• Tuberculosis and Respiratory Diseases
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• v.54 no.4
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• pp.403-414
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• 2003

Background : Proteasome inhibitors can promote either cell survival or programmed cell death, depending on both the specific type and proliferative status of the cell. However, it is not well known whether inhibition of proteasome activity is related to apoptosis in lung cancer cells. In addition, the exact mechanisms responsible for apoptosis induced by proteasome inhibition are not well understood. In the present study, we have examined the effect of proteasome inhibition on lung cancer cells and tried to test the mechanisms that may be associated with the apoptosis of these cells. Methods : We examined the effect of proteasome inhibition with MG132 or PS-341 on cell survival in A549 and NCI-H157 lung cancer cells using MTT assay, and analyzed the cleavage of PARP by Western blot analysis to find evidence of apoptosis. Next, we evaluated the activation of caspase 3 by Western blot analysis and the activity of JNK by immunocomplex kinase assay. We also examined the changes in anti-apoptotic pathways like ERK and cIAP1 by Western blot analysis after inhibition of proteasome function. Results : We demonstrated that MG132 reduced cell survival by inducing apoptosis in A549 and NCI-H157 cells. Proteasome inhibition with MG132 or PS-341 was associated with activation of caspase 3 and JNK, reduced expression of activated ERK, and downregulation of cIAP1. Conclusion : Apoptosis induced by proteasome inhibition may be associated with the activation of pro-apoptotic pathways like caspase 3 and JNK and the inactivation of anti-apoptotic pathways in lung cancer cells.

• Tuberculosis and Respiratory Diseases
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• v.53 no.3
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• pp.275-284
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• 2002

Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.

• Korean Journal of Plant Resources
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• v.25 no.2
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• pp.232-239
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• 2012

A laboratory experiment was conducted to determine the content of phenolics and flavonoids, antioxidant activity and cytotoxicity from methanol extracts of different plant parts of $T.$ $officinale$ F. H. Wigg. Total phenolics [mg chlorogenic acid equivalents (FAE) $kg^{-1}$ DW] was highest in flower extracts (72.0 mg $kg^{-1}$), followed by leaf, root, and stalk extracts of $T.$ $officinale$ ($p$ < 0.05). The result of total flavonoid level [mg naringin equivalents $kg^{-1}$ DW] had same tendency to differential total phenolics contents among plant parts, but showed lower ranges of amount. The antioxidant activity of the methanol extracts from all the plant parts dose-dependently increased. DPPH (1,1-diphenyl-2-picryl hydrazyl radical) free radical scavenging activity was highest in flower extracts ($IC_{50}$ value = 624.3 mg $kg^{-1}$ ), and followed by leaf, root, and stalk extracts of $T.$ $officinale$ ($p$ < 0.05). By means of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, cell viability of Calu-6 for human pulmonary carcinoma and SNU-601 for human gastric carcinoma showed the lowest $IC_{50}$ value in the flower extracts ($IC_{50}$ value = 85.7 and 311.4 mg $kg^{-1}$, respectively), indicating the highest cytotoxicity. The results suggested that total phenolics content and total flavonoids level in different plant parts of $T.$ $officinale$ were highly correlated with antioxidative ($r^2$=0.7280 to 0.9971) or with cytotoxic activities ($r^2$=0.5795 to 0.9515).

• KOREAN JOURNAL OF CROP SCIENCE
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• v.57 no.1
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• pp.51-59
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• 2012

Contents of phenolics and flavonoids, antioxidant activity and cytotoxicity were investigated in the methanol extracts of three different $Taraxacum$ species, $Taraxacum$ $coreanum$, $Taraxacum$ $mongolicum$, and $Taraxacum$ $officinale$. Total phenolics content at $1000mg\;kg^{-1}$ was more present in shoot parts than in roots, and was highest in $T.$ $mongolicum$ shoot and root extracts (76.8 and $40.0mg\;kg^{-1}$, respectively), followed by $T.$ $coreanum$ and $T.$ $officinale$ ($p$ < 0.05). Total flavonoid level had same tendency to total phenolics among $Taraxacum$ species, showing lower amounts ($6.5{\sim}36.4mg\;kg^{-1}$) than total phenolics. The antioxidant activity of the methanol extracts from all the species dose-dependently increased. DPPH free radical scavenging activity at $1,000mg\;kg^{-1}$ was highest in shoot and root extracts from $T.$ $mongolicum$ by 89.6 and 83.4%, respectively. According to MTT assay, cell viability of Calu-6 (human pulmonary carcinoma) was lowest in the $T.$ $mongolicum$ shoot and root extracts ($IC_{50}$ values=83.4 and $66.4mg\;kg^{-1}$, respectively), and followed by $T.$ $coreanum$ and $T.$ $officinale$ (lowest). Calu-6 was more sensitive to the extracts than SNU-601 (human gastric carcinoma). Antioxidative and anticancer activities in three different $Taraxacum$ species was more correlated with total phenolics content ($r^2$=0.0097 to 0.6213) than with total flavonoids level ($r^2$=0.0027 to 0.4627). The results showed total phenolics content and total flavonoids level were highly correlated with anticancer activity and antioxidant activity, and their content and activities were different depending on species.

• Korean Journal of Microbiology
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• v.48 no.1
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• pp.52-56
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• 2012

The aim of this study was to evaluate the antimicrobial effect of carvacrol against periodontopathic and cariogenic bacteria and its cytotoxicity in human oral tissue cells. We tested their antibacterial properties against mutans streptococci and five major periodontopathic bacterial species involved in periodontal disease. The antimicrobial activity was evaluated by the minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The cell viability of carvacrol on normal human gingival fibroblast (NHGF) cells was tested by metyl thiazolyl tetrazolium assay. The data showed that carvacrol had remarkable antimicrobial effect on tested bacteria with a MIC and MBC values ranged from 16 to $128{\mu}g/ml$ and from 32 to $128{\mu}g/ml$, respectively. In cell toxicity studies, carvacrol had significantly decreased cell viability when NHGF cells were treated at $128{\mu}g/ml$. These findings suggest that carvacrol has a strong antimicrobial activity against periodontopathic and cariogenic bacteria. However, in order to use it as a component of gargling solution or toothpaste, its concentration should be below $64{\mu}g/ml$ and other compounds having an antimicrobial activity against periodontopathic and cariogenic bacteria should be used together.

• Food Engineering Progress
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• v.22 no.4
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• pp.315-320
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• 2018

This study examined the effects of drying methods of Aronia melanocarpa on contents of total polyphenols and flavonoids, antioxidant activity and anti-inflammatory activity. As a result, freeze-dried aronia extract (FDAE) significantly exhibited higher contents of total polyphenols and flavonoids (155.76 mg GAE/g and 105.70 mg QE/g) than hot-air dried aronia extract (HDAE) (134.93 mg GAE/g and 82.29 mg QE/g). Also, FDAE showed greater antioxidant activity than HDAE in both DPPH and ABTS. For anti-inflammatory activity, NO production from lipopolysaccharide activated RAW264.7 cell reduced at a dose-dependent manner in both FDAE and HDAE. However, reduction rate of NO production is higher in FDAE (62.7%) than in HDAE (33.5%). These results suggest that comparatively, freeze-drying is a better method for preserving the antioxidant and anti-inflammatory activities and of aronia.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.3
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• pp.219-229
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• 2018

The purpose of this study was to investigate the role of the Moringa oleifera (M. oleifera) extract as a cosmetic additive. The tyrosinase and elastase inhibitory effects showed 47% and 39% at $1,000{\mu}g/mL$ concentration, respectively. Also, the collagenase inhibition effect was 31% at $500{\mu}g/mL$ concentration. A cell viability test, measured on macrophage cell (RAW 264.7) and melanoma cell (B16F10) by ethanol extract of M. oleifera, showed 94.2% and 94.8% at $100{\mu}g/mL$ concentration, respectively. In order to confirm anti-inflammatory activity, we examined the inhibitory effects on the production of lipopolysaccharides (LPS)-induced NO in RAW 264.7 cells by Griess assay. As a result, the M. oleifera extract showed a concentration-dependent inhibition of NO production. The protein expression inhibitory effects of M. oleifera extract were measured by western blot at 25, 50, $100{\mu}g/mL$ concentration and the ${\beta}-actin$. Results showed that the expression inhibition rates of the iNOS, COX-2, MITF, TRP-1, TRP-2, tyrosinase protein were decreased by 85.8%, 57.5%, 80.7%, 30%, 29.9%, 23.6% at $100{\mu}g/mL$ concentration, respectively. It was concluded that M. oleifera extracts had the anti-inflammatory and whitening effects and thus could be applied for cosmetics as a natural ingredient.

• Journal of the Korean Crystal Growth and Crystal Technology
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• v.22 no.2
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• pp.92-98
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• 2012

The co-precipitation technique has been applied to synthesize biphasic calcium phosphate (BCP). $Ca(NO_3)_2{\cdot}4H_2O$, $(NH_4)_2HPO_4$, TEOS and $Mg(NO_3)_2{\cdot}6H_2O$ as the starting materials was used. After the heat treatment of powder crystalline phases HAp and ${\beta}$-TCP analysis showed a mixed phase. The overall spectra appear to have mainly two modes corresponding to characteristic $PO^{3-}_4$ and $OH^-$ groups. After immersion in Hanks' Balanced Salt Solution (HBSS) for 1 week a precipitation started to be formed with individual small granules on the specimen surface. An MTT assay indicated that ionic substituted BCP powders had no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.215-225
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• 2005

Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of CTF-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). ResuIts : It has no cytotoxic effects on HepG2 cells in all concentrations (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$). More than twofold up-regulated genes were 19 genes. The number of more than twofold down-regulated genes was 13. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

• Clinical and Experimental Pediatrics
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• v.46 no.1
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• pp.56-66
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• 2003

Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.