• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.88-88
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• 2003

본 연구는 배란후 48시간째 회수된 난소로부터 황체세포를 체외배양 후 progesterone 분비에 대한 LH, IGF-1 및 TGF$\beta$첨가효과에 대해 검토하였다. 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithiothreitol)으로 37$^{\circ}C$의 진탕수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM 용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세정하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석한 후 24 well culture plate(Coming, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 12시간 간격으로 배양액을 회수하였으며 48시간까지 배양하였다. 배양액 1$m\ell$당 LH, IGF-I 및 TGF$\beta$1을 단독 혹은 공배양하여 회수된 배양액내의 progesterone 농도를 RIA법으로 분석하였다. 돼지황체세포는 배양후 24시간째에 높은 농도의 progesterone이 측정되었으며 LH를 첨가한 대조구에 비해 유의적으로 높은 progesterone이 측정되었다. 또한 IGF-1 과 TGF$\beta$1을 첨가한 군에서도 높은 농도의 progesterone이 측정되었다 그러나 TGF$\beta$ 혹은 IGF-I 단독으로는 대조구의 결과와 큰 차이가 없었다 따라서 돼지 황체세포에서의 progesterone 분비는 TGF$\beta$ 및 IGF-I 그리고 LH의 황체세포의 progesterone 분비기능을 촉진하는 것으로 나타났다.

• Journal of Veterinary Clinics
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• v.30 no.4
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• pp.305-309
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• 2013

Pyometra is a diestrual, chronic disease process with acute manifestations in the adult, ovary-intact bitch. Serosal inclusion cysts develop during postpartum involution as mesothelium becomes trapped during rapid uterine contraction. A 10-year-old golden retriever bitch presented with lethargy, anorexia, tachypnea, abdominal distention, and abnormal vaginal discharge. Radiographic, ultrasound, and laboratory examinations were performed. On ultrasound examination, the uterus was distended by fluid containing echogenic "snow storm" particles; cystic structures containing anechoic fluid were found adjacent to the body of the uterus. Leukocytosis, neutrophilia, and anemia were diagnosed by a complete blood cell count. The initial diagnosis was pyometra, and an ovariohysterectomy was performed. Macroscopically, the uterine body and horns were expanded and partially adhered to the abdominal wall; numerous cysts containing clear fluid protruded over the entire surface of the uterus. Escherichia coli that was sensitive to enrofloxacin, was cultured from the lumen of the uterus. Histopathological assessment confirmed a final diagnosis of pyometra and serosal inclusion cysts of the uterus.

• Journal of the Mineralogical Society of Korea
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• v.18 no.4 s.46
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• pp.277-288
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• 2005

Perlite, a hydrated volcanic glass, occurs mainly as a bed-like body, and is distributed intermittently along the unconformity surface between the Beomgockri Group and its lower formations, viz. Janggi Group. The perlite is intimately associated with surrounding pumiceous welded tuff and rhyodacites in space and time. Compared to the typical perlite, the perlite is rather silica-poor and impure, and thus, includes lots of phenocrysts and rock fragments. Nearly the perlite is compositionally rather close to a pitchstone than a perlite in water contents. Petrographic comparison between perlite and associated volcanic to volcaniclastic rocks indicates that pumiceous welded tuff and rhyodacite seem to be Protolith of the Perlite. A Zr/$TiO_{2}$-Nb/Y diagram and field occurrence of perlite and their protolithic rocks also conforms the above interpretation. Kn addition, remnant vesicles in perlite strongly reflect that the precursor of perlitic glass appeared to be pumice fragment as well as volcanic glass. The perlite was diagenetically formed by way of a pervasive water-rock interaction at the deposition of the Manghaesan Formation in lacustrine environment. During perlitization, $SiO_{2}$ and alkali tend to be consistently depleted. Preexisting system of the Beomgockri Group based on the perlite formation should be corrected, because the perlite was formed diagenetically without lateral persistence in its occurrence.

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.157-161
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• 2003

The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of cats oocytes and development of IVM/IVF embryos. The results were summarized as follows : 1. The fertilization and developmental rate of fresh and salts-stored oocytes with and whithout cumulus cells were 65.7%, 17.1% and 28.6%, 8.6% and 57.1%, 13.3%, 23.3%, 3.3%, respectively. The rate of oocytes with cumulus cells(13.3%∼65.7%) was higher than that of denuded oocytes(3.3%∼28.6%). 2. The fertilization and developmental rate of oocytes recovered from ovaries collected at different stages of the reproductive cycle were 68.9%, 44.4%, 48.9% and 17.8%, 8.9%, 12.8%, respectively. 3. The fertilization and developmental rate of oocytes in vitro cultured at different time of incubation(24, 36 and 48 h) were 66.7%, 46.7%, 48.9% and 17.8%, 11.1%, 8.5%, respectively. respectively. The rate of oocytes incubated 24 h(66.7%) was higher than that oocytes incubated 36 and 48 h(46.7%∼48.9%). 4. The fertilization and developmental rate of oocytes treated activation and non-activation oocytes were 57.4%, 31.4% and 22.9%, 11.4%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.175-180
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• 1999

Blood chemical values and sex steroid hormone concentration of repeat breeder and reproductive disorder in Hanwoo, which were bred in Chunchon areas, were examine to establish the basic physiological maker. The concentrations of albumin, blood urea nitrogen(BUN), calcium(Ca), cholesterol, creatinine, glucose, phosphorous, total protein and triglycerides in repeat breeder and reproductive disorder were 3.28$\pm$0.05 and 3.27$\pm$0.04 g/㎗, 11.07$\pm$0.61 and 12.69$\pm$0.88 mg/㎗, 9.98$\pm$0.10 and 9.56$\pm$0.11mg/㎗, 105.75$\pm$3.57 and 126.78$\pm$5.66mg/㎗, 1.62$\pm$0.05 and 1.65$\pm$0.09 mg/㎗, 67.46$\pm$9.07 and 76.97$\pm$3.76mg/㎗, 6.13$\pm$0.36 and 6.11$\pm$0.26mg/㎗, 5.82$\pm$0.08 and 6.18$\pm$0.08 g/㎗ , 12.82$\pm$1.46 and 15.19$\pm$1.61mg/㎗, respectively. Among the blood chemical values, Ca, cholesterol and inorganic phosphorous levels in repeat breeder and reproductive disorder were slightly higher than those in normal cows. The progesterone levels of pregnancy(0.18~6.56 ng/$m\ell$) and non-pregnancy(0.15~5.46ng/$m\ell$) cows at estrus cycle were showed similar trends from 0 day to 18 days. Those of pregnancy cows were increased gradually from 18 days to 180 days(5.90~7.51 ng/$m\ell$), while those of non-pregnancy cows were dramatically drop at 21 days (0.18$\pm$0.03mg/㎗). The concentrations of estrogen and progesterone in repeat breeder and reproductive disorder were 57.42$\pm$5.03, 56.76$\pm$8.73 pg/$m\ell$ and 6.26$\pm$0.83, 9.41$\pm$1.85 ng/$m\ell$ . respectively. These results indicate that blood chemical values in repeat breeder and reproductive disorder were not greatly difference in normal cows except for Ca, cholesterol, BUN and inorganic phosphorous. Since the progesterone levels in repeat breeder and reproductive disorder was maintained to the luteal phase levels of normal cows, it may cause of failing of conception and next estrus cycle.

• 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.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.65-65
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• 2001

본 연구는 칡소 귀세포를 공여핵으로 이용한 체세포 복제송아지 생산에 있어서 배양방법이 배발생 및 배반포 발달율에 미치는 영향과 체세포 복제란의 이식후 수태율에 미치는 영향을조사하여 복제송아지의 생산 효율을 제고하고자 실시하였다. 실험에 공시된 공여핵은 칡소 의 귀세포를 회수하여 10%FBS가 첨가된 DMEM배지에서 3-4일 동안 배양하여 monolayar Confluent 형성 후 0.25% trypsin을 처리하여 준비하였으며 공여세포는 적어도 passage가 5회 이상의 세포만을 사용하였다. 복제수정란의 생산은 18-20시간 동안 체외성숙 된 난자의 핵을 제거하고 공여핵을 주입하여 2.2kv/cm, 10$\mu\textrm{s}$의 전압으로 2회 자극함으로 융합하였으며, 융합된 난자는 5$\mu\textrm{g}$/$m\ell$ ionomycin에서 4분간, 1.9mM 6-dimethyl aminopurine에서 4시간동안 배양하여 활성화처리를 하였다. 핵이식수정란의 배양은 39$^{\circ}C$, 5%$CO_2$ incubator에서 처리구 I은 CRlaa에서 4일간 배양 후 CR2aa배지에서 배양, 처리구II는 CRlaa에 4일간 배양후 CR2aa배지에 cumulus cell과 공배양, 처리구III은 CR2aa 배지에 camulus cell과 함께 배양하였다. 수정란이식은 발정발현 7일째에 비외과적 방법으로 젖소 미경산우에 이식하였으며 이식란수는 2~4개의 핵이식된 수정란을 이식하였다. 임신진단은 45~60일 사이에 직장검사 및 초음파 진단기를 이용하여 실시하였다. 배양방법에 따른 배발생율은 처리구 I에서 92.2 %(83/90)으로 처리구II와 III의 62.4%(63/101)와 77.8%(144/185)에 비하여 높게 나타났으나 배반포 발달율은 처리구II와III에서 65.1%(41/63)와 50.0%(72/144)로 처리구 I의 30.1%(25/83)보다 높게 나타났다. 각 처리구에 따른 수정란 이식후 수태율은 처리구II와 III에서 공히 20%의 수태율을 나타낸 반면 처리구 I에서는 수태가 되지 않았다. 따라서 체세포 복제수정란의 생산에 있어서 배반포 발달율과 수태율을 높이기 위해서는 단순배양보다 공배양이 더 효과적인 것으로 사료되지만 이런 결과가 복제송아지 생산효율에 있어서도 효과적일지는 향후 더 많은 연구가 있어야 할 것으로 사료된다.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.195-199
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• 2006

This study was carried out to investigate the effect intra-uterine infusion of lipopolysaccharides(LPS) on the reproductive performance of dairy cows with retained placenta, The LPS isolated from Bacteroids helcogenes and Fusobacterium varium was injected at the rate of 100 ${\mu}g$ with 30 ml of phospahte buffer saline in each cow(n=33) at day 20 postpartum. Twelve cows with retained placenta were not heated with LPS and served as a control. Conception rate after first service was 0% and 33,3% for control and LPS groups, respectively. Service period was $149.6{\pm}34.3\;and\;53.0{\pm}12.5$ days for control and LPS groups, respectively(p<0.05). Services per conception was $3.6{\pm}0.8\;and\;2.1{\pm}0.3$ for control and LPS groups, respectively(p<0.05). In conclusion, intra-uterine infusion of LPS in cows with retained placenta can reduce the service period and services per conception. Reduced service period can improve the dairy farm profitability by decreasing cost un feeding and management of cows.

• Journal of Veterinary Clinics
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• v.31 no.4
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• pp.267-271
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• 2014

Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

• Development and Reproduction
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• v.12 no.3
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• pp.243-250
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• 2008

Mast cells containing a variety of mediators in their cytoplasmic granules are widely distributed in connective tissues and mucosal surfaces of skin, airways, and guts. Within these tissues, mast cells are involved in the pathophysiological conditions such as inflammation, self-defense, tissue-remodeling, and autoimmunity. In order to understand the functional roles of master cells in the uterus, we histologically examined the distribution and density of uterine mast cells in the different aged mice. Until 6 weeks mast cells were sparsely detected in the uterus. But at 7 weeks after birth, when estrous cycle begins, the number of mast cells within uterine tissues increased dramatically and the increment of mast cell density continued up to 32 weeks-age. After then, uterine glandular tissue degenerated gradually and density of uterine mast cell decreased. Uterine mast cells were mainly found in the myometrium and they were closely associated with smooth muscle cells, fibroblasts, and collagens, which contents were changed according to the uterine development in the myometrium. These results suggest that uterine mast cells could be involved in myometrial contractions mediated by smooth muscle cells and tissue reconstitution or remodeling during estrous cycle and parturition including the various immunological functions.