• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.3
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• pp.685-689
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• 2006

In order to examine the injury of vascular endothelial cells related with oxidative stress of reactive oxygen species(ROS), mophological changes of vascular endothelial cells were observed by light microscope after bovine pulmonary vascular endothelial cell line (BPVEC) was treated with 15 uM of hydrogen peroxide. In addition, the effect of vitamin E against ROS-induced oxidative stress was examined by light microscope. In this study, the cell number of BPVEC treated with ROS has significantly decreased than that of control, and the loss of cytoplasmic processes and cell swelling were observed in BPVEC treated with ROS. Whereas, cell number of BPVEC treated with vitamin E has significantly increased than that of BPVEC treated with ROS and also, cytoplasmic processes of BPVEC treated with vitamin E were preserved as control. These findings suggested that not only did ROS induce damage of BPVES by decrease of cell number, loss of cytoplasmic processes and cell swelling, but vitamin E also has protective effect against ROS-induced oxidative stress in cultures of BPVEC.

• Applied Microscopy
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• v.29 no.4
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• pp.427-435
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• 1999

Spermatogenesis and fine structure of the spermatozoon of the marbled sole, Limanda yokohamae were examined by means of the scanning and transmission electron microscopy. The process of spermatogenesis of the marbled tole is similar to that of other teleost with external fertilization. During the spermiogenesis, chromatin that has been became fine]y granular progressively condenses into many large globules and that homogeneously condensed in the spermatozoan head. A spermatozoon consists of head and tail, and the acrosome is absent. The cytoplasmic collar contained eight mitochondria is observed in the posterior part of the head. The well -developed axonemal lateral fins are observed in the tail. In the TEM observation, the cross section of the axial filament shows '9+2' axonemal structure of microtubules, and the numerous vesicles are observed in the cytoplasm.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.09a
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• pp.36-39
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• 2002

PKB activation may contribute to resistance to antiproliferative signals and breast cancer progression in part by impairing nuclear import and action of p27. PKB transfection caused cytoplasmic p27 accumulation and cytokine resistance. The nuclear localization region of p27 contains a PKB/Akt consensus site at threonine 157 and p27 phosphorylation by PKB impaired its nuclear import in vitro. PKB/Akt phosphorylated wild type p27 but not p27T157A. PKB activation led to cytoplasmic mislocalization of p27WT but p27T157A remained nuclear. In PKB activated cells, p27WT failed to cause Gl arrest, while the antiproliferative effect of p27T157A was not impaired. Cytoplasmic p27 was seen in 41% (52/128) of primary human breast cancers in association with PKB activation. Thus, we show a novel mechanism whereby PKB impairs p27 function that is associated with an aggressive phenotype in human breast cancer.

• Bulletin of the Korean Chemical Society
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• v.34 no.4
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• pp.1120-1126
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• 2013

The syndecan family consists of four transmembrane heparan sulfate proteoglycans present in most cell types and each syndecan shares a common structure containing a heparan sulfate modified extracellular domain, a single transmembrane domain and a C-terminal cytoplasmic domain. To get a better understanding of the mechanism and function of syndecan-4 which is one of the syndecan family, it is crucial to investigate its three-dimensional structure. Unfortunately, it is difficult to prepare the peptide because it is membrane-bound protein that transverses the lipid bilayer of the cell membrane. Here, we optimize the expression, purification, and characterization of transmembrane, cytoplasmic and short extracellular domains of syndecan4 (syndecan-4 eTC). Syndecan-4 eTC was successfully obtained with high purity and yield from the M9 medium. The structural information of syndecan-4 eTC was investigated by MALDI-TOF mass (MS) spectrometry, circular dichroism (CD) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. It was confirmed that syndecan-4 eTC had an ${\alpha}$-helical multimeric structure like transmembrane domain of syndecan-4 (syndecan-4 TM) in membrane environments.

• IMMUNE NETWORK
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• v.6 no.2
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• pp.67-75
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• 2006

Background: Cytotoxic function of killer cells is inhibited by specific recognition of class I MHC molecules on target cells by inhibitory killer Ig-like receptors (KIR) expressed on NK cells and some cytotoxic T cells. The inhibitory effect of KIR is accomplished by recruitment of SH2-containing protein tyrosine phosphatase (SHP) to the phosphotyrosine residues in the cytoplasmic tail. Methods: By in vitro coprecipitation experiments and transfection analysis, we investigated the association of KIR with an adaptor protein Shc in Jurkat T cells. Results: The cytoplasmic tail of KIR appeared to associate with an adaptor protein Shc in Jurkat T celilysates. Similar in vitro experiments showed that phosphorylated KIR cytoplasmic tail bound SHP-1 and Shc in Jurkat T cell lysates. The association of KIR with Shc was further confirmed by transfection analysis in 293T cells. Interestingly, however, Shc appeared to be replaced by SHP-2 upon engagement of KIR in 293T cells. Conclusion: Our data indicate that KIR associate with an adaptor protein Shc in Jurkat T cells, and suggest that KIR might have an additional role which is mediated by this adaptor protein.

• Korean Journal of Microbiology
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• v.9 no.2
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• pp.69-73
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• 1971

On June in 1970 the authors discovered a pathogenecity, cytoplasmic polyhedrosis virus, of the Smithia virus in the larvae of Liparis dispay L. appeared on quercus forest in Chung-Neung district and had carried out a experiment to detect the pathogenecity of Smithia virus through the inoculation of it into the larvase, such as Liparis dispay L. Hyphantrea cunea DRURY, and Dendrolinus spectabilis BUTLER. The results obtained were as follows ; 1) Death rate of L.dispay and D.spectabilis treated by 10$^{6}$ /ml cytoplasmic polyhedrosis virus of Smithis virus were 88.0% and 85.5% respectively, when the larvaes of these insects are big enough. But there were none of pathogenecity in case of Hyphantrea cunea DRURY. 20 Dead larvae caused by the injection of Smithia virus had begum to find out about on 10 days after inoculation. Miximum death rate of L. didpay and D. spectabilis appeared on 20-25days nad on 25-30days, respectively, after the incoulation. 3) In the cytoplasm of Mid-gut cylindrical cells of both of these insects, polyhedrosis, such s hexagonal (0.5-2.0-6.0 micron) were found out and in these insects, polyhedrosis, such as hexaginal (0.5-2.0-6.0 mivton) were found out and in case of D.spectabilis were a few polyhedrosis, such as tetragonal, trianglar polyhedrosis. 4) Diluted concentration of `0$^{6}$ /ml cytoplasmic polyhedrosis virus of Smithia virus were spread out in the field conditions. The corrected mortality was confirmed as about 87.8%.

• Proceedings of the Korean Magnetic Resonance Society Conference
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• 2002.08a
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• pp.90-90
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• 2002

Syndecans, transmembrane heparan sulfate proteoglycans, are coreceptors with integrin in cell adhesion process. It forms a ternary signaling complex with protein kinase C and phosphatidylinositol 4,5 bisphosphate (PIP2) for integrin signaling. NMR data indicates that cytoplasmic domain of syndecan-4 (4L) undergoes a conformational transition in the presence of PIP2, forming oligomeric conformation. The structure based on NMR data demonstrated that syndecan-4L itself forms a compact intertwined symmetric dimer with an unusual clamp shape for residues Leu$^{186}$ -Ala$^{195}$ . The molecular surface of the syndecan-4L dimer is highly positively charged. In addition, no inter-subunit NOEs in membrane proximal amino acid resides (Cl region) has been observed, demonstrating that the Cl region is mostly unstructured in syndecan-4L dimmer. However, the complex structure in the presence of PIP2 induced a high order multimeric conformation in solution. In addition, phosphorylation of cytoplasmic domain induces conformational change of syndecan-4, resulting inhibition of PKC signaling. The NMR structural data strongly suggest that PIP2 promotes oligomerization of syndecan-4 cytoplasmic domain for PKC activation and further induces structural reorganization of syndecan for mediating signaling network in cell adhesion procedure.

• Molecules and Cells
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• v.27 no.4
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• pp.391-396
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• 2009

Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• Journal of Mushroom
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• v.13 no.2
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• pp.129-134
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• 2015

Hybrid strains was selected by crossing between dikaryotic strain ASI 0628 (Dagul) bred from ASI 2596 (Suhan No.3) and ASI 2782(Black pileus mutant) and monokaryotic strain ASI 2344-84. The strain 84 that shown the best cultural characteristics was selected to be a new cytoplasmic hybrid variety and named as 'Chunhwashim'. The 'Chunhwashim' shown incompatibility among them forming the confrontation growth line against parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho). Fruiting body produced about $161.4{\pm}4.8g$ per bottle. And also the individual generation of fruit body is multiple than chunchu2ho as 40.9. The 'Chunhwashim' was cytoplasmic hybrid included hybrid DNA bands of parental strains ASI 0628 (Dagul) and ASI 2344 (chunchu 2ho) by Random Amplified Polymorphic DNA (RAPD) primer, and mitochondria DNA band of monokaryotic ASI 2344 (chunchu 2ho) using mitochondria microsatellite DNA marker. This new cytoplasmic hybrid variety 'Chunhwashim' of oyster mushroom is characterized by multiple of individual generation and deeply grey color of pileus.