• Macromolecular Research
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• v.13 no.4
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• pp.314-320
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• 2005

Crosslinked poly(vinyl alcohol) (PVA) membranes were prepared at various crosslinking temperatures using poly(acrylic acid-co-maleic acid) (PAM) containing different PAM contents. The thermal properties of these PVA/PAM membranes prepared at various reaction temperatures were characterized using differential scanning calorimetry (DSC). The proton conductivity and methanol permeability of PVA/PAM membranes were then investigated as PAM content was varied from 3 to 13 wt%. It was found that the proton and methanol transport were dependent on PAM content in their function both as crosslinking agent and as donor of hydrophilic -COOH groups. Both these properties decreased monotonously with increasing PAM concentration. The proton conductivities of these PVA/PAM membranes were in the range from $10^{-3}\;to\;10^{-2}S/cm$ and the methanol permeabilities from $10^{-7}\;to\;10^{-6}cm^{2}/sec$. In addition, the effect of operating temperature up to $80^{\circ}C$ on ion conductivity was examined for three selected membranes: 7, 9 and 11 wt% PAM membranes. Ion conductivity increased with increasing operating temperature and showed and S/cm at $80^{\circ}C$, respectively. The effects of crosslinking and ionomer group concentration were also examined in terms of water content, ion exchange capacity (IEC), and fixed ion concentration. In addition, the number of water molecules per ionomer site was calculated using both water contents and IEC values. With overall consideration for all the properties measured in this study, $7{\sim}9\;wt%$ PAM membrane prepared at $140^{\circ}C$ exhibited the best performance. These characteristics of PVA/PAM membranes are desirable in applications related to the direct methanol fuel cell (DMFC).

• Polymer(Korea)
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• v.38 no.6
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• pp.702-707
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• 2014

Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Korean Journal of Food Science and Technology
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• v.32 no.1
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• pp.218-224
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• 2000

The antimicrobial activities of chitosan oligosaccharide(chitohexaose) and two types of chitosans M.W.(10,000 and M.W. 100,000) were examined against Escherichia coli O157 : H7(ATCC 43894), Staphylococcus aureus(ATCC 144458) and Candida albicans(KFRI 432). Chitosan with molecular weight of 10,000 showed the strongest antimicrobial activities to E. coil O157 : H7 and S. aureus, whereas chitohexaose acted most strongly against C. albicans. The most effective concentration of chitosan was measured to be 0.1 mg/mL for E. coil O157 : H7 and S. aureus, and that of chitohexaose to be 1 mg/mL for C. albicans. Antimicrobial activities of chitosans and chitohexaose were maintained for 60 min after their treatment. They were found to induce leakage of intracellular proteins and nucleic acids from treated microorganisms. The efflux determined by assaying the ${\beta}-galactosidase$ leaked from the lactose-induced E. coli O157 : H7 cells was observed to reach the highest level within 60 min after treatment with the antimicrobial agents and chitosan with 10,000 molecular weight gave the highest ${\beta}-galactosidase$ activity. Therefore, it is supposed that the antimicrobial activity of chitosan with its unique polycationic nature might be caused by its binding to anionic component(s) of the cell envelope and thereby inhibiting the membrane metabolism and/or leaking intracellular materials.

• Journal of Life Science
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• v.26 no.6
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• pp.651-656
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• 2016

Bee pollen is produced by honeybees and is considered one of the most balanced and nourishing nutritional supplements available. Historically, bee pollen has been prescribed for its healing properties and consumed for its high-energy supply. Recent research has provided evidence that bee pollen has diverse biological activities, such as anti-oxidant, anti-inflammatory, anti-bacterial, and even anti-cancer effects. However, the outer membrane of the pollen grain, exine, is highly resistant to most acidic solutions, high pressure, and even digestive enzymes, and the resulting low bioavailability limits its nutritional and clinical applications. This study applied a wet-grinding method to destroy the exine effectively, and it then examined the pollen's enhanced biological activity. First, microscopic observations provided strong evidence that wet grinding destroyed the exine time-dependently. In addition, the content of polyphenols, well-known ingredients of bee pollen and used as internal standards for the quality control of commercial pollen preparations, increased up to 11-fold with wet grinding. Further, the anti-oxidant activity demonstrated on the ABTS anti-oxidant assay, as well as the DPPH radical scavenging assay, was also dramatically increased. Together, the results presented here support a new technology by which bee pollen can be used as a resource for medical, nutritional, and cosmetic applications.

• Journal of Life Science
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• v.22 no.9
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• pp.1145-1151
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• 2012

Aspirin and its deacetylated form, sodium salicylate (NaSal), have been shown to exert chemopreventive activities against many human cancers including those of the colon, lung, and breast. Previously, we showed that combined treatment of NaSal and genistein synergistically induced apoptosis in A549 lung cancer cells, indicating that these two natural chemicals could be used in combination for cancer therapy. In this study, we examined effects of NaSal/genistein combined treatment on other cancer cells and in three-dimensional multicellular tumor spheroid (MTS) and in an in vitro solid tumor model. We found that the combined treatment induces apoptosis in the HCT116 cells and the A549 cells, but not in the MCF-7 cells. Interestingly, the MCF-7 cells responded to the NaSal/genistein combined treatment by undergoing cell death when they were cultivated as MTS. The combined treatment induced apoptosis at an earlier stage in the MCF-7 MTS culture. However, when the MCF-7 MTS was cultivated for a longer period, it induced necrosis rather than apoptosis. We further found that the apoptotic pattern observed in MCF-7 MTS was incomplete: the chromatins were condensed and fragmented, but the nuclear membrane was still intact. Taken together, these results demonstrate that the NaSal/genistein combined treatment induces incomplete apoptosis and necrosis in the MCF-7 MTS culture system.

• Molecules and Cells
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• v.40 no.8
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• pp.567-576
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• 2017

The $Na^+/H^+$ exchanger is responsible for maintaining the acidic tumor microenvironment through its promotion of the reabsorption of extracellular $Na^+$ and the extrusion of intracellular $H^+$. The resultant increase in the extracellular acidity contributes to the chemoresistance of malignant tumors. In this study, the chemosensitizing effects of cariporide, a potent $Na^+/H^+-exchange$ inhibitor, were evaluated in human malignant mesothelioma H-2452 cells preadapted with lactic acid. A higher basal level of phosphorylated (p)-AKT protein was found in the acid-tolerable H-2452AcT cells compared with their parental acid-sensitive H-2452 cells. When introduced in H-2452AcT cells with a concentration that shows only a slight toxicity in H-2452 cells, cariporide exhibited growth-suppressive and apoptosis-promoting activities, as demonstrated by an increase in the cells with pyknotic and fragmented nuclei, annexin V-PE(+) staining, a $sub-G_0/G_1$ peak, and a $G_2/M$ phase-transition delay in the cell cycle. Preceding these changes, a cariporide-induced p-AKT down-regulation, a p53 up-regulation, an ROS accumulation, and the depolarization of the mitochondrial-membrane potential were observed. A pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 markedly augmented the DNA damage caused by the cariporide, as indicated by a much greater extent of comet tails and a tail moment with increased levels of the p-histone H2A.X, $p-ATM^{Ser1981}$, $p-ATR^{Ser428}$, $p-CHK1^{Ser345}$, and $p-CHK2^{Thr68}$, as well as a series of pro-apoptotic events. The data suggest that an inhibition of the PI3K/AKT signaling is necessary to enhance the cytotoxicity toward the acidtolerable H-2452AcT cells, and it underlines the significance of proton-pump targeting as a potential therapeutic strategy to overcome the acidic-microenvironment-associated chemotherapeutic resistance.

• Molecules and Cells
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• v.41 no.5
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• pp.413-422
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• 2018

Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

• The Korean Journal of Physiology
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• v.30 no.1
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• pp.63-76
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• 1996

The aim of present study was to characterize phosphate uptake and to investigate the mechanism for the insulin and insulin-like growth factor(IGF) stimulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells. Results were as follows : 1. The primary cultured proximal tubule cells had accumulated $6.68{\pm}0.70$ nmole phosphate/mg protein in the presence of 140 mM NaCl and $2.07{\pm}0.17$ nmole phosphate/mg protein in the presence of 140 mM KCl during a 60 minute uptake period. Raising the concentration of extracellular phosphate to 100 mM$(48.33{\pm}1.76\;pmole/mg\;protein/min)$ induced decrease in phosphate uptake compared with that in control cells maintained in 1 mM phosphate$(190.66{\pm}13.01\;pmole/mg\;protein/min)$. Optimal phosphate uptake was observed at pH 6.5 in the presence of 140 mM NaCl. Phosphate uptake at pH 7.2 and pH 7.9 decreased to $83.06{\pm}5.75%\;and\;74.61{\pm}3.29%$ of that of pH 6.5, respectively. 2. Phosphate uptake was inhibited by iodoacetic acid(IAA) or valinomycin treatment $(62.41{\pm}4.40%\;and\;12.80{\pm}1.64%\;of\;that\;of\;control,\;respectively)$. When IAA and valinomycin were added together, phosphate uptake was inhibited to $8.04{\pm}0.61%$ of that of control. Phosphate uptake by the primary proximal tubule cells was significantly reduced by ouabain treatment$(80.27{\pm}6.96%\;of\;that\;of\;control)$. Inhibition of protein and/or RNA synthesis by either cycloheximide or actinomycin D markedly attenuated phosphate uptake. 3. Extracellular CAMP and phorbol 12-myristate 13 acetate(PMA) decreased phosphate uptake in a dose-dependent manner in all experimental conditions. Treatment of cells with pertussis toxin or cholera toxin inhibited phosphate uptake. cAMP concentration between $10^{-6}\;M\;and\;10^{-4}\;M$ significantly inhibited phosphate uptake. Phosphate uptake was blocked to about 25% of that of control at 100 ng/ml PMA. 3-Isobutyl-1-methyl-xanthine(IBMX) inhibited phosphate uptake. However, in the presence of IBMX, the inhibitory effect of exogenous cAMP was not significantly potentiated. Forskolin decreased phosphate transport. Acetylsalicylic acid did not inhibit phosphate uptake. The 1,2-dioctanoyl-sn-glycorol(DAG) and 1-oleoyl-2-acetyl-sn- glycerol(OAG) showed a inhibitory effect. However, staurosporine had no effect on phosphate uptake. When PMA and staurosporine were treated together, inhibition of phosphate uptake was not observed. In conclusion, phosphate uptake is stimulated by high sodium and low phosphate and pH 6.5 in the culture medium. Membrane potential and intracellular energy levels are also an important factor fer phosphate transport. Insulin and IGF-I stimulate phosphate uptake through a mechanisms that involve do novo protein and/or RNA synthesis and decrease of intracellular cAMP level. Also protein kinase C(PKC) is may play a regulatory role in transducing the insulin and IGF-I signal for phosphate transport in primary cultured proximal tubule cells.

• The Korean Journal of Physiology
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• v.14 no.2
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• pp.7-16
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• 1980

The effects of various inotropic interventions on the shape of the steady state length tension relation and the length-dependent activation process in cardiac muscle were studied. The influence of inotropic interventions upon the action potential was also observed. The range of varying muscle length was from the optimal length$(l_{max})$, where the active tension production is maximal, to 0.85 $l_{max}$. Changes in stimulus frequency or in external bathing Ca concentration constituted the inotropic interventions in this experiment. The papillary muscles were isolated from the rabbit right ventricles and perfused with $HCO-_3\;-buffered$ normal Tyrode solution which was aerated with $3%\;CO_2-97%\;O_2$ mixed gas and kept at $35^{\circ}C$. Resting Passive tension at $l_{max}$ was approximately 30% of the total tension and appeared from the muscle length of 0.90 $l_{max}$. The effect of stimulus frequency on the steady state level of developed tension was: As the stimulus frequency was increased from 0.1 to 0.5 Hz, there was little change in developed tension. As the frequency was increased further, to a value of about 3 Hz, tension increased steeply. Further increase of the frequency to 5 Hz had little additional effect on the developed tension. The length-tension curves for isometric peak tension became more steeper with the degree of potentiation by inotropic interventions. The relative steepness of the normalized length-tension curves where tension production was expressed as a percentage of maximal tension developed at $l_{max}$, varied inversely with the level of inotropic state and these curves were not superimposable one another. Thus at the stimulus frequency of 2 Hz or at the external Ca concentration of 8 mM, the relative decline in the developed tension for a given change in muscle length was considerably less than the decline observed at the frequency of 0.5 Hz or at the concentration of 2 mM Ca. Action potential duration was prolonged significantly as the frequency increased from 0.2 to 2 Hz, and this change in action potential duration was not observable on the changes in muscle length. There was a tendency of the hyperpolarization of membrane potential when the muscle length was shortened from $l_{max}$ to 0.95 $l_{max}$. These results support the hypothesis that there is a length-dependence of the activation process.