• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.637-650
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• 2006

Genomic DNA isolated from two crucian carp species obtained from Yesan (Carassius auratus) and Dangjin (Carassius cuvieri) in Korea were amplified at several times by polymerase chain reaction (PCR). The amplified products were separated by agarose gel electrophoresis (AGE) with oligonucleotides decamer primer and stained with ethidium bromide. The seven arbitrarily selected primers OPC-11, OPC-14, OPC-18, OPD- 02, OPD-11, OPD-15 and OPD-20 generated the shared loci by each species, the polymorphic and specific loci. The seven primers generated the total 458 loci that can be scored from the crucian carp obtained in C. auratus species. 358 fragments were generated from the species obtained in C. cuvieri species. The size of DNA fragments varies from 150 to 1,600bp. The complexity of the banding patterns varies dramatically between the primers and two locations. In this study, 458 loci were identified in the crucian carp species from Yesan and 358 in the crucian carp species from Dangjin: 84 polymorphic loci (18.3%) in the C. auratus species and 48 (13.4%) in the C. cuvieri species. 154 shared loci by each species, the average 22 per primer, were observed in the C. auratus species and 187 loci, the average 26.7 per primer, in the Dangjin species. Based on the average bandsharing (BS) values of all samples, the similarity matrix ranged from 0.434 to 0.868 in the C. auratus species and from 0.449 to 0.924 in the C. cuvieri species. The average BS value was 0.641±0.013 within the C. auratus species and 0.684±0.013 within the C. cuvieri species. The average BS value between two crucian carp species 0.484 ± 0.007, ranged from 0.307 to 0.682. The BS value between the individual No. 09 and No. 16 was 0.682, which was the highest between two crucian carp species. Compared separately, the BS value of individuals within the C. cuvieri species was higher than the C. auratus species. The dendrogram obtained by the seven primers, indicates three genetic clusters: cluster 1 (AURATUS No. 01, 02, 03, 04, 05, 06, 07, 08, 09, 10 and 11), cluster 2 (CUVIERI No. 12, 13, 14, 15, 16, 17, 18, 19, 20 and 21) and cluster 3 (CUVIERI no. 22). The shortest genetic distance displaying significant molecular difference was between the individual AURATUS No. 09 and AURATUS No. 08 from Yesan (genetic distance=0.064). The longest genetic distance displaying significant molecular differences was between the individual CUVIERI No. 17 and AURATUS No. 11 between two crucian carp species (0.477). RAPD-PCR analysis has revealed the significant genetic distance between two crucian carp species pairs.(Key words: Carassius auratus, Carassius cuvieri, Crucian Carp, DNA Polymorphism, Genetic Distance)

• The Korean Journal of Food And Nutrition
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• v.23 no.3
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• pp.352-360
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• 2010

This study was performed to develop a Korean natural cheese with traditional medical wine, making it different from foreign natural cheese. The effects of cheese with Sasam(Codonopsis lanceolate) wine(CLW) on the quality properties during the ripening period of natural cheese were investigated. The properties investigated were growth of lactic acid bacteria, characteristics of ripening, and sensory characteristics. Four vats of cheese were made on the same day from the same tank of fresh milk. Cheese samples were prepared with CLW at 2.0%, 4.0% and 6.0% of raw milk. Changes in gross composition, viable cell counts, pH, water soluble nitrogen(WSN), non casein nitrogen(NCN), non protein nitrogen(NPN), and proteolysis during maturation were measured. Polyacrylamide gel electrophoresis(PAGE) patterns were determined with control cheese. Viable cell counts of control and CLW cheese were not significantly different. The pH of CLW cheese increased gradually during maturation, and saponin levels and levels of NPN, NCN, and WSN were higher in CLW cheeses than control cheese. For most compositional data, the 4.0% CLW cheese was most similar to the control cheese. The PAGE pattern of cheese caseins indicated that the CLW cheeses degraded more rapidly than the control cheese. Control and 2.0% CLW cheese had good sensory scores, while scores for 4.0% and 6.0% CLW cheese were lower. However, sensory data depreciated with added levels of CLW, especially at a level of 4.0% or more. Further studies on levels of CLW and processing methods are required to improve sensory quality.

• Food Science of Animal Resources
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• v.24 no.4
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• pp.349-354
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• 2004

Stress-susceptible pigs have been known as the porcine stress syndrome (PSS), swine PSS, also known as malignant hyperthermia (MH), is characterized as sudden death and production of poor meat quality such as PSE (pale, soft and exudative) meat after slaughtering. PSS and PSE meat cause major economic losses in the pig industry. A point mutation in the gene coding for the ryanodine receptor (RYR1) in porcine skeletal muscle, also known calcium (Ca$^{2+}$) release channel, has been associated with swine PSS and halothane sensitivity. We used the PCR-RFLP(restriction fragment length polymorphism) and PCR-SSCP (single strand conformation polymorphism) methods to detect the PSS gene mutation (C1843T) in the RYR1 gene and to estimate genotype frequencies of PSS gene in Korean pig breed populations. In PCR-RFLP and SSCP analyses, three genotypes of homozygous normal (N/M), heterozygous carrier (N/n) and homozygous recessive mutant (n/n) were detected using agarose or polyacrylamide gel electrophoresis, respectively. The proportions of normal, carrier and PSS pigs were 57.1, 35.7 and 7.1％ for Landrace, 82.5, 15.8 and 1.7％ far L. Yorkshire, 95.2, 4.8 and 0.0％ for Duroc and 72.0, 22.7 and 5.3％ for Crossbreed. Consequently, DNA-based diagnosis for the identification of stress-susceptible pigs of PSS and pigs producing PSE meat is a powerful technique. Especially, PCR-SSCP method may be useful as a rapid, sensitive and inexpensive test for the large-scale screening of PSS genotypes and pigs with PSE meat in the pork industry.y.

• Food Science and Preservation
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• v.23 no.6
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• pp.811-818
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• 2016

The purpose of this study was to investigate changes in the amino acid content and physicochemical properties of Cheonggukjang prepared by using various soybean cultivars (Daewon, Deapung, Seadanbeak, and Taekwang) and a functional microorganism (Bacillus subtilis HJ18-9). These soybeans were conventional Cheonggukjang (control) and Cheonggukjang fermented with Bacillus subtilis HJ18-9 (treated). The moisture contents of steamed, control, and treated soybean were 62.45~67.12%, 63.28~67.14%, and 64.50~66.87%; amino-type nitrogen contents were 6.53~24.25 mg%, 27.63~122.09 mg%, and 37.29~133.48 mg%; and ammonia-type nitrogen contents were 26.92~47.95 mg%, 45.45~156.36 mg%, and 28.02~121.13 mg%, respectively. The umami taste associated with several amino acids (aspartic acid and glutamic acid) in Cheonggukjang was lower than that for steamed soybeans, while the bitter taste from amino acids (methionine, valine, isoleucine, leucine, and phenylalanine) was higher than that for steamed soybeans. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the molecular weight of steamed soybeans was less than 100 kDa, while control and treated groups showed low molecular weights below 34 kDa, confirming their protein hydrolysis to small molecular weight. These results are information for developing functional fermented soybean paste and diversification using soybean cultivars.

• The Journal of Korean Obstetrics and Gynecology
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• v.15 no.4
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• pp.45-60
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• 2002

Mouse calvarial osteoblast cells were isolated and cultured. To examine whether the cells produce active gelatinases in culture medium or not,the cells were analyzed using by zymograsphic analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase- A. Then, mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agent's, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of $IL-1{\alpha}$, indomethacin and dexametasone produced a rightward shift in the IL-1 dose response curve. The results of in vitro cytotoxicities showed that Taeyoungjon-Jahage water extracts(T.Y.J-J.H.G extracts) have no any cytotoxicities in concentrations of $1-200\;{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $300\;{\mu}g/ml$ on mouse calvarial bone cells. T.Y.J.-J.H.G. extracts had protective activity against PTH (2 units/mI), or MCM (5%, v/v), or $rhIL-1{\alpha}$ (1 ng/mI) or $1,25(OH)_2D3$ (10 ng/ml) , $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the T.Y.J.-J.H.G.extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore. the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resortion agents of PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ and $IL-1{\alpha}$, with strong protective effect in pretreatment with the extracts. T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1 stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by T.Y.J.-J.H.G. extracts treatment in the mouse calvarial tissue culture system. These results indicated that the T.Y.J.-J.H.G.-water extracts are highly stable and applicable to clinical uses in osteoporosis.

• Journal of Life Science
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• v.25 no.7
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• pp.826-832
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• 2015

This study describes the effects of activated charcoal on the removal of salts, detergents, and pigments from protein extracts of ginseng leaves and roots. Incubation of protein extracts with 5% (w/v) activated charcoal (100-400 mesh) for 30 min at 4℃ almost removed the salts and detergents including NP-40 as can be observed on SDS-PAGE. In addition, analysis of chlorophyll content showed significant depletion of chlorophyll (~33%) after activated charcoal treatment, suggesting potential effect of activated charcoal on removal of pigments too along with the salts and detergents. 2-DE analysis of activated charcoal treated protein samples showed better resolution of proteins, further indicating the efficacy of activated charcoal in clearing of protein samples. In case of root proteins, although not major differences were observed on SDS-PAGE, 2-DE gels showed better resolution of spots after charcoal treatment. In addition, both Hierarchical clustering (HCL) and Principle component analysis (PCA) clearly separated acetone sample from rest of the samples. Phenol and AC-phenol samples almost overlapped each other suggesting no major differences between these samples. Overall, these results showed that activated charcoal can be used in a simple manner to remove the salts, detergents and pigments from the protein extracts of various plant tissues.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.238-245
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• 1999

Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.4
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• pp.435-441
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• 2017

The present study was carried out to evaluate the applicability of Tenebrio molitor larvae (mealworm) as a health functional food material in order to contribute to the development of the domestic insect industry and health functional food industry. Protein hydrolysates were prepared from mealworm powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and the hydrolysates were then tested for their antioxidant activities. Based on available amino group contents and sodium dodecyl sulphate-polyacrylamide gel electrophoresis analyses, mealworms treated with alcalase ($4,781.39{\mu}g/mL$), flavourzyme ($5,429.35{\mu}g/mL$), or neutrase ($3,155.55{\mu}g/mL$) for 24 h showed high degree of hydrolysis (HD) value, whereas HD values of bromelain ($1,800{\mu}g/mL$) and papain-treated ($1,782.61{\mu}g/mL$) mealworms were much lower. Protein hydrolysates showing high HD values were further separated into > 3 kDa and ${\leq}3kDa$ fractions by a centrifugal filter system and then lyophilized, and the production yields of the low molecular weight protein hydrolysates (${\leq}3kDa$) by alcalase, flavourzyme, and neutrase were 42.05%, 26.27%, and 30.01%, respectively. According to the RC_{50} values of the protein hydrolysates (${\leq}3kDa$) obtained from three different antioxidant analyses, all three hydrolysates showed similar antioxidant activities. Thus, alcalase hydrolysates showing the highest production yield of low molecular weight protein hydrolysates were further tested for their inhibitory effects on peroxidation of linoleic acid by measuring thiobarbituric acid values, and the results show that peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation. However, pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited linoleic acid peroxidation in a dose-dependent manner over 6 days.

• Tuberculosis and Respiratory Diseases
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• v.43 no.6
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• pp.842-851
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• 1996

Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

• Food Science and Preservation
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• v.18 no.4
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• pp.604-611
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• 2011

Several wild yeasts were isolated from Korean grape varieties before and during spontaneous fermentation. Among them, four strains were isolated based on the alcohol content and flavor production in wine after fermentation of apple juice. In this study, the four yeast strains were identified and characterized. PCR-restriction fragment length polymorphism analysis of ITS I-5.8S-ITS II region with restriction endonuclease Hae III and Hinf I resulted in that all the strains showed a typical pattern of Saccharomyces cerevisiae. Pulse field gel electrophoresis showed three different chromosome patterns with a same band between strains SS89 and SS812. When ITS I-5.8S-ITS II sequences of the four strains were compared with one another, they were similar to those of Saccharomyces cerevisiae CBS 4054 type strain. Identity of the sequences was higher than 97% with those of the type strain. Phylogenetic analysis showed based on the sequences showed they were genetically closed to the type strain. The four identified strains were tested in a medium containing 200 ppm potassium metabisulfite, and the MM10 and WW108 inhibition rates resulted at up to 24 h. The four strains were tested at an incubation temperature of $30^{\circ}C$. The 30% sugar concentration in the medium (w/v) showed the highest growth in 36 h, especially in the case of SS89, which was close to growth 40. The four strains were tested in an 8% ethanol medium (v/v). Alcohol tolerance was initially kept in the incubation process. The strains began to adapt, however, to the exceeded resistance. The four strains showed the lowest inhibition rate at 24 h.