• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.163-168
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• 1992

This study was carried out to set up the ovum bank for ovum donation and to determine the best freezing method for human immature oocytes. Human immature follicular oocytes were cryopreserved by slow freezing and rapid thawing method. Immature follicular oocytes were treated by propanediol(PROH) solution by 2 and 4 step method in protocols A & B, respectively. In protocol C, immature oocytes were exposed to sucrose prior to treatment of PROH by 4 step method. We compared survival rate, maturation rate, and fertilization rate of immature oocytes among three protocols. Results were as follows. 1. Oocytes treated by the protocol C showed the highest survival rate( 70.3 %) and maturation rate(34.6%) after thawing. 2. Survival rate of oocytes treated by the protocol C was significantly higher than that of the protocol B after thawing(p<0.05). In conclusion, treatment of oocytes with sucrose prior to expose PROH was the best freezing method. Sucrose may have reduced the toxic effect of cryoprotectant to oocytes. We failed to induce fertilization of oocytes, which were treated by any protocols, by conventional insemination method, but obtained 28.8% fertilization rate by using partial zona dissection(PZD) method. This result suggests that micromanipulation(PZD) of the thawed oocytes before insemination will improve the fertilization rate.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.96-96
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• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• 한국가축번식학회지
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• 제20권1호
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• pp.1-8
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• 1996

돼지난포란의 체외성숙, 체외수정 및 체외 배양체계의 개발은 핵치환에 의한 복제동물 및 형질전환 동물생산 등과 같은 첨단생명공학연구 추진에 크게 기여할 것이다. 그러나 돼지난자의 체외수정시 다정자 침입은 큰 문제점으로 지적되고 있는데, 그 원인 및 개선책은 많은 연구에도 불구하고 정확히 밝혀져 있지 않다. 따라서 본 연구는 돼지 난포란으로부터 채취한 난자를 체외성숙 후, 체외수정시 체외수정 배양액에 난관액을 첨가하여 수정율 및 다정자 침입율을 조사하고, 또한 체외수정 후 체외 배 발달율을 조사할 목적으로 실시하였다. 수정용 배양액에 1과 5% 난관액을 첨가하였을 때 정자침입율과 수정시 난자에 침입한 정자의 평균 수가 감소하였으며, 또한 투명대에 부착된 정자의 수도 감소하였다. 난관액이 첨가된 수정용 배양액에서 정자를 1.5와 3시간 동안 전 배양을 실시하였을 때 정자의 수정능 획득과 첨체반응이 대조군에 비해 증가되었다. 그리고 1% 난관액이 포함된 배양액에서 체외수정된 난자의 배반포까지 발달율은 18.9%로 대조구의 12.4%보다 유의성 있게 높았다. 이러한 연구결과는 난관 분비액의 어떤 인자가 정자의 첨체반응을 유도해서 정자침입율 및 다정자 침입을 감소시키고, 이어 배발달율을 증가시킨다고 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.501-508
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• 2013

The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode's albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2-4 cell, 8-16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.

• 한국토양비료학회지
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• 제46권2호
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• pp.136-141
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• 2013

노동력 절감을 위해 감나무의 수고 낮추기가 일반화되고 있지만 수고를 심하게 낮춘 해에는 나무 세력이 강해져 과원 관리가 어려워지는 것이 문제가 되고 있다. 본 시험은 동계전정 때에 강전정을 하여 3 m 이상의 수고를 약 2 m로 낮춘 12년생 '상서조생' 단감나무를 대상으로 수세 안정을 위한 시비량 조절 효과를 검토하였다. 관행 시비량을 기초로 4월 기비 (주당 N 224 g, $P_2O_5$ 172 g, $K_2O$ 172 g), 6월 추비(주당 N 112 g, $K_2O$ 99 g), 10월 추비 (주당 N 112 g, $K_2O$ 99 g)를 시용하는 관행시비구와 각 시기마다 관행시비구의 1/3, 2/3씩 주는 시비구 및 무시비구 등 4처리구를 두었다. 처리 당년에 무시비구는 관행시비구에 비해 신초장이 작고 불필요한 2차생장지 발생이 39% 줄었으나 수량과 과실 크기, 색도, 경도, 당도는 감소하지 않았다. 또한 시비구들간의 신초 및 과실의 반응에도 유의적인 차이가 없었다. 이듬해 3월에 채취한 휴면 가지의 가용성 당, 전분, 질소, 칼륨농도는 전년도 시비량에 따른 차이가 명확하지 않았다. 시비량이 동일했던 이듬해 신초장, 도장지 및 2차 생장지 발생, 수량 및 과실 특성은 전년도 시비량에 따른 차이가 없었다. 그러므로 수세 관리 및 시비 비용 절감 측면에서 강전정에 의한 수고 낮추기 당년에는 시비량을 관행의 1/3 이하로 줄여도 될 것으로 판단되었다.

• 환경생물
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• 제31권2호
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• pp.69-77
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• 2013

말똥성게 (Hemicentrotus pulcherrimus)의 생식세포 및 pluteus 유생을 이용하여 중금속인 Arsenic (As)와 Chromium (Cr)이 정상 수정률 및 배아 발생률에 미치는 독성 영향을 조사하였다. H. pulcherrimus의 수정률 및 배아 발생률에 미치는 As와 Cr의 독성은 6.25, 12.5, 25, 50, 100 ppb의 농도에서 조사하였다. 0.5 M KCl 용액을 이용하여 방란 및 방정을 유도하였고, 정상 수정률 및 배아발생률은 수정 후 각각 10분 및 64시간째 관찰하였다. As와 Cr을 첨가하지 않은 대조구에서 정상 수정률과 배아 발생률은 각각 94%와 93% 이상을 나타냈다. 이들 중금속 첨가에 의해 수정률은 아무런 변화가 나타나지 않았지만 배아 발생률은 농도 의존적 감소하는 것으로 나타났으며, As의 첨가에 의해 배아 발생률은 6.25 ppb에서 유의적으로 감소하였으며 (P<0.01), Cr의 경우는 25 ppb에서 유의적인 감소를 나타냈다 (P<0.05). H. pulcherrimus의 정상 배아 발생률에 대한 LOEC는 As의 경우는 6.25 ppb를 Cr은 25 ppb를 나타냈다. 이들 연구결과로 해양생태계 내에서 As가 6.25 ppb, Cr이 25 ppb를 초과하는 농도일 때는 H. pulcherrimus와 같은 무척추동물의 정상 부화율은 급격히 감소할 것으로 판단된다. 본 연구결과를 바탕으로, H. pulcherrimus의 정상 배아 발생률을 이용한 생물학적 평가방법은 중금속과 같은 유해물질에 대한 해양생태계의 영향을 판단하기 위한 시험방법으로 유용하게 이용될 수 있을 것으로 판단된다.

• 한국작물학회지
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• 제50권1호
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• pp.16-21
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• 2005

This study was conducted to establish the elaborate nitrogen fertilization method to enhance N use efficiency in direct-seeded rice on flooded paddy. The nitrogen uptake by rice plants was insignificant until 25 days after seeding, and increased gradually thereafter. During this early growth stage, rice plants absorbed only the $4\%$ of basal applied N, while the $45\%$ of N fertilizer remained in the paddy soil. The absorption of basal N by rice plants was almost completed at 46 days after application. Nitrogen top-dressed at 5-leaf stage was well matched to crop nutrient demand, so it could be absorbed so actively in 8days after application. As a result, we could cut down the amount of N fertilizer to $36\%$ of the basal N level without significant difference in yield. Plant recoveries of fertilizer $^{15}N$ applied with different application timings were $7.8\%$ for basal, $9.4\%$ for 5-leaf stage, $17.1\%$ for tillering stage, and $23.4\%$ for panicle initiation stage, respectively. When urea was applied with nitrogen fertilization practice based on basal incorporation (BN), plant recovery of $^{15}N$ at harvest was $31.0\%$, which was originated from $13.7\%$ for grain, and $21.3\%$ of the fertilizer $^{15}N$ remained in the soil, and the rest could be uncounted. Plant recovery of fertilizer $^{15}N$ applied with nitrogen fertilization practice based on topdressing at 5-leaf stage (TN), where N rate was reduced by $18\%$ compared with BN, was $35.1\%$ (grain $15.6\%$), and $19.9\%$ of the fertilizer $^{15}N$ remained in the soil, and the rest could be uncounted. TN showed a higher $^{15}N$ recovery than BN because it was to apply N fertilizer at a time to well meet the demand of rice plant direct-seeded on flooded paddy. We concluded that TN would be the nitrogen fertilization method to enhance N use efficiency in direct-seeded rice on flooded paddy.

• 한국작물학회지
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• 제58권1호
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• pp.8-14
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• 2013

벼 재배시 본답의 기비와 분얼비를 대체하여 묘판에 1회 시비하는 용출제어형 입상배합(Bulk Blending) 피복비료를 시용 후 벼 생육과 수량, 품질, 경제성 등 완효성비료의 처리효과를 검토한 결과는 다음과 같다. 1. 개발된 육묘상비료의 T-N용출량은 온도가 높을수록 빨라졌고 온도에 관계 없이 분얼비 시용 시점인 14일경부터 높아져 21일경 최고에 달했다. 2. 이앙 후 60일에서 질소흡수이용율은 표준시비 28.6%에 비하여 개발된 육묘상 비료가 높았으며 200 g시비구에서 73.1%, 300 g시비구에서 66.1%로 나타났다. 3. 생육단계별(최고분얼기, 유슈형성기 등) 초장, 경수, SPAD값 생육상황은 개발된 육묘상비료 300 g/상자 시비에서 표준과 대등하였다. 4. 성숙기에도 간장, 수장, 단위면적당 수수는 개발된 육묘상비료 300 g/상자 이상 시비구에서 표준과 같거나 많았다. 5. 10a당 백미수량은 개발된 육묘상비료 시용량이 많을수록 증가하였다. 시비량별로 보면 표준시비 528 kg 대비 개발된 육묘상 비료 300 g/상자 이상 시비구에서 525~552 kg으로 대등하거나 많았다. 6. 백미 품질은 개발된 육묘상비료 시용량이 많을 수록 단백질함량은 증가하였으며 완전미율, 아밀로스함량, 백도, 투명도는 비료량이 적을수록 증가하는 경향이었다. 7. 따라서 표준시비량 이상의 수량과 품질 및 경제적인 측면을 고려한 개발된 육묘상 비료의 적정 시비량은 300 g/상자(실비. 9 kg/10 a)이었다. 8. 시비량은 육묘상비료가 표준시비에 비해 49.3% 절감되었고, 시비노동력도 49.2% 절감되었다.

• Reproductive and Developmental Biology
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• 제29권3호
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• pp.141-144
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• 2005

This experiment was to investigate whether the mitochondria function assessment can be used for the prediction of sperm fertility through examining the correlation between mitochondria fluoromicroscopic frequency of frozen-thawed eight Hanwoo bull semen using rhodamine123 (R123) and in vitro embryo development following fertilization. Individual sperm were stained in 5 ${\mu}g/mL$ R123-added calcium-free Sp-TALP for 30 min at 0 h, 6 h, 12 h and 24 h after thawing and examined their mid-piece under an epifluorescence microscope using 495 nm excitation filter (x1,000). Three replications were taken, and at least 300 sperm per individual were examined. When semen samples were separated into two groups (good and poor) by sperm motility and fluorescent frequencies at just after thawing, average fluorescent frequencies were remarkably reduced as time going (0 h; $53.29{\~}72.94\%$, 6 h; $21.40{\~}58.90\%$, 12 h; $8.26{\~}25.93\%$, 24 h; $1.00{\~}13.78\%$, irrespective of selected group, and there were no differences at 6 h or 12 h after thawing between selected groups but indicated significant difference at 24 h after thawing (p<0.05). In vitro fertilization rates in good and poor groups ranging $70.8{\~}77.8\%$ and $52.1{\~}84.5\%$, respectively, were not significantly different. However, in vitro development rates of the same groups ranging $25.7{\~}40.0\%$ and $12.9{\~}1.8\%$, respectively, were significant different (p<0.05). These results demonstrate that mitochondria fluoromicroscopic assessment of frozen-thawed bovine sperm may be used as a criterion to select more fertile sperm.

• 여성건강간호학회지
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• 제22권2호
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• pp.78-85
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• 2016

Purpose: The purpose of this study was to evaluate the effect of abdominal massage on alleviating pain caused by the injection of recombinant gonadotropin for In Vitro Fertilization (IVF) among infertile women. Methods: This study employed a nonequivalent control group non-synchronized design. A total sample of 149 infertile women who never experienced in vitro fertilization was recruited at C fertility center. Seventy women were assigned into experimental group and 79 into controls. The experimental group had been informed to do abdominal massage prior to the injection and to record their subjective pain using visual analogue scale. The control group had been informed to record their subjective pain in the same way just after the injection. Results: Compared to the pre-tested pain scores, the pain scores in experimental group were significantly reduced by 0.7 points, whereas the control group increased by 0.9 points (t=-4.55, p=.001). Conclusion: This study confirms that the use of abdominal massage prior to the injection is an effective way to alleviate pain on injection site. This massage may be a useful intervention for infertile women about pain alleviation.