• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.581-596
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• 1994

This in vitro study was undertaken to obtain optimal tetracycline concentration that aids proliferation and spreading of human periodontal ligament cells, for clinical application in root surfaces of periodontally diseased teeth. Periodontal ligament cells used in this study were obtained from explants of periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment. The cells were cultured in Dulbecco's Modified Eagle Medium(DMEM) supplemented with 100 U/ml penicillin, $100\;{\mu}g/ml$ streptomycin and 10% FBS at $37^{\circ}C$, 100% humidity, 5% $CO_2-95%$ air. Cells were used between the third to 4th passage. After root planing of periodontally extracted teeth, the root slabs were cut with carborundum disk. In the cell proliferation experiment, experimental groups were root planing only group, immersed groups in 25, 50, 75, 100, 150mg/ml aqueous solution of Tetracycline HCl followed by a vigorous rinse in PBS. Human PDL cells at concentration of $1{\times}10^5\;cells/ml$ were seeded in each culture well which contained root slabs and incubated for 6 hours. Then, all of the root slabs were moved into new 24 culture well and incubated 24, 48 and 72 hours. The cell counting was done by inverted phase contrast microscope after trypsinization. The following results were obtained. The cell number was increased in order root planing only group, 25, 150, 50, 75, 100mg/ml of Tetracycline HCl treated group in 24, 48 and 72 hours. The maximal cell number was obtained when the root slabs were immersed in solution with 100mg/ml of Tetracycline HCl. There were statistically significant between the root planing only group and 75, 100 mg/ml of Tetracycline HCl treated group in 24 hours, between the root planing only group and 100mg/ml of Tetracycline HCl treated group in 48 hours, between the root planing only group and 50, 75, 100mg/ml of Tetracycline HCl treated group, between 25 and 100mg/ml of Tetracycline HCl treated group in 72 hours(p<0.05). In the cell spreading experiment, after 30 minutes of incubated, in the root planing only group, the cells were generally round in shape. The cell surface was mostly covered with blebs. The cells started to attach to root surface by cytoplasmic extension in 50, 100mg/ml of Tetracycline HCl treated groups, more numerous cells attached to root surface than root planing only group. Many orifices of dentinal tubule were exposed, cells showed radially spreaded cytoplasm and unspreaded central region of the cell was covered with blebs. After 6 hours of incubation, in the root planing only group, cells showed radially spreaded cytoplasm and were attached flat appearance. In 50, 100mg/ml of Tetracycline HCl treated groups, cellular margin was concaved and cytoplasm showed elongated appearance with polarity. After 24 hours of incubation, in the root planing group, cells showed characteristic polarity. In 50, 100mg/ml of Tetracycline HCl treated groups, cells showed more elongated and spindle - like appearance.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.883-890
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• 2015

The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

• Journal of Life Science
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• v.25 no.12
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• pp.1408-1414
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• 2015

In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.169-176
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• 2005

Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

• 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.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.437-442
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• 1996

Two strains of Lactobacillus(L.) casfi and one strain of L. Pentosus, which were isolated from pickles, were used to investigate in studing their characteristics of ${\beta}-galactosidase$. The preferable carbon sources and pH of the MRS media for enzyme production from L. casei No.10 was found to be 1.0% lactose and pH 7.5, from L. Pentosus No.63 was 1.0% galactose and pH 7.5, and from L. casei No.36 was 1.0% lactose and pH 6.5, respectively. The maximum enzyme production from each strain was found after 48 hours culture at $30^{\circ}C$ in a medium with preferable carbon source. The optimum reaction temperature with substrate for ${\beta}-galactosidase$ activity was found at $60^{\circ}C$ for all three strains . The stability of enzyme from L. casei No.36 was found to be at $45^{\circ}C$, from L. Pentosus No.63 was found at $55^{\circ}C$. This stability from L. casei No.36 was found at $40^{\circ}C$, but it was reduced to 60% at $55^{\circ}C$. These stabilities of enzymes remained about 90% at $40^{\circ}C$ for all three strains. The optimal pH for enzyme activities was found to be pH 6.5 for all three strains. Enzyme activity remained over 90% for L. casei No.10 at $pH\;5.0{\sim}6.0$, for L. casei No.36 at $pH\;5.0{\sim}8.0$, and for L. pentosus No.63 at $pH\;6.0{\sim}7.0$.

• Journal of Life Science
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• v.19 no.7
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• pp.994-1002
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• 2009

A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

• Korean Journal of Heritage: History & Science
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• v.44 no.4
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• pp.172-205
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• 2011

Go bears significant meanings in terms of cultural and entertaining functions in Asia Eastern such as China and Japan. Beyond the mere entertaining level, it produces philosophical and mythic discourse as well. As a part of effort to seek an identity of Korean traditional garden culture, this study traced back to find meanings of rock-go-board and taste for the arts which ancestors pursued in playing Go game, through analysis and interpretation of correlation among origin of place name, nearby scenery, carved letters and vicinal handed-down place name. At the same time, their position, shape and location types were interpreted through comprehensive research and analysis of stone-go-boards including rock-go-board. Particularly, it focused on the rock names related to Sundoism(仙道) Ideal world, fixed due to a connection between traces of Sundoism and places in a folk etymology. Series of this work is to highlight features of the immortal sceneries, one of traditional landscaping ideals, by understanding place identity and scenic features of where the rock-go-boards are carved. These works are expected to become foundation for promotion and preservation of the traditional landscaping remains. The contents of this study could be summarized as follows; First, round stone and square board for round sky and angled land, black and white color for harmony of yin and yang and 361paths for rotating sky are symbols projecting order of universe. Sayings of Gyuljungjirak(橘中之樂), Sangsansaho(商山四皓), Nangagosa(爛柯故事) formed based on the idea of eternity stand for union of sky and sun. It indicates Go game which matches life and nature spatiotemporally and elegant taste for arts pursuing beauty and leisure. Second, the stone-go-boards found through this research, are 18 in total. 3 of those(16.1%), Gangjin Weolnamsaji, Yangsan Sohanjeong and Banryongdae ones were classified into movable Seokguk and 15(83.9%) including Banghakdong were turned out to be non-movable rock-go-boards carved on natural rocks. Third, upon the result of materializing location types of rock-go-boards, 15 are mountain stream type(83.9%) and 3 are rock peak type(16.1%). Among those, the one at Sobaeksam Sinseonbong is located at the highest place(1,389m). Considering the fact that all of 15 rock-go-boards were found at mountainous areas lower than 500m, it is recognizable that where the Go-boards are the parts of the living space, not far from secular world. Fourth, there are 7 Sunjang(巡將) Go with 17 Hwajeoms(花點), which is a traditional Go board type, but their existences, numbers and shapes of Hwajeom appear variously. Based on the fact, it is recognizable that culture of making go-board had been handed down for an extended period of time. Among the studied rock-goboards, the biggest one was Muju Sasunam[$80(82)cm{\times}80(82)cm$] while the smallest one was Yangsan Sohandjeong Seokguk ($40cm{\times}40cm$). The dimension of length and breadth are both $49cm{\times}48cm$ on average, which is realistic size for actual Go play. Fifth, the biggest bed rock, an under-masonry with carved Go-board on it, was one in Muju Sasunam[$8.7m{\times}7.5m(65.25m^2)$], followed by ones in Hoengseong Chuiseok[$7.8m{\times}6.3m(49.14m^2$] and Goisan Sungukam[$6.7m{\times}5.7m(37.14m^2)$]. Meanwhile, the smallest rock-go-board was turned out to be one in Seoul Banghak-dong. There was no consistency in directions of the Go-boards, which gives a hint that geographical features and sceneries of locations were considered first and then these were carved toward an optimal direction corresponding to the conditions. Sixth, rock-go-boards were all located in valleys and peaks of mountains with breathtaking scenery. It seems closely related to ancestors' taste for arts. Particularly, rock-go-boards are apprehended as facilities related to taste for arts for having leisure in many mountains and big streams under the idea of union of sky and human as a primitive communal line. Go became a medium of hermits, which is a traditional image of Go-game, and symbol of amusement and entertainment with the idea that Go is an essence of scholar culture enabling to reach the Tao of turning back to nature. Seventh, the further ancient time going back to, the more dreamlike the Go-boards are. It is an evident for that Sundoism, which used to be unacceptable once, became more visible and realistic. Considering the high relation between rock-go-boards and Sundoism relevant names such as Sundoism peak in Danyang Sobaeksan, 4 hermits rock in Muju and Sundoism hermit rock in Jangsu, Sundoism hermit rocks and rock-go-boards are sceneries and observation spots to express a communication of worship and longing for Sundoism. Eighth, 3 elements-physical environment such as location type of the rock-go-boards, human activities concentrated on 8 sceneries and Dongcheongugok(洞天九曲) setup and relevancy to Confucian scholars, as well as 'Sangsansaho' motif and 'Nangagosa' symbolic meaning were used as interpretation tools in order to judge the place identity. Upon the result, spatial investigation is required with respect to Sunyoodongcheon(仙遊洞天) concept based on enjoyment to unify with the nature rather than Dongcheongugok concept of neo-Confucian, for Dongcheon and Dongmoon(洞門) motives carved around the rock-go-boards. Generally, places where mountain stream type rock-go-boards were formed were hermit spaces of Confucianism or Sundoism. They are considered to have compromised one other with the change of times. Particularly, in the rock-go-board at the mountain peak, sublimity-oriented advent of Sundoism is considered as a significant factor to control place identity. Ninth, including where the rock-go-boards were established, the vicinal areas are well-known as parts of Dongcheongugok and Palkyung(八景) mostly. In addition, many of Sundoism relevant expressions were discovered even in the neighboring carvings written by scholars and nobility, which means sophisticated taste based on longing for Sundoism world played a significant role in making go-board. The rock-go-board is an integration of cultural phenomena naturally managed by seclusion of scholars in the Joseon Dynasty as well as remains and essence of Korean traditional landscaping. Some rock-go-boards out of 17 discovered in South Korea, including ones in Sobaeksan Sinsunbong, Banghak-dong, Chungju Gongili, Muju Sasunam, Yangsan Eogokdong Banryongdae Seokguk, are damaged such as cracks in rocks or fainted lines by hardships of time and hand stains. Worse yet, in case of Eunyang Bangudae Jipcheongjeong board, it is very difficult to identify the shape due to being buried. Rock-go-boards are valuable sculptures in terms of cultural asset and artwork since they reflect ancestors' love for nature and longing for Sundoism world. Therefore, they should be maintained properly with right preservation method. Not only rock-boards itself but also peripheral places are excellent cultural heritages and crucial cultural assets. In addition, vicinal sceneries of where rock-goboards and pavilion spots are the representative remains of embracing prototype of Korean traditional landscaping and major parts of cultural properties.