• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.395-406
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• 2000

The present study was conducted to examine the developmental capacity of mouse oocytes within prenatal follicles cultured various concentrations of FSH and LH and the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc) and cytochrome P450 17 $\alpha$ -hydroxylase (P450)$_{17{\alpha}}$ mRNA, as luteinization and atretic marker, in these culture conditions. In addition, we investigated the concentrations of progesterone and testosterone in culture medium. The developmental potential up to blastocyst of the oocytes grown in vitro was higher in the FSH alone (30.2%) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (28.0%) groups than in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (22.0%). And the mean numbers of cell per blastocyst was higher in the FSH alone (50.9$\pm$26.1) and 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated (51.0$\pm$21.1) groups when compared to the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group (45.2$\pm$15.1). The expressions of P450scc and P450$_{17{\alpha}}$ mRNA in the oocyte -cumulus complexes were increased with increasing of LH concentration, and also the secretions of progesterone and testosterone were increased. Especially, in the 100 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH treated group, the expression of P450scc and P450$_{17{\alpha}}$ were significantly increased, and the secretion of progesterone and testosterone were significantly increased. Therefore, these data show that gonadotrophins are essential for the in vitro culture of preantral follicles, but that increasing of LH concentration is reduced the developmental capacity of oocytes. The cause of these findings may be due to increasing of progesterone and testosterone secretion by the enhance of P450scc and P450$_{17{\alpha}}$ mRNA expressions, as markers of luteinization and atresia. Conclusively, this study suggest that supplementation of 100 $m\ell$U/$m\ell$ FSH or 10 $m\ell$U/$m\ell$ LH and 100 $m\ell$U/$m\ell$ FSH may be optimal condition for the culture of mouse pre antral follicles.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.74-83
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• 2016

In order to improve the sour taste and foul odor of fermented soymilk, bacteria were isolated from kimchi and identified. Of the 89 bacterial strains isolated from kimchi, 3 isolates produced fermented soymilk with a sour taste and foul odor. The selected bacterial strains R53, R83, and R84 were identified by morphological, biochemical, and 16S rRNA analyses as Weissella koreensis. The strain R83, which produced fermented soymilk having the mildest sour taste and foul odor, was selected for further investigation and named W. koreensis KO3. The optimum culture condition for the fermentation of soymilk by W. koreensis KO3 was at $30^{\circ}C$ for 12 h. When soymilk was fermented under the optimum culture conditions, the viable cell count reached up to $8.71{\times}10^8CFU/ml$ and pH and acidity reached as low as 6.02 and as high as 0.33%, respectively. Twenty-seven amino acids and their derivatives were detected in fermented soymilk. The amounts of serine, glycine, threonine, alanine, and aspartic acid, which contribute to a sweeter taste, increased during fermentation. Orinithine, which was not detected before fermentation, increased during fermentation. Sensory evaluation showed that W. koreensis KO3-fermented soymilk has improved bean, roasted nut, and sour flavors as well as an enhanced mouthfeel, appearance, preferability, and overall acceptability compared with those of standard fermented soymilk. With further study and development, soymilk fermented by W. koreensis KO3 could serve as a health-promoting food with favorable sensory qualities.

• Applied Biological Chemistry
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• v.48 no.2
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• pp.132-139
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• 2005

The major obstacle in the popularization of Chungkookjang is the short shelf-life of $2{\sim}3$ months and some problems concerning storage including the growth of molds even in the products even within shelf-life. To solve these problems we conducted a research to improve its storage by using the vacuumed packaging and sanitary method through seed culture, innoculation and sterilization. For the optimization of storage time, temperature and sterilization temperature, we measured viable cell numbers of bacteria and fungi, amount of gas outbreak and contents of amino type nitrogen and monitored these experimental results by response surface methodology of SAS program, so that we could observe the quality changes of Chungkookjang during shelf-life. Especially fungi, which are the biggest troublemaker in Chungkookjang shelf-life, couldn't be detected from the generally and vacuum-packed samples; also, viable cell numbers were highly influenced by sterilization temperature and in vacuum-packed samples. In the case of vacuum-packed samples, amount of gas outbreak was highly influenced by sterilization temperature of its storage conditions and it was higher in generally packed samples as compared to vacuum-packed samples even at any storage conditions. The changes of pH in generally and vacuum-packed samples were highly influenced by the storage temperature. As the temperatures of storage and sterilization were higher and the storage time was longer, so the amount of gas outbreak was accordingly lower. These results showed that amino type nitrogen contents in generally and vacuum-packed samples were systematically influenced by the temperature, storage time and sterilization temperature. Also the result showed that the change of amino type nitrogen contents during storage was less in vacuum-packed samples than in general ones. Based on the above results, we can produce Chungkookjang products with extended shelf-life of as far as 6 months without any quality change using sanitary manufacturing method, vacuumed packaging condition, sterilization in $70^{\circ}C$ for 60 minutes and storage under $10^{\circ}C$ during shelf-life. According to this research, we have the possibility to greatly increase the goods value of Chungkookjang by developing the manufacture processing and packaging.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.231-237
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• 2009

This paper aimed to develop a solvent extraction and purification process to recover high-purified ${\beta}$-carotene from recombinant Escherichia coli. Cells harvested from the culture broth were treated through numerous steps: dehydration, solvent extraction, crystal formation and separation. To optimize the extracting condition, experiments were carried out to investigate the effect of cell disruption, temperature, organic solvents, solvent-biomass ratio on the yield of ${\beta}$-carotene extracted from cells. The result indicated that no significant differences of extraction yield were observed from cells with or without step of cell disruption. Among different extracting solvents, the highest extraction yield of ${\beta}$-carotene, 30.3 mg-${\beta}$-carotene/g-dry cells, was obtained with isobutyl acetate at solvent-biomass ratio 25 mL/g-dry cells at $50^{\circ}C$. Notably, in case of acetone, the extraction yield was quite low when using acetone itself, but increased almost up to the highest value when combining this solvent and olive oil. The purity of ${\beta}$-carotene crystals obtained from crystallization and separation was 89%. The purity degree was further improved up to 98.5% by treating crude crystals with additional ethanol washing.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.31 no.4
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• pp.281-286
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• 2009

Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

• Journal of radiological science and technology
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• v.33 no.2
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• pp.143-148
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• 2010

This study is to analyze effects of the growth of Chunhyang Young Radish (CYR) and Altari Radish (AR) according to the exposure for 31 days at low dose ${\beta}$-rays. This test has one contrast sample and eleven test samples each as to AR and CYR. The seeds from contrast and test sample were planted in the culture soil after 8 seeds were chosen from each with identical condition. The accumulated dose of test samples has been measured at consistent time on a daily basis for 31 days. The growing process and germination have been measured twice at consistent time in each week. The number of leaves, length of first leave and weight have been acquired average value by measuring for 20 and 25 days, respectively after being planted. The result of test sample in case of 25 days shows that 5% increase in length and 36% increase in weight for AR each at accumulated dose 0.01 Gy compared to the contrast sample. And the length of CYR has increased by 13~17% and 1% at accumulated dose 0.01~0.08 Gy and 0.3 Gy compared to the contrast sample. For the weight at accumulated dose 0.05 Gy and 0.23 Gy has increased by 36% and 2% compared to contrast sample. As to the number of leaves, AR has increased by 0~50% at accumulated dose 0.01-0.32 Gy compared to contrast sample. It also shows that the CYR has increased to 0~67% at accumulated dose 0.01-0.62 Gy compared to contrast sample. As a result of this study, it indicates that both AR and CYR has generally increased in their length, weight, and the number of leaves at low level accumulated dose part 0.01~0.2 Gy. The size of cell, area of nucleus and density of cell for test sample has been observed quite similar to the ones from contrast sample through microscope. In conclusion, AR and CYR irradiated by ${\beta}$-rays have estimated that they are achieved a rapid growth at low level accumulated dose region corresponding to its radiation hormesis theory. Further studies need to confirm the correlation between the radiation hormesis and the growth of the plants.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.7
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• pp.926-934
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• 2006

The base for preparing oyster hydrolysate-added yogurt was consisted of whole milk (1,000 mL), skim milk (44.05 to 42.05 g), enzymatic oyster hydrolysates powder (OHP, 0 to 2.0 g) and pectin. The yogurt base was fermented with 7 kinds of starter cultures (3% based on yogurt volume), such as Lactobacillus acidophilus, lactobacillus bulgaricus, lactobacillus casei, Lactobacillus fermentum, Lactobacillus pentosus, Streptcoccus thermophilus and the mixed starters (L. bulgaricus and S. thermophilus) at optimal temperature. Processing condition and quality characteristics of the yogurt were evaluated by analyzing pH, titratable acidity, viscosity, viable cell count, functional properties and sensory evaluation. The results suggested that the optimal conditions for preparing the good quality yogurt revealed the mixed starters (L. bulgaricus and S. thermophilus) for starter culture, 1.0 g of 3 kDa hydrolysate for amount, and 5.5 hrs for fermentation time. The good quality yogurt showed 4.31 for pH, 1.07% for titratable acidity, 469 cps for viscosity and $4.9{\times}10^8\;CFU/mL$ for viable cell count. The hydrolysate-added yogurt was 2 times higher in ACE inhibitory and antioxidant activities than commercial yogurt, and kept good quality during storage of 15 days at $5^{\circ}C$.

• The Korean Journal of Pesticide Science
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• v.14 no.2
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• pp.148-156
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• 2010

S59-4 isolate was evaluated as a potential biocontrol agent using in vivo wounded garlic bulb assay. When the spore suspension ($10^5$ spores/$m\ell$) of Penicillium hirsutum was co-inoculated with cell suspension of S59-4 isolate on wounded garlics, the isolate showed high suppressive effect to disease development. The isolate was identified as Pantoea agglomerans S59-4(Pa59-4) through Biolog system. Furthermore, soaking garlic bulbs in the suspension of Pa59-4 significantly reduced garlic decay caused by P. hirsutum. The optimal concentration of Pa59-4 for controlling garlic blue mold was $10^7\sim10^8$ cfu/$m\ell$. And suppressive effect of Pa59-4 on garlic storage decay reduced as inoculation concentration of Penicillium hirsutum increased. In addition in order to investigate population dynamics of Pa59-4 on application site of garlic cloves, two antibiotic markers, pimaricin and vancomycin were selected. Bacterial density of Pa59-4 on the wounded garlic cloves increased continuously both under room temperature condition and low temperature condition until 30days after application of Pa59-4, meanwhile that of Pa59-4 on intact garlic cloves increased until 15days after application of Pa59-4 and thereafter decreased continuously. Two culture media for mass-production of Pa59-4, LB medium and TSB medium, were selected. By-product of bio-fungicide formulated by mixing white carbon and bacterial suspension of Pa59-4 suppressed by 40 to 50% garlic blue mold. Above results suggest that Pa59-4 be a promising control agent against garlic blue mold.

• Applied Biological Chemistry
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• v.27
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• pp.29-46
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• 1984

To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.