• Journal of Embryo Transfer
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• v.28 no.3
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• pp.169-175
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• 2013

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

• Journal of Aquaculture
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• v.8 no.4
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• pp.307-316
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• 1995

Jicon scallop (Chlamys farreri), as one of new culture species, has been taken a growing interest in Korea recently. Artificial seeds are needed since seeds in natural waters are absolutely short. To develop the better methods for the artificial spawning and the rearing larvae on a large scale, experiments were carried out from July 1993 to May 1994. Mother shells were collected from the area near Tong-yeong and Geoje-do, the southern part of the Korean Penninsula. It was estimated that a period of 2-years was needed to be mother shells after the fertilization. Generally, spawning occourred intermittently from March to August and the maximum occurred in July. A best way for the artificial spawning was a chemical incentive, a 'serotonin injection' in parallel with the exposure and the temperature stimulation. Densities of the fertilized eggs for the normal development was from 30 to 40 per mililiter. It took 27-days from the fertilization to the settling larva averaging $155{\mu}m$ in size with water temperature 11.5\~13.0^{\circ}C$. Isochrysis galvana and Chaetoceros calcitrans with a density of $1,500\times10^4$ cells per ililiter were supplied for rearing the larvae. A proper density of larvae was 10 individuals per mililiter, and the survival rate of larvae from the flowing water system was much better than that of larvae from the still water.

• Journal of Aquaculture
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• v.9 no.2
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• pp.133-140
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• 1996

Hybird and allotriploid between female rainbow trout (Oncorhynchus mykiss) and male coho salmon (O. kisutch) were produced by artificial fertilization and heat shocks. Hatching and survival rate of allotriploid at 2 month after hatching was $77.6\%$ and $54.5\%$ respectively, and these rates clearly exceeded those of their hybrid. Cell and nuclear sizes of the erythrocyte of hybrid were intermidiate of their parents and those of allotriploid were larger than thier hybird. The somatic chromosome number of viable hybrid was 2n = 60 and that of allotriploid was $90\~93$ with chromosomal polymorphism. Allotriploid karyotpe was constituted by two sets of rainbow trout chromosome and one set of coho salmon chromosome.

• Development and Reproduction
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• v.18 no.3
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• pp.139-143
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• 2014

Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.281-287
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• 2009

As a simple and economical method for in vitro produced embryos, we have used BSA instead of serum for the production and embryo transfer of Hanwoo in vitro fertilized (IVF) embryos and obtained the following results: 1) When using serum (FBS; fetal bovine serum) or BSA-containing culture media as the initial culture media for immature oocytes, it is regarded as inappropriate to add only BSA to the culture solutions from maturation of the immature oocytes to development stage culture, but serum still needs be added though there is no significant difference in the concentration, with a change from 5% to 10%. 2) The results of culturing IVF embryos after development (4 cell stage) in the Medium199 solutions containing BSA instead of serum (FBS) showed that 0.3% BSA concentration is not optimal and 0.5% or higher BSA concentration has no significant difference among 0.5%, 0.7%, 1% and 2% (p > 0.05). 3) The post-freezing survival ratio after development in 5% FBS-Medium199 showed that 1% BSA concentration of the culture solution is the most suitable in the BSA concentrations of 0.3% (51%), 0.5% (67%), 0.7% (69%), 1% (77%) and 2% (75%). 4) The pregnancy rates of the transplanted fresh(not frozen) blastocyst had no significant concentration dependency (p > 0.5), and the average pregnancy rate was 63.8%. 14% of overweight calves were found among the calves given birth to by the transfer of IVF blastocysts cultured in the serum-added culture solution, but none was found in the experimental groups in which BSA was added instead of serum.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.2
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• pp.137-147
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• 1994

Controlled ovarian hyperstimulation(COH) for in vitro fertilization and embryo transfer(IVFET) often results in the production of more embryos than can be efficaciously transferred at one time. However, embryo cryopreservation provides a mechanism by which additional embryos can be stored for later thawing and transfer. From November, 1990 to October, 1992, we completed 42 transfer cycles of cryopreserved pronucleus(PN) l-cell embryos using the fixed protocol of hormonal replacement therapy in a physiological manner regardless of individual ovarian function. Artificial endometrial stimulation was performed with only exogenous estradiol and progesterone(E-P) in 36 transfer cycles (Group I) and with gonadotropin-releasing hormone agonist(GnRHa) and exogenous estradiol and progesterone(GEEP) in 6 transfer cycles(Group II ). The results were as follows. 1. The Survival rate of total cryopreserved-thawed embryos was 64.9%(198/305): 64.9% (172/265) in Group I and 65.0% (26/40) in Group II. 2. Total 168 embryos were transferred with an average of 4.7 per ET in Group I and total 26 embryos were transferred with an average of 4.3 per ET in Group II. 3. The pregnancy rate(PR) per cryopreserved-thawed ET and the implantation rate was 33.3 %(14/42) and 6.7%(13/194), respectively. The PRs per cryopreserved-thawed ET were 30.6% (11/36) in Group I and 50.0% (3/6) in Group II without significant difference. 4. The take home baby rate was 11.1%(4/36) in Group I and 33.3% (2/6) in Group II.

• Korean Journal of Agricultural and Forest Meteorology
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• v.22 no.2
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• pp.57-67
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• 2020

Pinus densiflora is the most widely distributed tree species in South Korea. Its ecological and socio-cultural attributes makes it one of the most important tree species in S. Korea. In recent times however, the distribution of P. densiflora has been affected by dieback. This phenomenon has largely been attributed to climate change. This study was conducted to investigate the responses of growth and physiology of P. densiflora to drought and nitrogen fertiliz ation according to the RCP 8.5 scenario. A Temperature Gradient Chamber (TGC) and CO₂. Temperature Gradient Chamber (CTGC) were used to simulate climate change conditions. The treatments were established with temperature (control versus +3 and +5℃; aCeT) and CO₂ (control: aCaT versus x1.6 and x2.2; eCeT), watering(control versus drought), fertilization(control versus fertilized). Net photosynthesis (P_n), stomatal conductance (g_s), biomass and relative soil volumetric water content (VWC) were measured to examine physiological responses and growth. Relative soil VWC in aCeT significantly decreased after the onset of drought. P_n and g_s in both aCeT and eCeT with fertiliz ation were high before drought but decreased rapidly after 7 days under drought because nitrogen fertilization effect did not last long. The fastest mortality was 46 days in aCeT and the longest survival was 56 days in eCeT after the onset of drought. Total and partial biomass (leaf, stem and root) in both aCeT and eCeT with fertiliz ation were significantly high, but significantly low in aCeT. The results of the study are helpful in addressing P. densiflora vulnerability to climate change by highlighting physiological responses related to carbon allocation under differing simulated environmental stressors.

• Clinical and Experimental Pediatrics
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• v.50 no.8
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• pp.740-745
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• 2007

Purpose : There is a dominant opinion that in vitro fertilization (IVF) leads to an increased incidence of twins, low birth weight (LBW) infants, prematurity and mortality. On the other hand, technical development of IVF and improvement of neonatal intensive care have increased the survival rate of neonates. The purpose of this study was to verify the tendency by comparing the clinical aspects of IVF and spontaneously conceived twins, and to establish methods to increase the survival rate of neonates after IVF. Methods : Retrospective reviews were performed on all twin infants who were admitted to the nursery and NICU at Kangnam Sacred Heart Hospital, Hallym University from January 1, 2000 to December 31, 2006. Medical records of IVF twins (study group, n=92) and spontaneously conceived twins (control group, n=265) were analyzed and compared. Neonatal outcomes and complications, as well as obstetric outcomes, were analyzed and compared. Results : Mean gestational age and birth weight of the study group ($34.6{\pm}3.5$ weeks, $2,203.9{\pm}617.2g$) were considerably lower than those of the control group ($36.3{\pm}2.4$ weeks, $2,367.0{\pm}517.9g$). The frequency of prematurity less than 37 weeks (68.5% vs 51.3%) and extremely LBW (15.2% vs 6.4%) were also significantly higher in the study group. Other neonatal outcomes were all insignificant. The obstetric characteristics, maternal age ($32.6{\pm}3.3$ years vs $30.3{\pm}3.9$ years) and the frequency of cesarean delivery(95.7% vs 79.9%) were significantly higher in the study group. Other obstetric outcomes were insignificant except for the frequency of incompetent internal os of cervix (36.2% vs 3.6%) and cerclage operation (38.3% vs 4.3%). Conclusion : Based on the above results, clinical outcomes of twin infants will be further improved by careful attention and thorough antenatal care of the IVF twins.

• Journal of Aquaculture
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• v.10 no.2
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• pp.179-187
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• 1997

Early development growth and sexual phenomena of the hybrid between male spotted flounder (Verasper variegatus) and female olive flounder (Paralichthys olivaceus) were investigated. The hybrid produced was newly called the "Bumnupchi" in Korean name, "Mottled flounder" in English name. Fertilization rate, hatching rate and early survival rate of the mottled flounder were 56.9~72.3%, 86.7~92.5% and 89.1~96.0%, respectively. At 50th day and 240th day after hatching, they were 2.1$\pm0.2cm\;and\;22.9\pm2.1cm\;in\;total\;length, \;0.10\pm0.002g\;and\;179.0\pm38.5g$ in body weight, respectively. Eye position of the mottled flounder was mixed with both dextral (77.7%) and sinistral type (22.3%). Gonads were differentiated into ovarian type and testis type at 80th day after hatching, and its ratio was 1:1 (P>0.05). Most Gonads seemed to be sterile without having germ cell, whereas only 3.6% of investigated indiciduals turned out to have oocytes in the gonad. When the induction of sex reversal were attempted by daily oral administration of $17\beta$-methyltestosterone (0.5mg/kg fish) andestradiol-17$17\beta$(0.5mg/kg fish)from 40th to 100th day after hatching, both testis and ovary type were induced 81.9% and 87.7% of each hormone-treated fish, respectively.fish, respectively.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.34 no.4
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• pp.408-421
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• 1989

The effects of seeding and clipping termination dates, and seeding and fertilizer rates on forage and grain yields were evaluated in 1989 harvest year for dual production of forage and grain of naked and malting barleys in Cheju province. In the experiment I, 'Saessalbori' (naked barleY)1 and 'Doosan 22' (malting barley) were seeded on Sept. 21, Oct. 12 and Nov. 2, respectively. An unclipped(grain-only) treatment that was planted on Nov. 2 was included for check. In the experiment II, the two above cultivars were planted at seed rate of 14, 20 and 26 and kg/l0a. Fertilizer rates were 9-6-12, 17-13-8 and 22-16-11 (N-P$_2$O$\sub$5/-K$_2$O) kg/l0a. Seeding dates were Oct. 13 and Nov. 7 for clipped treatments and grain -only treatment(seeding rate: 14kg/l0a, N-P$_2$O$\sub$5/-K$_2$O=9-6-12 kg/l0a), respectively. All treatments in both experiments were harvested for grain yield. In Saessalbori, survival following forage removal was 100％ regardless of seeding and clipping termination dates, and seeding and fertilizer rates. In Doosan 22, survival percenage decreased with increasing seeding and fertilizer rates when plants was clipped in January to Febuary. Forage yield of two cultivars increased with early planting and delays in clipping termination and tended to increase as seeding and fertilizer rates were increased. Crude ash and fat were not affected by seeding and fertilizer rates, and clipping date. Crude protein increased with an increase in fertilizer rate and decreased with delays in clipping date. N free extract tended to increase with delaying clipping date and decreased with increasing fertilizer rate. Earlier planting resulted in earlier heading and maturity. Clipping slightly delayed heading of Saessalbori and greatly delayed that of Doosan 22. Maturity of Doosan 22 was delayed 3 to 5 days by increased fertilization. Clipping shortened culm length more severely in Doosan 22 than in Saessalbori. The later the clipping termination, the shorter the culm length. The number of spikes per m$^2$ and the number of kernels per spike were not affected by clipping in Saessalbori while those of Doosan 22 were decreased with delays in clipping termination. Delaying forage harvest resulted In a reduction in grain yield. However, final clipping on Feb. 27 reduced grain yield of Saessalbori by only 6-11 ％ compared to the grain-only treatment. In Doosan 22, forage harvest after Dec. 28 resulted in 6 to 66％ reduction of grain yield. The data indicate that grazing of naked and malting barleys until late Feb. and late Dec., respectively, might not reduce grain yields when planted on mid-Sept. to mid-Oct.