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The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration (혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구)

  • Lee, Seung-Yeol;Seol, Yang-Jo;Lee, Yong-Moo;Lee, Ju-Yeon;Lee, Seung-Jin;Kim, Suk-Young;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.1-23
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    • 2001
  • Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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Early Life History and Spawning Behavior of Chinese Minnow, Rhynchocypris oxycephalus Reared in the Laboratory (실험실에서 사육한 버들치의 산란습성(産卵習性) 및 초기생활사(初期生活史))

  • Han, Kyeong-Ho;Noh, Byeong-Yul;O, Sung-Hyun;Park, Joon-Taek;Cho, Jae-Kwon;Seong, Ki-Baik
    • Korean Journal of Ichthyology
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    • v.11 no.2
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    • pp.177-183
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    • 1999
  • This study was conducted to observe the spawning behavior and early life history of chinese minnow, Rhynchocypris oxycephalus reared in the laboratory. The spawning period of Rhynchocypris oxycephalus is from May to June in nature. The eggs of Rhynchocypris oxycephalus were spawned on the sand and surface of the gravel. The fertilization eggs were demersal in shape and adhesive, released as a clump forming a thin layer and their diameter were 1.70~1.90mm (mean 1.80 mm, n=20). Hatching of eggs was started in 88 hour 45 minute after fertilization at water temperature $19{\pm}0.5^{\circ}C$ and finished in 90 hour. Newly-hatched larvae were measured 4.87~5.02 mm in total length (TL, mean 4.94 mm), mouth and anus were not opened. 6 days old larvae were 6.32~6.56 mm in TL (mean 6.44 mm). Yolk sac was almost absorbed, mouth and anus was began to open. 13 days old larvae were 6.74~6.91 mm in TL (mean 6.82 mm). Part of Dorsal fin was began to rising and myomere number was 15+23=38. 25 days after hatching, total length of larvae was 8.45~8.60 mm (mean 8.52 mm). Dorsal and anal fin rays became differentiated, and also caudal part of the notocord flexion was achieved at $45^{\circ}C$. In the time, growth rate was higher than the other stage. Aggregate numbers of all fin rays were completed at 16.39~16.57 mm in TL (60 days after hatching), at which time the larvae reached the juvenile stage, but fin-fold on ventral was remained yet. External features of adult specimens were almost completed at 80 day old juveniles (18.69~18.87 mm in TL).

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Amylase Activity and Characterization of Microorganism Isolated from in Aquacultural Effluents Sediment Layer (양식장 배출수 퇴적층에서 분리된 미생물의 다당분해효소 활성 및 특성)

  • Kim, Man-Chul;Jang, Tae-Won;Harikrishnan, Ramasamy;Moon, Young-Gun;Song, Chang-Young;Kim, Gi-Young;Heo, Moon-Soo
    • Journal of Life Science
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    • v.19 no.3
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    • pp.366-372
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    • 2009
  • In the course of screening of useful enzyme-producing microorganisms from marine sedimentary layers, we isolated 2 amylase producing strains and tested their amylase producing activities. Analyses of 16S rDNA sequences and biochemical methods (BIOLOG) of two isolates showed that they were confirmed to be a gram positive Bacillus sp. and gram negative Pseudoalteromonas sp., respectively. Excellent amylase producing strains were termed Bacillus sp. ST-63 and Pseudoalteromonas sp. ST-140, and further studies were conducted on their amylase producing characteristics. Optimum conditions for cell growth in amylase activity were obtained when the isolate (Bacillus sp. ST-63 and Pseudoalteromonas sp. ST-140) was cultured at $30^{\circ}C$ and pH $7{\sim}8$.

THE MORPHOLOGICAL OBSERVATION OF HUMAN GINGIVAL FIBROBLASTS ATTACHMENT AND SPREADING ON THE MECHANICAL TREATED TITANIUM PLASMA SPRAYED IMPLANT SURFACE (기계적 표면 처리된 TITANIUM PLASMA SPRAYED IMPLANT에 대한 치은섬유아세포전개양상의 형태학적 관찰)

  • Whang, Yun-Hi;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.741-755
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    • 1995
  • Currently titanium is the material of choice for implants because of its biological acceptance. This high degree of biocompatibility is thought to result, in part, from the protective and stable oxide layer that presumably aids in the bonding of the extracellular matrix at the implant-tissue interface. Endosseous dental implants are interfaced with bone, connective tissue, and epithelium when implanted into the jaw bone. The soft tissue interface including connective tissue and epithelium is one of the most critical factors in the determination of implant maintenance and prognosis. For maintenance of failing or failed implants, it is essential to treat the implant fixture surface to remove bacterial endotoxins and make a surface tolerated by surrounding soft and hard tissues. In this study, the effect of mechanical treatment on titanium plasma sprayed implant on adhesiveness and proliferation of human gingival fibroblasts and changed surface characteristics were studied. titanium plasma sprayed discs manufactured by Friedrichsfeld company were treated with loaw speed stone bur, a rubber point and a jetpolisher. Its surface components were analyzed with Energy dispersive X-ray spectroscopy to evaluate whether the surface characteristics were altered or not. To observe the spreading pattern of the human gingival fibroblasts which attached to the all specimens author used the scanning electron microscope. The results were as follows : Pure titanium and plasma sprayed titanium, stone polished titanium showed titanium peak and small amout of aluminum, so there was no alteration on surface characteristics. Under the scanning electron microscopic examination in the initial attachment of human gingival fibroblast, there was a slight enhancement in pure titanium, stone polished titanium than plasma sprayed titanium. After 6 hours, the pure titanium and stone polished titanium showed human gingival fibroblasts were elongated and connected with numerous processes. Human gingival fibroblasts were more intimately attached on the pure titanium discs than on the other discs. The human gingival fibroblasts attached on the plasma sprayed titanium by thin and elongated processes. After 24 hours, the human gingival fibroblasts connected with each other via numerous processes and compeletly covered the pure titanium and stone polshed titanium discs. Human gingival fibroblasts had multiple point contacts with more long and thin lamellopodia and showed a little bare surface on plasma sprayed titanium discs.

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INFLUENCE OF A SODIUM HYPOCHLORITE GEL ON MICROLEAKAGE OF COMPOSITE RESIN RESTORATIONS (차아염소산 나트륨의 사용이 복합레진 수복물의 미세누출에 미치는 영향)

  • Yang, Kye-Sik;Kim, Dae-Eop;Lee, Kwang-Hee;Jeong, Young-Nam
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.54-60
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    • 2003
  • This study evaluated the influence of chemomechanical caries removal agent $Carisolv^{TM}$(MediTeam, Sweden) for composite resin adhesion to sound human permanent and primary dentin. The buccal/labial surfaces of 80 permanent molars and 80 primary incisors were used. Four types of adhesives and one composite resin were used; AQ Bond(Sun Medical, Japan), Clearfil SE Bond(Kuraray, Japan), Single Bond(3M, USA), Scotchbond Multi-Purpose(3M, USA) and Z100(3M, USA). One drop of $Carisolv^{TM}$(MediTeam, Sweden) was pretreated on the dentin for 0 second(control) and 60 seconds. The specimens were thermocycled for 1,000 times in baths kept 5 degrees C and 55 degrees C with a 30 seconds dwell time. Shear bond strengths were tested and the data was statistically analyzed using one-way ANOVA with subsequent post hoc Scheffe test at p<0.05. $Carisolv^{TM}$ treatment significantly decreased the shear bond strength. Shear bond strength of permanent dentin was significantly higher than that of primary dentin. Clearfil SE Bond treatment groups showed the highest shear bond strength and AQ Bond treatment groups showed the lowest shear bond strength.

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On the penecontemporaneous deformation structures of the Sinri area at the mid western boundary of the Jinan Basin (진안분지 서변 중앙부 신리지역의 준퇴적동시성 변형구조)

  • Lee Young-Up
    • The Korean Journal of Petroleum Geology
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    • v.6 no.1_2 s.7
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    • pp.8-19
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    • 1998
  • In the Sinri area located at the mid western boundary of the Jinan basin, the Manduksan Formation which mainly consists of coarse sandstone narrowly intercalated with shale and the alternation of sand and shale and the Dalgil Formation mainly of shale are distributed. It consists of four lithofacies, such as coarse sandstone, interbedded sandstone/shale, shale and volcanic rock lithofacies. All sediments are interpreted to be deposited by turbidity currents and free fallouts in a lacustrine basin. In these rocks many penecontemporaneous defomation structures are observed such as fold and thrust fault at large scale, and swelling, boudin structure, flame structure, load structure, ptygmatic fold and convolute bedding at small scale. All these structures are developed between upper and lower undisturbed sedimentary strata. Two large folds are similar folds, but lower one gradually developed into concentric shape. The swelling structures by convergence of the sediments are observed in the hinge area and the boudin structures are developed in the limb. The thrust faults including minor folds and sandstone lobes show duplex structure with asymmetric and kink fold on and below in front of the detached sandstone layer. Development of the swellings, boudins and lobes indicates the flexbility of the sediments during deformational episodes. The folds and thrust faults rarely contain fractures relative their scales and lithologies. This feature also indicates the retrievability of sediments during deformation. At the flanks of the thrust faults the normal faults are formed contemporaneously. The deformation structures at small scale such as flame structures, load structures, ptygmatic folds and convolute beddings are syndepositional and penecontemporaneous, which show the effects of tectonic movements. All these deformed sedimentary structures of the Sinri area suggest the continuing tectonic movements during and/or after deposition.

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Nematicidal Activity of Streptomyces flavogriseus KRA15-528 to Meloidogyne incognita (Meloidogyne incognita에 대한 Streptomyces flavogriseus KRA15-528의 살선충활성)

  • Oh, Mira;Han, Jae Woo;Choi, Jung Sup;Choi, Yong Ho;Jang, Kyoung Soo;Choi, Gyung Ja;Kim, Hun
    • Research in Plant Disease
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    • v.22 no.4
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    • pp.227-235
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    • 2016
  • Plant disease caused by root-knot nematode is a major problem in crop production. Using of chemical pesticides, one of the most efficient methods to control nematodes, have raised issues in toxicity to humans and animals and environmental pollution. In this study, to select actinomycete strains that have potential to serve as a microbial agent for control of nematodes, we investigated nematicidal activity of culture broth from 670 Streptomyces isolates. A culture filtrate of KRA15-528 isolate that was identified as S. flavogriseus on the basis of 16S rRNA sequence analysis, showed strong nematicidal activity against second stage of juveniles of Meloidogyne incognita and inhibited egg hatching; exposure to 10% of culture filtrate resulted in 71% juvenile mortality at 48 hours afters treatment and suppressed egg hatching by 54% at 9 days after treatment. When the KRA15-528 culture filtrate was partitioned with ethyl acetate and butanol, ethyl acetate layer exclusively showed strong activity; 91%, 53%, 30% of mortality at 1,000, 500, $250{\mu}g/ml$, respectively. Additionally, the culture filtrate suppressed gall formation on cucumber plant by M. incognita with no phytotoxicity. These results suggest that S. flavogriseus KRA15-528 has potential to serve as a microbial nematicide for the control of root-knot nematode disease.

Application of Geophysical Methods to Detection of a Preferred Groundwater Flow Channel at a Pyrite Tailings Dam (황철석 광산 광미댐에서의 지하수흐름 경로탐지를 위한 물리탐사 적용)

  • Hwang, Hak Soo
    • Economic and Environmental Geology
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    • v.30 no.2
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    • pp.137-142
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    • 1997
  • At the tailings dam of the disused Brukunga pyrite mine in South Australia, reaction of groundwater with the tailings causes the formation and discharge of sulphuric acid. There is a need to improve remediation efforts by decreasing groundwater flow through the tailings dam. Geophysical methods have been investigated to determine whether they can be used to characterise variations in depth to watertable and map preferred groundwater flow paths. Three methods were used: transient electromagnetic (TEM) soundings, direct current (DC) soundings and profiling, and self potential (SP) profiling. The profiling methods were used to map the areal extent of a given response, while soundings was used to determine the variation in response with depth. The results of the geophysical surveys show that the voltages measured with SP profiling are small and it is hard to determine any preferred channels of groundwater flow from SP data alone. Results obtained from TEM and DC soundings, show that the DC method is useful for determining layer boundaries at shallow depths (less than about 10 m), while the TEM method can resolve deeper structures. Joint use of TEM and DC data gives a more complete and accurate geoelectric section. The TEM and DC measurements have enabled accurate determination of depth to groundwater. For soundings centred at piezometers, this depth is consistent with the measured watertable level in the corresponding piezometer. A map of the watertable level produced from all the TEM and DC soundings at the site shows that the shallowest level is at a depth of about 1 m, and occurs at the southeast of the site, while the deepest watertable level (about 17 m) occurs at the northwest part of the site. The results indicate that a possible source of groundwater occurs at the southeast area of the dam, and the aquifer thickness varies between 6 and 13 m. A map of the variation of resistivity of the aquifer has also been produced from the TEM and DC data. This map shows that the least resistive (i.e., most conductive) section of the aquifer occurs in the northeast of the site, while the most resistive part of the aquifer occurs in the southeast. These results are interpreted to indicate a source of fresh (resistive) groundwater in the southeast of the site, with a possible further source of conductive groundwater in the northeast.

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An Initiative Study on Relationship between Algal Blooms and Asian Dust for Regulation of Algal Blooms (조류 성장 억제를 위한 녹조 및 적조 발생과 황사의 상관관계 초기적 연구)

  • Kim, Tai-Jin;Jeong, Jaechil;Seo, Rabeol;Kim, Hyung Moh;Kim, Dae Geun;Chun, Youngsin;Park, Soon-Ung;Yi, Sehyoon;Park, Jun Jo;Lee, Jin Ha;Lee, Jay J.;Lee, Eun Ju
    • KSBB Journal
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    • v.29 no.4
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    • pp.285-296
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    • 2014
  • Although the problems of the algal blooms have been world-widely observed in freshwater, estuary, and marine throughout the year, it is not yet certain what are the basic causes of such blooms. Consequently, it is very difficult to predict when and where algal blooms occur. The constituents of the Asian dust are in a good agreement with the elements required for the algal growth, which suggests some possible relationship between the algal blooms and the Asian dust. There have been frequently algal blooms in drinking water from rivers or lakes. However, there is no any algal blooms in upwelling waters where the Asian dust cannot penetrate into the soil due to its relatively weak settling velocity (size of particles, $4.5{\pm}1.5{\mu}m$), which implies the possible close relationship of the Asian dust with algal blooms. The present initiative study is thus intended firstly in Korea to illustrate such a relationship by reviewing typical previous studies along with 12 years of weekly iron profiles (2001~2012) and two slant culture experiments with the dissolved Asian dust. The result showed bacterial suspected colonies in the slant culture experiment that are qualitatively in a good agreement with the recent Japanese studies. Since the diatoms require cheap energy (8%) compared to other phytoplankton (100%) to synthesize their cell walls by silicate, the present results can be used to predict algal blooms by diatoms if the concentrations of iron and silicate are available during spring and fall. It can be postulated that the algal blooms occur only if the environmental factors such as light, nutrients, calm water surface layer, temperature, and pH are simultaneously satisfied with the requirements of the micronutrients of mineral ions supplied by the Asian dust as enzymatic cofactors for the rapid bio-synthesis of the macromolecules during algal blooms. Simple eco-friendly methods to regulate the algal blooms are suggested for the initial stage of blooming with limited area: 1) to cover up the water surface with black curtain and inhibit photosynthesis during the day time, 2) to blow air (20.9%) or pure oxygen into the bottom of the water and inhibit rubisco for carbon uptake and nitrate reductase for nitrogen uptake activities in algal growth during the night, 3) to eliminate the resting spores or cysts by suction of bottom sediments as deep as 5 cm to prevent the next year germinations.

Effect of Microbial Additives on Metabolic Characteristics in Sheep and Milking Performance of Lactating Dairy Cows (미생물제제의 첨가가 면양의 반추대사 및 젖소의 유생산성에 미치는 영향)

  • Kim, G.L.;Choi, S.K.;Choi, S.H.;Song, M.K.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.819-828
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    • 2007
  • Two experiments were conducted to observe the effects of direct fed microbials on metabolic characteristics in sheep and milking performance in dairy cows. A metabolic trial with four ruminally cannulated sheep(60±6kg) was conducted in a 4×4 Latin square design to investigate the supplementation effects of Saccharomyces cerevisiae, Clostridium butyricum or mixed microbes of S. cerevisiae and C. butyricum on ruminal fermentation characteristics and whole tract digestibility. Sheep were fed 1.25 kg of total mixed ration(TMR, DM basis) supplemented with S. cerevisiae (2.5g/day), C. butyricum (1.0g/day) or its mixture(S. cerevisiae 1.25g/day+C. butyricum 1g/day), twice daily in an equal volume. But control sheep were fed only TMR. A feeding trial with 28 lactating Holstein cattle was also conducted for 12 weeks to investigate the effects of the same microbial supplements as for the metabolic trial on milking performance. The cows were fed the TMR(control), and fed S. cerevisiae(50g/day), C. butyricum(15g/day) or its mixture (S. cerevisiae 25g/day + C. butyricum 7.5g/day) with upper layer dressing method. Total VFA concentration and the digestibility of whole digestive tract in the sheep increased by supplementation of S. cerevisiae, C. butyricum or their combined microbials compare to control group. The proportion of propionic acid at 1h(P<0.039) and 3h(P<0.022) decreased by supplementation of S. cerevisiae while tended to increase acetic acid proportion at the same times. Daily dry matter intake(DMI) was not influenced by the microbial treatments, but milk yield(P<0.031) and feed efficiency(milk yield/DMI, P<0.043) were higher for the cow received C. butyricum than those for other treatments. The milk fat content was higher (P<0.085) when cows fed S. cerevisiae(4.11%) than that fed the control (4.08%), the diets with C. butyricum (3.85%) and the microbial mixture. Based on the results obtained from the current experiments, supplementation of C. butyricum or mixture with S. cerevisiae might be increased milk fat content and milk productivity of lactating daily cows. (Key words:Saccharomyces cerevisiae, Clostridium butyricum, Fermentation characteristics,