• Parasites, Hosts and Diseases
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• v.60 no.5
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• pp.357-360
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• 2022

Excretory-secretory products (ESP) of T. vaginalis have been shown to inhibit sperm motility, viability, and functional integrity, leading to a decreased fertilization rate in vitro. This study investigated whether T. vaginalis induce apoptosis and ultrastructural changes of sperm using flow cytometry and electron microscopy. Incubation of sperm with T. vaginalis ESP increased phosphatidylserine externalization and DNA fragmentation, and decreased mitochondrial membrane potential. Transmission electron microscopy of sperm incubated with ESP revealed abnormal features such as distorted heads, broken necks, and acrosomes exocytosis. This is the first report that demonstrates a direct impact of T. vaginalis ESP on sperm apoptosis and architecture in vitro.

• Journal of Embryo Transfer
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• v.13 no.3
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• pp.277-289
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• 1998

To establish a system for sperm swim-up separation through sucrose layer, indiscreet sperm migration should sufficiently to block but movement of sperm shouldn't inhibit. Thus, the effects of sucrose levels in sucrose layer, incubation times and types of sucrose layer on sperm separation were examined. And the results obtained were as follows; 1. Layer of 10mM sucrose inhibited sperm swim-up migration through sucrose layer. 2. Incubation for 25 minutes without sucrose layer significantly increased sperm swim-up migration. However, incubation for 10 minutes to induce swim-up through sucrose layer significantly stimulated sperm migration and maintained sperm movement. 3. There was no significant difference between Type I and Type II in barrier effect of sucrose layer. However, sucrose layer of Type II with shorter distance of barrier was efficient for sampling. To elucidate a function of follicular fluid on sperm chemotaxis using in vitro system of sucrose layer of Type II and incubation for 10 minutes, the effects of dilution, heat treatment, and protein and lipid extracts of follicular fluid on sperm swim-up separation were examined. And the results obtained were as follows; 4. Follicular fluid stimulated sperm migration and movement, and significantly-attracted capacitated-sperm at 10% level. 5. Follicular fluid heated at 55$^{\circ}C$ for 30 minutes maintained the effect of follicular fluid stimulating sperm migration and movement. 6. Follicular protein stimulated sperm movement that was reduced by filtration of the protein. 7. Follicular lipid didn't significantly stimulate sperm migration and movement. 8. Both of follicular protein and lipid reduced the effect of follicular fluid stimulating sperm migration and movement. In conclusion, sucrose layer could be used for a barrier against indiscreet sperm migration by swim-up. And follicular fluid stimulated migration and movement of sperm and attracted capacitated-sperm through sucrose layer. Especially, heat-resistant protein of follicular fluid stimulated sperm migration.

• Development and Reproduction
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• v.1 no.1
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• pp.29-36
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• 1997

Experiments were performed to study the effects of diluents, cryoprotectant, equilibration time, thawing temperature and addition of BSA and egg yolk. Among the various diluents, Alsever's solution was the best for sperm cryopreservation. A combination of Alsever's solution and 15% ethylene glycol showed the better results than others did. Sperm activity indection and survival rate gradually decreased with the equilibration time. The appropriate thawing temperature was 30 ${\pm}1^{\circ}$C. These results indicate that sperm cryopreservation methods can be developed in tiger puffer.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.41-42
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• 1998

The changes in calcium binding protein(CBP) of mouse epididymal sperm during their post-testicular differentiation were analyzed by two-dimensional SDS-PAGE. According to dpididymal maturation, capacitation and acrosome reaction of spermatozoa, both quantitative and qualitative changes of CBPs in the epididymal sperm was detected. It suggested that the development of fertilizing ability of epididymal sperm was closely related to the changes in the CBPs profiles of sperm during epidiyaml transit.

• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.141-146
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• 1990

12 crossbreed boars received 4 rations containing varying levels of lysine and DL-methionine. The results obtained from this study are summarized as follows ; 1. Semen volume, total sperm number of the treatment B, C and D were increased significantly (p<0.05) as compared with the treatment A but abnormal sperm percent of treatment B and C was decreased significantly (p<0.05) as compared with A. Sperm number and sperm mortility were not different from treatments. 2. Amino acids contents of sperm plasma were not different from treatments.

• Korean Journal of Animal Reproduction
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• v.15 no.1
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• pp.7-13
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• 1991

This study was carried out to investigate the effects of pH of sperm-washing solution, sperm-washing frequency and individual bull on concentration of hydrogen ion in sperm-washed solution and sperm acrosome reaction. The results obtained were as follow. 1. When bovine sperm was washed at 4 times with SHP solution and incubated, the difference of light absorbance between sperm-washing and sperm-washed solution(${\Delta}$Ao-${\Delta}$At) was higher in 2nd sperm-washed solution than that in the other washed solutions. 2. When sperm was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, ${\Delta}$Ao-${\Delta}$At was significantly increased at pH7.69 to 8.83, and ${\Delta}$Ao-${\Delta}$At in 1st sperm washed solution was significantly higher than that in 2nd and 3rd sperm washed solution. 3. When sperm of Holstein, KNC and Hereford was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, Holstein showed higher ${\Delta}$Ao-${\Delta}$At of sperm Washed solution than KNC and Hereford, and ${\Delta}$Ao-${\Delta}$At in sperm washed solution was significantly increased at 7.69 for Holstein and at 8.15 for KNC and Hereford, respectively. 4. When sperm was thrice with SHP solution of pH 6.8, 7.1 and 7.4, and then incubated in mTALP of pH 7.4 for 15 minutes, in 1st and 2nd sperm washing ${\Delta}$Ao-${\Delta}$At of sperm washed solution was significantly higher at pH 7.1 and 7.4 than at pH 6.8, and the sperm acrosome reaction of pH 6.8, 7.1 and 7.4 was 49.1, 68.8 and 72.9%, respectively. The sperm acrosome reaction of pH 7.1 and 7.4 was higher than that of pH 6.8.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.1
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• pp.53-60
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• 2006

Objective: We tested the usefulness of swim-down technique using human follicular fluid (hFF) in sperm preparation. Methods: Semen samples were obtained from twelve male partners showing asthenozoospermia (sperm motility < 50%) at the time of routine andrologic evaluation in Seoul National University Bundang Hospital. After dividing into two aliquots, each samples were processed either by swim-down using 100% hFF or density gradient using SpermGrad. Sperm quality was assessed by computer-assisted semen analyzer (CASA). Results: Motility, Rapid motility, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement), and hyperactivated sperms were significantly increased, and LIN (mean linearity) was decreased significantly after sperm preparation in both groups. Motility was significantly higher after swim-down using 100% hFF when compared with density gradient using SpermGrad ($81.2{\pm}4.7$ vs. $67.6{\pm}2.3$, p=0.02) The other parameters assessed by CASA were not different between the two methods. Conclusion: Swim-down method with 100% hFF may be a useful method in preparation of sperm from asthenozoospermia.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.299-305
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• 2016

Although cryopreservation of sperm is routinely used for clinical requirement, it has some problems, such as high generation of reactive oxygen species (ROS) and cold-shock. To reduce the detrimental damage in sperm, anti-oxidants were added to cryoprotectant for sperm. Curcumin is one of anti-oxidants, which are added in cryoprotectants. However, recent studies have demonstrated that curcumin decreases sperm viability and motility. This study was performed to identify the effect of curcumin on hydrogen peroxide ($H_2O_2$)-exposed bovine sperm, which were cryopreserved-thawed. In $H_2O_2$-exposed bovine sperm, reactive oxygen species (ROS) were significantly reduced by treatment with curcumin in a dose-dependent manner (p < 0.05). Among tested concentrations of curcumin (1 to $50{\mu}M$), 30 and $50{\mu}M$ curcumin showed anti-oxidant effect on $H_2O_2$-induced ROS generation. On the other hand, combination of 30 or $50{\mu}M$ curcumin with anti-oxidant $H_2O_2$ increased the percentage of apoptotic sperm compared to only $H_2O_2$ treatment. Sperm viability was also decreased in the combination of 30 or $50{\mu}M$ curcumin with $H_2O_2$ as judged by FDA/PI staining. $H_2O_2$-induced decrease in sperm progressive motility was recovered by treatment with $1{\mu}M$ curcumin. These results show that high concentration of curcumin has anti-oxidant effect, but it has also cytotoxic effect on bovine sperm. Sperm viability and motility might be more affected by cytotoxic signals of curcumin compared to antioxidant signals.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1713-1720
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• 2015

The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II ($p{\leq}0.05$). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower ($p{\leq}0.05$), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors that confirm the accuracy of insemination male selection can include a low rate of sperm motility decrease during the storage of diluted semen, low and contained incidence of secondary morphological changes in spermatozoa during semen storage and a high frequency of spermatozoa with undamaged cell membranes.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.119-126
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• 2003

Objective: This study was carried out to compare the clinical outcomes of intracytoplasmic sperm injection (ICSI) in patients with obstructive azoospermia according to sperm retrieval site and technique; microsurgical epididymal sperm aspiration (MESA), percutaneous epididymal sperm aspiration (PESA), testicular sperm extraction by open biopsy (TESE). Methods: The outcomes of ICSI and IVF-ET were evaluated and compared among 3 groups. Seventy three men suffering from infertility due to obstructive azoospermia had 107 ICSI cycles using MESA (21 cycles in 15 patients), PESA (26 cycles in 17 patients) and TESE (60 cycles in 41 patients). Results: In the clinical outcomes in patients undergoing ICSI with epididymal or testicular sperm, there were no significant differences in fertilization rate (66.1% vs. 60.5%), cleavage rate (94.9% vs. 97.6%), cumulative embryo score (CES) (51.3 vs. 58.8), implantation rate (7.9% vs. 6.1), and clinical pregnancy rate per ET (30.4% (14/46) vs. 25.4% (15/59)) between both groups. Also, in the clinical outcomes in ICSI patients using MESA, PESA, TESE, there were no significant differences in fertilization rate (61.8%, 69.4%, 60.5%), cleavage rate (92.1%, 97.3%, 97.6%), CES (38.1, 52.0, 58.8), implantation rate (9.5%, 6.6%, 6.1%), and clinical pregnancy rate per ET (35% (7/20), 26.9% (7/26), 25.4% (15/59)) among 3 groups. Conclusion: When compared with MESA or TESE, PESA, the clinical outcomes were similar in ICSI patients with obstructive azoospermia whatever the origin or the technique of sperm retrieval. However, we considered PESA is more time-saving and cost effective for ICSI in patients with obstructive azoospermia.