• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.186-186
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• 1994

Many agonists have been known to activate the hydrolysis of membrane phospholipids through the bindings with corresponding receptors on the various cells. Diacylglycerol and inositol 1,4,5-trisphosphate(IP3) generated by the action of phosphoinositide-specific phospholipase C (PI-PLC) are well known second messengers for the activation of protein kinase C and the mobilization of Ca2+ in many cells. Three types of PI-PLC isozyme (${\alpha}$,${\gamma}$, and $\delta$) and several subtrpes for each type have been identified from mammalian sources by purification of enzymes and cloning of their cDNAs. Each type PI-PLC isozyme is coupled to different receptors and mediators, for example, ${\beta}$-types are coupled to the seven-transmembrane-receptors via Gq family of G-proteins and ${\beta}$-types directly to the receptor tyrosine kinases. Specific modulators for the signaling pathway through each type of PI-PLC should be very useful as potential potential candidates for lend substances in developing novel drugs. To establish the sensitive and convenient screening systems for searching modulators on PI-PLC mediated signaling, two kinds of approaches have been tried. (1) Establishment of in vitro assay condition for each type of PI-PLC isozyme: Overexpression by using vaccinia virus and purification of each isozyme was carried out for the preparation of large amounts of enaymes. Optimum and sensitive assay condition for the measurements of PI-ELC activities were established. (2) Development of the cell lines in which each type of PI-PLC is permanently overexpressed: A fibroblast cell line (3T3${\gamma}$1-7) in which PI-PLC-${\gamma}$1 was overexpressed by using pZip-neo expression vector was developed and used for the measurement of PDGF-induced IP3 formation. The responses for IP3 formed in 3T3${\gamma}$1-7 cells by the treatment of PDGF is 8 times more sensitive than those in control cells. 3T3${\gamma}$l-7 cell is useful for the screening of the inhibitors on the PDGF-induced cellular responses from large number of samples in a small volume(50 ${\mu}$l) and short time(5-15 min). Using these systems, we screened hundreds of herb-extracts for the inhibition of PDGF-induced IP3 formation and selected several extracts that showed the inhibition as the candidates for isolation and characterization of active substances. The determination of the acting point of selected extracts or fractions in the PDGF signaling pathway has been analyzing.

• Toxicological Research
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• v.25 no.4
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• pp.181-188
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• 2009

Uncontrolled cell growth and increased cell proliferation are major features of cancer that are dependent on the stable structure and dynamics of the cytoskeleton. Since stable cytoskeleton structure and dynamics are partly regulated by myosin light chain kinase (MLCK), many current studies focused on MLCK inhibition as a chemotherapeutic target. As a potent and selective MLCK inhibitor, ML-7 [1-(5-iodonaphthalene-1-sulfonyl)-1 H-hexahydro-1,4-diazapine hydrochloride] is a promising candidate for an anticancer agent, which would induce apoptosis as well as prevents invasion and metastasis in certain types of cancer cells. This study assessed cytotoxic effects of ML-7 against HL-60 cells and therapeutic efficacy of ML-7 as a potential antileukemia agent. Trypan-blue exclusion assays showed dose- and time- dependent decreases in ML-7 treated HL-60 cells (p<0.05). Comet assays revealed a significant increase in DNA damage in HL-60 cells after treatment with $40{\mu}M$ ML-7 for 2h. Sub-G1 fractions, analyzed by flow cytometry increased in a dose-dependent manner, suggesting that ML-7 can induce apoptotic cell death in HL-60 cells. ML-7 was selectively cytotoxic towards HL-60 cells; not affecting normal human lymphocytes. That selective effect makes it a promising potential anti-leukemia agent. In addition, anticancer efficacy of ML-7 in combination with flavonoids (genistein or quercetin) or anticancer drugs (cisplatin or Ara-C) against HL-60 cells was assessed. Combination of ML-7 with flavonoids increased the anti-cancer effect of ML-7 to a greater extent than combination with the anticancer drugs. This implies that ML-7 in combination with flavonoids could increase the efficacy of anticancer treatment, while avoiding side effects cansed by conventional anticancer drug-containing combination chemotherapy.

• Korean Journal of Food Science and Technology
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• v.50 no.5
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• pp.543-548
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• 2018

The current study examined antioxidant and neuronal cell protective effects of the crude polysaccharide fraction in Cudrania tricuspidata fruits (CTP). The radical scavenging activities of (1,1-diphenyl-picrylhydrazyl and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)) and reducing power and FRAP of CTP were increased dose-dependently. In addition, the expression of neuroprotective effect of CTP was tested in HT22 mouse hippocampal cells. CTP treatment exhibited non cytotoxicity at dose levels below $500{\mu}g/mL$. Within this optimal concentration range, CTP treatment significantly increased cell viability in $H_2O_2-treated$ HT22 cells. Furthermore, CTP treatment increased superoxide dismutase (SOD) activity and decreased malonaldehyde (MDA) levels in HT22 cells. Therefore, these results indicate that the crude polysaccharide fraction from Cudrania tricuspidata fruits (CTP) possesses antioxidant activities and displays therapeutic potential as a useful source material in the development of brain disorder treatments targeting oxidative stress in neuronal cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.1
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• pp.35-41
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• 2017

Melanin synthesis inhibition activities were investigated for the extracts prepared from the leaves of Carpinus turczaninowii (C. turczaninowii) by using B16F10 melanoma cells. As a result, the ethanol extract ($100{\mu}g/mL$) showed 72.2% inhibition activities without cell toxicities in MTT assays. For the solvent fractions (n-hexane, ethyl acetate, n-butanol, water), the most potent activities were observed at the ethyl acetate fraction. To isolate the active constituents, the ethyl acetate fraction was further purified to afford four compounds; ethyl gallate (1), quercetin rhamnose (2), kaempferol rhamnose (3) and quercetin galloylrhamnose (4). The identification of the isolates was made by spectroscopic data including NMR spectra, and all of the compounds 1-4 were isolated for the first time from the leaves of C. turczaninowii. Anti-melanogenesis activities were studied for the isolates 1-4, and the compound 4 was determined to decrease the melanin synthesis dose-dependently without causing cell toxicities. ELISA measurement indicated that the isolate 4 decreased the contents of cell tyrosinase, a critical enzyme in melanogenesis. Based on these results, the extracts of C. turczaninowii were found to be applicable as whitening ingredients in cosmetic formulations.

• The Korean Journal of Zoology
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• v.7 no.2
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• pp.6-12
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• 1964

The present paper deals with the changes in serum protein fraction, total serum protein, hematocrit, red blood cell, haemoglobin, and weight of liver or kidney : body weight ratio of irradiated and non-irradiated snake head, Ophicephalus argus(CANTOR) . Irrardiation doses are 2537$\AA$-7 minutes (7M) and 2537$\AA$-15 minutes (15M). Serum electrophoretic patterns showed a marked decrease in albumin fraction at 1, 3, 9, 12 and 15 days on 7M group and 15 M group. On both experimental groups percentage increases in $\alpha$1 and $\beta$ fractions occurred at different time periods in general but are interpreted as only apparent changes accompanying the greater albumin fall. ${\gamma}$-Globulin decreased at 1, 6 and 15 days on 7M group. and at 1, 3, 6 and 9 days on 15M group. Also, A/G ratio was significantly low in groups subjected to above conditions as compared to the controls. A/G ratio decreased at 3 and 15 days remarkably on both groups. On the average, the reductions in the A/G ratio were not proportional to themagnitude of ultraviolet-light. Total protein of serum changed according to suggestive changes in electrophoretic patterns of serum . Total protein of serum declined at 1, 3, 6, 9 and 12 day periods on 7M groups and at 1, 3, 9, 12 and 15 day-periods on 15 M group, and increased approaching control values at 15 and 18 day-periods on both groups. Hematocrit increased remarkably at 1, 3, 6 and 12 days and decreased at 9 and 15 days on 7M group, and increased throughout the sampling period on 15M . Red blood cell decreased throughout the sampling period and increased slightly at 6 days on 7 M group and decreased at 1, 6, 9, 15 and 18 days and increased remarkably at 3 and 12 days on 15M group. Hemoglobin decreased remarkably at 1 day-period and increased at other days on both groups. The liver weight was not remarkably changed after whole body irradiation on both groups. and kidney was increased from 1st day on 7M group and 3 rd day on 15M group respectively. It appears that changes in electropphoretic patterns of serum, A/G ratio, total protein of serum, hematocrit, red blood cell, hemoglobin, and liver or kidney weight act to the detriment of the animal following non-ionizing irradiation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.2
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• pp.127-134
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• 2018

Arteriosclerosis is the major cause of coronary artery and cerebrovascular disease, which are leading causes of death. Pro-inflammatory cytokines induce injury to vascular endothelial cells by increasing cell adhesion molecules, leading to vascular inflammation, a major risk factor for the development of arteriosclerosis. In the current study, we investigated the inhibitory effect of enzymatic hydrolysate from Japanese mud shrimp Upogebia major on the inflammation of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$)-stimulated human umbilical vein endothelial cells (HUVECs). We first evaluated the antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities of eight U. major enzymatic hydrolysates: alcalase, papain, ${\alpha}$-chymotrypsin (${\alpha}-Chy$), trypsin, pepsin, neutrase, protamex and flavourzyme. Of these, ${\alpha}-Chy$ exhibited potent antioxidant and ACE inhibitory activities. The ${\alpha}-Chy$ hydrolysate was fractionated by two ultrafiltration membranes of 3 and 10 kDa. The ${\alpha}-Chy$ hydrolysate of U. major and its molecular weight cut-off fractions resulted in a significant reduction in NO production and a decrease in cell adhesion molecules [vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and endothelial-selectin (E-selectin)] and pro-inflammatory cytokines [interleukin-6 (IL-6), interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1)] in $TNF-{\alpha}$-stimulated HUVECs. These results suggest that enzymatic hydrolysate from U. major can be used in the control and prevention of vascular inflammation and arteriosclerosis.

• Biomedical Science Letters
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• v.6 no.1
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• pp.19-28
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• 2000

This study was performed to evaluate the activities of glutathione S-transferase (GST) and lactate dehydrogenase (LDH) in Maddin-Darby canine kidney (MDCK) cells infected with virus and/or treated with amantadine. On cell morphological findings, monolayer fractions in MDCK cells infected with virus were exfolated more than 80% in 1 TCID$_{50}$ group and that in 10 TCID$_{50}$ were completely exfolated after 3 days during infectious process. In proportion to the dose of amantadine, activities of GST and LDH of MDCK cells were significantly decreased and those of LDH in medium fraction were more significantly increased compared with control. According to in both dose and time of virus innoculation, activities of GST and LDH in MDCK cells were significantly decreased in 1 and 10 TCID$_{50}$ infected cells after 3 days. LDH activities in infectious medium were remarkably rised at 10 fold. In case of the cell line inoculated with type A 100 TCID$_{50}$ and additionally treated with amantadine, the decreasing rate to the control in activities of GST and LDH was lower than that in those in case of that infected with virus only. These results suggested that virus infection and amantadine treatment may effect the activity of the detoxicating enzyme in the target cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.9
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• pp.1302-1307
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• 2005

In this study, we investigated the anticarcinogenic and antioxidative activities of Rhodiola sachalinensis (RS). Hexane (RSMH), ethylether (RSMEE), ethylacetate (RSMEA), butanol (RSMB), aqueous (RSMA) fractions and methanol extract (RSM) were screened for their growth inhibition effects using 3- (4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on HepG2, HeLa, MCF-7 and HT-29 cells. The anticarcinogenic effects of RSMEE was most significant when tested on MCF-7 and HepG2 cell lines at the concentration of $500{\mu}g/mL$ which resulted about $84\%\;and\;90\%$ on MCF-7 and HepG2 cells, respectively. The quinone reductase (QR)-inducing activity of RSMH on HepG2 cells was 3.5 times higher compared with the control at the concentration of $200{\mu}g/mL$. Antioxidative activities of RSM, RSMEE, RSMEA and RSMB showed about $80\%$ of electron donating activity (EDA) which were very similar to that of vitamin C as a control. We observed morphological changes of shrinking and the blebbing of HepG2 cancer cell membranes depending on the concentration of RSMEE.

• Radiation Oncology Journal
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• v.37 no.3
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• pp.166-175
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• 2019

Purpose: This study aimed to investigate neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) as prognostic factors in patients with locally advanced non-small cell lung cancer (NSCLC) who received concurrent chemoradiotherapy (CCRT). Materials and Methods: We retrospectively analyzed 66 patients with locally advanced NSCLC treated with definitive CCRT. Among these patients, 95% received paclitaxel/carboplatin or docetaxel/cisplatin. The median radiation dose was 66 Gy in 33 fractions. The NLR and PLR before/after CCRT were evaluated. The maximally selected log-rank test was used to obtain the cutoff values related to the overall survival (OS). Results: Patients with high post-CCRT NLR (>3.12) showed worse OS, locoregional progression-free survival (LRPFS), and distant metastasis-free survival (DMFS) than those with low NLR (2-year OS: 25.8% vs. 68.2%, p < 0.001; 2-year LRPFS: 12.9% vs. 33.8%, p = 0.010; 2-year DMFS: 22.6% vs. 38.2%, p = 0.030). Patients with high post-CCRT PLR (>141) showed worse OS and LRPFS than those with low PLR (2-year OS: 37.5% vs. 71.1%, p = 0.004; 2-year LRPFS: 16.5% vs. 40.3%, p = 0.040). Patients with high NLR change (>1.61) showed worse OS and LRPFS than those with low NLR change (2-year OS: 26.0% vs. 59.0%, p < 0.001; 2-year LRPFS: 6.8% vs. 31.8%, p = 0.004). The planning target volume (hazard ration [HR] = 2.05, p = 0.028) and NLR change (HR = 3.17, p = 0.025) were the significant factors for OS in the multivariate analysis. Conclusion: NLR change after CCRT was associated with poor prognosis of survival in patients with locally advanced NSCLC. An elevated NLR after CCRT might be an indicator of an increased treatment failure risk.

• Applied Chemistry for Engineering
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• v.23 no.5
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• pp.474-479
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• 2012

Possibility of the selective removal of $Ca^{2+}$ ions from a mixed solution of $Na^{+}$ and $Ca^{2+}$ ions using membrane capacitive deionization (MCDI) was investigated. Adsorption equilibrium experiments were conducted to determine the selectivity of the CMX cation-exchange membrane toward $Ca^{2+}$ ions. In addition, desalination experiments for a mixed solution (5 meq/L NaCl + 2 meq/L $CaCl_{2}$) were performed using an MCDI cell. The adsorption equilibrium of CMX membrane showed that the equivalent fraction of $Ca^{2+}$ ions in the solution and the CMX membrane were 28.6 and 87.2%, respectively, which indicates the CMX membrane's high selectivity toward $Ca^{2+}$ ions. Desalination experiments were performed by applying a constant current to the MCDI cell until the cell potential reached 1.0 V. The amount of ions adsorbed did not significantly change as the applied current was changed. However, the equivalent fractions of $Ca^{2+}$ ions among the adsorbed ions were inversely proportional to the applied currents: 81.4, 78.4, 77.0, and 74.5% at 200, 300, 500, and $700\;A/m^{2}$ of applied current density, respectively. This result is attributed to the increased fraction of $Ca^{2+}$ ions adsorbed by the CMX membrane at lower applied current densities.