Kim, Jin-Woo;Yu, Mi-Kyung;Lee, Se-Joon;Lee, Kwang-Won
Restorative Dentistry and Endodontics
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v.28
no.1
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pp.80-88
/
2003
Object The purpose of this study were to evaluate the microtensile bond strength of resin fiber reinforced post to radicular dentin using resin cement according to various dentin surface treatment and to observe the inter face between post and root dentin under SEM Material and Method A total 16 extracted human single rooted teeth were used. A lingual access was made using a #245 carbide bur in a high-speed handpiece with copious air water spray. The post space was mechanically enlarged using H-file(up to #60) and Gates Glidden bures(#3). This was followed by refining of the canal space using the calbrating drill set provided in ER Dentinpost(GEBR, BRASSELER GmbH&Co. KG). The 16 teeth were randomly distributed into 4 group of 4 teeth. Group 1 teeth had their post space prepared using 10% phosphoric acid as root canal surface treatment agent during 20s. The canal was then rinsed with saline and dried with paper point. Group 2 teeth had their post space prepared using 3% NaOCl as root canal surface treatment agent during 30min. The canal was then rinsed with saline and dried with paper point. Group 3 teeth had their post space prepared using 17% EDTA as root canal surface treatment agent during 1min. The canal was then rinsed with saline and dried with paper point. Group 4 teeth had their post space prepared using 17% EDTA as root canal surface treatment agent during 1min. After rinsing with saline, the canal was rinced 10m1 of 3% NaOCl for 30min. After drying with paper point, the post(ER Dentinpost, GEBR, BRASSELER GmbH&Co. KG) was placed in the treated canals using resin cement. Once the canal was filled with resin cement(Super bond C&B sunmedical co. Ltd.), a lentulo was inserted to the depth of the canal to ensure proper coating of the root canal wall. After 24 hours, acrylic resin blocks($10{\cdot}10{\cdot}50mm$) were made. The resin block was serially sectioned vertically into stick of $1{\cdot}1mm$. Twenty sticks were prepared from each group. After that, tensile bond strengths for each stick was measured with Microtensile Tester. Failure pattern of the specimen at the interface between post and dentin were observed under SEM. Results 1. Tensile bond strengths(meen{\pm}SD$) ) were expressed with ascending order as follows group 4, $12.52{\pm}6.60$ ; group 1, $7.63{\pm}5.83$ ; group 2, $4.13{\pm}2.31$ ; group 3, $3.31{\pm}1.44$. 2. Tensile bond strengths of Group 4 treated with 17% EDTA +3%NaOCl were significant higher than those of group 1, 2 and 3 (p<0.05). 3. Tensile bond strengths of Group 1 treated with 10% phosphoric acid were significant higher than those of group 2 (p<0.05). Tensile bond strengths of Group 4 treated with 17% EDTA +3% NaOCl was significant higher than those of other groups.
Lee C.Y.;Ha S.H.;Lee H.P.;Baik K.H.;Jin S.K.;Sohn S.H.;Park M.J.
Proceedings of the KSAR Conference
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2005.06a
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pp.47-54
/
2005
In boars, unlike the cases in males of other species, gonadal hormones suppress voluntary feed intake. for this reason, barrows, compared with gilts or boars, eat too much feed resulting in excessive fat deposition. Two experiments were performed in the present study to investigate the effects of implantation of Revalor H[Experiment(Exp.) I: 140mg trenbolone acetate(a synthetic androgen) + 14mg estradiol-$17\beta(E_{2}\beta)$] and Compudose(Exp. II; 24mg $E_{2}\beta$) on growth efficiency, carcass characteristics and circulating concentrations of IGF-I and IGF-binding protein-3(IGFBP-3). In Exp. I, sixty-four cross-bred finishing barrows weighing approximately 60kg were randomly divided into eight pens under a 2[control vs Revalor implant] $\times$ 2(ad libitum vs $80\%$ ad libitum feeding) $\times$2[control($103\%$ NRC-recommended level) vs low-energy($87\%$ NRC recommendation) diet] arrangement of treatments. In Exp. II, effects of Compudose were studied using 80 finishing barrows(10 animals/pen). In both Exps., all the animals were slaughtered at 100- to 110-kg body weight. Both Revalor and Compudose implants caused a decrease in feed intake and backfat thickness without affecting major physicochemical characteristics of the carcass and an increase in circulating IGF-I concentration. Moreover, Revalor implant exhibited greater effects than restricted feeding, low-energy diet, or Compudose in these variables. In addition, Revalor implantation suppressed weight gain, but enhanced the feed efficiency without exhibiting any interaction with the diet or feeding. In summary, results suggest that 1) both androgen and estrogen suppress voluntary feed intake and backfat deposition and enhance IGF-I secretion and 2) these effects of the gonadal steroid hormones in growth are likely to be mediated, in part, by IGF-I in finishing barrows.
Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.
The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of $\beta$-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F$_2$ population composed of 214 individuals from an intercross between Korean Native Boars and Landrace sows. PCR products from two primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and HinfⅠ, revealed fragment length polymorphisms(RFLPs). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP/HinfⅠ was .38, .41 and .20, respectively, in the population. Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weights at 3, 5, 12, and 30 week of age (p〈.05 to .001). The ‘d’ allele was associated with gaining of body weight. In H-FABP/HinfⅠ locus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p〈.05 or p〈.001) and back fat thickness, body fat including abdominal and trimmed fat (p〈.001) and intramuscular fat(p〈.05) The ‘H’ allele was positively associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.
The leptin receptor gene(LEPR) produces a high affinity receptor that mediates the regulation of the leptin gene. Leptin secreted from adipose tissue plays an important role in regulating feed intake and energy balance. In this study, a microsatellite marker within LEPR was selected and genotyped for the F2 population composed of 354 individuals from an intercross between Korean Native boars and Landrace sows. Totally, six alleles (255, 259, 261, 263, 265 and 267bp) and nineteen genotypes were detected in the population, of which the CE (261/265), CC (261/261) and EE (265/265) types were observed by 20.0%, 10.1% and 9.6%, respectively. Relationships between their genotypes and economic traits were analyzed. We found specific genotypes associated with economic traits such as body weight at 12 weeks of age/body fat including abdominal and trimmed fat/shear force (P〈0.001), body weight of 30 weeks of age (P〈0.01) and body weight of 3 weeks of age/back fat thickness (P〈0.05). The DD (263/263) and DF (263/267) types were associated with body weight at 3, 5, 12 and 30 weeks of age. The DF (263/267) type showed a highly significant effect on back fat thickness and body fat including abdominal and trimmed fat. The DF (263/267) type showed positive effect on shear force, whereas the BB (259/259) and DD (263/263) types negatively affected on tenderness.
Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
Journal of Embryo Transfer
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v.20
no.2
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pp.105-112
/
2005
The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.
Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
Journal of Embryo Transfer
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v.20
no.2
/
pp.169-176
/
2005
Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.
The objective of this study was to supply excellent genetic resources to livestock farms by transferring embryos produced by genetically superior Korean cows (Hanwoo). Eighty Hanwoo donors were superovulated with gonadotropin ($Folltrpin^(R)\;or\;Antorin^(R)$) for 4 days combined with or without progesterone releasing intravaginal device (CIDR) insertion. The collected fresh or frozen-thawed embryos were transferred to 226 farm recipients. In this study, the effect of CIDR insertion in combination with gonadotropin ($Folltrpin^(R)$) treatments initiated at the random stage of estrous cycle on embryo production was evaluated and compared to conventional superovulation protocol. Moreover, the effect of gonadotropin ($Antorin^(R)$) dose in CIDR-treated Hanwoo donors on the embryo yield was determined. In addition, the effects of embryos (fresh vs. frozen-thawed), embryo transfer person, seasons and farms on the pregnancy rate were evaluated. In Hanwoo donors, CIDR insertion in combination with $Folltrpin^(R)$ treatments regardless of estrous detection resulted in increased numbers of total ova (6.5 vs. 5.8) and transferable embryos (3.9 vs. 3.2) compared to the conventional superovulation protocol (p<0.01). In CIDR-treated Hanwoo donors, the higher dose of $Antorin^(R)$ (36 vs. 28 mg) resulted in the increased number of transferable embryos (8.3 vs. 5.4, p<0.05). The embryos (fresh 43.9% vs. frozen-thawed 23.1%) and embryo transfer person (53.9 vs. $0{\sim}16.7%$) significantly affected the pregnancy rate after embryo transfer (p<0.01). These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos and, multiple ovulation and embryo transfer in Hanwoo might be effectively applied for livestock improvement if pregmancy rate with frozen-thawed embryos and embryo transfer skill would be improved.
It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.
The objective of this study was carried out to examine the polar body extrusion of in vitro matured porcine follicular oocytes as a non-invasive marker of oocyte quality to know the developmental competence in advance. The porcine oocytes matured for 48 hours were examined the polar body extrusion and some parts were stained. The examined oocytes were matured for additional $16{\sim}18$ hours and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 hours for diploid formation. The treated oocytes were washed and cultured for 7 days. The polar body extrusion and degeneration rates were varied with $9.9{\sim}52.4%$ and $21.4{\sim}61.8%$ by repetition. The polar body extruded oocytes were shown the polar body chromosome and metaphase II plate by staining. However the non-extruded oocytes were shown expanded nucleus with 39.1%, premature chromosome condensation with 19.6%, metaphase I plate with 10.9 %, metaphase II with 13%, condensed chromatin with 6.5%, and absent nuclear material with 8.7%. The oocytes that were not examined for the polar body extrusion were cleaved 45.0%, and developed to blastocyst stage with 11.3%. In examined oocytes for polar body extrusion,. the polar body extruded oocytes were cleaved 94.2% and developed with 42.5%. This result suggests that discarding of the degenerating oocytes and oocytes that not extruded polar body will be effective for experiments of culturing effect in porcine embryos and embryo biotechnology.
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