• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.233-240
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• 1994

This study was conducted to compare the endocrine milieu, and pregnancy rates in In Vitro Fertilization and Embryo Transfer(IVF-ET) program employing combined with gonadotropin releasing hormone agonist(GnRH-a) and pergonal(LH 75lU+FSH 75lU) when either human chorionic gonadotropin(HCG) or progesterone were used for luteal phase support. A total number of 40 IVF-ET treatment cycles were prospectively studied. Ovarian hyperstimulation method was modified ultrashort protocol using GnRH-a. All patients started Decapeptyl at menstrual cycle day # 2, and HMG was started at # 3 days. When leading follicle was ${\geqq}$18mm or at least two follicles were ${\geqq}$14mm in diameter, HCG 10000lU intramuscularly was injected. After 36 hours HCG administration, oocytes were retrieved as usual guided by transvaginal ultrasound. Embryo were transfered 36-48 hours later. The patient's cycles were prospectively randomized to receive HCG(20cycles) or Progesterone (20cycles) for luteal support. The progesterone group received 25mg 1M starting from the day of ET. The HCG group received 1500IU 1M. on days 0, +2, +5 after ET. Estadiol($E_2$) and Progesterone($P_4$) were measured on the day of oocyte aspiration, ET day, and every 6 days thereafter. Results were follows as; 1. Estradiol, progesterone and LH levels on the day of HCG trigger, retrieved oocytes and number of transfered embryo were not significantly different in both groups. 2. On the day of aspiration and embryo transfered day, $E_2$, $P_4$ level were significantly higher in progesterone group than HCG group(p<0.01). 3. $E_2$, $P_4$ level on 6 days after ET were significantly higher in progesterone group than HCG group(p<0.01). But, $P_4/E_2$ ratio was not different in both groups. 4. $E_2$, $P_4$ level 12 days after ET were decreased abruptly in both groups and higher hormonal level appeared in HCG group(P<0.01). 5. The total pregnancy rate in the HCG group was 40% (8/20) and in the progesterone group 15%(3/20). 6. Comparing the pregnant and nonpregnant cases progesterone group was not different the hormonal status. In HCG group, pregnant cases appeared in higher $P_4$, $P_4/E_2$ ratio than nonpregnanct cases(P<0.01).

• Reproductive and Developmental Biology
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• 제28권1호
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• pp.13-20
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• 2004

This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS, and primary fibroblast cultures were established in TCM-199 with 10% FBS. After maturation, expanded cumulus cells were removed by vigorous pipetting in the presence of 0.3% hyaluronidase. The matured oocytes were dipped in D-PBS plus 10% FBS＋7.5 $\mu\textrm{g}$/ml cytochalasin B and 0.05 M sucrose. The reconstructed oocytes were electrically fused with donor cells in 0.3 M mannitol fusion medium. After the electofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. On the other hand, the NT embryos with porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6∼8 day at $39^{\circ}C, 5% CO_2$ in air. In caprine-bovine NT embryos, the cleavage(2-cell) rate was 36.8% in confluence and 43.8% in serum starvation. The developmental rate of morula- and blastocyst-stage embryos was 0.0% in confluence and 18.8% in serum starvation. In caprine-porcine NT embryos, the cleavage(2-cell) rate was 76.7% in confluence and 66.7% in serum starvation. The developmental rate of morula and blastocyst stage embryos was 3.3% in confluence and 3.0% in serum starvation, and no significant difference was observed in synchronization treatment between donor cells. In caprine-bovine NT embryos, the cleavage(2-cell) rate of cultured donor cells was 30.8% and 17.6% in 5∼9 and 10∼14 passage(P＜0.05). The developmental rate of morula and blastocyst stage embryos were significantly higher(P＜0.05) in 5∼9 passage(23.1%) than in 10∼14 passage(0.0%) of cultured donor cells. In caprine-porcine NT embryos, the cleavage rate was significantly higher(P＜0.05) in 5∼9 passage(86.7%) than in 10∼14 passage(50.0%) of cultured donor cells. The developmental rate of morula and blastocyst stage embryos were 3.3 and 0.0% in 5∼9 and 10∼14와 passage of cultured donor cells. In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer, 33.9% in in vitro fertilization and 28.1% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P＜0.05) in interspecies nuclear transfer(5.1%) than in vitro fertiltzation(26.9%) and parthenotes(37.4%).

• 한국토양비료학회지
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• 제33권1호
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• pp.8-14
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• 2000

암모늄태질소를 질소의 급원으로 공급하기 위하여 요소와 유안을 질소조성물로 하여 2대1로 혼합한 후 다시 점결제 및 확산완충계와 혼합하여 비커형태의 용기안에 충진하여 제조된 시제품인 $NH_4$-Beaker Deeposit비료를 토마토유묘의 정식기에 유묘의 밑으로 1회 국지사용하여 토마토에 대한 생육 및 수량과 질소공급 효과를 일반 시비방법인 $NH_4NO_3$를 3회 전층 분배시용한 관행의 대구조와 비교하였다. 1년차 시험에서 질소의 용출을 조절하기 위하여 유안과 요소를 2:1로 혼합한 질소조성물에 7.5 g의 석고를 점결제로 사용하였을 때 토마토과실 수량은 1주당 6345 g으로 관행의 대조구의 수량인 5865 g보다 증수되었다. 토마토과실과 경엽의 총 질소흡수량 또한 7.5 g의 석고를 점결제로 사용한 $NH_4$-Beaker Deposit비료 시용구에서 8997 mg으로 관행의 7215 mg보다 통계적 유의성이 있었다. 2년차 시험에서는 C/N율이 10인 퇴비를 확산완충제로 비커에 충진한 시험구 5를 제외한 모든 $NH_4$-Beaker Deposit비료의 시용구에서의 질소흡수량이 높았다. 질소의 용출을 조절하기 위하여 7.5 g의 석고를 비커안에 충진하고, 그 윗부분으로 질소의 확산기울기를 조절하기 위하여 점질토양을 확산 완충제로 충진한 처리구에서의 토마토과실의 질소흡수량은 주당 8646 mg으로 가장 높았으며 관행의 대조구와 통계적인 유의성이 있었다. 토마토작물의 뿌리분포를 조사한 결과 확산완충제로 점질토양, C/N율이 16인 퇴비를 사용한 처리구의 경우 뿌리가 비커로 집중분포 되는 경향을 나타냈으며 굴지성에 반하여 비커안으로 자라는 현상을 나타냈다. 토마토 수량 및 질소흡수량을 고려할 때 7.5 g의 석고를 점결제로 사용하고, C/N율이 16인 부엽퇴비 및 점질토양을 확산완충제로 비커에 충진하여 제조한 시제품이 가장 좋은 처리로 판단되었다.

• 화훼연구
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• 제16권2호
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• pp.118-123
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• 2008

진한 붉은색을 띤 'Tiny Ghost'를 교배모본으로 Oriental group, Longiflorum group 및 Martagon group을 부본으로 하여 효율적인 종간잡종 생산과 상품성 있는 신품종을 획득하고자 실험을 수행하였다. 개화당일 채취한 화분의 발아율은 Oriental hybrid인 'Sorbonne'이 64%로 가장 높았다. 주두수분법과 화주 절단수분법을 이용하여 수분한 결과, 'Aktiva'를 부본으로 사용한 경우를 제외하고 모두 주두수분법을 사용한 교배에서 자방이 생성되었다. 또한 Longiflorum group인 'Norina'와 'Gelria' 중 화분발아율이 높은 'Norina' 가 모두 수정에 실패하여 같은 group 내에서도 종간의 유전적 친화성에 따라 수정률이 달랐다. 'Aktiva'를 부본으로 한 교배에서 1개의 embryo sac을 적출하여 1개의 구를 생성하였으며, 'Sorbonne'을 부본으로 한 교배에서는 4개의 ovule을 치상하여 1개의 구를 얻었다. 'Gelria'를 부본으로 한 교배에서는 1개의 자방에서 embryo 5개, embryo sac 15개, ovule 7개를 얻었으며 그 중 18개체가 발아하여 66.7%의 발아율을 나타내었다. Martagon group인 L. hansonii를 부본으로 한 교배에서는 5개의 ovule을 얻어 그 중 40%가 발아하였다.

• 한국작물학회지
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• 제64권3호
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• pp.234-245
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• 2019

본 연구에서는 천안지역의 논에서 밀-벼 이모작 작부체계 기술을 확립하여 천안지역 호두과자 원료곡인 국산밀 생산 증진과 경지이용률 증대에 기여하고자 최근 변화하고 있는 기상여건을 분석하여 이모작 재배여건에 대한 타당성을 검토코자 2015년부터 2017년에 걸쳐 수행하였으며 그 결과는 다음과 같다. 1. 밀 후작으로 벼는 출수 후 40일간의 적산온도 범위는 $840{\sim}920^{\circ}C$이었으며, 안전출수 한계 $800^{\circ}C$를 감안할 때 6월 29일, 7월 6일 이앙은 적합하였으나, 7월 13일 이앙은 필요적산온도에 미치지 못하였다. 일조시간은 전체적으로 최적 일조시간인 6.0~6.1.시간보다 약간 높은 6.3~6.5시간으로 조사되었다. 2. 웃거름 질소시비수준에 따른 밀의 간장 및 수장과 관련된 특성은 질소시비수준이 높을수록 간장 및 수장이 높았고, 특히 식물체의 질소농도는 고도의 상관성을 보여주었다. 3. 10a당 밀의 수량성은 질소시비량이 많을수록 증가하는 경향이었으며, 수당립수, $m^2$ 당 수수도 같은 경향이었다. 밀의 품종 간 10a당 평균수량은 수안 > 고소 > 금강 순으로 높았다. 4. 벼의 이앙~출수까지의 소요일수는 조평, 운광, 해담 순으로 짧았다. 10a당 백미수량은 이앙기가 빠를수록 증수하였으며, 6월 29일 이앙하였을 때는 3품종 모두 500 kg/10a 이상의 수량성을 보여주었다. 5. 본 시험을 통하여 천안지역 기후조건하의 논에서 이모작 적합품종은 고소밀-해담벼였다.

• 한국자원식물학회지
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• 제32권5호
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• pp.499-508
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• 2019

사과 '후지'/M.9 포트묘목 개발을 위해 질소시비농도에 따른 생장특성을 조사하고, 포트 우량묘목 생산에 적합한 질소시비농도와 잎의 무기영양성분 함량 및 토양 화학성의 안정성을 확인하였다. 질소시비농도가 높아질수록 묘목의 생장은 증가되었고, 특히16 mM 처리가 수체 생장에 가장 좋았으며, 우량묘목 판단기준에 부합하였다. 32 mM 이상의 고농도는 오히려 생장을 감소시켰다. 잎의 무기영양성분 함량은 8, 16 mM 처리구에서 기존 사과과원의 적정수준보다 높았고, 이러한 무기영양성분은 정식 후 수체 생장에 도움이 될 것으로 생각되었다. 토양 화학성 또한 8, 16 mM 처리구에서 안정적이었다. 따라서 수체생육, 잎의 무기영양성분, 포트 내 토양화학성을 고려한 결과, 사과 '후지'/M.9 우량 포트묘목 생산을 위한 적정 질소시비량은 16 mM로 판단되었다.

• 한국동물생명공학회지
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• 제34권1호
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• pp.10-19
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• 2019

Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: $79.9{\pm}3.8%$ vs G2: $57.5{\pm}4.6%$) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO ($0.1{\mu}M$, MT) treatment (G2: $68.4{\pm}3.2%$ vs G2 + MT: $73.9{\pm}1.4%$). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

• Animal Bioscience
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• 제34권4호
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• pp.546-557
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• 2021

Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

• 한국가금학회지
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• 제49권1호
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• pp.25-32
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• 2022

사료 내 인분해효소를 첨가급여한 산란계에서 사료섭취당 생계분 발생량의 차이는 없었다. 반면에 2,000 FTU/kg 인분해효소 처리구에서 발생한 생계분은 높은 함수율로 인하여 건계분 배출량이 유의적으로 감소하였고(P<0.05), 질소, 인산, 칼리의 배출량도 모두 감축되었다(P<0.05). 부숙한 계분으로 복합비료를 제조한 결과 인분해효소 급여에 따른 퇴비의 배합량과 비료의 양분 함량의 차이는 확인되지 않았다. 퇴비의 구성비는 25~30% 함수율의 퇴비를 사용 시에 비료 내 69.6~71.6%를 차지하였으며, 40~60% 함수율 퇴비는 비료 내 13.0~47.1%를 구성하였다. 따라서 계분처리량을 높이기 위해 계분의 함수율을 낮추기 위한 연구가 추가로 필요하다. 본 연구결과, 계분퇴비는 함수율 조절 및 다른 원료들과의 조합을 통해 양분 균형을 맞춘 복합비료의 주원료로 이용될 수 있음을 확인하였다.

• Journal of Veterinary Science
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• 제24권2호
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• pp.24.1-24.13
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• 2023

Background: Ergothioneine (EGT) is a natural amino acid derivative in various animal organs and is a bioactive compound recognized as a food and medicine. Objectives: This study examined the effects of EGT supplementation during the in vitro maturation (IVM) period on porcine oocyte maturation and subsequent embryonic development competence after in vitro fertilization (IVF). Methods: Each EGT concentration (0, 10, 50, and 100 μM) was supplemented in the maturation medium during IVM. After IVM, nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels of oocytes were investigated. In addition, the genes related to cumulus function and antioxidant pathways in oocytes or cumulus cells were investigated. Finally, this study examined whether EGT could affect embryonic development after IVF. Results: After IVM, the EGT supplementation group showed significantly higher intracellular GSH levels and significantly lower intracellular ROS levels than the control group. Moreover, the expression levels of hyaluronan synthase 2 and Connexin 43 were significantly higher in the 10 μM EGT group than in the control group. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and NAD(P)H quinone dehydrogenase 1 (NQO1) were significantly higher in the oocytes of the 10 μM EGT group than in the control group. In the assessment of subsequent embryonic development after IVF, the 10 μM EGT treatment group improved the cleavage and blastocyst rate significantly than the control group. Conclusions: Supplementation of EGT improved oocyte maturation and embryonic development by reducing oxidative stress in IVM oocytes.