• Journal of Veterinary Clinics
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• v.30 no.3
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• pp.178-185
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• 2013

This study evaluated the effects of milk yield on the postpartum health and reproductive performance of dairy cows. In total, data were collected from 1,060 cows on six dairy farms, including their milk production, body condition score (BCS), postpartum disorders, and reproductive performance. The lactation data were grouped based on the 305-day milk yield into control (< 10,000 kg, n = 445) and high milk yield (${\geq}$ 10,000 kg, n = 615) groups. The milk fat and protein, and BCS were lower during the first 5 months postpartum in the high milk yield group compared with the control group (p < 0.01). Ovarian cysts were more frequent (p < 0.0001) in the high milk yield group (28.6%) than in the control group (15.3%), whereas endometritis tended to be less frequent in the high milk yield group (29.6%) than in the control group (35.1%, p = 0.06). A higher proportion of cows tended to receive reproductive hormones (p = 0.06) in the high milk yield group (62.4%) than in the control group (56.6%). The probability of a pregnancy after first insemination tended to be lower (odds ratio = 0.78, p = 0.07) in the high milk yield group (30.2%) than in the control group (35.2%). Furthermore, the hazard of pregnancy by 210 days in milk was lower in the high milk yield group (hazard ratio = 0.84, p = 0.04) than in the control group, which resulted in a 20-day increase in the median interval to pregnancy. In conclusion, high milk yield was related to lower milk fat and protein, lower BCS, an increased incidence of ovarian cysts, and increased use of reproductive hormones, which resulted in decreased reproductive performance of dairy cows.

• Journal of Life Science
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• v.20 no.3
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• pp.381-387
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• 2010

To find offspring of Jeju Black cattle (JBC) produced by embryo transfer (ET) and artificial insemination (AI), a molecular genetic study was carried out in candidate cattle populations collected from cattle farms in Jeju Island, Korea. The genetic marker set was composed of 11 ISAG microsatellite (MS) markers, 11 SAES MS markers selected by our preliminary analysis for population diversity of JBC and two major coat color related genes: MC1R and ASIP. The results showed a combined non-exclusion probability for first parent (NE-P1) that was higher than that recommended by ISAG (above 0.9995), and a combined non-exclusion probability for sib identity of $5.3{\times}10^{-10}$. Parentage analysis showed that the cases identified the candidate's father only (77.0%), mother only (54.0%), and both parents (40.5%) in the candidate offspring population. The ET and AI calves were identified as 14.7% in the in vitro fertilized eggs provided and 32.4% in total population, respectively. However, the result from ISAG marker analysis showed 3 identical allele-combinations in 7 calves, and that from ISAG/SAES MS marker combination also showed 1 identical allele-combination in 2 calves. Data from MS and coat-color gene analyses provided information for complete identification of all animals tested. Because the present JBC population was mostly bred using small nuclear founders through bioengineering techniques such as AI and ET, the genetic diversity levels obtained from MS analysis in the JBC population were relatively lower than those of other cattle populations, including Hanwoo. The results suggested that the more efficient marker combinations, including coat color related genotypes, should be studied and used for constructing a system for identification and molecular breeding of JBC as well.

• Journal of Life Science
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• v.22 no.3
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• pp.366-372
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• 2012

This study was conducted to estimate the extent of linkage disequilibrium (LD) and effective population size using whole genomic single nucleotide polymorphisms (SNP) genotyped by DNA chip in Hanwoo. Using the blood samples of 35 young bulls born from 2005 to 2008 and their progenies (N=253) in a Hanwoo nucleus population collected from Hanwoo Improvement Center, 51,582 SNPs were genotyped using Bovine SNP50 chips. A total of 40,851 SNPs were used in this study after elimination of SNPs with a missing genotyping rate of over 10 percent and monomorphic SNPs (10,730 SNPs). The total autosomal genome length, measured as the sum of the longest syntenic pairs of SNPs by chromosome, was 2,541.6 Mb (Mega base pairs). The average distances of all adjacent pairs by each BTA ranged from 0.55 to 0.74 cM. Decay of LD showed an exponential trend with physical distance. The means of LD ($r^2$) among syntenic SNP pairs were 0.136 at a range of 0-0.1 Mb in physical distance and 0.06 at a range of 0.1-0.2 Mb. When these results were used for Luo's formula, about 2,000 phenotypic records were found to be required to achieve power > 0.9 to detect 5% QTL in the population of Hanwoo. As a result of estimating effective population size by generation in Hanwoo, the estimated effective population size for the current status was 84 heads and the estimate of effective population size for 50 generations of ancestors was 1,150 heads. The average decreasing rates of effective population size by generation were 9.0% at about five generations and 17.3% at the current generation. The main cause of the rapid decrease in effective population size was considered to be the intensive use of a few prominent sires since the application of artificial insemination technology in Korea. To increase and/or sustain the effective population size, the selection of various proven bulls and mating systems that consider genetic diversity are needed.

• Journal of The Korean Society of Grassland and Forage Science
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• v.38 no.4
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• pp.331-336
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• 2018

The aim of the present study was to identify the metabolic changes that have occurred in Hanwoo Native Cow. Analysis of serum metabolites was carried out to investigate relationship of pregnancy rate and feeding systems. Totally, 281 cows were used for grazing and barn feeding. Grazing was carried out for 5 months in the pasture. In barn feeding, concentrate 3.0 Kg(TDN 68%, CP 14%) and rice straw 6 kg(TDN 50%, CP 6.5%) were fed. Artificial insemination(AI) carried out by timed AI procedure. The level of glucose(mg/dl), cholesterol(mg/mL), BUN(mg/dl), AST(U/l), ALT(U/l), and NEFA(uEq/l) were analyzed. In Table 1, barn feeding with 120% group was showed high Cholesterol and BUN levels compared those with 80% feeding group(p<0.05). In Table 2, 60 pregnant cows in grazing herd showed low Cholesterol, AST and NEFA levels compared to 43 non-pregnant cows($149.8{\pm}4.9$, $99.1{\pm}2.6$, $326.7{\pm}15.7$ vs. $165.9{\pm}4.6$, $108.9{\pm}3.2$, $419.2{\pm}32.8$, respectively, p<0.05). In Table 3, 126 grazing feeding cows showed high Glucose, Cholesterol, AST, ALT, and NEFA levels compared to 22 barn feeding cows($84.8{\pm}1.3$, $142.5{\pm}2.5$, $97.7{\pm}2.3$, $34.3{\pm}0.5$, $317.8{\pm}13.6$ vs. $56.0{\pm}1.3$, $128.9{\pm}4.6$, $80.9{\pm}2.0$, $27.1{\pm}0.9$, $160.2{\pm}18.9$, respectively, p<0.05). Further study needed to obtain more accurate level of metabolites in serum for pregnant and non-pregnant cows.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.79-92
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• 1992

This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.49-56
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• 2010

Objective: To investigate assisted reproductive technology (ART) outcomes in women with WHO class I anovulation compared with control group. Design: Retrospective case-control study. Methods: Twenty-three infertile women with hypogonadotropic hypogonadism (H-H) who undertook ART procedure from August 2003 to January 2009 were enrolled in this study. A total of 59 cycles (H-H group) were included; Intra-uterine insemination with super-ovulation (SO-IUI, 32 cycles), in vitro fertilization with fresh embryo transfer (IVF-ET, 18 cycles) and subsequent frozenthawed embryo transfer (FET, 9 cycles). Age and BMI matched 146 cycles of infertile women were collected as control group; 64 cycles of unexplained infertile women for SO-IUI and 54 cycles of IVF-ET and 28 cycles of FET with tubal factor. We compared ART and pregnancy outcomes such as clinical pregnancy rate (CPR), clinical abortion rate (CAR), and live birth rate (LBR) between the two groups. Results: There was no difference in the mean age ($32.7{\pm}3.3$ vs. $32.6{\pm}2.7$ yrs) and BMI ($21.0{\pm}3.1$ vs. $20.8{\pm}3.1kg/m^2$) between two groups. Mean levels of basal LH, FSH, and $E_2$ in H-H group were $0.62{\pm}0.35$ mIU/ml, $2.60{\pm}2.30$ mIU/ml and $10.1{\pm}8.2$ pg/ml, respectively. For ovarian stimulation, H-H group needed higher total amount of gonadotropin injected and longer duration for ovarian stimulation (p<0.001). In SO-IUI cycles, there was no significant difference of CPR, CAR, and LBR between the two groups. In IVF-ET treatment, H-H group presented higher mean $E_2$ level on hCG day ($3104.8{\pm}1020.2$ pg/ml vs. $1878.3{\pm}1197.7$ pg/ml, p<0.001) with lower CPR (16.7 vs. 37.0%, p=0.11) and LBR (5.6 vs. 33.3%, p=0.02) and higher CAR (66.7 vs. 10.0%, p=0.02) compared with the control group. However, subsequent FET cycles showed no significant difference of CPR, CAR, and LBR between the two groups. Conclusion: H-H patients need higher dosage of gonadotropin and longer duration for ovarian stimulation compared with the control groups. Significantly poor pregnancy outcomes in IVF-ET cycles of H-H group may be due to detrimental endometrial factors caused by higher $E_2$ level and the absence of previous hormonal exposure on endometrium.

• The Journal of Korean Society for Radiation Therapy
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• v.33
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• pp.145-153
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• 2021

Purpose: The purpose of this study is to evaluate the ovarian dose during radiation therapy for breast cancer in women of childbearing age through an experiment. The ovarian dose is evaluated by comparing and analyzing between the calculated dose in the treatment planning system according to the treatment technique and the measured dose using a thermoluminescence dosimeter (TLD). The clinical usefulness of lead (Pb) apron is investigated through dose analysis according to whether or not it is used. Materials and Methods: Rando humanoid phantom was used for measurement, and wedge filter radiation therapy, 3D conformal radiation therapy, and intensity modulated radiation therapy were used as treatment techniques. A treatment plan was established so that 95% of the prescribed dose could be delivered to the right breast of the Rando humanoid phantom 3D image obtained using the CT simulator. TLD was inserted into the surface and depth of the virtual ovary of the Rando hunmanoid phantom and irradiated with radiation. The measurement location was the center of treatment and the point moved 2 cm to the opposite breast from the center of the Rando hunmanoid phantom, 5cm, 10cm, 12.5cm, 15cm, 17.5cm, 20cm from the boundary of the right breast to the center of treatment and downward, and the surface and depth of the right ovary. Measurements were made at a total of 9 central points. In the dose comparison of treatment planning systems, two wedge filter treatment techniques, three-dimensional conformal radiotherapy, and intensity-modulated radiation therapy were established and compared. Treatments were compared, and dose measurements according to the use of lead apron were compared and analyzed in intensity-modulated radiation therapy. The measured value was calculated by averaging three TLD values for each point and converting using the TLD calibration value, which was calculated as the point dose mean value. In order to compare the treatment plan value with the actual measured value, the absolute dose value was measured and compared at each point (%Diff). Results: At Point A, the center of treatment, a maximum of 201.7cGy was obtained in the treatment planning system, and a maximum of 200.6cGy was obtained in the TLD. In all treatment planning systems, 0cGy was calculated from Point G, which is a point 17.5cm downward from the breast interface. As a result of TLD, a maximum of 2.6cGy was obtained at Point G, and a maximum of 0.9cGy was obtained at Point J, which is the ovarian dose, and the absolute dose was 0.3%~1.3%. The difference in dose according to the use of lead aprons was from a maximum of 2.1cGy to a minimum of 0.1cGy, and the %Diff value was 0.1%~1.1%. Conclusion: In the treatment planning system, the difference in dose according to the three treatment plans did not show a significant difference from 0.85% to 2.45%. In the ovary, the difference between the Rando humanoid phantom's treatment planning system and the actual measured dose was within 0.9%, and the actual measured dose was slightly higher. This did not accurately reflect the effect of scattered radiation in the treatment planning system, and it is thought that the dose of scattered radiation and the dose taken by CBCT with TLD inserted were reflected in the actual measurement. In dosimetry according to the with or without a lead apron, when a lead apron was used, the closer the distance from the treatment range, the more effective the shielding was. Although it is not clinically appropriate for pregnancy or artificial insemination during radiotherapy, the dose irradiated to the ovaries during treatment is not expected to significantly affect the reproductive function of women of childbearing age after radiotherapy. However, since women of childbearing age have constant anxiety, it is thought that psychological stability can be promoted by presenting the data from this study.