• Journal of Life Science
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• v.17 no.10
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• pp.1447-1451
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• 2007

Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemo-sensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.

• Journal of Life Science
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• v.23 no.12
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• pp.1460-1470
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• 2013

To examine whether miscellaneous cereal grains have an antithrombotic effect, we investigated the anticoagulant activity of 80% ethanol extracts from eleven selected miscellaneous cereal grains. The 80% ethanol extract of hwanggeumchal sorghum (Sorghum bicolor) showed the highest anticoagulant activity, followed by that of green foxtail millet grains, in terms of thrombin time (TT). When the ethanol extract of hwanggeumchal sorghum was sequentially fractionated with n-hexane, methylene chloride, ethyl acetate, and n-butanol, the majority of the TT-inhibitory activity was detected in the hexane and methylene chloride fractions. Whereas aspirin (final conc. 480 ${\mu}g/ml$) prolonged TT by 2-fold, the ethanol extract, hexane fraction, and methylene chloride fraction in the same dose prolonged TT by 2.2-fold, 2.9-fold, and 2.5-fold, respectively. The ethanol extract of hwanggeumchal sorghum could delay activated partial thromboplastin time (APTT) as well as prothrombin time (PT). Although the APTT-inhibitory activity of the ethanol extract was mainly partitioned into the hexane and methylene chloride fractions, the PT-inhibitory activity of the ethanol extract was solely partitioned into the hexane fraction. The APTT- and PT-inhibitory activities of these organic solvent fractions were more potent than those of the control warfarin (final conc. 3.13 mg/ml). The TT-inhibitory activity of the ethanol extract was heat-stable and acid-stable. The ethanol extract, hexane fraction, and methylene chloride fraction of hwanggeumchal sorghum appeared to possess a direct fibrinolytic activity toward fibrin clotting. These results show that hwanggeumchal sorghum can exert anticoagulant and fibrinolytic effects and, thus, have the potential to be applicable as antithrombotic dietary sources.

• Journal of Life Science
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• v.27 no.9
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• pp.975-985
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• 2017

To investigate the beneficial effects of an agar gel mask (AGM) on UV-induced photoaging, SKH-1 hairless mice were treated with a topical application of AGM and an AGM dipped in essence (AGMdE). The mice were divided into an no radiation group, UV + AGM, UV + AGMdE, and UV + vehicle (PBS) treatment groups. Alterations in skin wrinkles, skin phenotype, histological structures, oxidative status, and toxicity were then evaluated during 4 weeks of exposure. The topical application of AGM and AGMdE inhibited wrinkle formation, suppressed the erythema index, prevented transepidermal water loss, and enhanced skin hydration. In addition, epidermal thickness recovered to a similar level as that in the no irradiation group in the UV + AGM and UV + AGMdE treatment groups compared with the UV + vehicle (distilled water) group. Furthermore, the expression levels of matrix metalloproteinase-1 (MMP-1) and tyrosinase were reduced in the UV + AGM and UV + AGMdE treatment groups, although the highest level varied. Moreover, superoxide dismutase (SOD) activity was significantly lower in the UV + AGM and UV + AGMdE treatment groups as compared with the UV + vehicle group. No significant alterations induced by most toxic compounds were measured in serum biochemical markers and liver and kidney histological features of the UV + AGM and UV + AGMdE treatment groups. These results suggest that AGM may protect against skin aging by regulating skin morphology, histopathological structures, and oxidative conditions.

• Tuberculosis and Respiratory Diseases
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• v.63 no.1
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• pp.42-51
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• 2007

Background: TRAIL is a cytokine that selectively induces apoptosis in various cancer cell lines. Gefitinib is new targeted drug applied in lung cancer that selectively inhibits EGFR tyrosine kinase. However, lung cancers have shown an initial or acquired resistance to these drugs. This study examined the effect of IGF-1R and its blockade on enhancing the sensitivity of lung cancer cell lines to TRAIL and gefitinib. Methods: Two lung cancer cell lines were used in this study. NCI H460 is very sensitive to TRAIL and gefitinib. On the other hand, A549 shows moderate resistance to TRAIL and gefitinib. The IGF-1R blockade was performed using adenoviruses expressing the dominant negative IGF-1R and shRNA to IGF-1R and AG1024 (IGF-1R tyrosine kinase inhibitor). Results: The adenovirus expressing dominant negative IGF-1R(950st) induced the increased expression of defective IGF-1R on the lung cancer cell surface, and the adenovirus-shIGF-1R effectively decreased the level of IGF-1R expression on cell surface. The genetic blockade of IGF-1R by the adenovirus-dnIGF-1R and AG1024 increased the sensitivity of A549 cells to TRAIL. The reduction of IGF-1R by transduction with ad-shIGF-1R also increased the sensitivity of the A549 cells to gefitinib. Conclusion: The blockade of IGF-1R through various mechanisms increased the sensitivity of the lung cancer cell line that was resistant to TRAIL and gefitinib. However, further studies using other cell lines showing acquired resistance as well as in vivo animal experiments will be needed.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.6
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• pp.784-789
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• 2005

The inhibitory effects of hot water extracts of Rhus verniciflua Stokes pith and peel roasted at 170, 200 and $220^{\circ}C$ on lipid peroxidation, formation of DPPH free radicals and growth of four human cancer cells such as HepG2 (liver cancer), SNU-1 (stomach cancer), MCF-7 (breast cancer) and Widr (colon cancer) were examined. The antioxidant activities and growth inhibitory effects on cancer cells of hot water extracts of peel were higher than those of pith, and the activities were dose-dependent. The roasting temperature showing the highest antioxidant activities and growth inhibitory effects on cancer cells was in the range of $170\~200^{\circ}C$ The lipid peroxidation and formation of DPPH free radicals of hot water extracts of roasted pith and peel were inhibited to 50.9, $56.5\%\;and\;79.0,\;78.4\%$ at the concentration of $500\mu g/mL$, respectively. The growth inhibitory effects of roasted pith and peel on cancer cells were in the order of Widr (41.5, $36.0\%$) > HepG2 (61.5, $44.0\%$) > MCF-7 (92.0, $69.2\%$)> SNU-1 (100, $100\%$) cells at the concentration of $1,000\mu g/mL$ as compared with the control, respectively. These results suggest that roasted Rhus verniciflua Stokes could be an useful natural medicinal plant for colon cancer.

• KSBB Journal
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• v.23 no.6
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• pp.513-518
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• 2008

This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.5
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• pp.535-540
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• 2009

The effect of cellulolytic (Viscozyme) and pectolytic (Pectinex) enzyme treatments on extraction of total polyphenol and antioxidant activity of extract from apple peel have been examined. Extraction was carried out with a dosage of 0.1, 0.25, 0.5, 1 and 2% (v/v) of Viscozyme, Pectinex and Viscozyme+Pectinex at $30{\sim}50^{\circ}C$ for $12{\sim}24$ hours. Total polyphenol contents (mg/mL) of extracts obtained with 2% of Viscozyme, Pectinex or Viscozyme+Pectinex treatment for 12 hours were $0.30{\pm}0.02$, $0.16{\pm}0.01$, and $0.33{\pm}0.02$ at $30^{\circ}C$, $0.34{\pm}0.01$, $0.19{\pm}0.01$, and $0.35{\pm}0.02$ at $40^{\circ}C$ and $0.34{\pm}0.01$, $0.22{\pm}0.01$, and $0.38{\pm}0.02$ at $50^{\circ}C$ respectively. The result shows that Viscozyme was more effective than Pectinex at all experimental temperatures, and Viscozyme+Pectinex resulted in the highest phenolic content at $50^{\circ}C$. Antioxidant activities determined by DPPH, ABTS and FRAP assays were increased with concentrations of extracts produced by 2% of Viscozyme+Pectinex treatment, which ranged from 0.10 to 0.40 vit. C eq mM for $5{\sim}25\;mg$ of dried matters, from 0.09 to 0.28 vit. C eq mM for $1{\sim}5\;mg$ of dried matters, and from 0.06 to 1.85 $FeSO_4$ eq mM for $1{\sim}5\;mg$ of dried matters, respectively.

• Journal of Gastric Cancer
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• v.6 no.4
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• pp.250-256
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• 2006

Purpose: Recently, interest in peroxisome-proliferator-activated receptors (PPAR) has increased, although clinical studies of the effect of $PPAR-{\gamma}$ expression on gastric cancer have not been reported yet. In this study, we investigated the role of $PPAR-{\gamma}$ expression in gastric cancer patients. Materials and Methods: One hundred twenty-eight (128) samples of both gastric cancer and normal tissues were obtained from 128 patients who had undergone at a curative gastrectomy at Seoul Medical Center from Jan. 2001 to Dec. 2005. $PPAR-{\gamma}$ expression was determined by using immunohistochemical staining, and the results were analyzed. The statistical analysis was based on clinicopathological findings and the differences in survival rates. Results: The mean age of the patients was 6n, and the male : female ratio was 1.9 : 1. $PPAR-{\gamma}$ expression was significantly higher in cancer tissues than in normal tissue (81.3% vs. 57.0%, p<0.001). There was insignificant difference between well and moderately differentiated types and poorly differentiated types in terms of the expression of $PPAR-{\gamma}$ (87.0% vs. 74.6%, P=0.074). In the univariate analysis the survival rate was significantly increased when $PPAR-{\gamma}$ was expressed in normal tissue (P=0.003). In the multivariate analysis, only the UICC TNM staging had significance related to the survival rate. Conclusion: The rate of $PPAR-{\gamma}$ expression was higher in cancer tissue than it was in normal tissue from gastric cancer patients. In the univariate analysis, $PPAR-{\gamma}$ expression in normal tissue had significance with respect to survival, but the multivariate analysis showed no such significance. Thus, we should further evaluate more cases to determine whether or not such a significance exists.

• Restorative Dentistry and Endodontics
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• v.36 no.4
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• pp.280-289
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• 2011

Objectives: The purpose of this study was to evaluate ${\mu}TBS$ (microtensile bond strength) of current dentin bonding adhesives which have different hydrophobicity with low-shrinkage silorane resin. Materials and Methods: Thirty-six human third molars were used. Middle dentin was exposed. The teeth were randomly assigned to nine experimental groups: Silorane self-etch adhesives (SS), SS + phosphoric acid etching (SS + pa), Adper easy bond (AE), AE + Silorane system bonding (AE + SSb), Clearfil SE bond (CSE), CSE + SSb, All-Bond 2 (AB2), AB2 + SSb, All-Bond 3 (AB3). After adhesive's were applied, the clinical crowns were restored with Filtek LS (3M ESPE). The 0.8 mm ${\times}$ 0.8 mm sticks were submitted to a tensile load using a Micro Tensile Tester (Bisco Inc.). Water sorption was measured to estimate hydrophobicity adhesives. Results: ${\mu}TBS$ of silorane resin to 5 adhesives: SS, 23.2 MPa; CSE, 19.4 MPa; AB3, 30.3 MPa; AB2 and AE, no bond. Additional layering of SSb: CSE + SSb, 26.2 MPa; AB2 + SSb, 33.9 MPa; AE + SSb, no bond. High value of ${\mu}TBS$ was related to cohesive failure. SS showed the lowest water sorption. AE showed the highest solubility. Conclusions: The hydrophobicity of adhesive increased, and silorane resin bond-strength was also increased. Additional hydrophobic adhesive layer did not increase the bond-strength to silorane resin except AB2 + SSb. All-Bond 3 showed similar ${\mu}TBS$ & water sorption with SS. By these facts, we could reach a conclusion that All-Bond 3 is a competitive adhesive which can replace the Silorane adhesive system.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.227-233
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• 2017

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.