• 농업과학연구
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• 제22권1호
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• pp.62-68
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• 1995

본 연구는 돼지 수정란의 2, 4, 8세포배 및 상실배로부터 분리된 분할배의 작성율, 발생능 및 급속동결에 미치는 영향을 구명하고자 실시되었으며, 그 결과는 다음과 같다. l. Pronase 처리로 분할배를 작성했을 때, 성공율은 2세포배에서 85.7% 이었고, 평균은 68.0% 이었다. Manipulator를 이용했을 경우에는 2세포배 76.6% 및 4세포배 74.3%의 성공율을 나타냈다. 2. 분할배를 체외배양했을 때 배반포기까지의 발달율은 2, 4, 8세포배 및 상실배에서 pronase 처리의 경우 각각 24.1%, 20.4%, 25.5% 및 26.2%이었고 manipulator를 이용하였을 경우 각각 36.4%, 39.5%, 36.1% 및 4l.9% 이었다. 3. 분할배를 동결 융해했을 때 배반포기까지의 발달율은 2세포배에서는 glycerol(16.l%), 4세포배에서는 DMSO(16.7%) 및 상실배에서는 ethyleneglycol(27.6%)의 내동제를 이용한 경우에서 각각 가장 좋은 성적을 나타냈다.

• BMB Reports
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• 제55권8호
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• pp.401-406
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• 2022

Ahnak, a large protein first identified as an inhibitor of TGF-β signaling in human neuroblastoma, was recently shown to promote TGF-β in some cancers. The TGF-β signaling pathway regulates cell growth, various biological functions, and cancer growth and metastasis. In this study, we used Ahnak knockout (KO) mice that underwent a 70% partial hepatectomy (PH) to investigate the function of Ahnak in TGF-β signaling during liver regeneration. At the indicated time points after PH, we analyzed the mRNA and protein expression of the TGF -β/Smad signaling pathway and cell cycle-related factors, evaluated the cell cycle through proliferating cell nuclear antigen (PCNA) immunostaining, analyzed the mitotic index by hematoxylin and eosin staining. We also measured the ratio of liver tissue weight to body weight. Activation of TGF-β signaling was confirmed by analyzing the levels of phospho-Smad 2 and 3 in the liver at the indicated time points after PH and was lower in Ahnak KO mice than in WT mice. The expression levels of cyclin B1, D1, and E1; proteins in the Rb/E2F transcriptional pathway, which regulates the cell cycle; and the numbers of PCNA-positive cells were increased in Ahnak KO mice and showed tendencies opposite that of TGF-β expression. During postoperative regeneration, the liver weight to body weight ratio tended to increase faster in Ahnak KO mice. However, 7 days after PH, both groups of mice showed similar rates of regeneration, following which their active regeneration stopped. Analysis of hepatocytes undergoing mitosis showed that there were more mitotic cells in Ahnak KO mice, consistent with the weight ratio. Our findings suggest that Ahnak enhances TGF-β signaling during postoperative liver regeneration, resulting in cell cycle disruption; this highlights a novel role of Ahnak in liver regeneration. These results provide new insight into liver regeneration and potential treatment targets for liver diseases that require surgical treatment.

• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.55-63
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• 2001

Objectives: This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Materials and Methods: Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Two-step 1,2-propanediol (PROH), dimethylsulfoxide (DMSO) and 4-step PROH DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. Results: As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in PROH and DMSO was significantly higher than 4-8 cell (64.5% versus 62.1 %,79.7% versus 73.2%) (p<0.01, p<0.01), but the development rates of 4-8 cell embryos in PROH and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4-8 cell embryos in PROH were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in PROH (74.4% versus 64.5%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The developmen1 rate from morular to hatching blastocyst, however, was significantly higher in PROH than in DMSO during 48 hr (p<0.01). The survival rate of 4-8 cell embryo was 62.1% in PROH and 73.2% in DMSO. The development rates of embryo in PROH were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. Conclusions: The survival rate of embryo in 2 cell stage was higher than in 4-8 cell stage, and PROH appears more effective cryoprotectant than DMSO because PROH showed better development rates of embryos in 2 and 4-8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

• 한국수정란이식학회지
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• 제29권2호
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• pp.141-148
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• 2014

Quercetin and genistein, plentifully present in fruits and vegetables, are flavonoid family members that have antioxidative function and plant-derived phytoestrogen activity. The antioxidative effects of quercetin and genistein on boar sperm characteristics and in vitro development of IVF embryo were investigated. The sperm motility was increased by addition of genistein $50{\mu}M$ for 6 hr incubation compared to control (p<0.05). The sperm viability was increased by addition of quercetin 1 and $50{\mu}M$ and genestein 1 and $50{\mu}M$ for 3 hr incubation. In addition, the sperm viability seemed to be increased dose-dependantly by addition of quercetin or genistein 1 and $50{\mu}M$, respectively (p<0.05). The membrane integrities were not increased by quercetin or genistein treatments for 3 hr or 6 hr incubation period except for quercetin $1{\mu}M$ for 3 hr incubation. In mitochondrial activities, addition of quercetin $50{\mu}M$ for 6 hr incubation increased mitochondrial activity but decreased at $100{\mu}M$ concentration compared with control (p<0.05). When porcine IVF embryos were cultured in PZM-3 medium supplemented with low concentrations of quercetin ($1{\sim}10{\mu}M$), the developmental rates to morula and blastocyst increased but significantly decreased at high concentrations of quercetin ($25{\sim}50{\mu}M$). The highest developmental rate to blastocysts among all concentrations of quercetin was shown at quercetin $10{\mu}M$ (p<0.05). The developmental rates to morula or blastocysts at low ($0.01{\sim}1{\mu}M$) and high ($5{\sim}10{\mu}M$) concentrations of genistein were not significantly different among all treatment group and genistein did not affect on IVF embryo development. These results suggest that quercetin and genistein seem to have positive effects at certain concentrations on sperm characteristics such as motility, viability and mitochondrial activity. In addition, low concentrations of quercetin (1, 5 and $10{\mu}M$) in this experiment, seem to have beneficial effect on porcine IVF embryo development but genistein did not affect on it at all given concentrations ($0.01{\sim}10{\mu}M$).

• 한국수정란이식학회지
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• 제13권3호
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• pp.235-243
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• 1998

적정 배양액의 선정, ITS의 첨가와 BSA의 농도조절 및 NaCl 농도의 조절을 통해 소 수정란의 무혈청, 체세포배제 배양체계를 확립하기 위하여 수행한 실험에서 다음과 같은 결론을 얻었다. 1. 배양액으로 CRlaa, TALP 및 SOF를 사용하여 발육률을 검토한 결과, 발육률의 유의적인 차이는 나타나지 않았다. 2. 배양액내의 고분자물질원으로 BSA, FBS 및 PVA를 첨가하여 사용한 결과 BSA 및 FBS 첨가군이 PVA 첨가군보다 유의적으로 높은 발육률을 나타내었다(p〈0.01). 3. 배양액내의 BSA 농도를 달리 하면서(1, 3, 8 mg/ml) 1% ITS를 첨가하여 실험한 결과 BSA의 농도가 증가할수록 후기배로의 발육률이 높았으며 모든 군에서 ITS 첨가군이 후기배로의 발육률이 높았으나 BSA가 1 mg/ml로 첨가된 군에서만 ITS 첨가에 따른 유의적인 차이가 인정되었다(p〈0.05). 4. 배양액내의 N3Cl 농도를 114 mM과 90 mM 로 나누어 소 수정란을 배양한 결과 90 mM 군의 후기배로의 발육률이 114 mM 군에 비해 유의적으로 높게 나타났다(p〈0.05).

• Reproductive and Developmental Biology
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• 제30권1호
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• pp.41-45
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• 2006

Electrical treatment has been widely used for porcine oocytes activation. However, developmental rates following electrical activation of porcine oocytes is relatively inefficient compared to other domestic animals. To investigate the effects of porcine oocytes on combined activation by both chemical and electrical treatment, in-vitro matured oocytes were activated by combined cycloheximide and electrical pulses treatment. Cumulus-free oocytes were exposed with NCSU-23 medium containing cycloheximide $(10{\mu}g/ml)$ for 0, 5, 10, 20, 30 min and then activated by electrical pulse treatment and cultured in PZM-3 for 8 days. Also effects of exposure to $6.25{\mu}M$ calcium ionophore for 2 min for cumulus-free oocytes were tested. The percentage of blastocyst formation in 10 min exposure to $10{\mu}g/ml$ cycloheximide and electrical pulse treatment was significantly increased (P<0.05) than in the control group. And exposure to $6.25{\mu}M$ calcium ionophore for 2 min with $10{\mu}g/ml$ cycloheximide for 10min and electrical pulse treatment significantly increased (P<0.05) the percentage of blastocyst developmental rates than the control group. In conclusion, activation by combined cycloheximide and electrical stimulation treatment promoted the subsequent development of porcine oocytes and improved the subsequence blastocyst development.

• 한국발생생물학회지:발생과생식
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• 제14권2호
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• pp.59-63
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• 2010

넙치 수정란의 발생에 미치는 수온의 영향을 조사하였다. 실험에 사용된 수정란은 실내 사육수조에서 자연산란된 것으로서, 6개의 수온별 실험구(5, 10, 15, 20, 25 및 $30^{\circ}C$)에 수용하여 발생과정을 관찰하였다. 수정란은 10, 15, 20 및 $25^{\circ}C$에서 부화 가능하였으며, 부화율은 각각 3, 12, 25 및 50%였다. 발생단계별 수온(T, $^{\circ}C$)과 소요 시간(1/t, hour)과의 상관관계식은 다음과 같다. 포배기: 1/t=0.0208T-0.0951 ($r^2$=0.8593) 쿠퍼씨포기: 1/t=0.0052T-0.0176 ($r^2$=0.9819) 근절기: 1/t=0.0034T-0.0172 ($r^2$=0.8508) 부화: 1/t=0.0016T-0.0068 ($r^2$=0.9915) 위 식을 이용하여 산정한 넙치 난 발생의 생물학적 영도는 $4.3^{\circ}C$였다.

• 한국수정란이식학회지
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• 제18권1호
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• pp.27-33
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• 2003

본 연구는 소형 개의 불임 해결과 체외수정란을 생산하기 위한 방안의 하나로써 난자의 형태, 번식주기, 배양시간 및 활성화 처리가 난포란의 체외성숙 및 체외발생에 미치는 영향을 조사하였다. 1. 신선, salt 및 4$^{\circ}C$에 보존한 난소로부터 채취한 난구세포부착 난자와 나화 난자로 각각 체외수정시켰을 때 16세포기로의 발생율은 14.3%, 5.0% 및 7.5%, 2.8%, 5.7% 및 0.0%로써 난구세포 부착난자군의 체외발생율이 나화 난자군에 비해 높게 나타났다. 2. 발정주기를 inactive, follicular, luteal 단계로 구분하여 채취한 난포란을 각각 체외배양시켰을 때 GV 및 MII로의 발생율은 11.3%와 9.4%, 50.7%와 26.7%, 16.9%와 13.8%였고, 16세포기로의 체외발생율은 0.0%, 10.7%, 1.5%였다. 3 신선한 난구세포 부착 난자를 각각 24, 32, 48시간 성숙배양 후 체외수정시켰을 때 분할율은 8.6%, 15.8%, 23.5%였으며, 16세포기로의 체외발생율은 각각 0.0%, 5.3%, 11.8%로써 48시간 배양군이 가장 높은 발생율을 나타냈다. 4. 활성화 처리 및 비활성화 처리 난자를 각각 체외수정시켰을 때 분할율은 각각 42.5%, 22.2%였고 16세포기로의 체외발생율은 각각 15.0%, 6.7%로써 활성화 처리 난자군이 비활성처리 난자군에 비해 높은 발생율을 나타냈다.

• 재활치료과학
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• 제13권1호
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• pp.87-97
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• 2024

목적 : 본 연구는 발달장애아동에게 활동분석과 전방 연쇄 방법을 이용하여 일상생활활동 기술을 습득시키고자 하였다. 연구방법 : 본 연구에서는 기준변동설계를 사용하여 발달장애아동의 일상생활활동을 단계별로 지도하였다. 중재는 캐나다 작업수행평가(Canadian Occupational Performance Measure, COPM)로 설정된 3가지 과제(신발 신기, 반바지 입기, 반팔 티셔츠)를 치료사가 활동분석한 후 전방 연쇄 방법으로 진행되었다. 독립 변인으로 사용된 3개 과제의 단계별 수행률과 중재 전과 후의 COPM 수행도와 만족도 점수를 종속 변인으로 설정하였다. 중재는 주 5일, 30분씩 이루어졌고, 기초선(3회)을 포함한 신발 신기는 총 18회기, 반바지 입기는 총 25회기, 반팔 티셔츠 입기는 총 18회기 실시되었다. 결과 : 연구 결과 아동의 신발 신기, 반바지 입기, 반팔 티셔츠 입기의 과제 수행률은 회기가 지날수록 향상되었고, 가정에서의 3가지 과제의 수행도와 만족도 역시 증가된 것을 알 수 있었다. 결론 : 본 연구 결과 발달장애아동의 수준에 따른 단계화되고 체계적인 지도 방법은 아동의 일상생활활동 기술을 학습시키는 데 효과적인 것을 확인할 수 있었다. 이를 기초로 과제 학습에 어려움이 있는 발달장애아동에게 일상생활활동뿐만 아니라 다양한 활동영역에 활동분석과 행동 연쇄 방법이 활용되기를 기대해 본다.

• 한국수정란이식학회지
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• 제6권2호
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• pp.1-17
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• 1991

It is widely recognized that the embryonic or fetal loss after breeding is common in the cattle and that it is an important factor affecting reproductive efficiency. The causes of this loss have been subject of extensive researches and the results indicate that the embryonic mortality may he primary factor responsible for low pregnancy rates in non-embryo transfer bovine populations as well as embryo transfer programs. However, it's causes are still not clearly understood. The embryonic mortality or pregnancy rate has been influenced by various embryonic and maternal effects related to genetic and environmental factors. The timing and extent of embryonic mortality vanes greatly according to authors and estimating methods, because it is difficult to make direct measurements. The major important factors that may influence the embryonic losses or pregnancy rates after embryo transfer can be summeirized. 1.When an embryo is transferred to unmated recipients, the contralateral transfer to corpus luteum results in a lower survival rate than ipsilateral deposition. When the embryos are transferred for the production of twin calves, their survivals and twin pregnancies have quite inconsistent according to the transfer methods either to the unmated-synchronized or already mated recipients and more works are needed to accurrately clarify the previous results. 2.Although embryos can be cultured in vitro some hours without the great declines in pregnancy rates, the rates differ markedly among culture times and media but may be improved by co-transfer systems. 3.Embryo developmental stages and quality grades clearly affect the survival rate following freezing and the pregnancy rate after transfer and the selection of embryos without chromosome abnormalities and of high fertile semen may also be considered to increase the pregnancy rates. 4.Many researches have attempted to relate the plasma progesterone levels to pregnancy rates and others have done either direct progesterone supplementation or luteal stimulation by hCG treatment in order to increase the pregnancy rates. However, these effects on pregnancy rates are inconsistent and also contradictory. 5.The asynchrony between donors or embryos and recipients may he a major cause of embryo death and low pregnancy rate and the sensitivity to uterine asynchyony differs in according to the quality and stages of embryos. 6.The extremes of poor or over nutrition during early pregnancy in the recipients are detrimental to the survival of embryos and the good body condition is required to prevent a reduejion of pregnancy rates. The uterine pathogens in embryonic mortality or fertility have been questioned but the infection of C.pyogenes and Campylobacter fetus is still important pathogens. 7.The heat stress during early pregnancy may reduce conceptus weight and possibly increase the embryonic mortality.