• The Korean Journal of Mycology
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• v.6 no.2
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• pp.1-10
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• 1978

In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

• Journal of Dairy Science and Biotechnology
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• v.27 no.2
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• pp.7-16
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• 2009

Appenzeller cheese samples were prepared by addition of 0.5, 1.0, and 2.0% green tea (Camellia sinensis, CS) powder and control cheese. We examined various quality characteristics of the novel cheese, such as viable-cell counts, pH, water-soluble nitrogen (WSN), non-casein nitrogen (NCN), non-protein nitrogen (NPN), and catechin level during maturation for 16 weeks at $14^{\circ}C$. To develop a Korean natural cheese containing green tea powder, we also analyzed the changes in the polyacrylamide gel electrophoresis pattern, chemical composition, and sensory qualities. The viable cell counts of the samples were not significantly different. Until the $3^{rd}$ week, the pH of the CS cheese decreased with an increase in the maturation time. However, the pH gradually increased by the $12^{th}$ week, while WSN, NCN, NPN also increased. The WSN, NCN, NPN, and catechin values for the CS cheese samples were significantly higher than the values for the control cheese. The polyacrylamide gel electrophoretic pattern of caseins for the CS cheese indicated that this cheese degraded more rapidly than the control cheese did. In the sensory evaluation, cheese with 1.0% CS powder showed the highest scores in taste and appearance and good scores in flavor and texture. These results indicate that 1.0% CS is the optimal value for addition to cheese, and cheese containing 1.0% CS shows good physiological properties and reasonably high overall sensory acceptability.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.21-34
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• 1982

One hundred and twenty-one strains each of Escherichia coli isolated from stools of 60 patients who received various antimicrobial drugs in hospital for more than one week and apparently healthy 60 students who have no history of taking antimicrobial drugs during recent one month, were tested for their resistance to 13 antimicrobial drugs. The frequency of resistance strains was highest to tetracycline with 69.2%, and followed by streptomycin(Sm), sulfisomidine(Su), chloramphenicol(Cm), ampicillin(Ap), and carbenicillin(Cb) in the decreasing order, ranging from 61.2% to 39.3%. Strains resistant to kanamycin(Km), cephaloridine(Cr), and trimethoprim(Tp) occupied about one-fourth of strains, and only four strains were resistant either one or more of nalidixic acid, gentamicin and amikacin, and no strain was resistant to rifampicin. The frequency of resistant strains to Cm, Ap, Km, Cr, and Cb was much higher among patient isolates than student strains, but strains resistant to the other drugs showed almost the same frequencies between patient and student isolates. There was a marked difference in average minimum inhibitory concentrations of between resistant and susceptible strains, suggesting that the resistance to drugs is the plasmid origin. Seventy-six percent of strains were resistant to one to 10 drugs tested, and no much difference was observed between strains from patients and students. However, strains resistant to four or more drugs were much more frequently found among patient isolates than student strains, with the increasing tendency of multiply resistant strains among patient isolates following the increase in the number of resistant drugs. The transfer of drug resistance by conjugation was tested and 98 strains(67.5%) among 145 which were resistant to two or more drugs were found to transfer their drug resistance to E. coli. Among 74 strains resistant to 7 or more drugs, all except one transferred the resistance, and the number of strains with transferable resistance decreased, as the number of resistant drugs decrease. A R plasmid from randomly selected p13 strain was tested for the incompatibility group, and the plasmid was classified into Inc F II. R plasmM DNA bands were identified by polyacrylamide gel electrophoresis.

• Korean Journal of Weed Science
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• v.17 no.1
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• pp.31-43
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• 1997

This study was carried out to investigate classfication of weedy rice (Oryza sativa) based on isozymes esterase and peroxidase, growth and developmental difference of weedy rices and rices grown under dry and water condition, and weedy rice control and tolerant difference of weedy rices in various herbicides using weedy rices collected from thirteen strains of Chonnam, one Chonbuk, two Kyeongki and two rice cultivars. 1. The collected weedy rices were classified into three groups based on isozyme esterase and peroxidase using polyacrylamide gel electrophoresis(PAGE) method. The classified groups were not same each other. 2. Plant height was taller in collected weedy rices than rice cultivars at 18 days after seeding under dry and water conditions, but number of leaves, shoot fresh weight, root fresh weight and root length were not significantly different between collected weedy rices and rice cultivars. In addition, growths of collected weedy rices were greater in dry- than water-condition. 3. After thiobencarb(S-4-chlorobenzyl diethythiocarbamate), molinate(S-ethyl hexahydro-1H-azepine-1-carbothioate) and oxadiazon(5-tert-butyl-3(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one) were applied at 6 days before seeding, the weedy rices controlled 100% by thiobencarb at 2.1kg ai/ha and 024kg ai/ha oxadiazon treatment but controlled 26% to 67% by molinate at 6.5kg ai/ha. Rice due to the herbicides was injured severely(25% to 100%) in flood condition at time of rice seeding after oxadiazon at 0.48kg ai/ha and 2.1kg ai/ha thiobencarb application, except for molinate which injured rice slightly(4% to 13%) in drain condition. The collected weedy rices to all experimented herbicides showed slight intraspecific variations. The intraspecific variations of weedy rices decreased in the order of thiobencarb>molinate>oxadiazon.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.79-88
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• 1998

This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1214-1221
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• 1998

The study was carried out to investigate the antioxidant effects of ethanol, methanol, and water extracts or fractions of ethanol extract of Ligularia fischeri on low density lipoprotein(LDL) and ethanol extract was further fractionated. The methanol extract and ethyl acetate fraction showed strong antioxidant effect with 71.7% (13.36 nmol/mg) and 95% (1.35 nmol/mg) inhibition in the presence of $15\;{\mu}g/mL$, respectively. In the value of malondialdehyde(MDA) with oxidation time, ethanol, methanol and water extract in the presence of $25\;{\mu}g/mL$ inhibited the oxidation up to 4h incubation and ethyl acetate fraction showed strong antioxidant effect up to 8h incubation. Also, the ethanol, methanol and water extract showed antioxidant effects in the agarose gel electrophoresis test. The conjugated diene formation by lipid oxidation with addition of $Cu^{2+}$ in the Ligularia fischeri extracts and their fractions was decreased approximately 1.1 to 2.8 times and 2.2 to 3.2 times, respectively compared to control. In the degradation of apo B-100 by oxidation using SDS-PAGE, ethanol, methanol and water extract showed similar degradation band pattern compared to that of native LDL band. Apo B-100 contents using densitometer were 77.8, 92.5% and 82.3% in the ethanol, methanol and water extract, respectively, compared to 100% of native LDL. In the meanwhile, apo B-100 contents in hexane, ethyl acetate and water fraction were 38.8, 94.5 and 65.5% respectively. This results indicated that ethyl acetate fraction showed the strongest antioxidant effect on MDA value or apo B-100 contents of LDL.

• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.716-723
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• 1995

The fucoidan purified from Korean brown seaweed, Ecklonia stolonifera was characterized on molecular structure and blood anticoagulant activities. Extraction was conducted at $100^{\circ}C$ with water and repeated twice. The crude fucodian was 151.1g out of 20.0 kg of Ecklonia stolonifera. The Fucoidan-1, which was purified from crude fucoidan using calcium chloride and cetyl pyridium chloride (CPC), was 35.2% against crude fucoidan. Fucoidan-5 was obtained approximately 28.1% from Fucoidan-1 through DEAE-Toyopearl 650 M ion-exchange column chromatography and showed one band by cellulose acetate electrophoresis. The molecular weight of Fucoidan-5 was estimated to be about 21,000∼23,000 dalton by Sephacryl S-300 gel filtration chromatography. Fucoidan-5 consists of 35.7% of fucose and 4.3% of galactose and the molar ratio of fucose and sulfate was about one to one. IR spectrum of Fucoidan-5 showed absorption at $1240\;cm^{-1}\;and\;850\;cm^{-1}$ and specific rotation value, $[\alpha]$, was $[\alpha]$. These results suggests that the sulfate maybe bind at $C_{4}$ carbon on ${\alpha}-L-fucose$. Gas chromatograph of methyl alditol acetate revealed that Fucoidan-5 is a fucose containing sulfated polysaccharide with $({\alpha}l-2)\;or\;({\alpha}l-2)$ glycosidic linkage. Anti-thrombin activity of the Fucoidan-5 was estimated as 1.4 time stronger than heparin. From above results, the purification methods using CPC and ion exchange chromatography is effective tools for obtaining highly purified fucoidan from Gompi, Ecklonia stolonifera.

• Journal of Life Science
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• v.19 no.11
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• pp.1581-1590
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• 2009

Hizikia fusiforme is a kind of brown edible seaweed that mainly grows in the temperate seaside areas of the northwest pacific, including Korea, Japan and China, and has been widely used as a health food for hundreds of years. Recently, H. fusiforme has been known to exert pharmacological activities including antioxidant, antimutagenic and anticoagulant activities. However, the molecular mechanisms of H. fusiforme in malignant cells have not been clearly elucidated yet. In this study, the effects of ethyl alcohol extract of H. fusiforme (EAHF) on the anti-proliferative effects of MDA-MB-231 and MCF-7 human breast cancer cells were investigated. EAHF treatment resulted in a concentration-dependent growth inhibition by including apoptosis in MDA-MB-231 cells and G1 phase arrest in MCF-7 cells, which could be proved by MTT assay, DAPI staining, agarose gel electrophoresis and flow cytometry analysis. In MDA-MB-231 cells, the increase in apoptosis induced by EAHF treatment correlated with up-regulation of pro-apoptotic Bax expression. EAHF treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant inhibition of poly (ADP-ribose) polymerase, $\beta$-catenin, phospholipase-${\gamma}1$ protein and DNA fragmentation factor 45/inhibitor of caspase-activated DNase. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. fusiforme.

• Journal of Life Science
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• v.20 no.3
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• pp.416-423
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• 2010

In this study, the properties and gene expression of the lactate dehydrogenase (EC 1.1.1.27, LDH) isozyme were studied in angelfish (Pterophyllum scalare) - known for their adaptation to the low oxygen environment of the tropics - which were acclimated to acute temperature change ($27{\pm}0.5{\rightarrow}18{\pm}0.5^{\circ}C$) and dissolved oxygen (DO) change ($6{\pm}1{\rightarrow}18\;ppm$) for 2 hours. The properties of the LDH isozymes were confirmed in the native-polyacrylamide gel electrophoresis, Western blot analysis and enzyme activity measurement. Liver- and eye-specific Ldh-C gene were expressed in liver, eye and brain tissues. Through Western blot analysis, the LDH $A_4$ isozyme was shown to have a more cathodal mobility relative to the $B_4$ isozyme. In the liver tissue, the LDH $A_4$ isozyme increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $C_4$ isozymes increased with DO increment, while the $B_4$ isozyme decreased. In the eye tissue, the LDH $A_4$ and B4 isozymse increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $B_4$ isozymes increased with DO increment, but the $C_4$ isozyme and isozymes including the subunit C decreased. In the heart tissue, LDH activity increased with DO increment, as well as the LDH $B_4$ isozyme. In the brain tissue, the LDH $A_4$ and $B_4$ isozymes increased with temperature drop. The LDH $B_4$ isozyme increased with DO increment. Accordingly, since the liver- and eye-specific Ldh-C are influenced by changes in DO and the LDH $B_4$ and $C_4$ isozymes are relatively controlled in the liver and eye tissues, the $C_4$ isozyme can be considered to have a lactate oxidase function.