• Title/Summary/Keyword: Hybrid Technology

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Evaluation of Potential of Mandarin Hybrid 'Shiranuhi' against inoculation of Bacterial Canker Disease Pathogen (Xanthomonas axonopodis pv. citri) in Citrus Field in Jeju Island

  • Hyun, Jae-Wook;Myung, Inn-Shik;Lee, Seong-Chan;Kim, Kwang-Sik;Lim, Han-Cheol
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.248-252
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    • 2003
  • This study was carried out to evaluate potential of Shiranuhi cultivar against inoculation of causal pathogen of citrus canker, Xanthomonas axonopodis pv. citri in Jeju Island by comparing degree of susceptibility of fruits and leaves/twigs, and analysis of incidence of canker disease. Progression of symptom, disease incidence, and percent area of lesion were surveyed for evaluation of resistance. In greenhouse condition, symptoms of bacterial citrus canker progressed more rapidly in sweet orange, a moderately susceptible cultivar, than in other four cultivars (satsuma mandarin, 'Kiyomi', 'Shiranuhi' and 'Yuzu'). At 20 days after inoculation, disease severity was the highest in sweet orange (5.0$\pm$0.0), and all tested leaves were distorted or had dropped. On the other hand, 'Yuzu' showed the lowest disease severity (2.6$\pm$0.47), followed by 'Kiyomi' (4.0$\pm$0.0), 'Shiranuhi' (4.0$\pm$0.82), and satsuma mandarin (4.3$\pm$0.47). Percent area of lesion per leaf 30 days after inoculation was the highest in sweet orange (8.31$\pm$1.78), followed by satsuma mandarin (1.51$\pm$1.25), 'Shiranuhi' (1.39$\pm$0.94), and 'Kiyomi' (1.1$\pm$0.9), while the lowest was in 'Yuzu' (0.26$\pm$0.17). Infield condition, percentage of diseased leaf in 'Shiranuhi' was very low, 5.2$\pm$2.9, compared with sweet orange, 71.0$\pm$ 11.5, while that of satsuma mandarin and 'Kiyomi' were 6.9$\pm$7.0 and 4.3$\pm$2.0, respectively. Percentages of diseased leaf was higher (17.4$\pm$7.1) than that of diseased fruit (3.2$\pm$2.5) in severely diseased trees of Shiranuhi cultivar, and the disease was not observed on twig in open field condition. Lesion sizes on leaves and fruits in open field condition were 4.1$\pm$2.2 mm2 and 5.1$\pm$5.6 mm2, respectively, while those in greenhouse condition were 8.7$\pm$5.7 mm2, 10.4$\pm$9.2 mm2 and 5.6$\pm$2.6 mm2 on leaves, fruits and twigs, respectively. The disease was observed in 5.6% out of total 107 farmers Shiranuhi fields under polyethylene film house, and average percentages of diseased tree in 31 fields of Shiranuhi cultivar and adjacent satsuma mandarin fields were 0.02% and 14.8%, respectively. Average percentage of diseased fruit was 1.6% in satsuma mandarin which was not observed in anyone of all the 31 Shiranuhi farmers fields. Therefore, it was concluded that 'Shiranuhi' cultivar is not potential against causal pathogen of citrus canker disease in Jeju Island because the cultivar has similar resistance as satsuma mandarin which occupies over 95% of total 25,000 ha in Jeju Island in polyethylene film houses protected from outside.

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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Allelism and Molecular Marker Tests for Genic Male Sterility in Paprika Cultivars (파프리카 시판 품종에 대한 유전자적 웅성불임성의 대립성 및 분자표지의 유용성 검정)

  • Lee, Jun-Dae;Do, Jae-Wahng;Han, Jung-Heon;An, Chul-Geon;Kweon, Oh-Yoel;Kim, Yong-Kwon;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.29 no.2
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    • pp.130-134
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    • 2011
  • Paprika (Capsicum annuum L.), a colored bell-type sweet pepper, is one of the most important money making vegetable crops in Korea. The cultivation area, total production, and exports of paprika are gradually getting increased, but the paprika cultivars used in Korea are all imported. It was well-known that the genic male sterility (GMS) is the main way to produce paprika hybrid seeds. However, it is little known that how many and what kinds of ms genes are used for breeding of paprika $F_1$ varieties. In this study, eight paprika cultivars ('Special', 'Debla', 'Plenty', 'Fiero', 'Boogie', 'Fiesta', 'Derby', and 'Minibell'), popularly cultivated in Korea and three different genic male sterile lines ('GMSP', 'GMS3', and 'GMSK') were used. For allelism test among the $F_1$ cultivars, half diallel crosses were performed. The result demonstrated that the most of the GMS in paprika cultivars except for 'Minibell' were same allele. To identify which GMS gene(s) were used for paprika $F_1$ cultivars, top crosses between previously known GMS lines and the $F_1$ cultivars were performed. As a result, we found that the $ms_k$ and the $ms_p$ genes were alleles for the GMS of 'Minibell' and for the other cultivars, respectively. We also confirmed that the GMS gene identification using GMSK-CAPS marker linked to the $ms_k$ gene and the PmsM1-CAPS marker linked to the $ms_p$ gene in $F_2$ progenies of 'Minibell' and 'Fiesta' and 'Derby' cultivars, respectively. In addition, we developed the PmsM2-CAPS marker for 'Plenty', 'Fiero', and 'Boogie' cultivars. We expect that these markers will be very useful for breeding new maternal (male sterile) line of paprika.

Structural change and electrical conductivity according to Sr content in Cu-doped LSM (La1-xSrxMn0.8Cu0.2O3) (Sr 함량이 Cu-doped LSM(La1-xSrxMn0.8Cu0.2O3)의 구조적변화와 전기전도도에 미치는 영향)

  • Ryu, Ji-Seung;Noh, Tai-Min;Kim, Jin-Seong;Lee, Hee-Soo
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.2
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    • pp.78-83
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    • 2012
  • The structural change and the electrical conductivity with Sr content in $La_{1-x}Sr_xMn_{0.8}Cu_{0.2}O_3$ (LSMCu) were studied. $La_{0.8}Sr_{0.2}MnO_3$ (LSM) and $La_{1-x}Sr_xMn_{0.8}Cu_{0.2}O_3$ ($0.1{\leq}x{\leq}0.4$) were synthesized by EDTA citric complexing process (ECCP). A decrease in the lattice parameters and lattice volumes was observed with increase of Sr content, and these results were attributed to the increasing $Mn^{4+}$ ions and $Cu^{3+}$ ions in B-site. The electrical conductivity measured from $500^{\circ}C$ to $1000^{\circ}C$ was increased with increase of Sr content in the $0.1{\leq}x{\leq}0.3$ composition range, and it was 172.6 S/cm (at $750^{\circ}C$) and 177.7 S/cm (at $950^{\circ}C$, the maximum value) in x = 0.3. The electrical conductivity was decreased in x = 0.4 because of the presence of the second phase in the grain boundaries. The lattice volume was contracted by increase of $Mn^{4+}$ ions and $Cu^{3+}$ ions in B-site according to increase of Sr content and the electrical conductivity was increased with increase of charge carriers which were involved in the hopping mechanism.

Developmental competence and Effects of Coculture after Crypreservation of Blastomere-Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis (착상 전 유전진단 기술 개발의 동물실험 모델로서 할구 생검된 생쥐 배아에서 동결보존 융해 후 배아 발생 양상과 공배양 효과에 관한 연구)

  • Kim, Seok-Hyun;Kim, Hee-Sun;Ryu, Buom-Yong;Choi, Sung-Mi;Pang, Myung-Geol;Oh, Sun-Kyung;Jee, Byung-Chul;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chae, Hee-Dong;Kim, Chung-Hoon
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.47-57
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    • 2000
  • Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

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Recent Progress in Air-Conditioning and Refrigeration Research : A Review of Papers Published in the Korean Journal of Air-Conditioning and Refrigeration Engineering in 2013 (설비공학 분야의 최근 연구 동향 : 2013년 학회지 논문에 대한 종합적 고찰)

  • Lee, Dae-Young;Kim, Sa Ryang;Kim, Hyun-Jung;Kim, Dong-Seon;Park, Jun-Seok;Ihm, Pyeong Chan
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.26 no.12
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    • pp.605-619
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    • 2014
  • This article reviews the papers published in the Korean Journal of Air-Conditioning and Refrigeration Engineering during 2013. It is intended to understand the status of current research in the areas of heating, cooling, ventilation, sanitation, and indoor environments of buildings and plant facilities. Conclusions are as follows. (1) The research works on the thermal and fluid engineering have been reviewed as groups of fluid machinery, pipes and relative parts including orifices, dampers and ducts, fuel cells and power plants, cooling and air-conditioning, heat and mass transfer, two phase flow, and the flow around buildings and structures. Research issues dealing with home appliances, flows around buildings, nuclear power plant, and manufacturing processes are newly added in thermal and fluid engineering research area. (2) Research works on heat transfer area have been reviewed in the categories of heat transfer characteristics, pool boiling and condensing heat transfer and industrial heat exchangers. Researches on heat transfer characteristics included the results for general analytical model for desiccant wheels, the effects of water absorption on the thermal conductivity of insulation materials, thermal properties of Octadecane/xGnP shape-stabilized phase change materials and $CO_2$ and $CO_2$-Hydrate mixture, effect of ground source heat pump system, the heat flux meter location for the performance test of a refrigerator vacuum insulation panel, a parallel flow evaporator for a heat pump dryer, the condensation risk assessment of vacuum multi-layer glass and triple glass, optimization of a forced convection type PCM refrigeration module, surface temperature sensor using fluorescent nanoporous thin film. In the area of pool boiling and condensing heat transfer, researches on ammonia inside horizontal smooth small tube, R1234yf on various enhanced surfaces, HFC32/HFC152a on a plain surface, spray cooling up to critical heat flux on a low-fin enhanced surface were actively carried out. In the area of industrial heat exchangers, researches on a fin tube type adsorber, the mass-transfer kinetics of a fin-tube-type adsorption bed, fin-and-tube heat exchangers having sine wave fins and oval tubes, louvered fin heat exchanger were performed. (3) In the field of refrigeration, studies are categorized into three groups namely refrigeration cycle, refrigerant and modeling and control. In the category of refrigeration cycle, studies were focused on the enhancement or optimization of experimental or commercial systems including a R410a VRF(Various Refrigerant Flow) heat pump, a R134a 2-stage screw heat pump and a R134a double-heat source automotive air-conditioner system. In the category of refrigerant, studies were carried out for the application of alternative refrigerants or refrigeration technologies including $CO_2$ water heaters, a R1234yf automotive air-conditioner, a R436b water cooler and a thermoelectric refrigerator. In the category of modeling and control, theoretical and experimental studies were carried out to predict the performance of various thermal and control systems including the long-term energy analysis of a geo-thermal heat pump system coupled to cast-in-place energy piles, the dynamic simulation of a water heater-coupled hybrid heat pump and the numerical simulation of an integral optimum regulating controller for a system heat pump. (4) In building mechanical system research fields, twenty one studies were conducted to achieve effective design of the mechanical systems, and also to maximize the energy efficiency of buildings. The topics of the studies included heating and cooling, HVAC system, ventilation, and renewable energies in the buildings. Proposed designs, performance tests using numerical methods and experiments provide useful information and key data which can improve the energy efficiency of the buildings. (5) The field of architectural environment is mostly focused on indoor environment and building energy. The main researches of indoor environment are related to infiltration, ventilation, leak flow and airtightness performance in residential building. The subjects of building energy are worked on energy saving, operation method and optimum operation of building energy systems. The remained studies are related to the special facility such as cleanroom, internet data center and biosafety laboratory. water supply and drain system, defining standard input variables of BIM (Building Information Modeling) for facility management system, estimating capability and providing operation guidelines of subway station as shelter for refuge and evaluation of pollutant emissions from furniture-like products.

Effects of Coculture on Development of Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis of Human Embryos (생쥐 모델을 이용한 배아의 할구 생검법과 할구가 생검된 배아의 배양시 공배양 효과에 관한 연구: 인간에서의 착상 전 유전진단 기술 개발을 위한 동물실험 모델의 개발)

  • Kim, S.H.;Ryu, B.Y.;Jee, B.C.;Choi, S.M.;Kim, H.S.;Pang, M.G.;Oh, S.K.;Suh, C.S.;Choi, Y.M.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.;Chae, H.D.;Kim, C.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.9-20
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    • 1999
  • The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.

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Comparative study of surface roughness between several finishing and polishing procedures on ormocer-based composite resin and nanohybrid composite resin (복합 레진에서 마무리 방법에 따른 표면 거칠기 비교)

  • Jeong, Suk-In;Oh, Nam-Sik;Lee, Myung-Hyeon;Lee, En-Jung;Cho, Jung-Hyeon;Ji, Sung-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.2
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    • pp.105-115
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    • 2008
  • Statement of problem: Proper finishing and polishing enhance both the esthetics and the longevity of restored teeth. Blade finishing technique would be suited for smoothing and finishing. Evaluation of this technique are necessary. Purpose: The purpose of this study was to evaluate the blade finishing and polishing procedures on the surface profile and roughness of ormocer-based composite resin and nanohybrid composite resin. Material and methods: The material included a ormocer-based composite resin ($Admira^{(R)}$ & $Admira^{(R)}$ Flow); a nanohybrid composite resin ($Grandio^{(R)}$ & $Grandio^{(R)}$ Flow). One hundred forty specimens of each group were prepared using a mylar strip and randomly divied into blade finishing and rubber polishing groups (n=10). The average surface roughness (Ra) in micrometers was measured and the surface profile was examined by scanning electron microscopy (SEM) (Magnification ${\times}$ 200). The data were analyzed by Mann-Whitney Test at 0.05 significance level. Conclusion: The results of this study indicated that the mylar strip produced the smoothest surface on all materials and among the finishing-polishing methods was not significanct difference (P>0.05). Ormocer-based flowable composite resin performedthe lowest variability in initial surface roughness among the tested materials.

Effects of Permeable Cryoprotectants on Viability of Mammalian Embryo Model (침투성 동결보호제가 포유류 초기배자의 생존성에 미치는 영향)

  • Kim, Hyun;Cho, Sang-Rae;Kim, Dong Kyo;Choe, Changyong;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.195-200
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    • 2015
  • The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female $F-{_1}$ mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG ($81.1{\pm}15.1$), 1,2-PROH ($88.0{\pm}21.1$) or the control ($99.9{\pm}21.3$) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO ($14.7{\pm}1.3$), EG ($12.1{\pm}1.1$), Glycerol ($15.2{\pm}1.8$), and 1,2-PROH ($11.5{\pm}1.3$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.5{\pm}0.7$, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

Development of Effective Cryopreservation Method for Mammalian Embryo (포유류 초기 배아의 효율적인 동결 보존 방법에 관한 연구)

  • Kim, Hyun;Cho, Young Moo;Ko, Yeoung-Gyu;Kim, Sung Woo;Seong, Hwan-Hoo;Yamanouchi, Keitaro
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.241-248
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    • 2014
  • This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4~8 cell embryos in EG were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG showed better development rates of embryos in 2 and 4~8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.