• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1763-1769
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• 2020

Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage. Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooled-storage at 17℃. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated. Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential. Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17℃.

• 대한수학교육학회지:수학교육학연구
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• 제8권2호
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• pp.745-752
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• 1998

컴퓨터의 급속한 보급으로 시각화는 수학 교육자사이의 논의에 자주 등장하는 소재가 되었다. 우리는 다양한 소프트웨어를 사용하여 준비한 수업에 학생들로 임하게는 하지만 거의 그들의 사고 발달과정에는 관심을 갖지 못하고 있다. 이 논문은 구성주의(Constructivism)와 정보처리체계(Information-Processing System)에 입각하여 수학의 시각화를 생각해보고 어떻게 시각화 환경을 준비해야하는지 논해보고자 하였다. 구성주의의 시각화에서는 반영적 추상(reflective abstraction), 반복되는 경험(repeated experience), 그리고 지식 위계성이 학습의 기능 체계를 이루므로 발견적 학습을 통해 학생 스스로 의미를 구성할 수 있도록 Thomas (1992)의 세 가지 제안을 이용하여 수업을 준비할 수 있다. 정보처리체계에서는 지식은 서술적인 것과 과정적인 것으로 나뉘어지고, 시각적 표상을 수록하고 삭제하는 과정과 조작 가능한(manipulative) 환경과의 상호작용으로 기호적 시각으로 표상을 변화하는 과정을 통해 개념을 습득하게된다. 시각화는 스키마와 개념상을 통해서 일어난다. 그래프, 애니메이션, 그리고 다른 시각적 표상 등은 이 개념상에 직접적 효과를 주므로 매우 중요하다. 이런 논란을 바탕으로 교사는 반영적 추상화를 위해 시간을 충분히 제공해야하고, 비슷한 문제를 가지고 여러번 시도를 할 수 있게 하며, 학생을 잘 관찰하여 그들의 지식 위계성을 이해하고 방향을 제시하며, 논리적이고 잘 연결된 시각적 표상을 제공하고, 상징적 사관으로 확장할 수 있게 조작할 수 있는 환경에서 시각화에 대해 학생과 많은 대화를 하도록 수업을 준비해야한다. 그한 예로 타원을 가르치기 위해 몇 가지 테크놀로지를 활용한 시각화 환경을 구성해보았다.ates of bisected bovine embryos by micromanipulator and micropipett were 29.2% and 19.1%, respectively. The rates of non-bisection embryos(46.7%) were significantly higher than those of bisection embryos. 2. The in vitro developmental rates of bisected bovine embryos by micromanipulator, micropipett and pipetting method were 32.4%, 19.4% and 25.6%, respectively.3. The in vitro developmental rates of with and without-zona pellucida of bisected bovine embryos by raicromanipulator were 30.8% and 25.0%, respectively. The rates of nonbisection embryos(53.1%) were significantly higher than those of bisection embryos.랑크톤 군집내 종 천이와 일차생산력에 크게 영향을 미칠 수 있음을 시사한다.TEX>5.2개)였으며, 등급별 회수율은 각각 GI(8.5%), GII(13.4%), GIII(43.9%), GIV(34.2%)로 나타났다.ments of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to

• 한국수정란이식학회지
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• 제4권1호
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• pp.1-13
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• 1989

This paper reviews the most important steps that have generated consistent progress in principles and developmental progress of embryo cryopreservation, and also study on freezing procedure and its application by conventional method and current improved method for freezing procedure and its appilcation of embryo cryopreservation in farm animals. Four were of particular interest: 1.The transport of water across the ccli membrane (zona pellucida) during freezing and thawing accordinglyplays a role in determing whether the celi survives. This movement of water is controlied mainly by extracellular phase changes and by the nature and concentration of any cryoprotective agent present. Therates of cooling, freezing and warming, and the intervals over which they are applied are further decisi've factors in determining whether a cryopreservation procedure allows survival after thawing. 2.The first successful deep freezing experiments with sheep morula and blastocysts during the seventies were based on the early procedures used for mouse embryos.Current research during the eighties is developed with the aim of simplifying and improving current procedures such as one-step dilution and rapid or ultra-rapid cooling by using the model of laboratory animals. 3.The conventional method for the embryo cryopreservation is described. An alternative to this method which may result in high survival and also in reducing of the freezing and thawing time is done by combing a permeable cryoprotectant such as glycerol, DMSO or propanediol and a non-permeable compound such as sucrose, trehalose, raffinose or lactose. 4.Finally a different approach to the preservation of embryos, named vitrification, is introduced. This procedure depends upon the ability of concentrated solutions of cryoprotective agents such as glycerol and propanediol to supercool to very low temperature (-196$^{\circ}C$) during rapid cooling before solidifying without formation of ice. However, more complete data are necessary for successful vitrification of blastocysts.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.247-252
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• 1994

Intracytoplasmic sperm injection(ICSI) was known as effective method in treatments of couples who unable to be helped by conventional in vitro fertilization. In 78 treatment cycles of 78 infertile couples using ICSI performed at our infertility clinic between May and August 1994 were analyzed. These patients were classified two groups, andrological factor(AF) and non-andrological factor(non-AF) group. The AF group, which had abnormal sperm physiology, included oligozoospermia, asthenozoospermia, oligoasthenoteratozoospermia(OATS) and microsurgical epididymal sperm aspiration(MESA) patients. The non-AF group, which had abnormal oocyte physiology, included abnormal zona pellucida, poor quality of oocyte and immune factor infertile patients. A single spermatozoon was injected into the ooplasm of 776 metaphase II oocytes. The fertilization rate was 44.6%(346/776) and 319 embryos were transferred. After 73 embryo transfers(93.6% of treatment cycles) 23 pregnancies were estabilshed, i. e. pregnancy rate of 29.4% per started cycle and 31.5% per embryo transfer. Fertilization rate of AF and non-AF group was 46.2% and 35.8%, pregnancy rate was 34.5%(20/58) and 20.0%(3/15), respectively. In order to increase the pregnancy rate, assisted hatching(AHA) has done after lCSl in 47 treatment cycles. Pregnancy rate of ICSI with AHA and without AHA group was 34. 0% (16/47) and 26.9%(7/26), respectively. ICSI was more effective in andrological factor infertility and the pregnancy rate was increased by ICSI with AHA procedure.

• 한국가축번식학회지
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• 제26권3호
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• pp.263-273
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• 2002

본 연구는 xanthine-xanthine oxidase system하에서 돼지 동결-융해 정자의 lipid peroxidation과 체외수정능력에 대한 $\beta$-mercaptoethanol ($\beta$-ME)의 영향을 검토하였다. 그 결과 돼지 동결-응해 정자가 X-XO system하에서 처리되었을 때, control구에서 높은 정자생존율이 관찰되었으나 처리구간의 유의차는 인정되지 않았다. 또한 첨체반응이 유기된 정자의 비율은 모든 처리구에서 $\beta$-ME 첨가시 보다 무첨가시가 유의적(P<0.05)으로 더 높았다. 한편, X-XO system하에서 체외수정시 난자에 대한 정자의 침입율은 모든 조건하에서 $\beta$-ME 첨가시가 무첨가시 보다 높은 경향을 나타냈지만, 유의적인 차이는 인정되지 않았다. 정자의 lipid peroxidation은 malondialdehyde (MDA)의 생성에 기초를 두고 평가하였는데, 모든 조건하에서 $\beta$-ME 첨가시 보다 무첨가시에 MDA의 생성이 높게 나타났지만, 유의적인 차이는 인정되지 않았다. 또한 동결-융해된 정자의 sulfhydryl (-SH) group의 함량을 측정한 결과 모든 처리구에서 $\beta$-ME 무첨가시 보다 첨가시에 높은 함량이 측정되었지만, 유의적인 차이는 인정되지 않았다. 한편, 체외에서 성숙시킨 난자의 투명대에 대한 동결-융해 정자의 접착 정도를 평가한 결과 모든 처리구에서 $\beta$-ME 첨가시 무 첨가시에 비해 다소 높은 경향을 보였으며, Control group의 경우 X+XO group에 비해 유의적(P<0.05)으로 높은 정자접착율이 관찰되었다 그렇지만, $\beta$-ME 첨가 유무에 따를 유의적인 차이는 인정되지 않았다. 된 연구의 결과는 X-XO system하에서 $\beta$-ME 첨가가 돼지에서의 체외수정능력 향상에 영향을 미치는 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1188-1195
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• 2001

A tetraparental chimeric bull was successfully produced by aggregating bovine IVF embryos of F1 (female Holstein${\times}$male Japanese Black) and F1(female Japanese Brown${\times}$male Limousin) and culturing in vitro without the zona pellucida at Yamaguchi Research Station in Japan. In the microsatellite genotyping, 12% (28/228) microsatellite primer sets ware potentially useful for this parentage analysis in the chimeric bull, 78.6% (22/28) of microsatellite present in the chimeric bull were uniquely contributed from the Japanese Black and 21.4% (6/28) from Limousin. This chimeric bull semen was used in producing IVF embryos. The chromosome preparations were made from peripheral lymphocytes. Based on chromosome analysis the Chimera had apparently normal chromosomes (29 acrocentric pairs, one large sub metacentric X chromosome and one small sub metacentric Y chromosome). The proportion of acrosome reacted spermatozoa after 1 h of incubation was higher (p<0.01) with the Chimera than with the Holstein and in Japanese Brown bulls. But did not differ from Japanese Black and Limousin bull sperm. Fertilization rates observed after 5 h of sperm-oocyte incubation with Chimera sperm were higher (p<0.05) than with Japanese Brown and (p<0.01) than with Holstein sperm, but did not differ from Japanese Black and Limousin sperm. The cleavage rates of IVF oocytes inseminated with Chimera sperm were also higher (p<0.001) compared with Holstein, (p<0.01) Japanese Brown and (p<0.05) Limousin, but did not differ from Japanese Black sperm. The blastocyst rates of IVM oocytes inseminated with sperm were higher (p<0.05) than in Limousin, Japanese Brown and Holstein, but did not differ from Japanese Black. Chimeric cattles were produced by aggregation of parthenogenetic (Japanese Brown) and in vitro fertilized (Holstein) bovine embryos at the Yamaguchi Research Station in Japan and by aggregation of parthenogenetic (Red Angus) and in vitro fertilized (Holstein) embryos at the St. Gabriel Research Station in Louisiana. The aggregation rate of the reconstructed demi-embryos cultured in vitro without agar embedding was significantly lower than with agar embedding. The aggregation was also lower when the aggregation resulted from a whole parthenogenetic and IVF-derieved embryos cultured without agar than when cultured with agar. The development rate to blastocysts, however, was not different among the treatment. To verify parthenogenetic and the cells derieved from the male IVF embryos in blastocyst formation, 51 embryos were karyotyped, resulting in 27 embryos having both XX and XY chromosome plates in the same sample, 14 embryos with XY and 10 embryos with XX. The viability and the percentage of zonafree chimeric embryos at 24 h following cryopreservation in EG plus T with 10% PVP were significantly greater than those cryopreserved without PVP. Pregnancies were diagnosed in both stations after the transfer of chimeric blastocysts. Twin male and single chimeric calves were delivered at the Yamaguchi station, with each having both XX and XY chromosomes detected. Three pregnancies resulted from the transfer of 40 chimeric embryos at the Louisiana station. Two pregnancies were Jost prior to 4 months and one phenotypically chimeric viable male born.

• 한국수정란이식학회지
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• 제16권2호
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• pp.139-144
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• 2001

본 연구는 숫소 개체별, 정자의 형태, 정자의 전처리 및 난자의 투명대부착과 미부착별로 현미조작에 의해 난자의 세포질내 단일정자를 주입했을 때 웅성전핵 형성율과 체외발생율을 조사하였다. 1. 숫소 개체별 정자를 이용하여 ICSI를 하였을 때 웅성전핵 형성율은 각각 73.9∼87.0%였고 체외발생율은 33.3∼60.9%를 나타냈다. 2. 신선 및 동결정자와 미부절단 정자와 두부, 미부손상 정자를 이용하여 ICSI를 하였을 때 웅성전핵 형성율은 각각 82.0%, 78.0% 42.2%, 51.1%였고, 체외발생율은 56.0%, 42.0%, 17.8%, 22.2%로서 신선정자를 이용했을 때가 다른 처리군에 비해 높은 전핵형성율과 체외발생율을 나타냈다. 3. 정자를 heparin, BFF, His, Ca Ionophore, I + caffeine 액으로 처리후 ICSI를 하였을 때 전핵 형성율은 66.7∼82.2%였고, 체외발생율은 33.3∼60.6%로서 I + caffeine 처리법이 다른 처리군에 비해 높게 나타났다. 4. 투명대 부착 난자 및 미부착 난자로 ICSI를 하였을 때 전핵형성율은 각각 80.0%, 72.0%였고, 체외발생율은 46.0%, 36.0%로서, 토명대부착 난자가 투명대 미부착 난자에 비해 높은 웅성전핵 형성율과 체외발생율을 나타냈다.

• 한국가축번식학회지
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• 제23권2호
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• pp.149-157
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• 1999

본 연구는 체외에서 돼지정자의 난자 내 침입에 있어서 난구세포의 기능적인 역할을 검토하기 위하여 실시하였다. 체외수정 시 정자침입율은 난구세포 부착 (61%) 난자가 제거 (25%) 된 난자에 비해 높았으나 (P＜0.001), 다정자침입에는 영향을 미치지 않았다. 한편 체외수정시 hyaluronidase 를 0, 0.01, 0.1 및 1.0mg/$m\ell$ 농도로 첨가된 배양액내에서 난구세포가 부착된 난자내의 정자침입률은 각각 61, 56, 66 및 39%로 이들 세포를 제거한 난자에서의 침입률 34, 35, 30 및 27%에 비해 높았다. 그러나 다정자침입률은 hyaluronidase의 농도에 관계없이 난구세포를 제거한 난자에서 낮았으며, hyaluronidase의 농도가 높아지면서 다정자침입률이 낮아지는 경향을 나타냈다. 또한 hyaluronidase를 첨가한 배양액내에서 수정후 16 및 24시간에서의 정자침입률은 난구세포를 제거한 경우 (25 및 31%) 보다 난구세포가 부착된 난자 (48 및 62%)에서 높았으며 (P＜0.05), 다정자침입률은 난구세포 제거 시 유의적으로 낮게 나타났다 (P＜0.05). 한편, 난자로부터 채취한 난구세포를 여러 농도로 첨가한 후 난구세포 제거난자를 이용하여 체외수정한 결과 hyal uronidase 첨가보다는 무첨가시 정자의 침입률과 다정자 침입률이 낮게 나타났다. 본 연구의 결과로부터 난구세포는 정자의 침입에 효과적으로 작용하였으며, hyaluronidase의 첨가와 난구세포수의 조절이 정자의 침입과 다정자침입에 영향을 미치는 것으로 추측되었다.

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.119-133
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• 1997

In spite of much progress in vitro fertilization and embryo transfer (IVF-ET) program, the pregnancy rate remains at 20-30%, and the endometrial implantation rate per embryo transferred at 10-15%. As a result, about 90% of embryos may fail to implant to the endometrium, and many attempts such as optimization of follicular development, improvement of in vitro culture system including coculture, and micromanipulation of zona pellucida have been made to improve embryonic implantation after IVF-ET. Recently, several procedures of assisted hatching (AH) using micromanipulation have been introduced, and pregnancies and births have been obtained after AH. To develop and establish AH as an effective procedure to improve embryonic implantation, AH with partial zona dissection (PZD) was performed in 116 cycles of 89 infertile couples who had previous repeated failures of standard IVF-ET more than two times (Group I: 71 cycles in 54 patients), or who had implantation failure of embryos with good quality (Group II: 15 cycles in 13), or who had undergone AH without specific indication (Group III: 30 cycles in 22) from January, 1995 to Februry, 1996, and the outcomes of AH were analyzed according to pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $9.9{\pm}7.1$ in Group I, $11.5{\pm}4.5$ in Group II, and $7.9{\pm}6.4$ in Group III. The number of embryos transferred after AH was $4.7{\pm}1.8$ in Group I, $5.3{\pm}1.3$ in Group II, and $3.5{\pm}2.4$ in Group III. The mean cumulative embryo score (CES) was $56.8{\pm}30.0$ in Group I, $76.1{\pm}35.9$ in Group II, and $38.5{\pm}29.9$ in Group III. The overall clinical pregnancy rate per cycle and per patient was 12.7% (9/71) and 16.7% (9/54) in Group I, 33.3% (5/15) and 38.5% (5/13) in Group II, and 6.7% (2/30) and 9.1% (2/22) in Group III, respectively. There were significant differences in the numbers of oocytes retrieved and embryos transferred, CES, and the clinical pregnancy rate per cycle among three groups. There was a significant inverse correlation between basal serum FSH level and CES, and no pregnancy occurred in patients with CES less than 20. In conclusion, AH of human embryos with PZD prior to ET has improved the implantation and pregnancy rates in IVF-ET patients with the past history of repeated failures, especially in spite of transfer of embryos with good quality, and AH will provide a range of novel techniques which may contribute much to effective management of infertile couples.

• 한국수정란이식학회지
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• 제14권1호
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• pp.17-22
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• 1999

도축장에서 채취한 돼지난자를 직경별로 $5\mu\textrm{m}$ 간격으로 나누어, 난자의 크기에 따른 체외성숙과 발육능을 구명코자 체외성숙과 체외수정 후의 배 발달율을 조사하였다. 채취한 난자의 투명대를 제외한 평균직경은 $114.4\pm5.45\;\mu\textrm{m}$ 이었으며, 직경별 분포는 < 105, 105~110, 110~115,115~120, 120~125, $125\;\mu\textrm{m}$<이 각각 3.0, 11.0, 31.2, 41.5, 13.0 및 0.4%로 72.7%가 110부터 $120\;\mu\textrm{m}$ 사이였다. 체외성숙율에 있어서 직경 $105\;\mu\textrm{m}$ 미만난자는 66.7%인 반면 $105\;\mu\textrm{m}$ 이상의 난자는 91.8~100%로 유의적(P<0.05)으로 높았다. 체외수정율도 직경 $105\;\mu\textrm{m}$ 미만인 난자는 50% 이었던 반면, $105\;\mu\textrm{m}$이상은 8106~85.5%로 유의적(P<0.05)으로 높았다.다정자침입율에 있어서 $110\;\mu\textrm{m}$이상의 난자는 17.8~27.7%로 $105~110\mu\textrm{m}$ 의 37.5% 보다 낮았다. 난자의 직경별 배반포기까지의 발달율은 $105\;\mu\textrm{m}$ 이하는 전혀 발달하지 않았고(0%), $105~110\mu\textrm{m}$는 701%, $110~11\;5\mu\textrm{m}$ 12.5%, $115~120\; \mu\textrm{m}$ 24.0%, $120~125\; \mu\textrm{m}$ 18.3%, $120\; \mu\textrm{m}$<0%로, $115~120\; \mu\textrm{m}$ 직경난자의 체외발달율이 유의적(P<0.05)으로 높았다.이상의 결과를 종합할 때, 돼지의 체외수정용 나자는 직경 $110\; \mu\textrm{m}$ 이상을 이용하는 것이 생산성을 높일 수 있는 방법으로 사료된다.