• Title/Summary/Keyword: MG-63 cells

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Bioactivity behavior of biphasic calcium phosphate powders prepared by co-precipitation method (공침법으로 합성된 biphasic calcium phosphate 분말의 생체활성 거동)

  • Kim, Tae-Wan;Kim, Dong-Hyun;Jin, Hyeong-Ho;Lee, Heon-Soo;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.2
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    • pp.99-104
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    • 2012
  • The co-precipitation technique has been applied to synthesize biphasic calcium phosphate (BCP). $Ca(NO_3)_2{\cdot}4H_2O$ and $(NH_4)_2HPO_4$ as the starting materials was used. X-ray diffraction (XRD) and Fourier transformed infrared (FT-IR) spectroscopy were used to characterize the structure of as-synthesized and calcined BCP powders. After immersion in Hanks' Balanced Salt Solution (HBSS), for 1 week a precipitation started to be formed with individual small granules on the specimen surface. An MTT assay indicated that BCP powders have no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

Evaluation of antibacterial activity and osteoblast-like cell viability of TiN, ZrN and $(Ti_{1-x}Zr_x)N$ coating on titanium

  • Ji, Min-Kyung;Park, Sang-Won;Lee, Kwangmin;Kang, In-Chol;Yun, Kwi-Dug;Kim, Hyun-Seung;Lim, Hyun-Pil
    • The Journal of Advanced Prosthodontics
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    • v.7 no.2
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    • pp.166-171
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    • 2015
  • PURPOSE. The aim of this study was to evaluate antibacterial activity and osteoblast-like cell viability according to the ratio of titanium nitride and zirconium nitride coating on commercially pure titanium using an arc ion plating system. MATERIALS AND METHODS. Polished titanium surfaces were used as controls. Surface topography was observed by scanning electron microscopy, and surface roughness was measured using a two-dimensional contact stylus profilometer. Antibacterial activity was evaluated against Streptococcus mutans and Porphyromonas gingivalis with the colony-forming unit assay. Cell compatibility, mRNA expression, and morphology related to human osteoblast-like cells (MG-63) on the coated specimens were determined by the XTT assay and reverse transcriptase-polymerase chain reaction. RESULTS. The number of S. mutans colonies on the TiN, ZrN and $(Ti_{1-x}Zr_x)N$ coated surface decreased significantly compared to those on the non-coated titanium surface (P<0.05). CONCLUSION. The number of P. gingivalis colonies on all surfaces showed no significant differences. TiN, ZrN and $(Ti_{1-x}Zr_x)N$ coated titanium showed antibacterial activity against S. mutans related to initial biofilm formation but not P. gingivalis associated with advanced periimplantitis, and did not influence osteoblast-like cell viability.

An Experimental Study on the Effect of Hypothermic Oxygenated Cardioplegic Solution on Myocardial Protection during Prolonged Aortic Cross-clamping (대동맥 차단시 저온 산소화 심정지액이 심근보호에 미치는 영향)

  • Wang, Yeong-Pil;Lee, Hong-Gyun
    • Journal of Chest Surgery
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    • v.18 no.4
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    • pp.759-770
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    • 1985
  • This study was experimentally undertaken to evaluate the effect of hypothermic oxygenated cardioplegic solution on myocardial protection during prolonged aortic cross clamping under cardiopulmonary bypass. Dogs were divided into two groups control group [received hypothermic unoxygenated cardioplegic solution] and experimental group [received hypothermic oxygenated cardioplegic solution]. Coronary sinus effluent was obtained at once and 30, 60, 90 minutes after cross-clamping for the determination of pH, PCO2,PO2 and lactate level during the infusion of cardioplegic solution and myocardial biopsies were obtained after cessation of 90 minutes of aortic cross-clamping. The results obtained were as follows: 1. There was no significant differences in the pH and PCO2 between the oxygenated and unoxygenated cardioplegic solution but the PO2 of the oxygenated solution was 4 times greater than unoxygenated solution, and also the oxygenated solution had a significantly greater oxygen content [2.020.05 ml 02/min] and had much more oxygen delivery than unoxygenated solution. 2. The myocardial oxygen consumption and the myocardial oxygen extraction in oxygenated group were 1.63 ml 02/100 ml and 67.32% respectively, which was greater than those in unoxygenated group. 3. Regarding to pH and PCO2 of coronary sinus effluent, there was no significant differences between two groups in early period of infusion of cardioplegic solution, but the pH shifted to acidosis from 60 minutes, PCO2 increased from 90 minutes of aortic cross-clamping, and PO2 markedly decreased from 90 minutes of aortic cross-clamping in unoxygenated group. 4. The lactate concentration of coronary sinus effluent revealed relatively normal in both groups, but showed slight increase up to 27.54.56 mg/100 ml at 90 minutes of aortic cross-clamping in unoxygenated group. 5. On electron microscopic study, the ultrastructural integrity of myocardial cells in oxygenated group was well preserved within 90 minutes. Slight swelling and deformity of mitochondria, interfibrillar widening, and disarrangement of myofibrils were observed at 90 minutes after aortic cross-clamping in unoxygenated group. From these results, the use of hypothermic oxygenated cardioplegic solution seemed to be effective and better method for the preservation of ischemic myocardium during the prolonged aortic cross-clamping.

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Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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Effect of Fusion Procedure on the Development of Embryos Produced by Somatic Cell Nuclear Transfer in Hanwoo (Korean Cattle) (한우에서 융합방법이 체세포 핵이식 수정란의 발달에 미치는 영향)

  • Im, G.S.;Yang, B.S.;Park, S.J.;Chang, W.K.;Park, C.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.365-373
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    • 2000
  • The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P<0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P<0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

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Analysis of Components in the Different Parts of Lythrum salicaria L. (털부처꽃의 부위별 성분 분석)

  • Kim, Hee-Young;Park, Yea-Jin;Lee, Ju-Yeon;Kim, Ki-young;Shin, Su;Choi, Min-Woo;Hong, Eun-Jin;Kim, Min-jeong;Yeo, Sujung;Park, In-hwa;Jerng, Ui Min;An, Hyo-Jin;Cha, Yun-Yeop
    • The Korea Journal of Herbology
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    • v.37 no.5
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    • pp.89-96
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    • 2022
  • Objectives : This research was performed to analyze the components in the different parts of Lythrum salicaria L. and to compare which parts of L. salicaria L. are appropriate for food development. Methods : L. salicaria L. was extracted in 20% EtOH at 100 ℃ for 4 hours. Cytotoxicity was investigated in 3T3-L1 cells after treatment of 10-500 ㎍/ml L. salicaria L. for 24 hours. Total polyphenol content (TPC) was estimated using 1 N Folin-ciocateu reagent. 2,2-Diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity was estimated using DPPH reagent and gallic acid. The chemical composition was analyzed by high-performance liquid chromatography (HPLC). 1) Results : The half maximal inhibitory concentration (IC50) in the extracts of the whole plant, aerial parts, and root parts was 350 ㎍/ml, over 500 ㎍/ml, and 150 ㎍/ml, respectively. The TPC in the extracts of the whole plant, aerial parts, and root parts was 527.1 mg/g, 422.6 mg/g, and 781.1 mg/g, respectively. The averages of vitexin contents in the aerial parts, and root parts were 256.7 ± 154.9 ㎍/g and 266.1 ± 63.2 ㎍/g, respectively. The averages of TPC in the leaves, roots, flower stalks and stems were 224.0 ± 53.7 tannin acid (TA) mg/g, 221.8 ± 70.2 TA mg/g, 249.8 ± 34.4 TA mg/g, and 67.7±8.9 TA mg/g, respectively. The averages of DPPH radical scavenging activity in the leaves, roots, flower stalks, and stems were 282.01 ± 43.3 gallic acid equivalent (GAE) 𝜇mole/g, 260.16 ± 44.1 GAE 𝜇mole/g, 288.0 ± 9.3 GAE 𝜇mole/g, and 97.6 ± 10.7 GAE 𝜇mole/g, respectively. Conclusions : There were no significant differences in the content of components or antioxidant activity in the aerial parts compared to those in the whole plant of L. salicaria L. Furthermore, the root parts had low extract yield, cytotoxicity, and quality control problems, therefore our results suggest that the use of the aerial part of L. salicaria L. would be the most appropriate for food development.

Effect of Fermented Ice Plant (Mesembryanthemum crystallinum L.) Extracts against Antioxidant, Antidiabetic and Liver Protection (아이스플랜트(Mesembryanthemum crystallinum L.) 발효추출물의 항산화, 항당뇨 및 간 보호효과)

  • Nam, Sanghae;Kang, Seungmi;Kim, Seonjeong;Ko, Keunhee
    • Journal of Life Science
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    • v.27 no.8
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    • pp.909-918
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    • 2017
  • Ice plant (Mesembryanthemum crystallinum L.) was fermented in brine in the form of mulkimchi (IPMB), and its contents of organic acid and cyclitols and biological activities were compared with those before fermentation. The pH of the IPMB continuously decreased until the sixth day of fermentation. The lactic acid yield was greatest on the fourth day. D-pinitol in ice plant mulkimchi solids (IPMS) decreased during fermentation. However, myo-inositol and D-chiro-inositol increased. The radical scavenging activities of ABTS and DPPH, in addition to the activity of FRAP, of the IPMS extract were generally higher after fermentation, with the activities highest on the fifth ($79.09{\pm}0.69%$), fourth ($87.55{\pm}1.21%$), and sixth ($78.72{\pm}0.99%$) days of fermentation, respectively, when treated with 1 mg/ml of the extract. As shown by a lipid/MA assay, antioxidant activity was generally higher after fermentation. The viability of BNL CL.2 cells damaged by t-BHP, $H_2O_2$, and ethanol was $14.19{\pm}0.98$, $13.80{\pm}2.25$, and $25.89{\pm}2.90%$, respectively. When treated with $200{\mu}g/ml$ of IPMS extract, the cell viability was $57.06{\pm}4.52%$ on the first day, and $66.06{\pm}1.36%$ on the fourth day, and $50.07{\pm}04.85%$ on the sixth day of fermentation. Hepatocyte protective effects did not increase significantly after fermentation. ${\alpha}-glucosidase$ inhibitory activity was quite high, with a range of $83.52{\pm}2.69$ to $92.79{\pm}2.16%$, and the activity increased gradually in all the groups over the fermentation period. There was no clear correlation between ${\alpha}-amylase$ inhibitory activity and fermentation.

Synthesis of L-threo-3,4-Dihydroxyphenylserine(L-threo-DOPS) with Thermostabilized Low-Specific L-Threonine Aldolase from Streptomyces coelicolor A3(2)

  • Baik, Sang-Ho;Yoshioka, Hideki;Yukawa, Hideaki;Harayama, Shigeaki
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.721-727
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    • 2007
  • Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

Quality and fermentative characteristics of yogurt added with hot water extract of Welsh onion root (파뿌리 열수추출물을 이용한 요구르트의 발효 및 품질특성)

  • Kim, Min-Jeong;Lee, Shin-Ho
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.387-393
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    • 2017
  • The objective of this study was to investigate the effect of hot water extract of Welsh onion root (HEWO) on growth of lactic acid bacteria and fermentative characteristics of yogurt. The physiochemical characteristics of HEWO such as pH, soluble solid, reducing sugar, total polyphenol content and DPPH radical scavenging activity were studied. The lactic bacterial count in brain heart infusion (BHI) broth with HEWO was about 1 log cycle higher than in control for 24 h at $37^{\circ}C$. The pH of yogurt prepared with HEWO (WY100) and 50% HEWO (WY50) was gradually decreased significantly but increased the viscosity of yogurt with increasing HEWO concentration during fermentation. The viable cells of lactic acid bacteria after fermentation for 24 h were 8.03 (control), 8.77 (WY50), 8.84 (WY100) log CFU/mL, respectively. The DPPH radical scavenging activity of yogurt increased with increasing HEWO concentration. Sensory quality of yogurt prepared with HEWO was higher than that of control. The pH and lactic acid bacteria of all tested yogurts decreased during storage for 10 days at $4^{\circ}C$ but lactic bacterial count of yogurt prepared with HEWO maintained $10^8CFU/mL$ during storage. These results indicated the potential use of HEWO as a valuable resource to improve fermentation and functionality of yogurt.

Quality Properties of Soy-paste Soybean Cultivar for Fermented Soybean Products (장류용 콩 품종별 발효물의 품질 특성)

  • Shin, Dong-Sun;Park, Chang Hwan;Choi, In Duck;Lee, Seuk Ki;Park, Ji Young;Kim, Nam Geol;Choi, Hye Sun
    • The Korean Journal of Food And Nutrition
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    • v.32 no.2
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    • pp.114-121
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    • 2019
  • This study evaluates the quality properties of soy-paste soybean cultivar for fermented soybean products. The six soybean varieties that include Jinpung, Saedanbaek, Daepung 2, Pyeongwon, Cheonga and Saeolkong were used in the experiment. The range of water uptake ratio, hardness after soaking and hardness after steaming were 117.00~131.33%, 1.65~3.30 kg and 0.05~0.14 kg, respectively. The physicochemical analysis indicated the following: Moisture content, 63.27~68.72%; pH, 6.43~6.60; total acidity, 0.27~0.45%. Color values for L value (lightness), a value (redness), and b value (yellowness) ranged from 39.07~67.92, 7.64~11.79, and 7.48~20.67, respectively. The amylase and protease activities of the Saedanbaek samples were the highest among all cultivars. The amount of viscous substance in the fermented soybean products by cultivars ranged from 5.93 to 8.37%, and Saedanbaek was the highest. The total viable cells counts for soybean fermented products were 9.11~9.42 log CFU/g. The amino-type nitrogen contents of all samples were in the range of 401.07 to 524.47 mg% and Saedanbaek cultivars showed the highest content (524.47 mg%). Based on the results, Saedanbaek will be suitable as a soy-paste soybean cultivar and the quality standards for the fermentation process of the fermented soybean products.