co-cultured human adipose tissue derived mesenchymal stem cells (ASC) with cancer cell lines H460 and U87MG. Then, expression data of ASCs co-cultured with cancer cells and cultured alone were obtained via microarray. Comparative expression analysis was carried out using DAVID (Database for Annotation, Visualization and Integrated Discovery) and PANTHER (Protein ANalysis THrough Evolutionary Relationships) in divers aspects including biological process, molecular function, cellular component, protein class, disease, tissue expression, and signal pathway. We found that cancer cells alter the expression profile of MSCs to cancer associated fibroblast like cells by modulating its energy metabolism, stemness, cell structure components, and paracrine effect in a variety of levels. These findings will improve the clinical efficacy and safety of MSCs based cell therapy.
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