• The Journal of Advanced Prosthodontics
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• v.6 no.2
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• pp.96-102
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• 2014

PURPOSE. This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS. Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS. From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION. The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.

• Ocean and Polar Research
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• v.31 no.4
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• pp.349-357
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• 2009

Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

• Molecules and Cells
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• v.44 no.7
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• pp.500-516
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• 2021

Cardiac hypertrophic signaling cascades resulting in heart failure diseases are mediated by protein phosphorylation. Recent developments in mass spectrometry-based phosphoproteomics have led to the identification of thousands of differentially phosphorylated proteins and their phosphorylation sites. However, functional studies of these differentially phosphorylated proteins have not been conducted in a large-scale or high-throughput manner due to a lack of methods capable of revealing the functional relevance of each phosphorylation site. In this study, an integrated approach combining quantitative phosphoproteomics and cell-based functional screening using phosphorylation competition peptides was developed. A pathological cardiac hypertrophy model, junctate-1 transgenic mice and control mice, were analyzed using label-free quantitative phosphoproteomics to identify differentially phosphorylated proteins and sites. A cell-based functional assay system measuring hypertrophic cell growth of neonatal rat ventricle cardiomyocytes (NRVMs) following phenylephrine treatment was applied, and changes in phosphorylation of individual differentially phosphorylated sites were induced by incorporation of phosphorylation competition peptides conjugated with cell-penetrating peptides. Cell-based functional screening against 18 selected phosphorylation sites identified three phosphorylation sites (Ser-98, Ser-179 of Ldb3, and Ser-1146 of palladin) displaying near-complete inhibition of cardiac hypertrophic growth of NRVMs. Changes in phosphorylation levels of Ser-98 and Ser-179 in Ldb3 were further confirmed in NRVMs and other pathological/physiological hypertrophy models, including transverse aortic constriction and swimming models, using site-specific phospho-antibodies. Our integrated approach can be used to identify functionally important phosphorylation sites among differentially phosphorylated sites, and unlike conventional approaches, it is easily applicable for large-scale and/or high-throughput analyses.

• Journal of Oriental Neuropsychiatry
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• v.20 no.3
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• pp.1-13
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• 2009

Objectives : The glial cell, located in between the blood vessel and nerve cell, takes charge of the cell support, nutrition supply, elimination of body waste, and cell action. GagamSohabhwangwon(GGSH), a chinese traditional medicinal prescription has been used orally for the treatment of seizures, infantile, convulsion, stroke and so forth. This paper examines the effect of the GagamSohabhwangwon(GGSH) essential oil on cell activity and anti oxidation. Methods : MTT assay methods were employed to measure the cell activity based on the amount of the GagamSohabhwangwon(GGSH) essential oil by using primarily cultivated glial cell. In addition, this paper measured a viability of the glial cell after a protein active retarder control to confirm the multiplication of the cell and examined the cell extinction by the active oxygen, an extinction shielding effect with different amount of the GagamSohabhwangwon(GGSH) essential oil to observe anti oxidation. Furthermore, this paper measured a viability of the cell and phosphorylation(phosphorylation) of the protein which affects the multiplication of the glial cell. Results : When controlling the amount of the GagamSohabhwangwon(GGSH), there was a multiplication effect of the primary glial cell, the multiplication of the cell was dependent on the density of the GagamSohabhwangwon. The multiplication power of the primary glial cell was suppressed by PKA inhibiter (H89). In compliance with the active oxygen the extinction of the primary glial cell was dependent on the density of the GagamSohabhwangwon, there was a shielding effect of the cell extinction when GagamSohabhwangwon(GGSH) was preprocessed. When inducing the multiplication of the primary glial cell, phosphorylation of the Akt, BDNF, CREB, ERK and ERM were increased. Conclusions: Based on the results, GagamSohabhwangwon essential oil will have the effect which activates the nervous system cell and protects the cell through anti oxidation.

• Journal of Nutrition and Health
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• v.36 no.3
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• pp.255-261
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• 2003

Food irradiation has been steadily increasing in many countries in line with increasing international trade and concerns about naturally occurring harmful contaminants in food. Although irradiation provides an excellent safeguard for the consumer by destroying almost 100% of harmful bacteria, it is necessary to ensure the safety of irradiated foods. This study was performed to investigate the effect of an irradiated diet on lipid peroxidation in the plasma, liver, small intestinal mucosa, and lymphocyte DNA damage in mice. Eight-week old ICR mice were assigned to two groups to receive either non-irradiated or irradiated (10 kGy) diets containing 20.38% fish powder and 6.06% sesame seeds for 4 weeks. The resulting changes in the degrees of lipid peroxidation were evaluated based on the level of plasma and liver thiobarbituric acid reactive substance (TBARS), transmission electron micrograph of jejunal mucosa, and free radical-induced oxidative DNA damage in lymphocytes, as measured by alkaline comet assay (single cell gel electrophoresis). The peroxide values of the gamma irradiated diet were measured every week, and the sample for comet assay was taken at the end of the four week experimental period. There was no significant difference in food efficiency ratio between the two groups. The peroxide values of the diet were immediately increased to 35.5% after gamma irradiation and kept on increasing during storage. After 4 weeks, no differences in tissue or plasma TBARS value were observed between the two groups, but epithelial cells of jejumum showed osmiophillic laminated membranous structures, considered as myelin figures,. The oxidative DNA damage expressed as tail moment (TM) increased 30% in the blood lymphocytes of the mice fed the irradiated diet. In conclusion, the comet assay sensitively detected differences in lymphocyte DNA damage after feeding with the irradiated diet for 4 weeks. However, in order to ensure the safety of irradiated foods, it would be more useful to conduct a long-term feeding regimen using an irradiated diet and examine the level of lipid peroxidation and the state of oxidative stress in a greater range of organs.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.385-394
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• 2000

When porcine primordial germ cells (PGCs) isolated from the genital ridge and placed in culture to establish EG cells, a large proportion of PGCs are lost during the early period of culture. To characterize the in vitro death of porcine PGCs, PGCs were cultured in suspension, and apoptosis analyzed using a fluorescent activated cell sorter-based DNA fragmentation assay. The results from flow cytometric analysis showed an increase in apoptosis in cultured cells. However, the cells isolated from the genital ridges are a mixture of PGCs and somatic cells. To detect apoptotic signals specific from porcine PGCs, quantitative TUNEL assay was performed at different time of culture (0 ∼ 24 h). The proportion of apoptotic porcine PGCs determined by double staining with alkaline phosphatase activity and in situ TUNEL assay increased as the time of culture progressed and continued at least 24 h. These results demonstrate that one of the causes of loss of porcine PGCs in vitro is apoptosis.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.153-169
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• 2011

Objectives : This study was performed to investigate the anti-fibrogenic effect and the effect on cell growth and apoptosis in YJCGT, YJCGT YSO and YJCGT YSCO on thioacetamide-induced rat liver tissue and the immortalized human hepatic cell line LX2. Materials and Methods : LX2 cells were treated with various concentrations (0, 50, 150, 300 ug/ml) of YJCGT, Y+YSO, and Y+YSCO extract for 24, 48 and 72 hours. After the treatment, cell viability was measured by using MTT assay. Caspase inhibitor assay, and cell viability were determined by a colorimetric assay with PMS/MTS solution. Rat liver fibrosis was induced by intraperitoneal thioacetamide injection 150 mg/kg 3 times a week for 5 weeks. After the treatment, body weight, liver & spleen weights, liver function test, the complete blood cell count and the change of portal pressure were studied. After YJCGT, Y+YSO, and Y+YSCO treatment, percentages of collagen in thioacetamide-induced rat liver tissue were measured. Results : The viability of the LX2 cell decreased in a dose- and time-dependent manner. Exposure of LX2 cells to YJCGT, YJCGT+YSO and YJCGT+YSCO induced caspase-3 activation, but co-treatment of YJCGT, YJCGT+YSO and YJCGT+YSCO with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. There was no difference in rat body weight between the thioacetamide only group and the YJCGT, YJCGT+YSO and YJCGT+YSCO groups. In the YJCGT, YJCGT YSO and YJCGT YSCO groups, the serum level of GPT significantly went down compared with the thioacetamide only group. In the YJCGT, Y+YSO, Y+YSCO groups, white blood cell elevated by thioacetamide injection decreased but RBC, Hgb, and Hct increased. In the Y+YSO group, the portal pressure elevated by thioacetamide injection significantly decreased. In the histological finding, thioacetamide injections caused severe fibrosis, but YJCGT, Y+YSO, and Y+YSCO treatment significantly reduced the amounts of hepatic collagens. Conclusions : YJCGT, Y+YSO, and Y+YSCO inhibit the growth of LX2 cells by inducing apoptosis through caspase activity. YJCGT, Y+YSO, and Y+YSCO have beneficial effects on the treatment of cirrhotic patients as well as patients with chronic hepatitis.

• Journal of Life Science
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• v.31 no.10
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• pp.929-936
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• 2021

In the present study, we screened candidate natural compounds which possess the strong anti-proliferative effects on human colorectal HCT116 cells using the commercial natural product library (Selleckchem, L1400) based on cell viability assay. Human colorectal cancer HCT116 cells were incubated with 50 μM of each compound from the natural product library, and then cell viability was measured by MTT assay. From the first screening, five different kinds of natural products (chrysin, diosmetin, emodin, piperlongumine, and tanshinone I) were selected based on cell viability assay in HCT116 cells and commercial availability. All selected natural products significantly decreased cell viabilities in HCT116 cells, whereas pro-apoptotic protein NAG-1 is strongly induced by chrysin or emodin treatment. Chrysin and emodin decreased cell viability in a dose-dependent manner. Moreover, chrysin and emodin increased the expression of pro-apoptotic NAG-1 protein in a dose- and time-dependent manner. In addition, PARP cleavage induced by chrysin or emodin was recovered in part by the transfection of NAG-1 siRNA indicating that NAG-1 may be one of the genes responsible for apoptosis induced by chrysin or emodin. Overall, our findings may provide basic screening data on natural products which possess anti-proliferative activities and may help to understand the molecular mechanisms of anti-proliferative and pro-apoptotic activities mediated by chrysin and emodin.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.115-118
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• 2000

A microfabricated cell chip was developed to evaluate drug effect on mouse B16-F1 melanoma cell line. The cell chip system consists of 8-well culture cartridge incorporated with interdigitated array gold electrodes on each well, lock-in amplifier, 8-well cell scanner and computer as a system controller. Impedance of an electrode is increased according to adherent cell growth on the electrode surface. In order to verify investigated. The change of impedance was measured differentially between a control electrode containing only media and cell-cultured sample electrodes with time.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.11
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• pp.4917-4920
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• 2016

For advanced non-small-cell lung cancer (NSCLC) cases, a platinum-based regimen is the first-line chemotherapy treatment. The excision repair cross-complementing group 1 (ERCC1) plays an important role in DNA repair and has been related to resistance to platinum chemotherapy. This study aimed to investigate the effects of the ERCC1 (C118T) polymorphism on treatment response in 26 Thai advanced NSCLC patients receiving first line platinum-based chemotherapy during January to July 2015 at King Chulalongkorn Memorial Hospital (KCMH). DNA was extracted from peripheral blood lymphocytes and the single nucleotide polymorphism of ERCC1 was genotyped using a real-time PCR method with the TaqMan assay. The distribution of C/C, C/T and T/T genotypes was 57.7 %, 34.6 % and 7.7 %, respectively. The response rate to platinum-based chemotherapy in the wild type (C/C) of ERCC1 (C118T) was better than with the variant types (C/T and T/T) but the difference was not statistically significant (29.7% vs 9.1%, P=0.274). The results showed that a genetic polymorphism in ERCC1 might influence patient response to platinum-based chemotherapy. Further multicenter studies are now required to confirm the results of our study.