• Journal of Embryo Transfer
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• v.20 no.2
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• pp.79-87
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• 2005

The ovaries from Hanwoo and Holstein were collected from labattoir and transferred to laboratory. Oocytes were aspirated and incubated in $CO_2$ incubator for 24 hours for maturation. Oocytes were coincubated with the sperms for 5 hours. Cleaved oocytes were selected 48 hours after coincubation and half of the medium was changed newly every 48 hour until blastocyst formation. Cleavage rate and blastocyst rate were investigated according to different breeds and different status of cumulus cells surrounding the oocytes. Blastocysts were either transferred to the recipients or frozen until use. The result of embryo transfer with fresh or frozen embryos was investigated. The rate of male offspring following embryo transfer was also investigated. The rate of cleavage was $66.4\%$ for Hanwoo and $62.4\%$ for Holstein oocytes. The rate of cleavage according to status of oocyte was shown highest in the oocytes completely surrounded with cumulus cells and lowest in denuded oocytes for both Hanwoo and Holstein oocytes. The rate of blastocyst from cleaved oocytes was $40.6\%$ for Hanwoo and $36.9\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with fresh IVF embryos was $57.2\%$ for Hanwoo and $53.3\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with frozen IVF embryos was $40.9\%$ for Hanwoo and $36.4\%$ for Holstein. The rate of male calf produced by embryo transfer was $63.6\%$ for Hanwoo and $50.0\%$ for Holstein.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.105-112
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• 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.

• 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.20 no.2
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• pp.185-190
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• 2005

This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

• 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 The Korean Society of Grassland and Forage Science
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• v.24 no.3
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• pp.217-224
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• 2004

This study was carried out to select the promising forage crops for forage production in paddy field of HARI, RDA, Iksan from 1999 to 2001. The species of forage crop used in this experiment were barley(5 varieties), wheat(2), rye(3) and Italian ryegrass(IRG, 3). Stage of heading, milk and yellow ripe of barley were 26 $April{\sim}l$ May, middle May and late May, respectively. In milk to yellow ripe stage, dry matter(DM) yield of barley was $6.64{\sim}10.47$ MT per ha, and averaged ADF and NDF contents were $31.6\%\;and\;62.2\%$ respectively. Albori was the most promising variety among the barley). The growth of wheat was a little late compared to that of barley, but it seemed to be crop having higher forage yield md nutritive value. In milk to yellow stage, DM yield of wheat was $7.51{\sim}11.96$ MT per ha, and nutritive value of wheat was high(ADF $30.6\%$ and NDF $60.1\%$). Heading stage of rye was $25{\sim}27$ April, and it seemed to be suitable to harvest at heading to flowering stage because of heavy lodging after flowering. In flowering stage, DM yield of rye was 17.26 MT per ha, and nutritive value of rye was the lowest among 4 species(ADF $38.6\%$ and NDF $65.5\%$). Heading stage of early maturing IRG was 30 April and it seemed to be suitable for harvest at flowering stage of middle May in cropping system. In flowering stage, DM yield of early type IRG was 10.77 MT per ha, and averaged ADF and NDF contents were $31.6\%\;and\;59.2\%$, respectively. The results demonstrated that the promising forage crops for forage production in paddy field were rye, barley, early maturing IRG and wheat in Honam region. In Southern region, rye harvesting at early May was recommended when considered middle May of rice transplanting period, and barley(Albori) and early maturing IRG harvesting at middle May were recommended when considered late May of rice planting period. And also, wheat with high nutritive value and possibility for harvesting at late May were recommended in case of transplanting period at early June.

• 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.

• Tuberculosis and Respiratory Diseases
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• v.51 no.5
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• pp.426-436
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• 2001

Objective : This study was performed to determine the smoking status and its associated factors in military life among young Korean men to provide data for smoking prevention in military areas as a part of the health promotion program for soldiers. Methods : 1,033 infantrymen serving in 17 military units, which are located in suburbs of K city, were enrolled in this study. A questionnaire on the soldiers smoking status and other various characteristics was obtained from October 2nd to 12th, 2000. Logistic regression analysis was performed on the various variables in order to determine the factors related to smoking. Results : 1) 830(80.3%) out of 1033 subjects had smoked cigarettes and 759 persons(73.4%) were regular smokers. 2) Among the 734 soldiers who started smoking before their military service, 714 soldiers(97.2%) continued to smoke after, while 45 soldiers(15.0%) among the 299 non-smokers started smoking after joining the army. 3) Through the logistic regression analysis. the education level. specialty in the army. smoking amongst the friends they met before military service, smoking amongst soldiers in the army. dating girl friends, drinking, satisfaction with their army lives and the presence of convenient smoking places at the barracks were significant factors related to the prevalence of smoking in soldiers. Conclusion : The factors related with incidence of smoking in the army are the complicated interactions among the individuals personal background, general characteristics of the soldier and the special environment in the barracks. Therefore, to improve national soldiers' health, an active non-smoking campaign and education should be pushed to discourage smoking in the barracks.

• Korean Journal of Plant Taxonomy
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• v.45 no.1
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• pp.96-107
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• 2015

This study presents a comprehensive account of the flora of hydrophytes and hygrophytes on Jeju Island. This approach aims to contribute to fundamental information about the conservation of plant diversity in wetlands. Field surveys were carried out from May of 2010 to September of 2014 in various types of wetlands on Jeju Island. A total of 189 taxa, including 52 families, 98 genera, 181 species, 6 varieties, 2 forma and 189 taxa, were found. Among them, 99 taxa (38 families, 60 genera, 96 species and 3 varieties) were hydrophytes and 90 taxa (23 families, 45 genera, 85 species, 3 varieties and 2 forma) were hygrophytes. In addition, Eleocharis${\times}$yezoensis H. Hara (Cyperaceae) was newly added to the flora of Korea. The protected wild plants designated by the Ministry of the Environment were 5 taxa, including Brasenia schreberi J. F. Gmel. and Dysophylla yatabeana Makino. The endangered species found included one Critically Endangered Species (i.e., Mankyua chejuense B.-Y. Sun, M. H. Kim & C. H. Kim), 5 Endangered Species (e.g., Lysimachia leucantha Miq.) and 7 Vulnerable Species (e.g., Tillaea aquatica L.). The floristic regional indicator plants found in this area were 44 taxa comprising 10 taxa of grade V, 7 taxa grade IV, 15 taxa of grade III, 5 taxa of grade II, and 7 taxa of grade I. The identified naturalized plants were 10 taxa; 8 families, 8 genera, 9 species and 1 variety. In the wetlands on Jeju Island, high plant species diversity was shown, and a number of rare plants and phytogeographically important plants were found. For these reasons, we consider that the wetlands on Jeju Island are very important areas from a botanical point of view. It is necessary to establish specific protection and maintenance policies in an effort to conserve these wetlands and species.