• Journal of Life Science
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• v.24 no.10
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• pp.1078-1084
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• 2014

Here, we showed that Acanthocoris sordidus extract inhibited both cell and DNA damage caused by oxidative stress. In a radical scavenging assay, the scavenging activity of the A. sordidus extract against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals was 48.9% and 37.8%, respectively, that of ascorbic acid, which was used as a positive control. The ferrous iron chelating activity of the A. sordidus extract was 80.0% compared to that when ethylenediaminetetraacetic acid (EDTA) was used a control. To verify the inhibitory effect of the extract on oxidative cell damage induced by reactive oxygen species (ROS), a lipid peroxidation assay was performed. The results showed that peroxidation was completely inhibited in an extract-treated group compared to a radical-treated group. The level of p21 protein expression was 68.1% that of a control sample. The DNA cleavage-inhibiting property of the A. sordidus extract-treated group was 53.3% that of a control group. Moreover, the phosphorylation of the H2AX protein was reduced to 39.0% of that treated with radical agents, indicating that the extract might inhibit the DNA damage that causes radical oxidation. Taken together, our findings suggest that the A. sordidus extract is effective not only in repressing oxidation by free oxygen radicals and hydroxyl radicals but also in decreasing cell and DNA damage caused by oxidative stress.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.318-325
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• 1993

The optima] conditions for the formation, the regeneration. and the spheroplast fusion of Flavimonas aryz/habitans spheroplasts were investigated. Cells were transformed to spherop]asts effectively by treatment of 0.5% volume (v/v) of 0.] M EDTA and ]00 flg/ml lysozyme at $37^{\circ}C$ for 30 min without shaking. Magnesium chloride and calcium chloride were effective on the stabilization of spheroplasts. and 20 mM calcium chloride in the rich regeneration medium improve the yield of regenerants as much as 3.5-fo]d. Addition of 0.8% bovine serium albumine (BSA) in dilution buffer for spheroplast formation improved the stabilization of spheroplasts over extended periods (4-6 hr) at room temperature. and thus increased the yield of recombinants to 4.5-fold. The spheroplast formation frequency and regeneration frequency of F aryzihabitans strain was 90.10% and 3.800/." respectively. The first regenerated cell of F. aryzihabitans spheroplasts were appeared 6 hours after plating. By I I hours after plating, 80% of spheroplasts were regenerated on thc rich regeneration medium containing 0.5 M sucrose. The intraspeci11c spheroplast fusion of F urvz/habitans was carried out and the properties of obtained fusants were investigated. Formation of fusion products was effective when the Flav/munas spheroplast mixture was treated with 40%(w/v) PEG6000 and 20 mM CaCl, for 10 min at room temperature. and thc formation of frequency of recombinants were $2.0{\times}10^{-5}~3.6{\times}10^{-5}$. All tested recombinant clones were very stable on further propagation.

• Journal of Life Science
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• v.23 no.1
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• pp.31-37
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• 2013

The present study aimed to investigate effects of ethanol extracts from Hydropsyche kozhantschikovi on cell and DNA damage caused by oxidative stress. In a radical scavenging assay, compared with ascorbic acid used as a control, the level of DPPH (1,1-diphenyl-2-picrylhydrazyl) and that of hydroxyl radicals in H. kozhantschikovi extracts were 60.0% and 43.7%, respectively. The ferrous iron chelating level was 37.5% compared to the chelating value of EDTA (ethylenediaminetetraacetic acid) as a positive control at the same concentration. To verify inhibitory effects of oxidative cell damage induced by reactive oxygen species (ROS), the relative level of lipid peroxidation and the expression level of the p21 protein were compared in extracts-treated and untreated groups. Lipid peroxidation was completely inhibited in the extracts-treated group compared with the radical-only treated group. The level of p21 protein expression was restored to 92.2% of p21 protein expression in the control sample. In addition, DNA cleavage inhibition in the H. kozhantschikovi extracts was 74.1% compared with that of the control group, suggesting that H. kozhantschikovi extracts repress DNA cleavage induced by ROS. Moreover, the phosphorylation ratio of the H2AX protein was 16.7% in the radical-treated group, indicating that the ethanol extracts inhibited 83.3% of DNA damage. Our findings suggest that ethanol extracts from H. kozhantschikovi are effective not only in repressing the oxidation of free radicals and highly toxic hydroxyl radicals, but also in decreasing cell and DNA damage caused by oxidative stress.

• Applied Biological Chemistry
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• v.34 no.3
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• pp.225-230
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• 1991

In this study, several combined effect of KCl addition into bring solution, microwave heating of Chinese radish for 3 minutes and addition of two salt mixtures into half fermented Kakdugi were investigated to compare their improving effects of storage stability of Kakdugi, Kakdugi was prepared by salting in 15% NaCl solution for 2 hours and fermented at $25^{\circ}C$ and determined for the changed in pH, total acidity and reducing sugars. It was found from the result that the combined method of salts addition and heating treatment improved the storage stability by 6.4 folds based on pH change where salts mixture of phosphated, Ca EDTA, and Sod. citrate contributed the most effect. Changed in reducing sugar were greatly reduced by salt mixture addition while microwave heating and KCl addition affected little. Storage of Kakdugi at $4^{\circ}C$ after preparation with combined method could extend the storage life more than 8 folds of those Kakdugi stored at $25^{\circ}C$. However total acidity showed higher values for those prepared by combined method and stored at $4^{\circ}C\;or\;25^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.39 no.2
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• pp.209-216
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• 2007

The present study investigated anti-oxidative and anti-inflammatory activity of glycoprotein isolated from Morus Indica Linne (MIL glycoprotein). We found that MIL glycoprotein has a molecular weight of 32 kD and consists of carbohydrate (40.03%) and protein (59.97%), and that it has a strong scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical $({\cdot}OH)$, and superoxide anion $(O_2{\cdot}\;^-)$ radicals. In addition, MIL glycoprotein had a stable character and an optimal DPPH radical scavenging activity in the alkaline and neutral pH solution, and up to at 105. However, the results indicated that it has a minimal scavenging activity in the metal ionic solution ($Ca^{2+}$, $Mn^{2+}$, and $Mg^{2+}$) in the presence of EDTA. In addition, we further investigated whether MIL glycoprotein scavenges oxygen radicals and blocks inflammation-related signals in the bisphenol A (BPA)-stimulated Raw 264.7 cells. The results in this study showed that it has a character to scavenge the productions of reactive oxygen species (ROS) and nitric oxide (NO) dose-dependently. Also it blocked the activities of inflammation-related signals such as nuclear factor-kappa B ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS). For example, it had an inhibitory effect on the activation of $NF-{\kappa}B$ (p50) and iNOS proteins at 200 ${\mu}g/mL$ MIL glycoprotein. Here, we speculate that MIL glycoprotein is one of natural antioxidants and of modulators of the BPA-induced inflammation.

• Journal of Soil and Groundwater Environment
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• v.14 no.2
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• pp.26-32
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• 2009

In this study, the degradation of BTEX (benzene, toluene, ethylbenzene, xylene) was tested in both (Fe$^{3+}$+chelating agent)/H$_2$O$_2$system [Fe(III) 1 mM, oxalate 6 mM, H$_2$O$_2$ 3%, and pH 6] and UV/(Fe3++ chelating agent)lHzOz system [UV dose 17.4 kWhlL, Fe(III) 1mM, oxalate 6 mM,H$_2$O$_2$ 1%, and pH 6]. The types of chelating agents used in experiments were catechol, NTA, gallic, acetyl acetone, succinic, acetate, EDTA, citrate, malonate, and oxalate and the optimum chelating agent for BTEX degradation was determined. The results showed that acetate was the optimum chelating agent for BTEX degradation in both (Fe$^{3+}$+chelating agent)/H$_2$O$_2$ and UV/(Fe$^{3+}$+chelating agent)/H$_2$O$_2$ system, and UV radiation enhanced the degradation of BTEX with any types of chelating agents. Moreover, UV/(Fe$^{3+}$+chelating agent)/H$_2$O$_2$ system, which chelating agent was acetate, removed effectively mixtures of BTEX and MTBE (methyl tert-butyl ether) when the concentration of both BTEX and MTBE was 200 mg/L, respectively. In this system, BTEX was degraded completely and 85% of MTBE was degraded at the reaction time of 180 min. Therefore, UV/((Fe$^{3+}$+chelating agent)/H$_2$O$_2$ system with acetate as a chelating agent removed not only BTEX but also BTEX and MTBE, effectively.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.225-236
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• 2012

In this study, the cellular protective effects on HaCaT cells and human erythrocytes and antioxidative effects of P. tricuspidata stem extracts were investigated. The ethyl acetate ($50{\mu}g/mL$) and aglycone fraction ($25{\mu}g/mL$) of P. tricuspidata stem extracts doesn't show any characteristics of cytotoxicity. When HaCaT cells were treated with 10 mM $H_2O_2$ and $30{\mu}M$ rose bengal, the ethyl acetate ($6.25{\sim}50{\mu}g/mL$) and aglycone ($6.25{\sim}25{\mu}g/mL$) fraction protected the cells against the oxidative damage in a concentration dependent manner. The P. tricuspidata stem extracts showed more prominent cellular protective effect than (+)-${\alpha}$-tocopherol, known as lipid antioxidant at $10{\mu}g/mL$. The ethylacetate fraction of P. tricuspidata stem extracts ($18.5{\mu}g/mL$) showed more free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC5_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. tricuspidata stem extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate ($1.72{\mu}g/mL$) and the aglycone fraction ($1.53{\mu}g/mL$) showed similar ROS scavenging activity of L-ascorbic acid ($1.50{\mu}g/mL$). These results indicate that extract/fractions of P. tricuspidata stem extracts can function as natural cytoprotective agents and antioxidants in biological systems, particularly skin exposed to UV radiation by protecting cellular membrane against ROS.

• Clinical and Experimental Pediatrics
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• v.45 no.10
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• pp.1263-1272
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• 2002

Purpose : Rett syndrome(RTT) is an X-linked dominant neurodevelopmental disorder affecting 1 per 10,000-15,000 female births worldwide. It was initially described by Andreas Rett in 1966. RTT involves developmental regression characterized stereotypic hand movements, tremors, gait apraxia, seizures, deceleration of head growth after the age of 6-18 months. The disease-causing gene was identified as MECP2 on chromosome Xq28. We carried out mutational analysis of MECP2 genes in RTT patients. Methods : Whole blood(5 cc) of 34 sporadic RTT patients was collected in EDTA-anticoagulated tubes. Genomic DNA was extracted from peripheral blood using the E.Z.N.A. blood DNA kit. Four exons of the MECP2 gene were amplified by PCR in 34 Korean with RTT. We carried out PCR divided the exon three into two parts and the exon four into five parts. Primer sequences designed by Amir et al. in 1999 were almost used(AF030876). Sequencing primers used were the same as PCR. DNA sequencing reactions were performed using an ABI 377 DNA sequencer and ABI PRISM dye terminator cycle sequencing reaction kit(Perkin-elmer). The results were compared with the normal DNA sequence(X99686). To confirm the change of sequence on novel mutations, RFLP analysis was performed. Results : The MECP2 mutations were detected in 23(67.6%) of the 34 patients. The mutations consisted of 12 different types including nine missense and three nonsense mutations. Of these, three (L100V, G161E and T311M) mutations were newly identified. Most of the mutations discovered are located within MBD(39.1%) and TRD(39.1%). In this study, three(T158M, R270X, R306C) mutations were identified high frequency. Conclusion : MECP2 gene was also an important cause of Korean RTT patients. MECP2 gene study is an important tool for diagnosis of Korean RTT patients.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.59-66
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• 2003

Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.3
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• pp.307-312
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• 1984

The dried pure shells deprived of soft tissues were subjected to analysis of chitin-protein complexes from 5 species of marine crustaceans, including 2 species of crabs and 3 species of shrimps. The protein fractions were obtained from chitin-protein complexes under the varying conditions of extractions and the crude chitin was prepared from the shells by the sulfurous acid process. The crude chitin was purified through the extraction with several organic solvents such as dimethyl-acetamide, N-methylpyrrolidone. The purified chitin was also examined using the phase contrast microscope. Total protein contents of the shells were diverse, showing 9.6% for Portunus trituberculatus, 3.1%, Charybdis bimaculata, 9.4%, Penaeus japonicus, 10.9%, Metapenaeus intermedius and 5.8%, Squilla oratoria. Covalently bound protein varied with species from 2.1% for Charybdis bimaculata to 9.9% for Metapenaeus intermedius. The puified chitin contents of the shells were shown to 21.1% for Portunus tritube rculatus, 6.2%, Charybdis bimaculata, 20.2%, Penaeus japonicus, 27.1%, Metapenaeus intermedius and 25.5%, Squilla oratoria. Exceptionally low analytical value obtained with Charybdis bimaculata are supposed to be due to the very young subjects. The ratios of chitin to covalently bound protein in the shells were various such as 2.7 to 1 for Portunus trituberculatus, Penaeus japonicus and Metapenaeus intermedius, 3.1 to 1, Charybdis bimaculata and 6.1 to 1, Squilla oratoria. The microscope finding of the purified chitin showed the filamentous form in all the specimen.