• Restorative Dentistry and Endodontics
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• v.22 no.1
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• pp.76-92
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• 1997

Porpilyromonas endodontalis is specifically involved in endodontic infections. The bacterium can be isolated almost exclusively only from infected rool canals. P. gingivalis also has been implicated in endodontic infection. Pathogemcity of P. gingival is is attributed to a variety of virulence factors, especially proteases, produced by the bacterium. Importance of P. endodontalis in endodontic infection has been revealed. However, the pathogenic property of P. endodontalis has not been extensively studied. The present study was undertaken to characterize the proteolytic activity of P. endodontalis and compare the activity with that of P. gingivalis which has the most potent and diverse proteases among oral bacteria. For this purpose, culture supematants(SUP) and cell extracts(CE) were obtained from these two bacteria and were subjected to zymography using 15% polyacrylamide gel copolymerized with gelatin, type I, IV collagens or albumin. Hydrolysis of the collagens was further investigated by the cleavage assay using native type I and IV collagens in solution-phase. The results were as follows: 1. P. endodontalis apparently has a proteolytic activity that is comparable with that of P. gingivalis. 2. SUP and CE obtained from P. endodontalis and P. gingival is showed the strongest activity for gelatin, followed by type I and IV collagens, and albumin. 3. In the zymography, no noticeable difference in proteolytic activity for gelatin and albumin between the SUP and CE was observed, but in the cleavage assay using native collagens, the SUP showed a stronger collagenolytic activity than the CE. 4. The gelatinolytic activity of both the SUP and CE from these two bacteria was diminished in the presence of $CaCl_2$ or reducing agents such as ${\beta}$-mercaptoethanol and dithiothreitol(DTT). 5. Type I(calf skin and human placenta) collagenolytic activity of P. endodontalis and P. gingivalis was reduced by DTT but not affected by $CaCl_2$. The inhibitory effect of DTT, however, was reduced to some extent by $CaCl_2$. 6. Type IV collagenolytic activity of these two bacteria was not affected by $CaCl_2$ but increased to some extent in association with the reducing agents. 7. Hydrolysis of albumin by P. endodontalis and P. gingivalis was demonstrated only in the presence of the reducing agents. The overall results indicate that with respect to proteolytic activity, P. endodontalis appears to be as potent as P. gingivalis, or maybe more, and its proteolytic characteristic is similar to that of P. gingivalis. This suggests that P. endodontalis has so potent proteolytic activity that can participate by itself in endodontic infections and apical periodontitis, causing tissue destruction.

• Applied Microscopy
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• v.28 no.1
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• pp.107-119
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• 1998

This study was designed to investigate the appearence and the characteristics of the apoptotic cells and the process of the joint cavity formation in mouse knee joint. Fetal mouse knee joints from 15 to 19 days of gestation were used. Paraffin-embedded serial sections, stained with H & E for light microscopic observation, Epon 812 embedded thin sections for electron microscopic observation and Lowicryl HM 20 embedded thin sections for immune-electron microscopic observation were prepared. Monoclonal antibodies to $\beta-tubulin$ and polyclonal antibodies to tissue transglutaminase were used for immune-electron microscopic study. The results obtained were as follows. 1. At 15 days of gestation, blood vessels, which have invaded in the mesenchymal cells, were present in the synovium, to form the joint cavity in the future. 2. At 16 days of gestation, the joint cleft was first appeared and several RBCs were present in the joint cleft. The invasion of blood vessels into the joint cleft was continuing, and apoptotic cells were present in the inner cell layer, adjacent to the joint cleft. Necrotic cells were also present in the outer cell layer; they were present 18 days of gestation, but apoptotic cells did not appear after 17 days of gestation. 3. In the apoptotic cells, transglutaminase were localized around vacuoles and the marginal site of the cytoplasm. 4. In the apoptotic cells, tubulin was around the endoplasmic reticulum and the marginal site of the cytoplasm. In the late stage of apoptotic cells, tubulin was localized diffusely in the cytoplasm. Tubulin was also strongly labeled around in the cytoplasm of the neighboring cell at which the apoptotic body was phagocytosed. Tubulin labeled particles were apparently increased in the seperated apoptotic bodies. On the basis of the above findings, it is proposed that during the development of the mouse knee joint, blood vessel invasion first occurs and then apoptosis and cell necrosis follow it. In the apoptotic cell, present in the synovium of the developing knee joint of the mouse. it is suggested that the redistribution of tubulin is associated with apoptotic process. And transglutaminase overexpressed in the apoptotic cell.

• Journal of Embryo Transfer
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• v.11 no.1
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• pp.15-26
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• 1996

This study was conducted to improve the in vitro maturation(JVM), in vitro fertilization (IVF) and in vitro developmental capacity of oocytes derived from slaughtered Korean native cattle. The recoverd oocytes, obtained from a local slaughter house, were used completely surrounded by at least 3 layers of cumulus cells in combination with a homogeneous cytoplasmic pigmentation. In vitro maturation was induced in TCM-199 or Ham's F-10 supplemented with LH(1O $\mu$g/rnl), FSH(35 $\mu$g/ml), estradiol-17$\beta$(1 $\mu$g/ml) at 39$^{\circ}C$ under 5% $CO_2$ in air for 24 hours. Sperm from caudal epididyrnis and previously matured cumulus-oocytes complexes were cultured for 24 hours in 100 $\mu$l droplets of fertilization media under paraffin oil. The zygotes were cultured with media(TCM-199 with bovine oviductal epithelial cells or CRlaa) for 7 to 10 days. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following maturation using Ham's F-10 (59.9%) than TCM-199 (51.6%). Development to the blastocysts among cleaved embryos was not signficantly different between maturation media: Ham's F-10 (16.0%) and TCM-199(11.9%). However, the hatching rate was affected significantly (P<0.05) on rnaturation media as 62.9% in Ham's F-10, compared with 41.2% in TCM-199. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following IVF using m-TALP medium (80.1%) than BO medium (51.6%). The percentage of in vitro developed blastocysts among cleaved embryos was not signficantly different between fertimization media: BO (11.7%) and m-TALP (17.6%). The cleavage and the developmental rate to the blastocysts after IVF in m-TALP or condition medium(CM) with or without oviduct epithelial cell monolayer(OECM) was similar(80.1% and 17.6% in m-TALP, 83.8% and 19.4% in M-TALP with OECM. 82.9% and 18.9% in CM, 87.6% and 16.0% in CM with OECM, respectively). The percentage of in vitro developed blastocysts among cleaved embryos was significantly (P<0.05) higher in TCM-199 medium co-cul tured with bovine oviduatal epithelial cell monolayers(35.2%) than CRlaa medium(1.9%). These results stggest that the most transferable IVF embryos could be produced from Ham's F-10, m-TALP and TCM-199 medium with bovine oviductal epithelial cell monolayers for IVM, IVF and IVC, respectively.

• Korean Journal of Food Science and Technology
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• v.47 no.4
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• pp.517-524
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• 2015

The objectives of the present study were to investigate the protective effects of Angelica keiskei juice and Oenanthe javanica juice against oxidative damage in LLC-$PK_1$ renal epithelial cells and to evaluate their free radical-scavenging activities. Both A. keiskei juice and O. javanica juice showed a strong in vitro antioxidant activities such as ${\alpha}$, ${\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH), nitric oxide (NO), $O_2{^-}$, and ${\cdot}OH$ radical-scavenging activities. The LLC-$PK_1$ cells showed significant lipid peroxidation and cell death due to oxidative stress when it was induced by 2, 2'-azobis (2-amidinopropane) dihydrochloride (AAPH), sodium nitroprusside (SNP), pyrogallol, and 3-morpholinosydnonimine (SIN-1). Treatment with both A. keiskei juice and O. javanica juice significantly increased cell viability and inhibited lipid peroxidation. These results suggest that A. keiskei juice and O. javanica juice are potential natural antioxidants.

• Journal of Life Science
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• v.20 no.6
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• pp.940-947
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• 2010

The effects of fermented products from 40 medicinal herbals commonly available in Korea were examined according to concentrations of polyphenolic compound and kojic acid, and the activities of DPPH ($\alpha,\alpha$'-diphenyl-$\beta$-picrylhydrazyl) free radical scavenging and tyrosinase. The polyphenolic compound concentrations were 0.24 by Corydalis turtschaminovill ~ 11.42% (dry matter basis) by Syringa velutina in the extracts and 0.18 by Poria cocos ~ 12.27% by S. velutina in the fermented products. Kojic acid concentrations were 0.02 by Poria cocos Sclerotium ~ 9.67 mM by S. velutina in the extracts and 0.33 by P. cocos ~ 10.32 mM by S. velutina in the fermented products. Syringa velutina contained the highest polyphenolic compound and kojic acid concentrations, which were higher in the fermented product than in the extract. Higher DPPH free radical scavenging activity (>60%) was observed in the extracts of A. sessiliflorum, Citrus nobillis, and Angelica gigas and the fermented product of A. sessiliflorum compared to the other medicinal plants. Higher tyrosinase inhibition activity (>50%) was observed in the extracts of Morus alba, Glycyrrhiza glabra, and Rubus coreanus and the fermented products of G. glabra, Cnidium officinale, and S. velutina. Based on the above results, G. glabra, C. officinale, and S. velutina possessed high tyrosinase-inhibitive activities and kojic acid concentrations, which could be definitely enhanced by the fermentation of Phenillus linteus mycelium.

• Korean Journal of Food Science and Technology
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• v.26 no.2
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• pp.147-151
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• 1994

Hydrogen donating activities(HDA) of solvent extracts of Hololeion maximowiczii roots which were treated with freeze drying, hot air drying and hot air drying after steam blanching were investigated by the method of ${\alpha},{\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl(DPPH). Hydrogen donating activity of hot air drying after steam blanching group showed the highest activity among three drying methods. Among the extracts of freeze dried root extracted with various solvents, color intensity at 420 nm of aqueous extract was highest, but hydrogen donating activity was shown in order of 50%, ethyl alcohol, methyl alcohol, water extracts. By heating the powder of freeze dried root at $100^{\circ}C$, color intensity of its water extract was decreased until 20 min, thereafter it was increased slowly but hydrogen donating activity was rapidly increased until 30 min. Hydrogen donating activity at the concentration of 6.05 mg/ml of 50% alcohol extract was similar to that of ${\alpha}-tocopherol$ at the concentration of $2{\times}10^{-4}M$. Comparison of hydrogen donating activity of the inner and outer solution after dialysis of water extract showed that the outer solution had a stronger hydrogen donating activity than those of the inner one.

• KSBB Journal
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• v.24 no.3
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• pp.267-272
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• 2009

During a dilute acid hydrolysis, degradation products are formed or liberated by pre-treatment of lignocelluloses depend on both the biomass and the pretreatment conditions such as temperature, time, pressure, pH, redox conditions, and addition of catalysts. In lignocellulosic biomass, sugars can be degraded to furfural which is formed from pentoses and 5-hydroxymethulfurfural (HMF) from hexoses. 5-HMF can be further degraded, forming levulinic acid and formic acid. Acetate is liberated from hemicellulose during hydrolysis. Some decomposed compounds hinder the subsequent bioconversion of the solubilized sugars into desired products, reducing conversion yields and rates during fermentation. In the present work, samples of rapeseed strawwere hydrolyzed to study the optimal pretreatment condition by assessing yields of sugars and decomposed products obtained under different reaction conditions ($H_2SO_4$ 0.5-1.25% (w/w), reaction time 0-20 min and temperature range 150-220 C). A careful analytical investigation of acid hydrolyzate of rapeseed straw has not yet been undertaken, and a well-closed mass balance for the hydrolyzate in general is necessary to verify the productivity and economic predictions for this process.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.733-744
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• 2002

The fibroblasts are the principal cells in the periodontal ligament of peridontium. As the periodontal ligament fibroblasts (PDLF) show similar phenotype with osteoblasts, the PDLF are thought to play an important role in alveolar bone remodeling. Cell-to-cell contacted signaling is crucial for osteoclast formation. Recently it has been reported that PDLJ enhance the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The aims of this study were to $clarify\;^{1)}$ the mechanism of PDLF-induced osteoclastogenesis $and\;^{2)}$ whether we can use preosteoclast cell line instead of primary hematopoietic preosteoclast cells for studying the mechanism of PDLF-induced osteoclastogenesis. Osteoclastic differentiation of mouse macrophage cell line RAW264.7 was compared with that of mouse bone marrow-derived M-CSF dependent cell (MDBM), a well-known hematopoietic preosteoclast model, by examining, 1) osteoclast-specific gene expression such as calcitonin receptor, M-CSF receptor (c-fms), cathepsin K, receptoractivator nuclear factor kappa B (RANK) ,2) generation of TRAP(+) multinucleated cells (MNCs), and 3) generation of resorption pit on the $OAAS^{TM}$ plate. RAW264.7 cultured in the medium containing of soluble osteoclast differentiation Factor (sODF) showed similar phenotype with MDBM-derived osteoclasts, those are mRNA expression pattern of osteoclast-specific genes, TRAP(+) MNCs generation, and bone resorbing abivity. Formation of resorption pits by osteoclastic MNCs differentiated from sODF-treated RAW264.7, was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for ODF, to the sODF-containing culture me야um. The effects of PDLF on differentiation of RAW264.7 into　the TRAP(+) multinucleated osteoclast-like cells were examined using coculture system. PDLF were fxed with paraformaldehyde, followed by coculture with RAW264.7, which induced formation of TRAP(+) MNCs in the absence of additional treatment of sODF. When compared with untreated and fixed PDLF (fPDLF), IL-1 ${\beta}$-treated, or lipopolysaccha-ride-treated and then fixed PDLF showed two-folld increase in the supporting activity of osteoclastogenesis from RAW264.7 coculture system. There were no TRAP(+) MNCs formation in coculture system of RAW264.7 with PDLF of no fixation. These findigs suggested that we can replace the primary hematopoietic preosteoclasts for RAW264. 7 cell line for studying the mechanism of PDLF-induced osteoclastogenesis, and we hypothesize that PDLF control osteoclastogenesis through ODF expression which might be enhanced by inflammatory signals.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.26-32
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• 1977

The multiple 7S globulins composed of two fractions (A and B) in the electrophoresis with Davis' method were isolated at different stages of the soybean seed development. Electrophoresis of their subunits liberated in PAWU solvent [phenol-acetic acid-water (2 : 1 : 1) solution plus 5M urea] yielded 4 major bands. Observation of both the electrophoretic bands of the multiple 7S fractions(7S-A and 7S-B) and those of their subunits was suggestive of a similarity of the subunit pattern between two 7S fractions. The two fractions in multiple 7S globulins were isolated with DEAE-Sephadex A-50 column$(2.0{\sim}100cm)$ chromatography. They were separated into 2 fractions in a linear gradient concentration of 0.28 to 0.40M NaCl with phosphate buffer (pH 7.8) containing 10mM ${\beta}-mercaptoethanol$(ME). The isolated protein was dissociated into subunits with two different solvent systems; in PAWU solvent and in Tris-HCl buffer(pH 8.0) containing 1% sodium dodecyl sulfate (SDS) and 40mM ME. The dissociated subunits were subjected to electrophoresis in PAWU-treated 7.5% acrylamide gel and in 1% SDS-treated 5.6% acrylamide gel. In PAWU gel electrophoresis, total 7S globulin was separated into 5 major bands, two of which were occupied in common by two 7S fractions(7S-A and 7S-B). In SDS gel electrophoresis, total 7S globulin was separated into 7 major bands, three of which were overlapped with the subunit of the two 7S fractions. The above results alluded us to the presence of a common and/or similar subunit between the multiple 7S globulins.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 1994.07a
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• pp.23-24
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• 1994

본 연구는 성숙과의 저장중에 세포벽분해효소가 세포벽을 분해해서 물성의 변화를 유발함으로 일어나는 과실의 연과가 품질과 저장성의 저하 뿐만 아니라 영양적, 경제적 손실을 초래한다는 점을 고려하여 대추의 성숙중에 일어나는 연화현상을 연구하고자 성숙에 따른 경도, 세포벽분해효소의 활성, 세포벽 다당류 pectin질, 비섬유성 중성당 및 조직의 변화를 조사하였다. 경도는 대추의 숙성에 따라 감소하였고, polygalacturonase와 $\beta$-galactosidase의 활성은 각각 변색기와 완숙기에 나타난 이후 급격히 증가하였다. 세포벽 다당류인 pectin질과 알칼리 지용성 hemicellulose는 완숙기가지 증가햐였으나 cellulose는 완숙기에 산가용성 hemicellulose와 cellulose를 제외한 세포벽 다당류의 함량은 다소 감소하였다. 대추의 세포벽 비섬유성 중성당으로 rhamnose, arabinose, xylose, mannose, galactose, glucose가 동정되었고, 성숙동안에 pectin질에서는 arabinose, mannose, galactose와 총 비섬유성 중성당의 함량이 감소하였고, 산가용성 hemicellulose에서는 xylose와 mannose가 뚜렸하게 증가하였으나 중성당은 변화없었으며, 알칼리 가용성 hemicellulose에서는 성숙에 따른 변화가 거의 없었다. pectin질의 경우 수용성 pectin, EDTA 용해성 pectin 및 총 pectin은 성숙중에 증가하는 경향이있으나 불용성 pectin은 감소하는 경향이였으며 과숙기에는 불용성 psctin EDTA용해성 pectin 및 총 pectin의 함량은 모두 현저히 감소하였다. 대추의 성숙중 조직에서는 pectin질로 구성된 중충의 붕괴현상이 뚜렸하게 나타났다.발이 절실히 필요한 실정이다. 이러한 배경으로 본 강연에서는 효소적갈변 저해제의 개발과 그들의 식품가공에의 적용 현환 및 화장품, 의약품으로의 응용에 대해 설명하고자 한다.L주에 비해 S주는 수정후 용과가 더 심하다. 9) 화분관의 행동은 수정력과 완전히 일치된다. 즉 L-selfing, $L{\times}L$, S-selfing, $S{\times}S$등의 부적법 수분에서는 화분관은 화주의 미중에서 정지되지만 $L{\times}S$, $S{\times}L$,에서는 수분 약 40-50분 후이면 화분관은 자방까지 도달된다. 10) S주는 웅본으로 오인되어 있지만 인위적법수분을 하면 수정력이나 화분관의 행동은 L주에서와 동일하다. 11) S화분은 완전하지만 L화분은 약 70%가 내용공허한 Adortive pollen 이다. 12) L화분중 나머지 30%도 S화분에 비해서 염색도가 낮은것이 많고 S화분 같이 농염되는 것은 극히 소수이다. 13) 본장물은 분화가 고도로 진행된 전형적인 이형예작물이여 마치 Dimorphism 에서 Dioecious 에로 이행되는 수가 있다는 것을 표시하는 증거가 되는 것 같다. 다소 높은 산소농도 3%~5% 이산화탄소 농도 5~8%에서 저장하는 것이 효과적일 것으로 판단되었다.철쭉군목으로 대표되나 군단이 하의 군목들은 다소 차이를 보이는 것으로 나타났다. 중간상인이론의 수정이 필요하다고 본다.가\ulcorner 본 논문에서는 표면적 형태에도 불구하고 [-wh]의미의 겹의문사는 병렬적 관계의 합성어가 아니라 내부구조를 지니지 않은 단순한 단어(minimal $X^{0}$ elements)로 가정한다. 즉, [+wh] 의미의 겹의문사는 동일한 구성요 소를 지닌 병렬적 합성어([