• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.105-109
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• 2003

Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

• Clinical and Experimental Reproductive Medicine
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• 제29권1호
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• pp.57-66
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• 2002

Objective : This study was designed to investigate the interrelationship and clinical usefulness of sperm morphology by strict criteria (SM), acrosome reaction following ionophore challenge test (ARIC) and sperm penetration assay (SPA) using zona-free hamster ova as prognostic factors in in vitro fertilization. Materials and Methods: Semen samples were provided by 83 patients undergoing IVF. We first evaluated the differences between normal fertilization group and poor fertilization group on three andrologic tests. Secondly, we analyzed the relationship between the three andrologic tests and in vitro fertilization on IVF settings. Finally, we evaluated the effectiveness of the three andrologic tests as the prognostic indicators for fertilizing ability. Results: The fertilization rate of all men in the poor fertilization group was less than 30%; but there was no evidence that this poor fertilization was due to oocyte defects. The results of three andrologic tests were significatly higher in normal fertilization group. Fertilization rate (%) in vitro was highly correlated (p<0.001) with % normal sperm by SM, ARIC value (%), and SPA result. By using Receiver-Operator-Characteristic curve (ROC), we evaluated the effectiveness of these three tests. The sensitivity and specificity of SM, ARIC test and SPA in predicting fertilization potential in IVF setting were 76% and 75%, 84% and 90%, and 76% and 95%, respectively. Conclusion: Our data suggest that the three andrologic tests can be reliable tools as prognostic factors of sperm fertilizing ability. Among these test, ARIC test and SPA gave more accurate information on fertilizing capacity. ARIC test was shown to have a predictive value for fertilizing ability comparable to that of SPA that appears to be a simple and cost-effective addition to current andrology laboratory. Combined application of these three tests may give more information on predicting sperm fertilizing capacity.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.251-260
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• 1998

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol $(E_2)$ were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with $10{\mu}l$ of 10% DMSO was induced spontaneously in $10.1{\pm}9.8%$, and acrosomal reaction with calcium ionophore A 23187 was induced in $27.4{\pm}18.1%$, and the ARIC value was $17.4{\pm}16.2%$. There were no significant correlation between the ARIC value and the fertilization rate ($r^2$=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos ($r^2$=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the micro assisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.

• 한국발생생물학회지:발생과생식
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• 제22권4호
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• pp.297-307
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• 2018

The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on cumulus expansion, nuclear maturation, fertilization capacity and subsequent development in porcine oocytes. The oocytes were incubated with 0, 25, 50, and $100{\mu}M$ ALA. Cumulus expansion was measured at 22 h, and gene expresison and nuclear maturation were analyzed at 44 h after maturation. Then, mature oocytes with ALA were inseminated, and fertilization parameters and embryo development were evaluated. In results, both of cumulus expansion and nuclear maturation were increased in $50{\mu}M$ ALA groups compared to control groups (p<0.05). However, expression of gap junction protein alpha 1 (GJA1, cumulus expansion-related gene), delta-6 desaturase (FADS1, fatty acid metabolism-related gene), and delta-5 desaturase (FADS2) mRNA in cumulus cells were reduced by $50{\mu}M$ ALA treatment (p<0.05). Cleavage rate was enhanced in 25 and $50{\mu}M$ ALA groups (p<0.05), especially, treatment of $50{\mu}M$ ALA promoted early embryo develop to 4 and 8 cell stages (p<0.05). However, blastocyst formation and number of cells in blastocyst were not differ in 25 and $50{\mu}M$ ALA groups. Our findings show that ALA treatment during maturation could improve nuclear maturation, fertilization, and early embryo development through enhancing of cumulus expansion, however, fatty acid metabolism- and cumulus expansion-related genes were down-regulated. Therefore, addition of ALA during IVM of oocytes could improve fertilization and developmental competence, and further studies regarding with the mechanism of ALA metabolism are needed.

• 한국초지조사료학회지
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• 제8권1호
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• pp.40-46
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• 1988

This study was carried out to determine the effects of nitrogen (N) fertilization levels (0, 100 and 200kg/ha) and formaldehyde (0.4,0.8 and 1.2%: w/w CM) on the chemical composition, dry matter (DM) yield and in-vitro dry matter digestibility (IVDMD) of forage rape (Brassica napus Subsp. oleifera) silage. The results obtained are summarized as follows : 1. Forage rape was a catch crop which was highly digestible and low concentration in NDF, ADF, cellulose and lignin. 2. The concentration of the water soluble carbohydrate (WSC) and buffering capacity (PK) was 17.9% and 6.77g/100g, respectively, provided the WSC to PK ratio was 2.65. 3. DM content of silage decreased as the rate of N fertilization increased, but concentration of ammonia-N decreased. IVDMD was not affected by the rate of N fertilization. 4. By the addition of formaldehyde, the pH and IVDMD were increased, but the DM, lactic acid, total organic acid and ammonia-N content of rape silage were decreased, that was effective as a silage additive for forage rape.

• Clinical and Experimental Reproductive Medicine
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• 제18권1호
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• pp.73-80
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• 1991

We compared fertilizing potential measurements by the zona-free hamster egg penetration assay with the in vitro fertilization and embryo transfer program was evaevulated for their ability to fertilize zona free hamster egg. Spermatozoa from 12 presumeably fertile donors and from the male partners of 56 infertile couples were evaluated for their ability to fertilizing potentials. Penertration rates of fertile donors were $36.2{\pm}27.7%$ ; Fertilization rates of infertile couples between with normal semen parameters and with abnormal semen parameters were $28.7{\pm}19.1$, $5.7{\pm}8.9%$, respectively. Sperm motility of couples with penetration rates between on 15-30% and on 30> were $54.1{\pm}4.6$, $55.5{\pm}8.3%$ respectively. Hamster penetration rates of couples participating in an in vitro fertilization and embryo transfer program was $38.9{\pm}29.9%$. But in one case, a positive fertility assessment was obtained in the absence of fertilization of the wife's eggs attributable to egg immaturity. This method may have potential value as a diagnostic tool in evaluation human sperm fertilization capacity which avoids the ethical and logistical problems associated with fertilizing of human eggs in vitro.

• 생물환경조절학회지
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• 제20권2호
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• pp.123-133
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• 2011

본 연구에서는 E. pellita와 A. mangium 용기묘를 대상으로 시비 처리에 따른 광합성, 엽록소 형광반응, 엽록소 함량 등의 생리적 특성 및 생장 특성 변화를 조사 분석하여 열대림 두 수종에 대한 시설양묘과정에서의 최적 시비 조건을 구명하고자 연구를 실시하였으며, 다음과 같은 결론을 얻을 수 있었다. 시비처리에 따른 E. pellita의 광합성 능력, 광화학 효율 및 엽록소 함량은 $1g{\cdot}l^{-1}$ 처리구에서 가장 우수하였으며, $2g{\cdot}l^{-1}$에서는 과량 시비로 인한 생육저하 현상으로 오히려 감소하는 경향을 나타냈다. A. mangium은 $2g{\cdot}l^{-1}$ 처리구에서 가장 우수한 광합성 능력, 광화학 효율 및 엽록소 함량을 보이면서 시비수준이 높을수록 우수한 능력을 나타냈다. 두 수종 모두 적정 시비 조건에서 높은 엽록소 함량과 광화학 효율에 의해 활발한 광합성 활동이 이루어졌다. 즉, 양분 조건에 따라 광합성 기구가 유동적으로 변하는 것을 알 수 있다. 근원경과 간장 생장 및 물질생산량 또한 E. pellita는 $1g{\cdot}l^{-1}$, A. mangium은 $2g{\cdot}l^{-1}$ 처리구에서 가장 우수하였으며, 생리적 특성과 같은 경향을 보였다. H/D율과 T/R율은 시비 처리구가 무시비 처리구보다 높은 값을 보였지만, 시비 처리 간 유의적 차이는 나타나지 않았다. 묘목품질지수는 두 수종 모두 위 결과와 같이 수종별로 적정 시비 처리구에서 가장 높았으며, 적정 시비 처리에 의해 생산 된 묘목 우수한 형질을 나타낸 것이다. 본 연구 결과를 종합해 보면, 열대 수종인 E. pellita와 A. mangium의 용기묘 양묘 시 시비 처리에 따른 양분 조건에 의한 생리 및 생장 특성의 변화를 볼 수 있었다. 양분 부족은 광합성 기구의 활동 감소에 의한 생장 저하가 초래되어 불량한 묘목이 생산되는 과정을 볼 수 있다. 즉, 양묘과정에서 수종별 양분 요구도에 맞는 생육환경조절은 시설양묘시업에 의한 건전한 묘목을 대량 생산함과 동시에 조림과정에서도 높은 활착과 생장으로 우수한 조림성과를 기대할 수 있을 것으로 판단된다. 또한 일률적인 시비를 실시하는 것이 아니라, 기간별 상대생장량을 조사하여 수종별 생장패턴에 따라 기간별로 양분 요구량에 맞는 집중, 효율적인 시비를 실시할 수 있을 것으로 기대된다. 이에 띠라 양묘과정에서의 환경적 측면과 함께, 비용 측면에서 이점이 있어 경제적으로도 묘목 생산비를 절감할 수 있을 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권2호
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• pp.191-200
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• 2000

Objective: This study was carried out to compare the effects of the stepwise exposure treatments on the morphological normality, fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing and to use as a fundamental data for the cryopreservation of human oocytes. Materials and Methods: The morphological normality and fertilization rates of the vitrified and ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were observed. After choosing the 3step exposure treatment groups, we observed the morphological normality and fertilization, blastocyst formation rate of the vitrified and ultra-rapid frozen mouse mature oocytes. Results: The morphological normality and fertilization rates of the vitrified mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 75%, 85%, 88% and 58%, 61 %, 54% respectively. There were no significant differences among treatments(p>0.05). The morphological normality and fertilization rate of the control was 92% and 65%. There were no significant differences in fertilization rate among control and treatments (p>0.05). The morphological normality and fertilization rates of the ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 83%, 83%, 84% and 75%, 63%, 56% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rate of the control was 95% and 67%. There were no significant differences among control and treatments (p>0.05). The morphological normality and fertilization rate of the vitrified or ultra-rapid frozen mouse mature oocytes after 3step exposure treatment were 69% and 75%, respectively. The blastocyst formation rate was 60% and 57%. The results did not differ significantly between vitrification and ultra-rapid freezing (p>0.05). Conclusion: As known in the above results, there were no significant differences in the fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing among the control and treatments. It is suggested that vitrification and ultra-rapid freezing method were effective for the cryopreservation of mouse mature oocytes.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1501-1508
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• 2004

In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.85-91
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• 2011

The objective of this study was to evaluated the efficiency on sperm cryosurvival and ability of in vitro fertilization using Triladyl and Lactose Egg-Yolk(LEY) as extenders for cryopreservation of separated sperm by 65% percoll in miniature pig. Sperm viability was measured with SYBR-14/PI double stained sperm by flow cytometry. Ability on embryo cleavage rate and blastocyst development were observed by in vitro fertilization after frozen-thawing of sperm separated by 65% percoll. The experimental groups were designed that separated sperm by 65% percoll with Triladyl (ST) or LEY(SL) and unseparated sperm with Triladyl(UT) or LEY(UL) for cryopreservation. As a results, the viability was significantly(p<0.05) higher in ST(55.1%), SL(63.1%), UL(58.8%) than UT(38.2%) group. Sperm viability in SL(63.1%) group was significantly(p<0.05) higher than other experimental groups. On the other hand, embryo cleavage rate was significantly(p<0.05) higher in ST(79.1%), SL(83.2) than UT(74.1) and UL(75.7%) groups at 96h after in vitro fertilization. Blastocyst development was also significantly(p<0.05) higher in ST(21.5%), SL(20.9%) than UT(17.0%) and UL(18.8%) groups. In conclusion, cryopreservation of miniature boar sperm separated by 65% percoll were beneficial to viability and capacity on in vitro fertilization.