• Proceedings of the Korean Society of Life Science Conference
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• 2001.06a
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Korean Journal of Ecology and Environment
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• v.42 no.2
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• pp.161-171
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• 2009

A 'continuous removal of organic matters (CROM) system' using a native freshwater bivalve in Korea Anodonta woordiana, was developed to determine its potential of controlling various sestons in eutrophic lake system, and to evaluate its effect on water quality improvement under consideration of sediment addition as habitat. We designed CROM experiments with four treatments: no mussels and no sediment (W, negative control), no mussels and sediment (WS, positive control), mussels and no sediment (WM), and mussels and sediment (WMS). The experiments were performed at the condition of 18${\sim}$25 L $h^{-1}$ of inflow, mussel density of 486.1 indiv. $m^{-2}$, and temperatures between 15 and $22^{\circ}C$ for 13 consecutive days. Physicochemical and biological parameters were measured at daily (10:00 am) intervals after the mussel addition. Results indicated that mussel stockings without addition of sediment effectively removed sestons (suspended solids and chlorophyll-a) at nearly same level over 80 percentage of the control during the study, while there were no differences in removal activities of sestons between with and without sediment (P>0.5). Therefore, it clearly suggests that CROM system using A. woordiana has a strong potential to control the seston in surface water of eutrophic lake.

• Current Research on Agriculture and Life Sciences
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• v.14
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• pp.111-122
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• 1996

S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% com steep liquor, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/ml) when bacteria were cultured in 2.5 l jar fermentor for 48hrs at $30^{\circ}C$ and pH 7.0. Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires $Mg^{2+}$ for activity, $Ca^{2+}$, $Co^{2+}$ is not an inhibitor but inhibit by $Fe^{3+}$, $Hg^{2+}$, and $Cu^{2+}$, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.491-501
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• 2002

A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.

• Korean Journal of Soil Science and Fertilizer
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• v.18 no.4
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• pp.336-347
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• 1985

Soils and rice plants subject to smoke from the Jang-hang Smelter were samples at two depths to assess the nature and extend of Cd, Cu, Pb, and Zn contamination, particularly with respect to distance from pollution source. Soils at east site within 1km of the center of the smelter were highly contaminated with Cd, Cu, Pb, and Zn. Metal levels in the smoke-polluted area were higher in surface soil than in subsurface soil, and decreased rapidly with distance reaching nearly background levels at 5km from pollution source. A position correlation was found between contents of total and 0.1 N HCl-extractable, or 1.0N $CH_3COONH_4$- extractable heavy metals in surface soils of pollution. Contents of heavy metals in soils were positively correlated with soil organic matter and cation exchange capacity. The contents of heavy metals in brown rice collected from the paddy field around the Jang-hang Smelter were in the range of 0.23-1.33 ppm for Cd, 2.39-6.25 for Cu, 0.95-8.32 ppm for Pb and 14.60-27.31 for Zn. The contents of Cd, Cu, and Zn in rice straw were positively correlated with those in the brown rice. Negative correlation was found between the contents of Cd, Cu, Pb, and Zn in brown rice and the distance from the source.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.1
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• pp.22-29
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• 2015

The objectives of this study were to monitor the changes in soil solution nutrients and to evaluate their effect on rice uptake and yield. The changes of chemical characteristics of paddy soil solution were examined from the 58th fertilization experiment in which the continuous rice cropping experiment started in 1954 at the National Academy of Agricultural Science. The treatments were no fertilization (No fert.), inorganic fertilization (NPK), inorganic fertilizer plus rice straw compost (NPKC) and inorganic fertilizer plus silicate and lime fertilizer as a soil amendment (NPKCLS). The fertilizers were added at rates of standard fertilizer application rate in which nitrogen (N), phosphate ($P_2O_5$), potassium ($K_2O$), and sililcate ($SiO_2$) were applied at rates of $75{\sim}150kg\;ha^{-1}$, $70{\sim}86kg\;ha^{-1}$, $75{\sim}86kg\;ha^{-1}$, and $7.5Mg\;ha^{-1}$ respectively and lime was applied to neutralize soil acidity until 6.5. Average Electrical Conductivity (EC) of soil solution in NPKCLS and NPKC ranged from 1.16 to $2.00dS\;m^{-1}$. The $NH{_4}^+$ and $K^+$ levels in NPKCLS and NPKC were higher than that of the other treatments, due to high supply power of rice straw compost. The content of $H_3SiO{_4}^-$ was higher in NPKCLS because of silicate application. The dominant ions in soil solution were $Ca^{2+}$, $Mg^{2+}$ and $Na^+$ among cations and $HCO{_3}^-$, $SO{_4}^{2-}$, and $Cl^-$ among anions in all treatments. The continuous application of inorganic fertilizers plus rice straw compost (NPKC) and silicate fertilizer (NPKCLS) led to the changes of various chemical composition in soil solutions. Also, they had a significant impact on the improvement of rice inorganic uptake and grain yield. Especially, inorganic uptake by rice in NPKC and NPKCLS significantly increased than those in NPK plot; 14~46% for T-N, 32~36% for P, 43~57% for K, and 45~77% for Si. Therefore, the combined application of inorganic fertilizers with organic compost as a soil amendment is considered as the best fertilization practice in the continuous rice cropping for the improvement of crop productivity and soil fertility.

• Journal of the Korean Dietetic Association
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• v.25 no.3
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• pp.217-228
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• 2019

There have been no published studies concerning the anti-inflammatory effects of corn silk on colon cancer cells. Thus, this study was conducted to investigate the effect of corn silk extract containing high levels of maysin on inflammation and its mechanism of action in colon cancer cells. SW 480 human colon cancer cells were treated with $1{\mu}g/mL$ of lipopolysaccharide (LPS) to induce inflammation, and next they were treated with different concentrations of corn silk extract (0, 5, 10 and $15{\mu}g/mL$). The concentrations of nitric oxide (NO) were determined. The mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), interleukin-1beta ($IL-1{\beta}$) and interleukin-6 (IL-6), were determined. Western blot analysis was performed to determine the protein expressions of nuclear factor-kappa B ($NF-{\kappa}B$) and mitogen-activated protein kinases, and the latter consists of extracellular signal-related kinase (ERK), c-jun NH2-terminal kinase (JNK) and p38 MAP kinase (p38). The concentration of NO and the mRNA expression of iNOS were significantly and dose-dependently decreased in the corn silk-treated groups (P<0.05). The mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were significantly increased in the LPS-treated group (P<0.05), but these expressions were significantly and dose-dependently decreased in the corn silk treated groups (P<0.05). The protein expressions of $NF-{\kappa}B$ (in a dose-dependent fashion), ERK (at 10 and $15{\mu}g/mL$), JNK (at $15{\mu}g/mL$) and p38 (at 10 and $15{\mu}g/mL$) were significantly decreased with corn silk treatments (P<0.05). In conclusion, corn silk extract containing high levels of maysin seems to inhibit the LPS-induced inflammatory responses in SW480 colon cancer cells via the $NF-{\kappa}B$ pathway.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.3
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• pp.375-386
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• 2019

Objective: We hypothesized that silage additives may alter the undigested neutral detergent fiber (uNDF) content through ensiling. Therefore, urea and formic acid were applied to corn, whole barley crop (WBC) and alfalfa to change uNDF content of the ensiled forages. Methods: Six experimental diets at two groups of high uNDF (untreated corn and alfalfa silages [CSAS] and untreated whole barley and alfalfa silages [BSAS]) and low uNDF (urea-treated corn silage+untreated alfalfa silage [$CS_UAS$], urea-treated whole barley silage+untreated alfalfa silage [$BS_UAS$], untreated corn silage+formic acid-treated alfalfa silage [$CSAS_F$], and untreated whole barley silage+formic acid-treated alfalfa silage [$BSAS_F$]), were allocated to thirty-six multiparous lactating Holstein dairy cows. Results: The untreated silages were higher in uNDF than additive treated silages, but the uNDF concentrations among silages were variable (corn silage0.05). Milk yield tended to increase in the cows fed high uNDF diets than those fed low uNDF (p = 0.10). The cows fed diet based on urea-treated corn silage had higher milk yield than those fed other silages (p = 0.05). The substitution of corn silage with the WBC silage tended to decrease milk production (p = 0.07). Changing the physical source of NDF supply and the uNDF content from the corn silage to the WBC silage caused a significant increase in ruminal $NH_3-N$ concentration, milk urea-N and fat yield (p<0.05). The cows fed diets based on WBC silage experienced greater rumination time than the cows fed corn silage (p<0.05). Conclusion: Administering additives to silages to reduce uNDF may improve the performance of Holstein dairy cows.

• Resources Recycling
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• v.31 no.4
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• pp.19-25
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• 2022

In this study, tungsten carbide (WC) powder was prepared using a novel recycling process for hard metal sludge that does not use ammonium paratungstate. Instead of ammonia, acid was used to remove the sodium and crystallized tungstate, resulting in the formation of tungstic acid (H₂WO₄). The WC powder was successfully synthesized by the carbothermal reduction of tungstic acid through H₂O decomposition, reduction of WO₃ to W, and formation of WC. The carbon content and holding time at the carbothermal reduction temperature were optimized to remove free carbon from the WC powder. As a result, most of the free carbon in the WC powder prepared from sludge was removed, and the content of free carbon in the synthesized WC powder was lower than that in commercial WC powder. Moreover, the crystallite size of WC prepared from H₂WO₄ was much smaller than that of commercial micron-sized WC powder produced from APT. The small crystallite size of WC induces grain growth during the sintering of the WC-Co composite; thus, a WC-Co composite with large WC grains was fabricated using the WC powder prepared from H₂WO₄. The large WC grains affected the mechanical properties of the WC-Co composite. Further, due to the large grain size, the WC-Co composite fabricated from H₂WO₄ exhibited a higher toughness than that of the WC-Co composite prepared from commercial WC powder.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.53-60
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• 2013

Objectives : DojukSan is known to be effective for treating a urinary diseases and stomatitis. However, there has been a lack of studies regarding the effects of Dojuksan on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. To elucidate the molecular mechanisms of Dojuksan water extract (DJS) on pharmacological and biochemical actions in inflammation, we examined the effect of DJS on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of MAPKs. Cells were treated with 200 ng/mL of LPS 1 h prior to the addition of DJS. Cell viability was measured by MTS assay. The investigation focused on whether DJS inhibited nitric oxide (NO) and prostaglandin E2 ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Results : We found that DJS inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, DJS suppressed the LPS-induced phosphorylation of p38 MAPK and c-Jun NH2-protein kinase (JNK). Conclusions : These results suggest that DJS has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.