• Journal of Korean Neurosurgical Society
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• v.66 no.6
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• pp.642-651
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• 2023

Objective : Endothelial colony-forming cells (ECFCs) have been reported to play an important role in the pathogenesis of moyamoya disease (MMD). We have previously observed stagnant growth in MMD ECFCs with functional impairment of tubule formation. We aimed to verify the key regulators and related signaling pathways involved in the functional defects of MMD ECFCs. Methods : ECFCs were cultured from peripheral blood mononuclear cells of healthy volunteers (normal) and MMD patients. Low-density lipoproteins uptake, flow cytometry, high content screening, senescence-associated β-galactosidase, immunofluorescence, cell cycle, tubule formation, microarray, real-time quantitative polymerase chain reaction, small interfering RNA transfection, and western blot analyses were performed. Results : The acquisition of cells that can be cultured for a long time with the characteristics of late ECFCs was significantly lower in the MMD patients than the normal. Importantly, the MMD ECFCs showed decreased cellular proliferation with G1 cell cycle arrest and cellular senescence compared to the normal ECFCs. A pathway enrichment analysis demonstrated that the cell cycle pathway was the major enriched pathway, which is consistent with the results of the functional analysis of ECFCs. Among the genes associated with the cell cycle, cyclin-dependent kinase inhibitor 2A (CDKN2A) showed the highest expression in MMD ECFCs. Knockdown of CDKN2A in MMD ECFCs enhanced proliferation by reducing G1 cell cycle arrest and inhibiting senescence through the regulation of CDK4 and phospho retinoblastoma protein. Conclusion : Our study suggests that CDKN2A plays an important role in the growth retardation of MMD ECFCs by inducing cell cycle arrest and senescence.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.76-85
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• 2004

Biodevices composed of biomolecular layer have been developed in various fields such as medical diagnosis, pharmaceutical screening, electronic device, photonic device, environmental pollution detection device, and etc. The biomolecules such as protein, DNA and pigment, and cells have been used to construct the biodevices such as biomolecular diode, biostorage device, bioelectroluminescence device, protein chip, DNA chip, and cell chip. Substantial interest has focused upon thin film fabrication or the formation of biomaterials mono- or multi-layers on the solid surfaces to construct the biodevices. Based on the development of nanotechnology, nanoscale fabrication technology for biofilm has been emerged and applied to biodevices due to the various advantages such as high density immobilization and orientation control of immoblized biomolecules. This review described the nanoscale fabrication of biomolecular film and its application to bioelectronic devices and biochips.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.15 no.1
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• pp.1-36
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• 1989

The protective effects of 24 flavonoids against singlet oxygen - induced photohemolysis of rabbit erythrocytes were investigated and compared with their antioxidant activities. 1. All the flavonoid aglycons investigated besides flavone and 7,8-benzoflavone showed tad dose-dependant Protective effects. Especially galagin showed the most Pronounced effect under the experimental condition. 2. The order of the radical-scavenging activities of flavonoids was as follow (-)-EGCG > flavonols > flavonols > flavones > flavanonols. The radical-scavenging activities of flavonoid glycosides were not different from those of their aglycons. 3. When added after irradiation, flavonoids did not alter the photohemolysis pattern of control. In this study, several antioxidant were found and it was newly found that Photohemolysis experiment is a convenient, reproductive diagnostic method in the screening and researching the compound which has antioxidant activity or protective effect on cell membrane against active oxygen species.

• BMB Reports
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• v.49 no.6
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• pp.319-324
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• 2016

RNA polymerase II C-terminal domain phosphatases are newly emerging family of phosphatases that contain FCPH domain with Mg⁺²-binding DXDX(T/V) signature motif. Its subfamily includes small CTD phosphatases (SCPs). Recently, we identified several interacting partners of human SCP1 with appearance of dephosphorylation and O-GlcNAcylation. In this study, using an established cell line with inducible CTDSPL2 protein (a member of the new phosphatase family), proteomic screening was conducted to identify binding partners of CTDSPL2 in nuclear extract through immunoprecipitation of CTDSPL2 with its associated. This approach led to the identification of several interacting partners of CTDSPL2. This will provide a better understanding on CTDSPL2.

• BMB Reports
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• v.48 no.5
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• pp.256-265
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• 2015

Cardiovascular and neurodegenerative diseases are major health threats in many developed countries. Recently, target tissues derived from human embryonic stem (hES) cells and induced pluripotent stem cells (iPSCs), such as cardiomyocytes (CMs) or neurons, have been actively mobilized for drug screening. Knowledge of drug toxicity and efficacy obtained using stem cell-derived tissues could parallel that obtained from human trials. Furthermore, iPSC disease models could be advantageous in the development of personalized medicine in various parts of disease sectors. To obtain the maximum benefit from iPSCs in disease modeling, researchers are now focusing on aging, maturation, and metabolism to recapitulate the pathological features seen in patients. Compared to pediatric disease modeling, adult-onset disease modeling with iPSCs requires proper maturation for full manifestation of pathological features. Herein, the success of iPSC technology, focusing on patient-specific drug treatment, maturation-based disease modeling, and alternative approaches to compensate for the current limitations of patient iPSC modeling, will be further discussed. [BMB Reports 2015; 48(5): 256-265]

• Animal cells and systems
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• v.13 no.3
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• pp.283-288
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• 2009

The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.

• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.321-326
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• 1996

In the course of screening for new bioactive compounds from oligotrophs in soil, a microorganism, designated as SA-246 and now identified as Micromonospora sp., has been shown to produce a strong antibacterial compound. The active compound was purified from broth filtrate by ethylacetate extraction, silica gel column chromatography, preparative TLC and HPLC, and was identified as crisamicin A based on mass and NMR spectral data. The compound SA- 246 exhibited not only strong antibacterial activity against Gram-positive bacteria but also cytotoxicity against cancer cell lines such as A549 (lung), SK-OV-3 (ovarian), SK-MEL-2 (melanoma), XF498 (central nervous system) and HCT15 (colon).

• Journal of environmental and Sanitary engineering
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• v.15 no.1
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• pp.34-38
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• 2000

In the screening of phenol-degrading bacteria, a strain showing good growth in media containing phenol was isolated by using enrichment culture from various sample and identified as genus Klebsiella pneumoniae. The optimal temperature and pH for cell growth of Klebsiella pneumoniae was $35^{\circ}C$ and 8.0, respectively. When phenol was added to the minimal media as a sole source of carbon and energy, the concentration of maximum and optimum for cell growth was 1,200ppm and 1,000ppm, respectively. It was observed that Klebsiella pneumoniae was able to degrade 98% of phenol (1,000ppm) after 40hr in culture. The isolated could utilize various kinds of aromatic compounds and showed good growth in presence of phenol, m-cresol and 3-methyl catechol.

• Journal of Microbiology
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• v.40 no.4
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• pp.267-273
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• 2002

Urinary tract infection (UTI) is one of the most common domiciliary and nosocomial bacterial infections prevalent in both males and females. UTI is diagnosed on the basis of clinical symptoms, microscopy and culture of urine. In order to evaluate the efficacy of microscopic detection for presumptive diagnosis of UTI we analyzed urine samples of Nepalese patients. We have conducted Gram staining and counting of pus cells, red blood cells (RBC) and epithelial cells. We observed that RBC and epithelial cell counts were not sensitive enough to be used for presumptive diagnosis of UTI. However, pus cell counts as well as Gram stain are sensitive and significant enough to presume UTI. When the Gram stain result was compared with the culture result, it was statistically significant. From this, we suggest that Gram stain of centrifuged urine is a very sensitive screening method to detect bacteriuria. In addition, we found that E. coli was the most predominant microorganism causing UTI and nitrofurantoin was the most effective antibiotic against the isolated urinary pathogens.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.41-49
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• 1994

The most common conventional method of discovering a drug involves a massive screening of a large number of compounds in chemical libraries or in the extracts from natural sources such as plants or microbial broths followed by chemical modification of one or more active compounds to improve their properties as a drug. When the three-dimensional structure of the target molecule for which the drug is searched is known the drug discovery process can be significantly simplified, This is especially true when the three-dimensional structure of a complex between the target and a lead compound is known. In this lecture our experience on the structure-based drug design for human CDK2(cyclin-dependent protein kinase 2) will be discussed with special emphasis on the strength and weakness of this approach of drug discovery. The regulation of the activity of CDK2 plays an important role in the cell proliferation of normal and cancer cells.