• Journal of Embryo Transfer
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• v.20 no.2
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• pp.169-176
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• 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.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.147-156
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• 2006

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.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.157-162
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• 2006

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.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.169-175
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• 2006

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.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.177-182
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• 2006

This study was conducted to investigate the effects of melengesterol acetate (MGA) and $PGF_{2{\alpha}}$ administrations on serum progesterone level, synchrony of estrus and conception rates in Han-woo. Firstly, ten heifers and one freematin were fed 0.5 mg MGA/day for 14 days in a grain carrier, and after 19 days of MGA feeding, a single injection of 25 mg $PGF_{2{\alpha}}$ were treated. Blood samples were collected to evaluate serum progesterone concentrations from the start of feeding of MGA until the end of feeding and subsequent estrous detection and artificial insemination (AI) at 3 days intervals, and on days of $PGF_{2{\alpha}}$ injection, estrous detection, AI, and 15th and 60th days after AI. The level of progesterone in the blood began to increase from 7 days after MGA feeding, and 9 days after feeding it became 5.4 ng/ml and maintained that level thereafter. On the 33th day when the $PGF_{2{\alpha}}$ was injected, it reached the peak level of 7.6 ng/ml. However, 2-3 days after $PGF_{2{\alpha}}$ injection, it dropped to 1.4 ng/ml drastically (p<0.05). Secondly, one hundred and ninety four Hanwoo heifers or cows were divided into two groups to compare estrous induction and conception rates: the one treated with MGA and $PGF_{2{\alpha}}$, (n=104) and the other with $PGF_{2{\alpha}}$ treatment (two injections at 11 days interval, n=90). The heifers or cows treated with MGA and $PGF_{2{\alpha}}$ were identical to those used as above. The percentages of heifers or cows showed estrus were higher in the $MGA+PGF_{2{\alpha}}$ treatment (91.3%) than in the $PGF_{2{\alpha}}$ treatment (72.2%, p<0.05). Conception rates were also higher in the $MGA+PGF_{2{\alpha}}$ treatment (94.2%) than in the $PGF_{2{\alpha}}$ treatment (88.9%, p<0.05). The results of this experiment indicate that estrus synchronization using $MGA+PGF_{2{\alpha}}$ is more effective than that using $PGF_{2{\alpha}}$ (two injections) in Hanwoo.

• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.3
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• pp.241-250
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• 1999

A field plot experiment was carried out on mixed pasture from October 1995 to October 1997 to examine the effect of weed control as affected by the difference of mixed pasture species, planning with spilt plot design which main plots of 2 treatments (companion crop Italian ryegrass Lolium multiflorum and Kentucky bluegrass Poa pratensis) are divided into subplots of 4 treatments (T1: single companion crop, T2: Tall fescue Festuca arundinasea + Orchardgrass Dactylis glomerata + Ladino clover Trifolium repens, T3 : Perennial ryegrass Lolium perenne + orchardgrass + ladino clover, T4 : orchardgrass + ladino clover). Average 2 year-dry matter yields of companion crops as whole plots, Italian ryegrass and Kentucky bluegrass, were 15,437kg/ha and 14,629kg/ha, respectively and those of T1, T2, T3 and T4 as sub plots were 14,815, 15,407, 14,629 and 15,187kg/ha respectively without statistical1y significant difference(P>0.05). Though there was no significant difference between the dry matter yields of Italian ryegrass and Kentucky bluegrass, as whole plots, at 1st harvest of second year, dry matter yields of T3 and T4, as 4,591kg/ha and 4,816kg/ha respectively, were higher than those of T1, 3,376kg/ha(P<0.05), significantly. There was a significant difference among 2nd year-average DM yields of sub plots(P<0.05), showing highest with T4 15,131kg/ha and least with T1 13,085kg/ha and T3 13,269kg/ha. Italian ryegrass, as the companion crop, was better for the control of weeds in the early spring than Kentucky bluegrass, but reached to the similar botanical composition level with Kentucky bluegrass after summer with rapid decrease of Italian ryegrass. Percentage of Kentucky bluegrass in botanical composition increased slowly, but did'nt become an important factor for the control of weeds at the end of this experiment. Using orchardgrass in mixed pasture such as T2, T3 and T4 made weeds more decreased in mixed pasture than one in single conpanion crop. There was no difference among treatments of whole or sub plots in crude protein contents of pasture taken in this study. Contents of NDF and ADF of companion crop. Italian ryegrass plots harvested in October 1997 were hither than those of Kentucky bluegrass plots(P<0.05). ADF contents of Kentucky blue grass-plots taken in July 1997 has shown to be high significantly as compared to those of Italian ryegrass plots(P<0.01). The results of this study support that the companion crop Italian ryegrass was good for weed controls only in spring of 1st year after sowing and orchardgrass in mixed pastures played good roles for increase of dry matter yield and weed control.

• Korean Journal of Ecology and Environment
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• v.44 no.2
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• pp.239-251
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• 2011

To compare the different water environments in the Jigsaw stream (st. 1, 2) and Jujin stream (st. 3, 4) crossing over the Byeonsan-Bando and Seonunsan parklands, the physico-chemical factors and the species compositions of epilithic diatoms were analyzed from March 2009 to February 2010. The mean values of conductivity (681.1 ${\mu}S\;cm^{-1}$), suspended solids (27.83 mg $L^{-1}$), biological oxygen demand (3.15 mg $L^{-1}$), total nitrogen (3.24 mg $L^{-1}$), total phosphorus (0.24 mg $L^{-1}$) and chlorophyll-${\alpha}$ (12.99 mg $m^{-3}$ ) in Jujin stream were significantly higher than each element in Jigsaw stream. Eighty taxa were classified into 73 species and seven varieties belonging to 24 genera, eight families, three suborders and two orders in Jigsaw stream. Eighty eight taxa were classified into 81 species and seven varieties belonging to 22 genera, seven families, three suborders and two orders in Jujin stream. Biological water quality assessments revealed that Jigsaw stream was ${\beta}$~${\alpha}$-oligosaprobic and Jujin stream was ${\beta}$-mesosaprobic. Dominant and diversity indices were indicated from 0.25~0.81 and from 1.73~4.14 in Jigsaw stream, respectively, and from 0.29~0.64 and from 2.72~4.02 in Jujin stream, respectively. Jujin stream was more eutrophic than those of Jigsaw stream. The different water environments between the two streams could be due to the different ecosystems of the neighboring basins of each stream crossing over in the mountain. Further studies should investigate the water environments of two streams by continuous and regular monitoring to analyze the different ecosystem mechanisms.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.701-707
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• 2008

Antidiabetic effect of Korean red ginseng (RG) processed by puffing in streptozotocin (STZ)-induced diabetic (DM) rats was investigated. Five week-old SD rats were divided into four groups; normal control (NC) group, DM group, red ginseng (RG) group and puffed red ginseng (PG) group. The RG and PG groups were orally provided with RG or PG dissolved in water (500 mg/kg) respectively for seven weeks after single injection of STZ (50 mg/kg, i.v.) followed by identification of DM. NC group received saline vehicle instead of STZ. At the end of feeding of RG or PG, the changes of fasting blood glucose, serum insulin and amylase level and serum lipid profiles were evaluated. Also, oral glucose tolerance test (OGTT), comet assay and histopathological examination were performed. At 7th week, the fasting blood glucose levels of the RG and PG groups were reduced compared to the DM group by 11.54% and 20.22%, respectively. The result of OGTT did not show significant differences among DM and two red ginseng groups. While serum insulin and TG levels were predominantly improved in PG group (p<0.05), serum amylase level was increased in RG group. Alkaline comet assay for checking the oxidative damage of DNA showed that TL (tail length, ${\mu}m$) and TM (tail moment) in the blood lymphocyte of PG group significantly decreased in contrast with DM group. Histopathological results of pancreas showed that destruction of exocrine as well as endocrine might be cured by the administration of RG and PG. These results suggest that PG could exert more protection against STZ-induced toxicity than RG group.

• Proceedings of the Korean Geotechical Society Conference
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• 1998.05a
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• pp.35-81
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• 1998

Distinct Element Method(DEM) has a great advantage to model the discontinuous behaviour of jointed rock masses such as rotation, sliding, and separation of rock blocks. Geometrical data of joints by a field monitoring is not enough to model the jointed rock mass though the results of DE analysis for the jointed rock mass is most sensitive to the distributional properties of joints. Also, it is important to use a properly joint law in evaluating the stability of a jointed rock mass because the joint is considered as the contact between blocks in DEM. In this study, a stochastic modelling technique is developed and the dilatant rock joint is numerically modelled in order to consider th geometrical and mechanical properties of joints in DE analysis. The stochastic modelling technique provides a assemblage of rock blocks by reproducing the joint distribution from insufficient joint data. Numerical Modelling of joint dilatancy in a edge-edge contact of DEM enable to consider not only mechanical properties but also various boundary conditions of joint. Preprocess Procedure for a stochastic DE model is composed of a statistical process of raw data of joints, a joint generation, and a block boundary generation. This stochastic DE model is used to analyze the effect of deviations of geometrical joint parameters on .the behaviour of jointed rock masses. This modelling method may be one tool for the consistency of DE analysis because it keeps the objectivity of the numerical model. In the joint constitutive law with a dilatancy, the normal and shear behaviour of a joint are fully coupled due to dilatation. It is easy to quantify the input Parameters used in the joint law from laboratory tests. The boundary effect on the behaviour of a joint is verified from shear tests under CNL and CNS using the numerical model of a single joint. The numerical model developed is applied to jointed rock masses to evaluate the effect of joint dilation on tunnel stability.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.1
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• pp.62-72
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• 2007

The purpose of present study was to determine whether different kinds of curing lights can alter microtensile bond strength(MTBS) of class I cavity pulpal and axial wall specimens in primary molar. Thirty clean mandibular 2nd primary molar's occlusal enamel were removed and class I cavity, size of $2{\times}4{\times}2mm$ was prepared. Dentin bonding agent was applied according to manufacturer's manual. Each group was cured with Halogen Curing Unit, Plasma Curing Unit and LED Curing Unit. Composite resin was bulk filled and photo cured with same curing unit. MTBS specimens which size is $0.7{\times}0.7{\times}4mm$ were prepared with low speed saw. Specimens were coded by their curing lights and wall positions (Halogen - Axial wall group, Halogen - Pulpal wall group, Plasma - Axial wall group, Plasma - Pulpal wall group, LED - Axial wall group, LED - Pulpal walt group). MTBS were tested at 1 mm/min cross Head speed by Universal Testing Machine. Fractured surface and bonding surface was observed with SEM. T-test between axial and pulpal specimens in each curing lights, one-way ANOVA among different curing light specimens in each wall positions were done. Weibull distribution analysis was done. The results were as follows : Mean MTBS of pulpal wall specimens were significantly greater than that of axial wall specimens at each curing units(p<.05). There was no significant difference in the MTBS among three curing units at axial wall and pulpal wall. In Weibull distribution, pulpal wall specimens were more homogeneous than axial wall specimens.