• Development and Reproduction
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• v.7 no.2
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• pp.105-112
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• 2003

Previously, we discovered a new MMP-2 isoform GA110, of which appearance in human follicular fluid(FF) and serum was increased by EDTA. The present study was conducted to investigate how GAI 10 can appear by EDTA. To examine possible involvement of protein disulfide isomerase(PDI), an enzyme responsible for the dimerization of protein via disulfide formation, effect of PDI inhibitor on the appearance of GA110 by EDTA was investigated. When PDI inhibitor added to FF before EDTA treatment, the gelatinolytic activity of GA110 was abolished in a concentration dependent manner. By contrast, the activity of 72 kDa gelatinase increased. However, the PDI inhibitor added to FF after EDTA treatment, the gelatinolytic activity of GA110 was unaffected. To find out the nature of the enzyme which converts 72 kDa gelatinase into GAI 10, chromatographic separation method of FF proteins was done. Using hydroxyapatite column, fractions rich in 72 kDa gelatinase were isolated and pooled. By using this pool as substrate for the 72 kDa converting enzyme, protein fractions containing the converting activity were obtained from chromatographic separation of FF onto glutathione sepharose fast flow column. When immunoblotting was performed on this enzymatically active protein fractions against polyclonal anti-PDI antibody, distinct immunoreactivity was observed, although appeared in smaller molecular weight region. Based on these observations, it is suggested that the appearance of GAI 10 in FF by EDTA treatment could be due to an activation of PDI-like enzyme, which dimerizes 72 kDa gelatinase into GAI 10 via the formation of disulfide bond between molecules.

• Korean Journal of Weed Science
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• v.32 no.3
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• pp.211-221
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• 2012

The fourth leaves (younger leaves) amongst extended 4-upper leaves in 18 squash cultivar were the highest tolerance to the paraquat application, followed by third, the second, and the first leaves (older leaves). The forth leaves in Joongangaehobak showed more than three times higher tolerance to the paraquat application than did the first leaves. When the combining of water extract from the fourth leaves with paraquat were applied to the leaves and stems of maize, the paraquat phytotoxicity in maize was reduced compared to the paraquat application alone. Therefore, this study continued to investigate if the phytotoxicity inhibitor exist in the fourth leaves. The water extract in the fourth leaves were isolated by silica gel column chromatography, Sephadex LH-20 column chromatography, TLC, and HPLC, and the substance in the extract was speculated as a malic acid by identifying through NMR. The mixture malic acid and paraquat were applied to the maize to verify the application effect of malic acid on paraquat toxicity. The 100 ${\mu}M$ of paraquat application alone showed 62% of paraquat toxicity to the corn leaves, while the combined application of 100 ${\mu}M$ paraquat with malic acid at 0.1, 0.3, 0.5, and 1.0% did not show the symptom.

• Journal of the Korea Concrete Institute
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• v.29 no.2
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• pp.209-216
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• 2017

One of the important considerations in structural designing the flat plate system is ensuring the resistance to punching shear caused by axial loads and the ductile ability to follow horizontal deformation under earthquake. In this study, the ECC (Engineered Cementitious Composite) has been placed in the critical section zone of punching shear at slab-column joint to improve ductility and the advanced details of shear reinforced area nearby critical section zone has been developed using stud and steel fiber. The shear performance tests were performed on the specimens with parameters of fiber type mixed with ECC, stud and steel fiber set into the shear reinforced area in which the failure pattern, joint strength, displacement and strain of the specimen were compared and analyzed. The test results showed that the strength and ductility of specimens with ECC applied to joint were better than those of RC flat plate system. Also, the shear reinforcement effect of stud and the ductility improvement of steel fiber concrete were confirmed in the shear reinforcement area.

• ALGAE
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• v.31 no.3
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• pp.277-287
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• 2016

Five fractions separated from Nannochloropsis oculata using solvent-solvent partition chromatography of 80% methanolic extract of N. oculata (NOM) followed by the open silica column chromatography of its hexane fraction (NOMH) for the anti-inflammatory on RAW 264.7 cells and anti-cancer activities on HL-60, A-549, HEP-3B, HCT-116, and SW-480 cancer cells. All the five fractions showed potential anti-inflammatory activities against lipopolysaccharide-stimulated RAW 264.7 macrophages cells with IC₅₀ values less than 6.25 μg mL⁻¹. Moreover, 90% n-hexane column elution of NOMH (NOMH90) down-regulated lipopolysaccharide-stimulated protein levels of inducible nitric oxide synthase and cyclooxygenase-2. Furthermore, NOMH90 showed marked cytotoxic effect on the HL-60 cells with IC₅₀ value of 23.58 ± 0.09 μg mL⁻¹. In addition, Hoechst 33342 cell permeable dye used to visualize the apoptosis nucleus and cell cycle analysis measured Sub-G1 DNA contents to confirm reduction of the cell viability in NOMH90 treated cells due to induction of apoptosis in HL60. These results are quite related to the phytosterol contents of the NOMH fractions and the results suggest N. oculata extracts might be useful as potential sources of natural anti-inflammatory and anti-cancer compounds. In conclusion, the sterol content in N. oculata might provide a promising role in future medicines in anti-inflammatory and anti-cancer.

• Journal of Korean Society of Forest Science
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• v.87 no.2
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• pp.260-269
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• 1998

This study was conducted to screen the biological activity of urushiol in the sap of lac tree(Rhus verniciflua STOKES) which has been used in traditional folk remedies. Cytotoxic activity of urushiol was screened with L1210(mouse luekemia cell), PC-9(human lung adenocarcinoma cell), A427(human lung adenocarcinoma cell) and KATO III (human stomach adenocarcinoma cell) The stepwise hexane : acetone eluent fractions of the urushiol were obtained by the silica gel adsorption column chromatography and added to the culture media containing L1210, PC-9. A427, and KATO III, respectively. A hexane : acetone(90 : 10, v/v) eluent fraction of them showed the lowest 50% inhibition concentration($IC_{50}$) of $0.018{\mu}g/m{\ell}$ for the cell line of A427. Much lower level of $IC_{50}$ of the hexane : acetone(90 : 10, v/v) eluent fraction of the urushiol showed the equal inhibition effect with tetraplatin(i.e., anti-cancer drug of platinum complexes) on the cancer cell lines as follows ; 3.4 times lower for L1210, 3.9 times lower for PC-9, and 105.5 times lower for A427. However, $IC_{50}$ of the hexane : acetone(90 : 10 v/v) eluent fraction for KATO III was exceptionally 3.9 times higher than that of tetraplatin.

• Research in Plant Disease
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• v.15 no.3
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• pp.222-229
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• 2009

A Pseudomonas strain SG3 producing biosurfactant and showing antifungal and insecticidal activities was isolated from agricultural soil severely contaminated with machine oils. The antagonistic bacterium inhibited mycelial growth of all of the tested fungal pathogens. The fermentation broth of SG3 also effectively suppressed the development of various plant diseases including rice blast, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew and red pepper anthracnose. An antifungal substance was isolated from the fermentation broth of SG3 by ethyl acetate partitioning, silica gel column chromatography and preparative HPLC under the guide of bioassay. The chemical structure of the antifungal substance was determined to be rhamnolipid B by mass and NMR spectral analyses. The antifungal biosurfactant showed a potent in vivo antifungal activity against gray mold and late blight on tomato plants. In addition, rhamnolipid B inhibited mycelial growth of B. cinerea causing tomato gray mold and zoospore germination and mycelial growth of P. infestans causing tomato late blight. Pseudomonas sp. SG3 producing rhamnolipid B could be used as a new biocontrol agent for the control of plant diseases occurring on tomato plants.

• Korean Journal of Pharmacognosy
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• v.36 no.2 s.141
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• pp.129-135
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• 2005

Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

• Korean Journal of Food Science and Technology
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• v.36 no.6
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• pp.1001-1007
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• 2004

Chloroform layer from methanol extract of Corni fructus (Cornaceae) showed strong antiproliferation effect on human cancer cell lines by SRB assay. Anticarcinogenic-active compound was isolated and purified by silica gel column and thin layer chromatograpies, and identified as ursolic acid ($3{\beta}$-hydroxyrus-12-ene-28-oic acid, MW:456) by mass and IR spectrophotometries, and $^1H-and\;^{13}C-NMRs$. The compound inhibited proliferation of A549 (human lung cancer cell line) and MCF-7 (human breast cancer cell line) cells in dose-dependant manner when treated for 48 hr. Inhibition rates of both cells were over 40% and 90% compared with control cells at the $30\;{\mu}g/mL\;and\;100\;{\mu}g/mL$, respectively. Morphology of cells treated with the compound for 15 hr at $10\;{\mu}g/mL$ was distorted with shrinked cell mass, and cell number was lower than that of control cells. Cell cycle analysis showed sub-G1 phase arrest in both cell lines following 15 hr exposure to the compound; % of cell phase increased to 11.7 and 11.2% compared to the control of 4.0% and 2.1% in A549 and MCP-7 cells, respectively.

• Journal of Applied Biological Chemistry
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• v.54 no.3
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• pp.159-165
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• 2011

Rosmarinus officinalis extracts had a significant antimicrobial activity against Helicobacter pyori. Total phenolic contents and inhibition zone of rosemary extracts were estimated to be 25.7 mg/g and 14 mm at $200{\mu}g/mL$ of phenolic contents, respectively. The Sephadex LH-20 and MCI-gel CHP-20 column chromatographic separations for the phenolic extracts from R. officinalis leaves led to isolation of five acids, whose structures were determined as protocatechuic acid (A), coumaric acid (B), caffeic acid (C), chlorogenic acid (D), and rosmarinic acid (E), from interpretation of spectroscopic data including nagative fast atom bombardment (FAB)-mass, $^1H$-NMR, $^{13}C$-NMR, and IR. All isolated compounds were tested for antimicrobial activity against H. pyori. The purified single compound showed less antimicrobial activity against H. pylori than the mixed purified compounds, which generate A+B, A+E, C+D, C+E (each $200{\mu}g/disc$) excellent as large clear zone by synergy effect. These results indicate rosemary extracts are preventive agents against H. pyori.

• Korean Journal of Medicinal Crop Science
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• v.24 no.5
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• pp.401-407
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• 2016

Background: Salvia has been widely cultivated for use in flavoring and folk medicines in many countries, including Korea and China. In this study, we investigated the anti-inflammatory activity and the underlying active compounds of Salvia extract and its fractions. Methods and Results: The anti-inflammatory activity was measured by assessing the inhibition of cysteinyl leukotriene production in rat basophilic leukemia (RBL)-2H3 mast cells. Salvia plebeia R. Br. was found to have the most potent inhibitory activity on leukotriene production than S. japonica and S. chanroenica had. Fifty percent ethanol extracts of S. plebeia R. Br. were successively partitioned with n-hexane, methylene chloride, ethyl acetate, 1-butanol and water. The ethyl acetate (EtOAc) fraction showed stronger anti-inflammatory activity than other solvent fractions did. The EtOAc fraction was subjected to silica gel column chromatography elution with a chloroform and methanol gradient system (100 : 1 ${\rightarrow}$ 1 : 1) yielding 10 fractions. Three kinds of fractions (chloroform:methanol = 20 : 1, 10 : 1 and 5 : 1) showed high inhibitory activity on leukotriene production. We confirmed the major compounds with anti-inflammatory activity from S. plebeia R. Br. Conclusions: In this study, the major components of S. plebeia that showed leukotriene production inhibitory activity were isolated using solvent extraction and silica gel column chromatography. Rosmarinic acid, hispidulin and luteolin were identified as the major compounds with anti-inflammatory effect.