• 생명과학회지
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• 제30권1호
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• pp.77-81
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• 2020

본 연구는 돼지정자의 동결보존에 있어 0.5 ml straw에 비해 5.0 ml straw가 가지고 있었던 제한적인 요소들을 극복하기 위하여 동결속도, 융해온도 및 융해 시간을 확립하기 위하여 실시되었다. 동결속도는 정자가 -140℃까지 도달하는 시간을 8분 30초인 동결법(FR-1)과 14분인 동결법(FR-2)으로 나누어 각각 0.5 ml 및 5 ml straw를 이용하여 동결하였으며, 동결-융해한 정자는 CASA 장비를 이용하여 정자 성상을 비교 분석하였다. 동결속도 만큼 융해 온도 및 시간 또한 매우 중요한 요소이기 때문에 융해 방법을 37℃, 50℃ 및 70℃에서 각각 115초, 45초 및 25초간 실시하여 정자 성상을 비교하였다. 그 결과 FR-2의 동결속도 보다 FR-1의 동결속도에서 높은 생존율과 운동성을 보였고, 50℃에서 45초간 융해 하였을 때 가장 높은 생존율과 운동성을 보였다(73.4±3.6; 74.5±2.2%, p<0.05). 정자의 첨체 충실성 및 형태학적 특징에서는 처리구간에 유의적인 차이가 없었다. 본 연구를 통해 얻은 데이터를 종합해볼 때, 그 동안 동결정액을 이용한 돼지 인공수정 시 5.0 ml straw가 0.5 ml straw 비해 저조한 정자 생존율 및 활력을 나타냄으로써 번식성적이 떨어져 사용을 기피하여 왔지만 개선된 프로토콜을 활용하면 동결정액을 이용한 돼지 인공수정에 있어 여러 straw를 융해해야 하는 불편함 대신에 인공수정 시 1 straw를(dose) 융해하여 사용함으로써 사용자의 편리성을 높일 뿐만 아니라 인공수정에 있어 집중력을 높여 번식성적 향상에도 기여할 것으로 사료된다.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.225-230
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• 2012

Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti-Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.127-133
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• 2008

The objective of this study was two folds: to investigate the relationship between paternal identification rate and sperm quality parameters such as motility and sperm chromatin structure assay after heterospermic insemination; to see if mutual complement between tests and development of useful technique to enhance the fertility in artificial insemination. In individual boar's fertilizing ability, 3 high fertility boars showed significantly high fertility (p<0.05) compared to 3 low fertility boars, but there was no difference in litter size between two groups. Sperm motility test in pooled and individual semen using computer assisted sperm analysis (CASA) revealed that no significant difference among boars. The high fertile boar showed tendency of low %Red (High red fluorescence/green+red fluorescence) in sperm chromatin structure assay (SCSA) but paternal identification rate from piglets did not differ after heterospermic insemination. The correlation coefficient between individual or pooled semen function test and farrowing rates were well correlated as follows: %Red with litter size (r= - 0.53, p=0.03); %Red with paternal identification rates (r=-0.51, p=0.03); paternal identification rates with litter size (r=0.57, p=0.02). These results indicate that sperm chromatin structure assay and sperm quality parameter test in pooled semen are useful method to predict and evaluate the fertilizing capacity after heterospermic insemination in boars.

• Clinical and Experimental Reproductive Medicine
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• 제49권4호
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• pp.277-284
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• 2022

Objective: Oxidative stress is a key player in the development of idiopathic male infertility (IMI), and various antioxidants have been used for the treatment of IMI with inconsistent results. Coenzyme Q10 (CoQ10) is a cofactor and an antioxidant that may improve semen parameters and reduce oxidative stress in patients with idiopathic oligoasthenospermia (OA). Therefore, this study aimed to explore the effect of CoQ10 on semen parameters and antioxidant markers in patients with idiopathic OA. Methods: Fifty patients with idiopathic OA and 35 fertile controls were enrolled in this prospective controlled study. All participants underwent a comprehensive fertility assessment. All patients received CoQ10 (300 mg/day) orally once daily for 3 months. Semen parameters, seminal CoQ10 levels, reactive oxygen species (ROS) levels, total antioxidant capacity (TAC), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were measured in patients and controls at the start of the study and after 3 months. Results: Treatment with CoQ10 resulted in increased sperm progressive motility (p<0.05), total motility (p<0.01), seminal TAC (p<0.01), SOD (p<0.05), GPx (p<0.001), and seminal CoQ10 (p<0.001) levels and reduced ROS (p<0.01) in patients as compared to baseline. Sperm concentration and motility were also significantly correlated with antioxidant measures and seminal CoQ10 levels (r=0.38-0.57). Conclusion: CoQ10 therapy (300 mg/day for 3 months) improved sperm motility and seminal antioxidant markers in patients with idiopathic OA. Therefore, CoQ10 could be a promising treatment for patients with idiopathic infertility and may improve their fertility potential.

• 한국수정란이식학회지
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• 제23권4호
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• pp.283-289
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• 2008

Lepidium meyenii, known as Maca, is traditionally employed in the Andean region for its supposed properties to improve energy and fertility. In the present study, we investigated the effects of gelatinized and fermented Maca on improvement of physical stamina and epididymal sperm counts, and on blood biochemical parameters related to fatigue and tissue injury: creatine phosphokinase, aspartate transaminase, lactate dehydrogenase, blood urea nitrogen, glucose, total cholesterol and total proteins. Adult male mice was divided at random into two main groups (resting and excercise groups). The excercise group was separated into three subgroups (exercise only, exercise with gelatinized Maca and fermented Maca-treatment groups). Gelatinized or fermented Maca (800 mg/kg) were orally administered for 30 days. All animals in exercise groups were subjected to daily 30-min swimming for 28 days 30 min after Maca treatment. Daily exercise decreased the body weight gain, and fermented Maca further attenuated the body weight increase. Gelatinized and fermented Maca significantly increased the maximum swimming time on 14 and 28 days of treatment (p<0.05), respectively, suggestive of a long-term stamina-enhancing effect of fermented Maca. Both Maca fully or significantly recovered blood parameters of energy as well as muscular and hepatocytic injuries changed by repeated exercise and maximum swimming performance (p<0.01). Moreover, gelatinized and fermented Maca increased epididymal sperm counts 22.0% and 32.0%, respectively. In conclusion, the results indicate potential benefits of Maca for improving both physical stamina by minimizing muscular and hepatic damage and preserving energy during swimming exercise and male reproductive function by increasing epididymal sperm counts.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.311-318
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• 1996

Mycoplasmas have long been suspected of contributing to involuntary infertility in couples. However considerable disagreement exits concerning the role of genital mycoplasma infection in human infertility. Several investigators have noted abnormalities in the semen analysis of men with positive mycoplasma cultures, and early epidemiologic studies indicated that Ureaplasma urealyticum was linked to human reproductive failure on the basis of higher frequencies of isolation from infertile versus fertile couples and successful pregnancies in infertile couples after doxycycline therapy. However, subsequent investigators have questioned these findings because there are many studies in which treatment for mycoplasma in the male or female did not demonstrate an improved pregnancy rate, and semen samples from unexplained infertile men containing ureaplasmas have not revealed poorer motility, fewer spermatozoa and more aberrant forms. The objective of this study were to investigate the incidence rate of mycoplasma in semen and to investigate whether the presence of mycoplasma in semen makes significant difference to the semen volume, sperm motility and sperm counts. The results were that the rate of isolation of mycoplasma species was 70.3%. Semen volume is $2.84{\pm}1.01ml$ for culture negative and $3.15{\pm}1.42ml$ for culture positive group. Sperm motility is $46.23{\pm}15.80%$ for culture negative and $50.09{\pm}15.69%$ for culture positive group, and sperm count is $95.47{\pm}47.14({\times}(P)10^6/ml)$ for culture negative and $86.73{\pm}47.59({\times}10^6/ml)$ for culture positive group. In conclusion, we suggest that the presence of mycoplasma in semen makes no significant differences to the sperm parameters.

• Clinical and Experimental Reproductive Medicine
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• 제43권2호
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• pp.90-96
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• 2016

Objective: Diabetes mellitus (DM) is known to cause many systemic complications as well as male infertility. Astaxanthin (ASTX) is a powerful antioxidant that is involved in a variety of biologically active processes, including those with anti-diabetes effects. The present study investigates the effect of ASTX on the spermatozoa function in streptozotocin (STZ)-induced diabetic rats. Methods: We divided 30 adult rats into three groups (10 rats per group), with a control group that received corn oil mixed with chow. DM was induced by intra-peritoneal injection of STZ. Eight weeks after the STZ injection, half of the diabetic animals were used as diabetic controls, and the rest were treated with ASTX for 56 days. Then the parameters and chromatin integrity of the epididymal sperm were analyzed using chromomycin A3, toluidine blue (TB), and acridine orange (AO) staining. Results: The count, viability, and motility of the epididymal sperm were decreased significantly in the STZ group in comparison with the control group (count and viability, p<0.001; motility, p<0.01). ASTX increased normal morphology and viable spermatozoa compared to the STZ group (morphology, p=0.001; viability, p<0.05). The percentage of abnormal chromatins in TB and AO staining was higher in the STZ group compared to the control group (p<0.001). The mean percentage of TB and AO positive spermatozoa in STZ rats was significantly lower in the STZ+ASTX group (TB, p=0.001; AO, p<0.05). Conclusion: This study observed that in vivo ASTX treatment partially attenuates some detrimental effect of diabetes. Conversely, ASTX improved sperm viability, normal morphology, and DNA integrity.

• Clinical and Experimental Reproductive Medicine
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• 제49권4호
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• pp.270-276
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• 2022

Objective: The present study assessed the biological characteristics of human spermatozoa at different time intervals (0, 1, 1.5, and 2 hours) after incubation at 37℃. Methods: Twenty-five normozoospermic semen samples were incubated at 37℃. Incubation was performed at four time intervals of 0 (after liquefaction), 1, 1.5, and 2 hours. The samples were evaluated for sperm parameters at each time interval. Results: The rate of sperm progressive motility decreased at 1.5 hours compared to 0 hours as well as 2 hours compared to 1 hour and 0 hours. The rate of non-motile spermatozoa also decreased after 2 hours compared to after 0 hours. No significant changes were observed in sperm viability (p=0.98) and non- progressive motility (p=0.48) at any time intervals. Abnormal sperm morphology increased at 1.5 hours of incubation time (p<0.001). No significant changes were observed in DNA fragmentation at 1 hour compared to 0 hours (median [interquartile range]: 19.5 [4] vs. 19 [4]), as well as at 1.5 hours compared to 1 hour (20 [5]). However, a significant increase in DNA fragmentation was observed at 1.5 hours compared to 0 hours. The mitochondrial membrane potential decreased remarkably after 1 hour of incubation time. No significant differences were observed in the acrosome reaction or malonaldehyde levels at any time point (p=0.34 and p=0.98, respectively). Conclusion: The incubation of normozoospermic samples before use in assisted reproductive technology should be less than 1.5 hours to minimize the destructive effects of prolonged incubation time on general and specific sperm parameters.

• Clinical and Experimental Reproductive Medicine
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• 제49권1호
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• pp.49-56
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• 2022

Objective: Oxidative stress and sperm DNA fragmentation (SDF) have been linked to idiopathic male infertility (IMI). Various antioxidants have been tried to improve semen parameters and fertility potential in IMI patients, but with inconsistent results. The study aimed to compare the effects of coenzyme Q10 (CoQ10) and Centrum multivitamins on semen parameters, seminal antioxidant capacity, and SDF in infertile men with idiopathic oligoasthenospermia (OA). Methods: This prospective controlled clinical study involved 130 patients with idiopathic OA and 58 fertile controls. The patients were divided randomly into two groups: the first group received CoQ10 (200 mg/day orally) and the second group received Centrum multivitamins (1 tablet/day) for 3 months. Semen parameters, CoQ10 levels, reactive oxygen species (ROS), total antioxidant capacity (TAC), catalase, SDF, and serum hormone levels (follicle-stimulating hormone, luteinizing hormone, testosterone, and prolactin) were compared at baseline and after 3 months. Results: Both CoQ10 and Centrum improved sperm concentration and motility, but the improvement was greater with Centrum therapy (p<0.05). Similarly, both therapies improved antioxidant capacity, but TAC and catalase improvement was greater (p<0.01 and p<0.001 respectively) with CoQ10, whereas ROS (p<0.01) and SDF (p<0.001) improvements were greater with Centrum administration. Centrum therapy was associated with reduced serum testosterone (p<0.05). Conclusion: In conclusion, both CoQ10 and Centrum were effective in improving semen parameters, antioxidant capacity, and SDF, but the improvement was greater with Centrum than with CoQ10. Therefore, Centrum-as a source of combined antioxidants-may provide more effective results than individual antioxidants such as CoQ10 in the treatment of infertile men with idiopathic OA.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.163-166
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• 2012

This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.