• Asian-Australasian Journal of Animal Sciences
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• 제23권1호
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• pp.33-40
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• 2010

In this study, the production of transgenic goats using sperm to integrate exogenous DNA and artificial insemination (AI) was carried out and the technical protocols for sperm-mediated gene transfer (SMGT) in the goat were optimized. The standard sperm parameters and the ability to bind foreign genes were assessed to select suitable sperm donor bucks. A total of 134 oestrous does were divided into 4 groups and inseminated using different methods and sperm numbers. The does of Groups I to III were inseminated with fresh semen ($1-2\times10^{7}$ and $10^{6}$ sperm) or frozen-thawed semen ($10^{6}$ sperm), respectively, through conventional intra-cervical AI, and the does of Group IV with frozen-thawed semen ($10^{6}$ sperm) through intrauterine AI. Total genomic DNAs were extracted from ear biopsies of the offspring. The presence of $pEGFP-N_{1}$ DNA was screened by PCR and then by Southern blotting analysis. A total of 76 live kids were produced and 8 kids were tested transgene positive on the basis of agarose gel electrophoresis of the PCR-amplified fragment. Southern blotting analysis of the samples showed 5 positive kids. A transgenic ratio of 10.53% was detected using PCR and 6.58% using Southern blotting. The positive kid rate assayed by PCR and Southern blotting of frozen-thawed goat semen was 3.61% and 9.27% higher than that of untreated semen. The results show that transgenic goats can be produced efficiently by the method of artificial insemination using sperm cells to integrate the exogenous DNA and intrauterine insemination allowed low numbers of DNA-transfected spermatozoa to be used, with satisfactory fertility.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.351-355
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• 1996

Objective: To analyze the direct effect of nitre oxide, generated from sodium nitroprusside, on sperm motility and reactive oxygen species. Design: Human sperm samples were treated to allow swim-up and washing. And the samples were devided into four aliquots. Each aliquot was incubated with either concentration at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside. Intervention: Samples were measured chemiluminosence for reactive oxygen species of each aliquot with concentrations at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside at allowing swim-up and washing of sperm. Main Outcome Measures: Percent motion parameters and viability were asse-ssed at 0, 3, 6, 12, 24 hours incubation. Results: The percent viablity was lower slightly in control group (50.2%) than that in sperm treated with 100nM of nitroprusside(57.5%) at 24 hours after incubation, while was reduced significantly in sperm with concentra-tion of $10{\mu}M(42.1%)$ and 1mM(21.3%)of nitroprusside at 6 hours after incubation. And the sperm treated with 1mM of nitroprusside was immotile totally at 6 hours after incubation. The straight line$(35.3{\pm}5.6%)$, the rapid forward$(37.2{\pm}6.4%)$ and the weak curvilinear velocity$(9.6{\pm}2.4%)$were more favorable comparing with those ($32.4{\pm}4.2%$, $30.0{\pm}7.8%$ and $18.0{\pm}4.6%$ respectively) in control group at 3 hours after incubation, but reduced significantly in sperm treated with $10{\mu}M$ and 1mM of nitroprusside. The levels of reactive oxygen species in control(700 c.p.m.) is lower significantly than that in each experimental groups of sperm treated with nitroprusside. And the levels of reactive oxygen species were 2200 c.p.m. in 100nM, 6200c.p.m. in $1{\mu}M$ and 12800c.p.m. in 1mM respectively. Conclusion: These results suggested that the concentration of 100nM of nitroprusside on sperm is beneficial to the maintanance of viablity and motile velocity, but detriment in high concentration of $10{\mu}M$ or 1mM of nitroprusside.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.139-143
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• 2007

본 연구는 체세포를 이용하여 생산된 복제 한우 수소의 번식능력을 검토하기 위해 실시하였다. 복제 한우 수소(C-38 및 C-39) 또는 일반 한우 종모우로부터 정액을 채취하여 정자의 수 및 동결 전 후의 생존성 등을 살펴보았으며, 정자의 운동성 등은 computer assisted sperm analysis(CASA)를 이용하여 측정하였다. 또한, 이들의 수정 능력을 확인하기 위하여 체외수정과 인공수정을 각각 실시하였다. 정액 성상에서는 복제 수소들과 일반 종모우 간에 정액의 양, 정자의 농도 및 동결융해 후의 생존성 등에서 차이가 나타나지 않았다. CASA를 이용한 분석에서 운동성, 곡선 운동 속도(VCL), 직선운동 속도(VSL) 및 평균 진행 속도(VAP) 등은 복제 수소의 정액이 일반 종모우의 정액에 비하여 유의적으로 높았다 (p<0.05).체외수정에 따른 수정란의 분화율 및 배반포로의 발달율은 복제 수소와 일반종모우 간에 차이가 나타나지 않았다. 복제소 정액(C-38)을 이용하여 인공수정을 한 5두의 체세포 복제 대리모에서 암 수 각각 한 두씩의 건강한 복제 후대 송아지 2두를 생산하였다. 이상의 결과를 종합하여 보면, 실험에 공시된 복제 수소 개체간의 차이가 나타나기는 하였지만, 복제 수소는 정액 성상과 정자의 운동성 등에서 일반 종모우와 차이가 없었으며, 또한 인공수정을 통해 송아지를 생산함으로써 정상적인 번식능력이 있음을 확인하였다.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.7-9
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• 1996

Here, A forty year old patient is presented, showing azoospermia following vasovasostomy. The bilateral testicular biopsies were performed to confirm whether ductal obstruction due to vasovasostomy or testicular failure existed. The finding of biopsy was spermatogenic arrest. After completion of medical testicular stimulation with clomiphene citrate and pentoxifilline for 3 months, repeat vasovasostomy was performed. Semenanaylsis revealed normal sperm parameters after operation. Necessity of testicular biopsy before deciding repeat vasovasostomy, accuracy of testicular biopsy, efficacy of testicular stimulation in the patient with spermatogenic arrest and effect of testicular biopsy on testicular fuction are discussed.

• Animal Bioscience
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• 제36권12호
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• pp.1796-1805
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• 2023

Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

• Clinical and Experimental Reproductive Medicine
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• 제34권3호
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• pp.159-166
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• 2007

목 적: 불임 부부에서 자궁강내 정자주입술 시행 후 연속하여 시행한 자궁강내 정자주입술 및 전환하여 시행한 체외수정시술이 정액검사소견에 미치는 영향을 알아보고자 하였다. 연구방법: 진단적 목적으로 컴퓨터정액분석기를 이용한 정액검사 (computer-assisted semen analysis (CASA))를 시행한 후 자궁강내 정자주입술을 시행하였으나 임신에 실패한 53예를 대상으로 하였다. 자궁강내 정자주입술 시행 후 38예의 환자에서는 두 번째 자궁강내 정자주입술을 시행하였고 (Group 1), 15예의 환자에서는 체외수정시술을 시행하였다 (Group 2). 이들에서 두 번째 불임시술을 위해 채취한 정액의 양 (volume), 정자의 농도 (concentration), 운동성 (motility), 총운동성정자수 (total motile sperm count) 등의 컴퓨터정액분석소견을 이전에 자궁강내 정자주입술 시 시행한 정액검사소견과 비교하였다 결 과: 대상 불임 부부간의 남편 연령, 첫 번째 시술과 두 번째 시술 사이의 시간 간격, 불임의 원인 등에는 유의한 차이가 없었다. Group 1에서 첫 번째 시행 시의 정액검사소견의 정상 여부와 관계없이, 정자의 운동성은 첫 번째 자궁강내 정자주입술 시행 시보다 두 번째 자궁강내 정자주입술 시행 시에 통계적으로 유의하게 감소하였다. Group 2에서는 첫 번째 자궁강내 정 자주입술 시보다 체외수정시술 시 정자의 농도, 정자의 운동성, 총운동성정자수에서 통계적으로 유의하게 감소하였다. Group 2의 하위그룹 분석에서는 자궁강내 정자주입술 시행 시 정상 정액소견을 보인 경우는 체외수정시술 시 정자의 농도와 총운동성정자수가 유의하게 감소하였으나, 자궁강내 정 자주입술 시행 시 비정상 정액소견을 보인 경우에는 체외수정시술 시 정자의 운동성이 유의하게 감소하였다. 결 론: 자궁강내 정자주입 술 시행 후 임신에 실패하여 체외수정시술을 시행하게 된 환자의 정액검사소견은 자궁강내 정 자주입술을 계속 시행 받은 환자군에 비해 더 악영향을 받는다. 그 이유를 설명하기 위해 심리적 스트레스에 대한 후속적인 연구가 필요할 것으로 사료된다.

• Advances in Traditional Medicine
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• 제9권4호
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• pp.339-349
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• 2009

An attempt has been made to assess whether the dose dependent effect of benzene extract of Ocimum sanctum leaves on the morphological changes in the cauda epididymal spermatozoa and sperm parameters in male albino rats. Scanning Electron Microscope observations illustrate the disturbance in plasma membrane as well as acrosomal membrane. Most of the sperms appear morphologically abnormal in the mid region of the tail; there is formation of balloon like cytoplasmic droplet. Sperm parametric study exhibits decrease in the total sperm count, sperm motility, forward velocity and increase in the percentage of abnormal sperms in dose dependent manner on treatment benzene extract of Ocimum sanctum leaves. The results suggest that the effects may have resulted from a general disturbance in the proteins and alteration in cauda epididymal milieu probably due to androgen deficiency consequent upon antiandrogenic property of Ocimum sanctum leaves.

• Clinical and Experimental Reproductive Medicine
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• 제20권1호
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• pp.1-7
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• 1993

The present study was designed to test the validity of the semen analysis(S/A) and the sperm penetration assay(SPA) as a prognostic indicator of male fertility in 123 patients undergoing in vitro fertilization(IVF). We attempted to correlate the traditional semen parameters or the extent of sperm penetration in SPA with the results of human IVF rate or cleavage rate. Poor correlation was found between the results of S/A and human IVF rate(sensitivity, 80.6% ;specificity, 46.7%; positive predictive value, 91.6%;negative predictive value, 25%). Conversely, good correlation was found between the results of SPA and human IVF rate(sensitivity, 100% ; specificity, 80% ;positive predictive value, 97.3% ;negative predictive value, 100%). Our results corroborate the conclusion that SPA can be a valuable tool as a prognostic indicator of male fertilizing ability.

• Natural Product Sciences
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• 제25권2호
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• pp.157-164
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• 2019

The study was conducted to investigate the acute and sub-acute toxicity effect of Aquilaria malaccensis leaves aqueous extract (AEAM) towards male ICR mice in terms of body weight, relative organ weight, mortality rate and sperm parameters. In acute toxicity study, a single dose at of 2000 mg/kg was performed. In sub-acute toxicity study, the mice were received normal saline (control group), 50, 100, 150, 200, 500, or 1000 mg/kg of AEAM orally for 21 days of treatment. In sub-acute toxicity study, the number of abnormal sperm were significantly decreased in AEAM 100, 150, 200, 500, and 1000 when compared to the control group. While, the motility of sperm were found to be significantly increased in AEAM 100, 150, 200, and 1000 as compared to the control group. No mortality was recorded in the control group and treated groups in both toxicity studies except for one mouse from AEAM 1000 group. However, the mild sedative effect in terms of the tendency to sleep was clearly noticeable in both toxicity studies. Results indicated that the AEAM can be one of the useful alternative medicine to enhance fertility rate by increasing healthy sperm production.

• Clinical and Experimental Reproductive Medicine
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• 제48권2호
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• pp.142-149
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• 2021

Objective: Bacteriospermia and urogenital infections are common problems in male infertility. This study aimed to evaluate the effects of bacteriospermia on sperm parameters and clinical outcomes in semen samples infected with two common bacteria (Staphylococcus saprophyticus and Escherichia coli) in northern Iran. Methods: Microbiological tests were performed to isolate and identify organisms from 435 semen samples from infertile couples. Semen samples were assessed according to the World Health Organization criteria. The protamine status, chromatin structure, chromatin condensation, and acrosome reaction of sperm and assisted reproductive outcomes were determined in couples with different male infertility factors. Results: Among the total cases, the two most prevalent pathogens were considered: S. saprophyticus (38.2%) and E. coli (52.9%). In the semen samples infected with E. coli, the spontaneous acrosome reaction and abnormal chromatin condensation were more common (p<0.05). Significant increases in abnormal chromatin condensation and deprotamination were seen in the presence of S. saprophyticus. In washed semen, tight adhesion between the sperm midpiece and S. saprophyticus was observed. There was also a significant decrease in the fertilization rate using semen samples infected with S. saprophyticus and E. coli during in vitro fertilization cycles (p<0.001). In addition, the presence of S. saprophyticus and E. coli in semen samples was associated with a lower likelihood of clinical pregnancy in couples with various factors of male infertility. Conclusion: Poor results of assisted reproductive techniques may be correlated with semen samples infected with two common bacteria in northern Iran.