• Journal of Embryo Transfer
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• v.28 no.1
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• pp.63-71
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• 2013

In this study, we used flow a cytometric assay to evaluate plasma membrane integrity and mitochondrial activity in post-thawed sperm that was supplemented with ginsenoside-$Rg_1$. Varying concentrations of ginsenoside-$Rg_1$ (0, 25, 50 and $100{\mu}M/ml$) were used in the extender during cryopreservation to protect the DNA of thawed sperm, thereby increasing the viability and motility rate as evaluated using a computer-assisted sperm analysis (CASA) method. The results derived from CASA were used to compare the fresh, control, and ginsenoside-$Rg_1$ groups. Sperm motility and the number of progressively motile sperm were significantly (p<0.05) higher in the $50{\mu}M/ml$ ginsenoside-Rg1 group ($61.0{\pm}4.65%$) than in the control ($46.6{\pm}7.02%$), $25{\mu}M/ml$ ($46.2{\pm}4.76%$), and $100{\mu}M/ml$ ginsenoside-$Rg_1$ ($52.0{\pm}1.90%$) groups. However, the velocity distribution of post-thawed sperm did not differ significantly. Membrane integrity and MMP staining as revealed using flow cytometry were significantly (p<0.05) higher ($91.6{\pm}0.82%$) in the $50{\mu}M/ml$ ginsenoside-$Rg_1$ group than in the other groups. Here, we report that ginsenoside-$Rg_1$ affects the motility and viability of boar spermatozoa. Moreover, ginsenoside-$Rg_1$ can be used as a protective additive for the suppression of intracellular mitochondrial oxidative stress caused by cryopreservation.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.1
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• pp.44-47
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• 2018

Onco-testicular sperm extraction is used to preserve fertility in patients with bilateral testicular tumors and azoospermia. We report the case of a testicular tumor in the solitary testis of a patient who had previously undergone successful contralateral orchiectomy and whose sperm was preserved by onco-testicular sperm extraction. A 35-year-old patient presented with swelling of his right scrotum that had lasted for 1 month. His medical history included a contralateral orchiectomy during childhood. Ultrasonography revealed a mosaic echoic area in his scrotum, suggesting a testicular tumor. The lesion was palpated within the normal testicular tissue along its edge and semen analysis showed azoospermia. Radical inguinal orchiectomy and onco-testicular sperm extraction were performed simultaneously. Motile spermatozoa were extracted from normal seminiferous tubules under microscopy and were frozen. Eventual intracytoplasmic sperm injection using the frozen spermatozoa is planned. Onco-testicular sperm extraction is an important fertility preservation method in patients with bilateral testicular tumors or a history of a previous contralateral orchiectomy.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.149-153
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• 1995

In IVF-ET program, intracytoplasmic sperm injection(ICSI) has been performed with testicular sperm extraction(TESE) in case of no normal spermatozoon could be retrieved from the epididymis. We wished to see whether the quality of testicular sperm affect the fertilization and pregnancy rate in TESE-ICSI cycles(n=40). These cycles were classified into three groups by the total number of normal motile spermatozoa(TNMS) in the TESE sample: i) good sperm(GS) group(n=12), TNMS > 10,000; ii) moderate sperm(MS) group(n=19), 1,000 < TNMS < 10,000; iii) poor sperm(PS) group(n=9), TNMS < 1,000. Among 423 injected oocytes, 307(72.6%) oocytes were normally fertilized and 43 zygotes were cryopreserved. The fertilization rates of GS group(79.3%) and MS group(75.9%) were significantly(p<0.005) higher than PS group(60.2%). After the embryo transfer(n=40), clinical pregnancy was obtained in 14 cycles(35.0%) and on-going pregnacy in 13 cycles(32.5%). The clinical and on-going pregnancy rates were similar in each group. From these results it can be concluded that testicular spermatozoa are successfully used with ICSI in IVF-ET program in spite of very poor quality of TESE sample.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.168-172
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• 2007

In the present study, attempts were made to find out the physico-chemical properties of milt and the sperm motilities in various osmotic conditions using Panther puffer, Takifugu pardalis. The average concentration of sperm in the milt was $12.1{\pm}3.2{\times}10^9/mL$. pH and osmolality of seminal plasma were $8.2{\pm}0.3$, $385.5{\pm}12.5mOsm/kg$, respectively. Spermatozoa were immotile when the milt was mixed with solutions (electrolyte or non-electrolyte) of lower osmolality than the average seminal plasma osmolality ($385.5{\pm}12.5mOsm/kg$), but became motile after mixing milt with hyperosmotic solutions.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.142-148
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• 2020

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode's albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.247-252
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• 2005

The purpose of this study was to assess sperm quality during in vitro storage of miniature-pig semen in order to determine which extender should be used and how extender can be diluted for in vitro storage of miniature-pig semen. Freshly ejaculated miniature-pig's semen was diluted with same volumes of Beltsville Thawing Solution (BTS), Androhep, Modena, Mulberry III and modified-Modena extenders. Sperm quality was evaluated by examining viability, motility, abnormality, acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining. Sperm motility decreased with storage period prolonged and differences among BTS, Androhep, Modena and Mulberry III were apparent On Day 1, approximately 80% of the sperm were motile, but motility decreased to $40\%$ at Day 7. During the 7 days of storage, sperm survival in modified-Modena B extender was higher than another extenders. However, it was not differ significantly among other extenders. The percentage of F and B patterns were not differ significantly among the extenders. However, F pattern in modified-Modena B extender was slightly higher until 3 days of storage than that of Modena extender, modified-Modena A extender and modified-Modena C extender. The percentage of AR patterns in modified-Modena B extender was slightly lower, but did not differ significantly among other extenders. The results of present study suggest that modified-Modena B was effective as new extender for in vitro storage of miniature-pig semen.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.1
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• pp.111-124
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• 2006

Purpose : This study was conducted to investigate the dose dependent effects of Panax Ginseng on the reproductive functions in mice. Methods : We used the 8-week-old mice, and administered 0.2ml extract solution of Panax Ginseng in the different concentration once a day for 60 days. The control group was administered 0.2ml normal saline in the same way and duration. We counted the total, motile and normal sperm number of the cauda epididymis and measured the activities of sperm hyaluronidase, peroxidase and catalase of the isolated testis tissues. And we observed histological changes of surgically isolated testis by histochemical methods. Results : All Panax Ginseng extract solution groups showed significantly dose dependent differences in the total number, the motility and normality of sperms compared with the control group, respectively. In the histological analysis of the testicular tissues, all Panax Ginseng extract solution groups showed the enlargement of testicular lobe diameter and apparent angiogenesis between seminiferous tubules. And the activity of typical sperm enzyme, hyaluronidase, was significantly increased in the Panax Ginseng extract solution groups compared to the control group. In the antioxidant activity analysis, the activities of peroxidase and catalase were significantly increased in the Panax Ginseng extract solution groups compared to the control group. Conclusion : This study shows that Panax Giseng has the beneficial effects on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase, testicular peroxidase and catalase. We can suggest that Panax Ginseng be useful for the treatment of male infertility.

• Journal of Embryo Transfer
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• v.31 no.2
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• pp.109-116
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• 2016

The recovery of epididymal sperm in animals is considered as one of the important tools to preserve high value or endangered species. However, there are no appropriate castrating indicators such as months of age in bull, sperm morphology, and motility, particularly in young Korean native bull (Hanwoo). Therefore, this study aimed to investigate sperm number, morphology, and motility of sperm in the epididymis tail of young Hanwoo bulls at 8 and 15 months of age. After castration, epididymal tails were collected and minced with blades to recover sperm. In experiments 1 and 2, sperm number, morphology, and motility were examined. Total number of sperm and percentage of normal sperm from bulls at 8 months of age was lower than that of bulls at 15 months of age after collection (P<0.05). Percentage of abnormal head, tail, proximal cytoplasmic droplet, dead and damaged acrosome of sperm from bulls at 8 months of age were higher than those of bulls at 15 months of age (P<0.05). In experiment 3, sperm motility from bulls at 8 and 15 months of age were examined before freezing and after thawing. Frozen-thawed sperm at 8 months of age showed low total motility and motile sperm with ${\geq}25{\mu}m/sec$ compared to those at 15 months of age and commercially-used sperm (P<0.05). In conclusion, sperm derived from the epididymal tail of bulls at 8 months of age showed high abnormal morphology and poor motility, which are not adequate for AI and IVF. On the other hand, sperm derived from the epididymal tail of bulls at 15 months of age showed high normal morphology and motility.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.87-94
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• 1992

From December 1991 to March 1992, 34 anejacuratory patients with spinal cord injury underwent 90 of electric stimulations with Seager NRH model 12. The average patient age was 43.5 years with a range of 23 to 48 years. The level of cord injury was cervical in 7, thoracic in 6, lumbar in 11, lumbosacral in 7 and conus medullaris in 3. The average number of electric stimulation per a patient was 2.65 with a range of 1 to 4. The average voltage and amplitude per a stimulation were 17.72 volts and 309. 89 mAmp with ranges of 5 to 25 volts and 50 to 500 mAmp. The total and motile sperm number were evaluated microscopically and analyzed statistically by paired t-test according to the frequency of electroejaculation, level of cord injury and voiding pattern. The results were obtained as follows. 1. An overall success rate of electroejaculation was 85.3% among 34 patients and 82.2% among 90 electric stimulations. 2. The total and motile sperm number per a stimulation were not correlated the frequency of electric stimulation, level of cord injury and voiding pattern. 3. Complications occured in 10 cases; severe low abdominal pain in 5, hypertension in 2, sweating in 1, headache in 1 and neck stiffness in 1. All the copmlications subsided spontaneously within 5 to 10 minutes after transient interruption of the electric stimulation. In summary, rectal probe electroejaculation is an accepted safe means of procuring sperm from spinal cord injury patients with ejaculatory incompetence. However very poor sperm motility was found and it was not related with the frequency of electroejaculation, level of cord injury and voiding pattern. Further investigation would be needed to conclude and to identify the reasons for impaired sperm motility.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.5
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• pp.809-815
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• 1997

A series of experiments were conducted to study effects of osmolality and $Ca^{2+}$ on sperm motility in marbled sole, Limanda yokohamae. Spermatozoa were immotile when the milt was mixed with solutions (electrolyte or non-electrolyte) of lower osmolality than the average seminal plasma osmolality (351 mOsm/kg), but became motile after mixing milt with an hyperosmotic solution. However, with the osmolality above 1,639 mOsm/kg spermatozoa ceased their movement. When the milt was suspended in the $Ca^{2+}$ free artificial seawater (ASW) containing ethylenglycoltriamine (EGTA), the percentage of motile spermatozoa decreased in proportion to the increase of EGTA. Most spermatozoa were quiescent in a medium containing 3 mM EGTA. The motility of spermatozoa prevented by 3 mM EGTA was recovered by the subsequent addition of $Ca^{2+}$ over the concentration of 0.25 mM. Effects of calmodulin antagonist, trifluoperazin (TFP) on spermatozoa were examined to elucidate the possible functions of calmodulin in sperm motility. TFP immobilized spermatozoa 5n ASW at the concentration of 0.5 M. These findings suggest that calcium is an effective external factor in the initiation process of sperm motility.