• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.26 no.1
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• pp.1-5
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• 2017

This study investigated the mechanical tearing of a cell membrane using a nanostructured alumina filter for easy and quick mechanical cell disruption. Nanostructured alumina filters were prepared by a multi-step aluminum anodizing process and nanopore etching process. Six different types of nanostructures were formed on the surface of the nanoporous alumina filters to compare the mechanical cell disruption characteristics according to the shape of the nanostructure. The prepared alumina filter was assembled in a commercial filter holder, and then, NIH3T3 fibroblast cells in a buffer solution were passed through the nanostructured alumina filter at a constant pressure. By measuring the concentration of proteins and DNA, the characteristics of mechanical cell disruption of the nanostructured alumina filter were investigated.

• BMB Reports
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• v.33 no.2
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• pp.103-111
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• 2000

Phosphorylation of the eukaryotic elongation factor 2 (eEF-2) blocks the elongation step of translation and stops overall protein synthesis. Although the overall rate of protein synthesis in mitosis reduces to 20% of that in S phase, it is unclear how the protein translation procedure is regulated during the cell cycle, especially in the stage of peptide elongation. To delineate the regulation of the elongation step through eEF-2 function, the changes in phosphorylation of eEF-2, and in activity of corresponding $Ca^{2+}$/calmodulin (CaM)-dependent protein kinase III (CaMK-III) during the cell cycle of NIH 3T3 cells, were determined. The in vivo level of phosphorylated eEF-2 showed an 80% and 40% increase in the cells arrested at G1 and M, respectively. The activity of CaMK-III also changed in a similar pattern, more than a 2-fold increase when arrested at G1 and M. The activity change of the kinase during one turn of the cell cycle also demonstrated the activation at G1 and M phases. The activity change of cAMP-dependent protein kinase (PKA) was reciprocal to that of CaMK-III. These results indicated: (1) the activity of CaMK-III was cell cycle-dependent and (2) the level of eEF-2 phosphorylation followed the kinase activity change. Therefore, the elongation step of protein synthesis might be cell cycle dependently regulated.

• BMB Reports
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• v.28 no.2
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• pp.87-93
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• 1995

To investigate the biological functions of the EGF-like domain of mouse betacellulin (BTC), mouse BTC(33-80), a 48-residue peptide corresponding to the EGF-like domain, was synthesized by stepwise solidphase methods using a 9-fluorenylmethoxycarbonyl (Fmoc) strategy. The homogeneity of synthetic mouse BTC(33-80) was confirmed by analytical reversed phase (RP)-HPLC, amimo acid analysis, and fast atom bombardment mass spectrometer (FAB-MS). Three disulfide bond pairings of synthetic mouse BTC(33-80) were established by amino acid analysis of cysteine-containing fragments derived from thermolytic digestion. These were consistent with the pairings of EGF and transforming growth factor ($TGF-{\alpha}$). The EGF-Iike domain of mouse BTC showed equipotent activity in both EGF-receptor binding on A-431 epidermoid carcinoma cells, and mitogenesis on NIH-3T3 fibroblast cells, as compared with authentic h-EGF. Results suggest that the EGF-Iike domain of BTC plays a significant role in mitogenic activity with an EGF-receptor mediated system.

• Journal of Pharmaceutical Investigation
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• v.39 no.1
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• pp.1-6
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• 2009

To overcome the stability problem of hydrophilic quantum dot (Q-dot), cellular uptake of hydrophobic instead of hydrophilic Q-dot was studied in the hope to find a simple method to use Q-dot as a cellular imaging probe. Hydrophobic Q-dot and poly-L-lactic acid (PLLA) were co-dissolved in chloroform to prepare stable films. Due to the cellular compatibility of PLLA, adherent cells were cultured on the film to observe the degree of Q-dot uptake and cytotoxicity of the prepared films. The results show that Q-dots were absorbed into NIH3T3 and EMT6 cells. Cellular uptake was also observed when hydrophobic Q-dots were coated directly on a glass plate. PLLA/Q-dot film and Q-dot coated on glass plate did not show major cytotoxicity. In vivo tumor model was also used to show the uptake of Q-dot from the PLLA/Q-dot film to the tumor site.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.96-96
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• 2003

Previously, wogonin (5,7-dihydroxy-8-methoxyflavone) was found to suppress proinflammatory enzyme expression including cyclooxygenase-2 (COX-2), contributing to in vivo anti-inflammatory activity against skin inflammation. However, the detailed effect on each skin cell type has not been understood. Therefore, present investigation was carried out to find the effect of wogonin on inflammation-associated gene expression from skin fibroblasts in culture using reverse transcriptase-polymerase chain reaction. As a result, it was found that wogonin (10 - 100 ${\mu}$M) clearly down-regulated COX -2 expression from NIH/3T3 cells treated with 12-O-tetradecanoylphorbol 13-acetate, interleukin-1${\beta}$ or tumor necrosis factor-a. But, the expression levels of COX-1, interleukin-1${\beta}$ and fibronectin were not significantly affected. This finding was well correlated with significant reduction of prostaglandin E$_2$(PGE$_2$) production by wogonin. As a comparison, NS-398 (selective cyclooxygenase-2 inhibitor) did not suppress COX -2 expression and other gene levels, while PGE$_2$production was potently reduced at 0.1 - 10 ${\mu}$M. All these results suggest that COX -2 down-regulation of skin fibroblasts may be, at least in part, one of anti-inflammatory mechanisms of wogonin against skin inflammation.

• YAKHAK HOEJI
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• v.51 no.5
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• pp.343-349
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• 2007

To investigate the effect of ethanol extract of Saxifraga stolonifera MEERBURGH on skin care, we measured anti-oxidant and anti-aging activity. S. stolonifera ethanol extract itself had anti-oxidant activity in a dose-dependent manner in DPPH radical scavenging. Silica dose-dependently increased the intracellular ROS generation in RAW 264.7 cells. S. stolonifera ethanol extract inhibited silica-induced intracellular superoxide anion generation, $H_2O_2$ and hydroperoxide generation in RAW 264.7 cells. S. stolonifera ethanol extract significantly inhibited both hyaluronidase and elastase activity, also significantly inhibited MMP-1(collagenase) activity as well. In NIH 3T3 fibroblast cells, S. stolonifera ethanol extract significantly increased collagen-like polymer synthesis, which suggesting the S. stolonifera ethanol extract might be used as hydration and anti-wrinkle agents. From the above results, it is suggested that the main ingredients of S. stolonifera ethanol extract play an important role in anti-oxidant and anti-aging activity.

• Molecules and Cells
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• v.28 no.1
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• pp.31-36
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• 2009

Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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• pp.87-94
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• 1996

한국산 동백나무겨우살이(Pseudixus japonicus) 추출물의 암치료를 위한 생약으로서 활성 유효성을 검증하기 위하여 메탄올, 석유에테르, 클로로포름, 초산 에틸 용매로 순차적으로 추출하여 겨우살이 추출물의 다섯 가지 분획을 얻어, 이에 대하여 in vitro로 1차와 2차 검색 시스템을 사용해 항암활성 성분을 체계적으로 검색하였다. 다섯 가지 분획 중 클로로포름 가용성 분획이 1차 검색 세포인 $P388D_1$에 대해 가장 높은 항암활성을 나타내어 MSB1, NIH/3T3, SNU-1, SNU-C2A 등 2차 검색 시스템에 대해 클로로포름 가용성 분획의 항암활성을 다양한 농도하에서 비교 검색하였다. 혈액암 세포중 특히 $P388D_1$의 생장이 클로로포름 추출물에 의해 강하게 저해되었으며, 형질전환된 생쥐의 태아 섬유아세포와 사람의 대장암, 위암세포들도 어느 정도의 생육저해를 나타내었다. 이 클로로포름 가용성 분획의 주성분은 원소분석, 발색시약과의 반응, IR, GC-MS, $^{13}C$-NMR의 스펙트럼의 결과로 세 종류의 알칼로이드 화합물로 확인되었고, 부성분으로는 지방산 메틸 에스테르와 프탈라이드 화합물이 MS 스펙트럼을 통해 동정되었다.

• Molecules and Cells
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• v.24 no.2
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• pp.288-293
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• 2007

The conversion of mitotic chromosome into interphase chromatin consists of at least two separate processes, the decondensation of the mitotic chromosome and the formation of the higher-order structure of interphase chromatin. Previously, we showed that depletion of BAF53 led to the expansion of chromosome territories and decompaction of the chromatin, suggesting that BAF53 plays an essential role in the formation of higher-order chromatin structure. We report here that BAF53 is associated with mitotic chromosomes during mitosis. Immunostaining with two different anti-BAF53 antibodies gave strong signals around the DNA of mitotic preparations of NIH3T3 cells and mouse embryo fibroblasts (MEFs). The immunofluorescent signals were located on the surface of mitotic chromosomes prepared by metaphase spread. BAF53 was also found in the mitotic chromosome fraction of sucrose gradients. Association of BAF53 with mitotic chromosomes would allow its rapid activation on the chromatin upon exit from mitosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.6
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• pp.1556-1560
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• 2006

The effects of the methanol subfraction from Pueraria Radix on hydrogen peroxide-induced lipid peroxidation and cadmium-induced cytotoxicity were investigated in NIH 3T3 fibroblast cells. After the methanol subfraction treatment, the content of MDA induced by 600 ${\mu}g$ $H_2O_2$ significantly decreased in proportion to the subfraction concentrations as well as 50 ${\mu}M$ $CdCl_2$-induced cytotoxicity. Especially, 200 ${\mu}g/mL$ concentration of methanol subfraction was strongly shown inhibition of lipid peroxidation and detoxification of cadmium. These results suggest that the methanol subfraction from Pueraria Radix retains a potential antioxidant and protective effect against cadmium.