• 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.

• Molecules and Cells
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• v.41 no.6
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• pp.582-590
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• 2018

Endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs) play a pivotal role in vascular regeneration in ischemic tissues; however, their therapeutic application in clinical settings is limited due to the low quality and quantity of patient-derived circulating EPCs. To solve this problem, we evaluated whether three priming small molecules (tauroursodeoxycholic acid, fucoidan, and oleuropein) could enhance the angiogenic potential of EPCs. Such enhancement would promote the cellular bioactivities and help to develop functionally improved EPC therapeutics for ischemic diseases by accelerating the priming effect of the defined physiological molecules. We found that preconditioning of each of the three small molecules significantly induced the differentiation potential of $CD34^+$ stem cells into EPC lineage cells. Notably, long-term priming of OECs with the three chemical cocktail (OEC-3C) increased the proliferation potential of EPCs via ERK activation. The migration, invasion, and tube-forming capacities were also significantly enhanced in OEC-3Cs compared with unprimed OECs. Further, the cell survival ratio was dramatically increased in OEC-3Cs against $H_2O_2$-induced oxidative stress via the augmented expression of Bcl-2, a pro-survival protein. In conclusion, we identified three small molecules for enhancing the bioactivities of ex vivo-expanded OECs for vascular repair. Long-term 3C priming might be a promising methodology for EPC-based therapy against ischemic diseases.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.324-331
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• 2009

MK1 strain, an obligate aerobic heterotrophic bacterium isolated from the rotten tissue of Neungee mushroom (Sarcodon aspratus), produces a copious amount of exopolysaccharide (EPS), which could evoke macrophage activation. Investigations on optimal culture conditions of MK1 and physical properties of MK1 EPS were made. Glucose, galactose, fructose, and sucrose supported well growth of MK1, but potato starch and dextrin did not. However, lactose seemed to be a less favorable carbon source. Optimal growth of MK1 was obtained at pH 7.0, $30^{\circ}C$, and 200 rpm with 2% glucose, and 0.2~0.05% $(NH_4)_2SO_4$. $EPS_{opt}$ obtained from an optimal growth condition constituted of carbon (37.1%), nitrogen (2.2%), oxygen (49.3%), and hydrogen (6.4%), but no sulfur. Paper chrogromatogram of the acid-hydrolysate of $EPS_{opt}$ suggested that MK1 EPS seemed to be hetropolysaccharide composed of a few number of monosaccharides including amino- and acidic-sugars. Its molecular mass determined by SDS-polyacrylamide gel electrophoresis varied from 14.8 to 47.9 kDa. Physical properties of $EPS_{glu}$ obtained from cell grown in glucose medium, such as relative viscosity ($_{rel}$) and crystalline morphology were rather affected by pH of the growth medium. Relative viscosity ($_{rel}$) of exopolysaccaride (0.1 g/ml) harvested from cells grown at medium pH ranging from 6.0 and 7.5 was 1.23 and 1.39, respectively. The freeze-dried exopolysaccharide obtained at low pH (6.0 and 6.5) was fine crystaloid and water-soluble, whereas those obtained at high pH (7.0 and 7.5) was rather gluey and less water-soluble.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1580-1588
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• 2016

The 70% ethanol extract from Adenophora triphylla showed a strong antioxidant effect and reduced cytotoxicity of $H_2O_2$ in SK-N-SH cells. The chloroform fraction from A. triphylla extract (AT-CH) among the six fractions showed strong DPPH radical and intracellular reactive oxygen species (ROS) scavenger effects and the highest protective effect against $H_2O_2$-induced SK-N-SH cell death. Bioactivity compounds were purified from AT-CH, and the chemical structures of the compounds were determined as ${\beta}$-sitosterol and daucosterol on the basis of $^1H-NMR$, $^{13}C-NMR$, and EI mass spectra. ${\beta}$-Sitosterol and daucosterol also had protective effects against oxidative stress in SK-N-SH cells. Phospho-p38 MAPK levels were elevated by $H_2O_2$ but were inhibited by treatment with AT-CH and phytosterols (${\beta}$-sitosterol and daucosterol) isolated from AT-CH. These results suggest that AT-CH has brain neuroprotective effects against oxidative stress ($H_2O_2$) by inhibiting activation of p38 pathways and scavenging intracellular ROS.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.44-45
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• 2010

We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.62-74
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• 1998

In feedlot cattle the abrupt change of diet from roughage to a large quantity of grain for the purpose to improve production often results in increased occurrence of rumen acidosis or acute carbohydrate encouragement enterotoxemia, bloats diarrhea liver abscess and laminitis or robot disease. The common management practice to control these problem is to increase the amount of concentrates in the diet in a stepwise manner until the animals are adapted to a high-grain ration. However this practice requires at least about 3 weeks adaptation period and specially prepared adaptation rations which contain various amount of concentrates. Present experiment was undertaken in order to findout the more simple and rapid adaptation method of cattle to a high grain ration. Nineteen Korean calves aging from four to six month were fed artifical hay (Youngchoun Chuk-Hyup, Korea) which contains 10% of concentrates or alfalfa and rye grass hays for two months and randomly alloted to three experimental groups and two control groups. The experimental group-1 was inoculated by stomach tube for two days with li500 ml/day of ruminal fluid fished from Korean beef cattle that had been previously adapted to a high-energy ration. The experimental group-2 was inoculated by trocalization for two days with the same ruminal fluid. The experimental group-3 was inoculated by trocalization with 1,500 ml/day of bacterial culture which contained 2$\times $10$^{9}$/m1 of Gram-negative bacteria derived from adapted luminal fluid. The two control groups were treated with normal saline solution by the same methods. All animals were fed high-energy ration that contained 80% of grain ad libitum for 30-74 days beginning on the third of the treatment. The effect of the inoculation on the adaptation was observed clinicopathologically with the following results; All of the experimental calves inoculated with the ruminal fluid or Gram-negative bacterial culture derived from adapted cattle did not show any signs of rumen acidosis or other related diseases, while most of the control calves did show diarrhea and bloat and a calf laminitis. The average daily weight gain and feed efficiency of experimental calves were slightly improved compared with control calves. Following the feeding of high-grain rational the pH of the ruminal fluid was lowered in both the experimental and control groups. However severe acidosis with the pH of below 5.0 was observed in only a control group-2. The protozoal number in ruminal fluid was markedly decreased during the high-grain feeding in both the experimental and control calves. However the decrease was mere severe in control calves compared with the experimental calves. The activation of the protozoa were completely disappeared within nine hours at the refrigerator temperature (4"C). No significant differences in heamatological and blood chemical values between the experimental and control calves were recognized. However in one control calf which showed clinically laminitis marked elevations of serum glutamic oxaloacetate transaminase and lactic dehydrogenase activities and a decrease of serum glucose level were observed. From these results it would be concluded the intraruminal transplantation of unadapted calves with the adapted ruminal fluid from cattle previously adapted to a high-energy ration prevents disease problem associated with high-grain feeding and improve weight gain and feed efficiency.ency.

• Resources Recycling
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• v.15 no.4 s.72
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• pp.27-35
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• 2006

In order to use alunite as an activator of blast furnace slag, we studied the hydration characteristics of the calcined alunite and the ground blast furnace slag. The alunite calcined at $650{\cire}C$ consists of KAl($KAl(SO_{4})_{2}$ and $Al_{2}O_{3}$. The calcined alunite reacts with $Ca(OH)_{2}$ and gypsum to form etrringite ($3CaO{\cdot}Al_{2}O_{3}{\cdot}3CaSO_{4}{\cdot}32H_{2}O$) as fellows:$2KAl(SO_{4})_{2}+2Al_{2}O_{3}+13Ca(OH)_{2}+5CaSO_{4}{\cdot}2H_{2}O+73H_{2}O{\rightarrow}3(3CaO{\cdot}Al_{2}O_{3}{\cdot}3CaSO_{4}{\cdot}32H_{2}O)+2KOH$. The $SO_{4}^{2-}$ ions from calcined alunite reacts with CaO in blast furnace slag to from gypsum, which reacts with CaO and $Al_{2}O_{3}$ to from ettringite in calcined alunite-blast furnace slag system. Therefore blast furnace slag can be activated by calcined alunite.

• Journal of Microbiology and Biotechnology
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• v.29 no.10
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• pp.1665-1674
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• 2019

Zika virus (ZIKV) is a mosquito-transmitted, emerging Flavivirus that causes Guillain-$Barr{\acute{e}}$ syndrome and microcephaly in adults and fetuses, respectively. Since ZIKV was first isolated in 1947, severe outbreaks have occurred at various places worldwide, including Yap Island in 2007, French Polynesia in 2013, and Brazil in 2015. Although incidences of ZIKV infection and dissemination have drastically increased, the mechanisms underlying the pathogenesis of ZIKV have not been sufficiently studied. In addition, despite extensive research, the exact roles of individual ZIKV genes in the viral evasion of the host innate immune responses remain elusive. Besides, it is still possible that more than one ZIKV-encoded protein may negatively affect type I interferon (IFN) induction. Hence, in this study, we aimed to determine the modulations of the IFN promoter activity, induced by the MDA5/RIG-I signaling pathway, by over-expressing individual ZIKV genes. Our results show that two nonstructural proteins, NS2A and NS4A, significantly down-regulated the promoter activity of IFN-${\beta}$ by inhibiting multiple signaling molecules involved in the activation of IFN-${\beta}$. Interestingly, while NS2A suppressed both full-length and constitutively active RIG-I, NS4A had inhibitory activity only on full-length RIG-I. In addition, while NS2A inhibited all forms of IRF3 (full-length, regulatory domain-deficient, and constitutively active), NS4A could not inhibit constitutively active IRF3-5D. Taken together, our results showed that NS2A and NS4A play major roles as antagonists of MDA5/RIG-I-mediated IFN-${\beta}$ induction and more importantly, these two viral proteins seem to inhibit induction of the type I IFN responses in differential mechanisms. We believe this study expands our understanding regarding the mechanisms via which ZIKV controls the innate immune responses in cells and may pave the way to development of ZIKV-specific therapeutics.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.1
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• pp.64-70
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• 2007

In this experiment we transformed the Ca-bentonite into Na-bentonite with two inorganic Na-chemicals under different temperatures. These two Na-chemicals were selected among five different Na-chemicals which carries Na as cation. The swelling capacity of the Na chemical-treated bentonite was increased with increasing Na concentration, while the maximum concentration of Na solution decreased with increasing temperature. $Na_2CO_3$ was most effective in exchanging Ca ions and resulting in the highest swelling index among the Na-chemicals. The swelling index was significantly increased with increasing temperature to $100^{\circ}C$. But the equilibration time reversely affected the swelling index due to a rapid increase in evaporation of water. Within same amount of Na treatment SI slightly decreased not only with increasing contacting time but also with increasing temperature. The adsorption for the transformed Na-bentonite was increased with increasing equilibrium concentrations of Pb and Cd ions for all the activated Na-B and indigenous Ca-B and Na-B while the adsorbability of $Pb^{2+}$ onto each Na-B sample is more than that of $Cd^{2+}$. And the maximum adsorption capacity sequence of Na-B samples for Pb and Cd has been found to be 5 % $Na_2CO_3.$ - 5 % $NaHCO_3$ > 3 % $NaHCO_3$ > 3 % $Na_2CO_3$ > 1 % $NaHCO_3$ > 1 % $Na_2CO_3$ > indigenous Na-B > indigenous Ca-B, showing that there are contradictory results about the relationship of cation adsorption to CEC.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1043-1052
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• 2019

Active lipase-producing bacterium Burkholderia gladioli Bps-1 was rapidly isolated using a modified trypan blue and tetracycline, ampicillin plate. The electro-phoretically pure enzyme was obtained by purification using ethanol precipitation, ion-exchange chromatography, and gel filtration chromatography. The molecular weight was 34.6 kDa and the specific activity was determined to be 443.9 U/mg. The purified lipase showed the highest activity after hydrolysis with $p-NPC_{16}$ at a pH of 8.5 and $50^{\circ}C$, and the $K_m$, $k_{cat}$, and $k_{cat}/K_m$ values were 1.05 mM, $292.95s^{-1}$ and $279s^{-1}mM^{-1}$, respectively. The lipase was highly stable at $7.5{\leq}pH{\leq}10.0$. $K^+$ and $Na^+$ exerted activation effects on the lipase which had favorable tolerance to short-chain alcohols with its residual enzyme activity being 110% after being maintained in 30% ethanol for 1 h. The results demonstrated that the lipase produced by the strain B. gladioli Bps-1 has high enzyme activity and is an alkaline lipase. The lipase has promising chemical properties for a range of applications in the food-processing and detergent industries, and has particularly high potential for use in the manufacture of biodiesel.