• Animal cells and systems
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• v.6 no.2
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• pp.171-180
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• 2002

Nitric oxide has high affinity for iron, and thus it can cause intracellular iron loss. We tested the idea that intracellular iron can be the primary target of NO toxicity by comparing the signaling mechanisms involved in cell death caused by iron depletion and that caused by NO. Treatment of HL-60 cells with a NO donor, S-nitroso-N-acetyl-DL-penicillamine (SNAP), decreased the intracellular iron level rapidly as that observed with the iron chelator deferoxamine (DFO). Iron chelators such as DFO and mimosine could induce death of human leukemic HL-60 cells by a mechanism requiring activation of p38 kinase, c-Jun N-terminal kinase, caspase-3 and caspase-8. DFO and SNAP also caused release of cytochrome c from mitochondria. Inhibition of p38 kinase by a selective inhibitor, SB203580, abolished the NO and DFO-induced cell death, release of cytochrome c, and activation of caspase-3 and caspase-8, thus indicating that p38 kinase lies upstream in the cell death processes. In a parallel situation, the cells that are sensitive to NO showed similar sensitivity to DFO. Moreover, simultaneous addition of ferric citrate, an iron-containing compound, inhibited the SNAP and DFO-induced activation of caspases and also blocked the NO-mediated cell cycle arrest at $G_1$ phase. Collectively, our data implicate that the NO-induced cell death of tumor cells including HL-60 cells is mediated by depletion of iron and further suggest that activation of p38 kinase lies upstream of cytochrome c release and caspase activation involved in this apoptotic process.

• Journal of Life Science
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• v.19 no.8
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• pp.1164-1169
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• 2009

Aberrant DNA methylation plays an important role in the development of cancer. It has been reported recently that DNA hypermethylation is involved in the maintenance of cancer stem cells. The present study was designed to test the hypothesis that the demethylating agent, 5-aza-2'-deoxycytidine (AZA), can inhibit the potential for maintenance of cancer stem cells. To validate this hypothesis, we used 4T1 syngeneic mouse models of breast cancer. The AZA pre-treated 4T1 cells showed a dramatic inhibition of tumorsphere formation, compared to their counterparts in vitro. In addition, the AZA treatment significantly suppressed the expression of stem regulator genes, such as oct-4, nanog and sox2, compared to counterparts in vivo. Therefore, selective inhibition of DNA methylation may be useful for stem-specific cancer therapy.

• Herbal Formula Science
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• v.7 no.1
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• pp.131-141
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• 1999

In order to eludidate the mechanism of oxidative stress in cultured spinal motor neurons damaged by oxygen free radicals, cytoxicity was assesed by MTT assay and NR assay after spinal motor neurons from mouse were cultured in media containing various concentrations of xanthine oxidase(XO) and hypoxanthine(HX) for 3 hours. In addition, neuroprotective effects of several herb extracts on oxidant-induced neurotoxicity were examined in these cultures, compared with nerve growth factors such as basic fibroblast growth factor(bFGF). XO/HX decreased cell viability in dose- and time dependent manners on cultured mouse spinal motor neurons, and MTT50 and NR50 values were measured at 20mU/ml XO and 0.1mM HX for 3 hours in these cultures. bFGF significantlt increased cell viability. In neuroprotective of herb extracts, Epimedium Koreanum Nakai(EK) and Alpinia oxphylla Mig(IJI) was very effective in the prevention of the neurotoxicity induced by XO/HX in cultured mouse spinal motor neurons. From the above results, it is suggested that XO/HX shows toxic effect in cultured mouse spinal motor neurons and selective herb extracts such as Epimedium Koreanum Nakai(EK) and Alpinia oxphylla Mig(IJI) were very effective in the increase of cell viability against the neurotoxicity induced by oxygen radicals in these cultures.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.3
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• pp.223-228
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• 2013

The calcium-activated $K^+$ ($BK_{Ca}$) channel is one of the potassium-selective ion channels that are present in the nervous and vascular systems. $Ca^{2+}$ is the main regulator of $BK_{Ca}$ channel activation. The $BK_{Ca}$ channel contains two high affinity $Ca^{2+}$ binding sites, namely, regulators of $K^+$ conductance, RCK1 and the $Ca^{2+}$ bowl. Lysophosphatidic acid (LPA, 1-radyl-2-hydroxy-sn-glycero-3-phosphate) is one of the neurolipids. LPA affects diverse cellular functions on many cell types through G protein-coupled LPA receptor subtypes. The activation of LPA receptors induces transient elevation of intracellular $Ca^{2+}$ levels through diverse G proteins such as $G{\alpha}_{q/11}$, $G{\alpha}_i$, $G{\alpha}_{12/13}$, and $G{\alpha}s$ and the related signal transduction pathway. In the present study, we examined LPA effects on $BK_{Ca}$ channel activity expressed in Xenopus oocytes, which are known to endogenously express the LPA receptor. Treatment with LPA induced a large outward current in a reversible and concentration-dependent manner. However, repeated treatment with LPA induced a rapid desensitization, and the LPA receptor antagonist Ki16425 blocked LPA action. LPA-mediated $BK_{Ca}$ channel activation was also attenuated by the PLC inhibitor U-73122, $IP_3$ inhibitor 2-APB, $Ca^{2+}$ chelator BAPTA, or PKC inhibitor calphostin. In addition, mutations in RCK1 and RCK2 also attenuated LPA-mediated $BK_{Ca}$ channel activation. The present study indicates that LPA-mediated activation of the $BK_{Ca}$ channel is achieved through the PLC, $IP_3$, $Ca^{2+}$, and PKC pathway and that LPA-mediated activation of the $BK_{Ca}$ channel could be one of the biological effects of LPA in the nervous and vascular systems.

• The Korean Journal of Pesticide Science
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• v.7 no.2
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• pp.75-82
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• 2003

We discuss that the growth inhibition activities against root and shoot of rice plant (Oryza sativa L.) and barnyard grass (Echinochloa crus-galli) by N-phenyl-3,4,5,6-tetrahydrophthalimide (A) and N-phenyl-3,4-dimethylmaleimide (B) derivatives with changing substituents can be explained and predicted using comparative molecular field analyses (CoMPA) method. And the results show that the cross-validation value, $q^2$ at three components and Pearson correlation coefficient, $r^2$ were rice plant: shoot; $r^2=0.987$, $q^2=0.387$ & root; $r^2=0.923$, $q^2=0.307$ and barnyard grass: shoot; $r^2=0.902$, $q^2=0.535$ & root; $r^2=0.900$, $q^2=0.450$, respectively. In addition, The activities of unknown compounds were predicted by CoMFA method. From the contour map of (A) derivatives, the selective factors to remove barnyard grass takes positive charge on the benzylic carbon atom (C27), negative charged carbon atom (C29) of meta position and steric bulky groups on the cyclic imino ring (C7-C8).

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.2
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• pp.155-162
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• 2007

The purpose of this study is to investigate the anti type I allergic effects and mechanisms of the phyto extract mixture (PEM381) which contains Camellia sinensis (leaf), Psidium guajava (leaf), and Rosa hybrida (flower). PEM381 was tested for its inhibitory effects on arachidonic acid cascade related enzymes (5-lipoxygenase and cyclooxygenase), the mast cell mediated allergic reaction and passive cutaneous anaphylaxis. $IC_{50}$ value of PEM381 against 5-lipoxygenase was $14.11{\pm}0.51ppm$ while that of positive control (nordihy-droguaiaretic acid) was $0.54{\pm}0.08ppm$. PEM381 also exhibited considerable selective inhibition of cyclooxygenase-2. PEM381 could inhibit both degranulation and histamine release in a dose dependent manner from rat peritoneal mast cells activated by compound 48/80. In addition, oral administration of PEM381 showed an inhibitory effect on passive cutaneous anaphylaxis reaction activated by anti-dinitrophenyl IgE antibody in mice. These results suggest that PEM381 may be useful for the prevention and treatment of type Ⅰ allergy related diseases.

• Journal of the Korean Geosynthetics Society
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• v.19 no.1
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• pp.25-33
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• 2020

To improvement the swelling characteristics of the existing cutoff wall against the moisture, the permeability of the sand, calcium bentonite and solidifier mixture according to the contact with trichloroethylene (TCE) was evaluated. Characteristics analysis and the permeability test of the research materials were performed. The permeability was decreased as the mixing ratio of the calcium bentonite was increased and it was increased as the mixing ratio of the solidifier was increased. In conclusion, when mixing 15% of calcium bentonite and more than 30% of solidifier, the permeability coefficient in the underground water movement was analyzed as more than α × 10^-4 cm/sec showing that it does not block the underground water movement. In addition, as the permeability coefficient of mixtures after TCE reaction was analyzed as less than α ×10^-7 cm/sec, it satisfied the condition of blocking layer (less than 1.0 × 10^-6 cm/sec). Therefore, the calcium bentonite and solidifier can be utilized as barrier that showing the characteristic of percolation ability conversion in soil and underground water contaminated with TCE.

• KSBB Journal
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• v.15 no.6
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• pp.664-669
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• 2000

Micellar aggregates are known to be useful for the selective isolation of biologically active materials such as amino acids, proteins, and enzymes from crude mixtures sparsely dispersed in water. In this study, the effects of pH, salt type and its concentration on the solubilization of $\alpha$-chymotrypsin into the organic micellar phase, which consisted of AOT (sodium 야(2-ethylhexy)sulfosuccinate) and iso-octane, were comprehensively examined. It was found that maximum extraction efficiency was attained at a pH below the isoelectric point of $\alpha$-chymotrypsin; at pH=5.0 for NaCl and KCl, and at pH=7.0 for $CaCl_2$and $MgCl_2$. In order to avoid complications stemming from the precipitationof protein at low pH interfaces, the protein concentrations in the organic and aqueous phases were directly measured. The size of the micelle water pool was estimated by measuring the molar ratio of the surfactant to the water, W(sub)o. The resulting values of W(sub)o were nearly constant at 30 and 19 for NaCl and KCl, respectively, and were independent of pH. The addition of 1:2 salts like $MgCl_2$and $CaCl_2$ led to much lower, but a constant value of, W(sub)o than the 1:1 salts.

• KSBB Journal
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• v.25 no.6
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• pp.527-532
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• 2010

A thiazolidinedione derivative, TD49 with the highly selective algicide to red tide was newly synthesized and its acute toxicity was examined in order to evaluate the effect on aquatic ecosystems of coast. Major three species having important role in the food chain of marine ecosystem, such as Skeletonema costatum of microalgae, Daphnia magna of crustacea, Paralichthys olivaceus of flatfish fingerling were employed for the acute toxicity assessment. $EC_50$ or $LC_50$ as the assessment criterion was investigated to each specie, and NOEC (No Observed Effect Concentration) and PNEC (Predicted No Effect Concentration) from most sensitive specie to toxicity of TD49 were further calculated. $EC_50$ of S. costatum in 96-hour, $EC_50$ of D. magna in 48-hour, and $LC_50$ of P. olivaceus in 72-hour for TD49 were $0.34\;{\mu}M$, $0.68\;{\mu}M$, and $0.58\;{\mu}M$, respectively. NOEC from the results of S. costatum was estimated to be $0.20\;{\mu}M$ and PNEC was estimated as 3.40 nM by applying factor value of 100 to $EC_50$ $0.34\;{\mu}M$ of S. costatum. In addition, it was revealed that Solutol used as the dispersing agent of TD49 had very little toxic influence under the concentration range of $0{\sim}0.4\;{\mu}M$ used in TD49 toxicity experiment. Although the estimated concentration of TD49 that will be sprayed onto the coastal field for the algicide is higher than NOEC value, it is considered that the spraying concentration would not be a considerable problem due to a dilution effect by tide at the opened coast.

• Molecules and Cells
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• v.38 no.2
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• pp.138-144
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• 2015

Raloxifene is a selective estrogen receptor modulator (SERM) that binds to the estrogen receptor (ER), and exhibits potent anti-tumor and autophagy-inducing effects in breast cancer cells. However, the mechanism of raloxifene-induced cell death and autophagy is not well-established. So, we analyzed mechanism underlying death and autophagy induced by raloxifene in MCF-7 breast cancer cells. Treatment with raloxifene significantly induced death in MCF-7 cells. Raloxifene accumulated GFP-LC3 puncta and increased the level of autophagic marker proteins, such as LC3-II, BECN1, and ATG12-ATG5 conjugates, indicating activated autophagy. Raloxifene also increased autophagic flux indicators, the cleavage of GFP from GFP-LC3 and only red fluorescence-positive puncta in mRFP-GFP-LC3-expressing cells. An autophagy inhibitor, 3-methyladenine (3-MA), suppressed the level of LC3-II and blocked the formation of GFP-LC3 puncta. Moreover, siRNA targeting BECN1 markedly reversed cell death and the level of LC3-II increased by raloxifene. Besides, raloxifene-induced cell death was not related to cleavage of caspases-7, -9, and PARP. These results indicate that raloxifene activates autophagy-dependent cell death but not apoptosis. Interestingly, raloxifene decreased the level of intracellular adenosine triphosphate (ATP) and activated the AMPK/ULK1 pathway. However it was not suppressed the AKT/mTOR pathway. Addition of ATP decreased the phosphorylation of AMPK as well as the accumulation of LC3-II, finally attenuating raloxifene-induced cell death. Our current study demonstrates that raloxifene induces autophagy via the activation of AMPK by sensing decreases in ATP, and that the overactivation of autophagy promotes cell death and thereby mediates the anti-cancer effects of raloxifene in breast cancer cells.