• Clinical and Experimental Reproductive Medicine
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• v.49 no.2
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• pp.110-116
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• 2022

Objective: Sperm vitrification leads to the production of reactive oxygen species (ROS) that can damage the functional parameters of sperm. The present study aimed to investigate the antioxidant effect of Nigella sativa extract on motility, plasma membrane function, mitochondrial membrane potential (MMP), DNA damage, and intracellular ROS production. Methods: A total of 20 sperm samples were used. Samples were divided into six experimental groups, including groups with aqueous extract from N. sativa seeds at concentrations of 1% to 6%, a cryopreserved control group, and a fresh control group. Results: Statistical analysis showed significantly higher total sperm motility at concentrations of 3% to 6% than in the vitrified semen control group. Additionally, progressive motility and all motion characteristics at all concentrations were significantly higher than in the vitrified semen control group. The presence of N. sativa seed extract also improved the quality of the sperm parameters assayed in all experimental groups (1%-6%; intracellular ROS production, DNA damage, MMP, and sperm membrane function) compared to the control group. Conclusion: Higher concentrations of N. sativa led to improvements in all sperm parameters and sperm quality. These findings indicate that N. sativa seed extract is effective for improving the quality of sperm after vitrification.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.9-16
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• 2013

Artificial insemination (AI) has been performed widely in swine industry using fresh liquid sperm instead of frozen type of sperm. However fresh sperm are not able to preserve more than three days with optimal motility and other sperm parameters for the successful fertilization, since in vitro stored sperm has an oxidative stress that resulted increase of abnormality and acrosome reation. To overcome these major problems, novel preservative formulation is needed to neutralize the oxidative stress and to provide suitable physiological environment for sperm in in vitro. In this study, naturally derived substances such as Poncirus trifoliate (Trifoliate orange), Garcinia mangostana (Mangosteen), pig placenta and testis extracts were tested as sperm preservative agents. Placenta extracts (PE), trifoliate orange extracts (TOE), testes extracts (TE) and mangosteen extracts (ME) were applied to analyze specific parameters for sperm motion characteristics individually and combinatorial. Each individual extract treatment can accelerate the sperm motility but noticeably TOE, TE and ME treatments exhibited the considerable and significant preservation of sperm motility. PE, TE and ME showed a significant (p<0.05) increase in ALH after one week. Further we evaluated the five different combinations of these extracts on sperm motility and its motion characteristics. Surprisingly even after one week ME, TOE and TE combination significantly preserved the sperm motility about 75%. It is noteworthy that unlike individual extract treatment, combination of ME, TOE and TE simultaneously protect the sperm motility and its motion characteristics. Taken together these data conclude that addition of ME, TOE and TE can be effective for preservation of pig sperm.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.2
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• pp.120-124
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• 2024

Objective: Seasonal variations in semen quality are known to occur in temperate regions, but results regarding tropical areas remain inconclusive. The aim of this study was to determine whether monthly variations in semen parameters are present among men in a tropical region. Methods: Data were retrospectively collected from semen analyses of 3,000 men over a 10-year period, from 2012 to 2022. Analysis of variance and the independent-samples t-test were employed to observe variations in semen parameters throughout the entire period and between months, respectively. Results: The mean±standard deviation sperm concentration was significantly lower in June, at 42.5±31.4 million/mL, compared to other months. The highest sperm concentration was found in March, at 57.8±42.6 million/mL, constituting a mean difference of 15.3 million/mL between the lowest and highest concentrations. The total sperm count displayed a similar pattern of monthly variation, with a difference of 47.2 million between the highest and lowest months. No significant monthly differences were observed in other parameters, such as sperm motility, morphology, and semen volume. Conclusion: Significant monthly variations in sperm concentration and total sperm count were evident in this Sri Lankan population. March, which displayed the highest sperm counts, is in the spring in temperate regions, while the month with the lowest counts, July, is part of the summer. Fluctuations in photoperiod appear to most strongly influence these variations.

• Animal Bioscience
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• v.34 no.12
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• pp.1912-1920
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• 2021

Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT^® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl₂ and MgCl₂ (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA. Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.61-68
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• 2021

Objective: This study was conducted to investigate the relationship of semen parameters in samples used for intracytoplasmic sperm injection (ICSI) with fertilization and pregnancy rates in infertile couples. Methods: In this prospective study of Infertile couples with male factor infertility that had undergone ICSI, fractions of the same semen samples obtained for microinjection (to ensure the best predictability) were evaluated to determine the semen parameters and sperm DNA fragmentation index (DFI) on the day of oocyte recovery. Results: In total, 120 couples completed the study and were subdivided into fertilized (n=87) and non-fertilized couples (n=33). The fertilized couples were further classified into pregnant (n=48) and non-pregnant (n=39) couples. Compared to non-fertilized and non-pregnant couples, fertilized and pregnant couples showed statistically significantly higher sperm viability and percentage of normal sperm morphology, as well as significantly lower sperm DFI values. A receiver operating characteristic curve analysis of data from the 120 ICSI cycles showed that sperm viability, normal sperm morphology percentages, and sperm DFI were significant prognostic indicators of fertilization at cutoff values of 40%, 7%, and 46%, respectively. A sperm DFI of 46% showed sensitivity and specificity of 95% and 90%, respectively, for predicting fertilization, and no clinical pregnancies occurred in couples with a sperm DFI above 46%. Conclusion: Semen parameters from the ICSI day sample, especially sperm viability, normal morphology, and DFI, had an impact on fertilization and pregnancy outcomes in ICSI cycles.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.2
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• pp.125-134
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• 2024

Objective: Vitamin D deficiency is a major problem for human health worldwide. The mechanisms of vitamin D in the male reproductive system are unknown. After coronavirus disease 2019 (COVID-19) vaccines were developed, doubts were raised about their possible effects on male fertility. Based on vitamin D's function in the immune system, its potential role as an adjuvant for COVID-19 vaccines is intriguing. The aims of this study were to assess the effects of vitamin D first on sperm parameters and sex hormones, and then as an immune adjuvant on sperm parameters and sex hormones after study participants had received their second doses of COVID-19 vaccines. Methods: Phase 1 (before the COVID-19 pandemic) included 72 men with idiopathic infertility, and phase 2 had 64 participants who received two doses of COVID-19 vaccines. Both groups were instructed to take 50,000 IU of vitamin D twice monthly for 3 months. Sperm parameters and sex hormones were assessed pre-and post-supplementation. Results: Regular vitamin D intake for 3 months significantly increased the participants' vitamin D levels (p=0.0001). Both phases showed a positive correlation between vitamin D intake and sperm parameters. Vaccination had no negative effects on sperm parameters and sex hormones. Vitamin D was associated with follicle-stimulating hormone (p=0.02) and testosterone (p=0.0001) in phase 2 after treatment. Conclusion: Our results support vitamin D supplementation as an immune adjunct to COVID-19 vaccination for improving sperm parameters and hormone levels. COVID-19 vaccination is not harmful for male fertility potential, and vitamin D is an effective factor for male fertility.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.1
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• pp.48-56
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• 2024

Objective: This study investigated the relationship of anthropometric and metabolic risk factors with seminal and sex steroidal hormone parameters in a screened population of healthy males. Methods: The participants were healthy young men without chronic or congenital diseases. The body composition parameters that we investigated were measured weight, height, and waist circumference (WC), as well as bioelectrical impedance analysis. Semen samples were analyzed for semen volume, sperm concentration, sperm motility and morphology, seminal pH, and liquefaction time. Biochemistry analysis, including glucose and lipid metabolism parameters, was conducted on fasting blood samples. Testicular volume was calculated separately for each testis using ultrasonography. Results: Body mass index exhibited an inverse association with total sperm count. WC showed negative correlations with numerous seminal parameters, including sperm concentration, total sperm count, sperm morphology, and follicle-stimulating hormone levels. The basal metabolic rate was associated with seminal pH, liquefaction time, and sperm motility. WC, fat mass percentage, and triglyceride levels exhibited negative correlations with sex hormone binding globulin. The measures of glucose metabolism were associated with a greater number of seminal parameters than the measures of cholesterol metabolism. C-reactive protein levels were inversely associated with sperm concentration and total sperm count. Conclusion: Anthropometric and metabolic risk factors were found to predict semen quality and alterations in sex steroidal hormone levels.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.262-269
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• 2023

Objective: The aim of this study was to compare semen parameters and sperm DNA fragmentation (SDF) and explore the relationship between semen parameters and SDF between 2 and 7 days of abstinence and a short abstinence period (within 4 hours) in oligozoospermic infertile patients. Methods: Two semen samples were collected from infertile oligozoospermic men (n=34) after an abstinence period of 2 to 7 days and within 4 hours, respectively. Sperm parameters were compared between the two abstinence duration groups, including semen volume, sperm concentration, total sperm count, sperm motility, total motile sperm count (TMSC), morphology, and SDF. Results: The semen volume, concentration, and total sperm count were significantly decreased after 4 hours of abstinence than after 2 to 7 days of abstinence, with median differences of 1.2 mL (p<0.001), 2×10⁶/mL (p=0.011), and 9.6×10⁶/ejaculation (p<0.001), respectively. TMSC was significantly lower after a short abstinence, with a median difference of 4.24×10⁶/ejaculate (p<0.001). However, there were no significance differences in the percentage of motility, the SDF, and the percentage of sperm with normal morphology. Interestingly, volume, concentration, total sperm count, sperm motility, and SDF, but not TMSC, exhibited significant linear correlations between the two abstinence groups in univariate regression analysis, except for TMSC. Conclusion: In oligozoospermic men, the volume, concentration, and total sperm count were significantly lower after a short abstinence period, but without adverse effects on sperm motility and SDF.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.4
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• pp.295-300
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• 2001

Objective : To assess the effects of progesterone and acetyl-L-carnitine used after treated with Isolate�� gradient before semen cryopreservation-thawing on sperm parameters and membrane integrity. Material and Methods : From April 2001 to July 2001, ten normal male partner of couples who were visited in vitro fertilization (IVF) clinics. the semens were treated with $Isolate^{(R)}$ gradient before cryopreservation, spermatozoa was incubated with progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$), or both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$) for 30 min. Results: There were no differences in sperm parameters and vital stain among isolate only treated group, progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$) and both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$). But, in high concentration of acetyl-L-carnitine ($10{\mu}M$) treated group, sperm parameters and vital stain were decreased. The statistical method was used ANOVA (Kruskal-Wallis test) and p value was <0.01. Conclusions : Neither progesterone nor acetyl-L-carnitine show to be protective effect on the cryodamage assessed by sperm parameters and vital stain (eosin-Y stain) in normal sperm. High concentration of acetyl-L-carnitine ($10{\mu}M$), however, was harmful effect on cryoprevention.

• Development and Reproduction
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• v.28 no.1
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• pp.13-19
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• 2024

Ras-related (Rab) proteins, integral members of the monomeric G-protein family, play a pivotal role in regulating intracellular vesicular transport. These proteins contribute to male reproductive processes, specifically in acrosome formation, exocytosis, and sperm motility. Although a prior study indicated a correlation between Rab3A and sperm motility, including motion kinematic parameters such as mean dance, this association has only been explored within a limited sample size. Therefore, further verification is required to confirm the correlation between Rab3A and sperm motility parameters. In the present study, Rab3A expression, sperm motility, and motion kinematic parameters were analyzed in 150 boar spermatozoa. Additionally, correlations between Rab3A expression and sperm kinematic characteristics were evaluated statistically. The results revealed significant associations between Rab3A protein expression levels and various motion kinematic parameters. Specifically, Rab3A levels exhibited positive correlations with average path velocity (p<0.05), mean amplitude of lateral head displacement (p<0.05), and curvilinear velocity (p<0.01). Consequently, it is proposed that Rab3A protein plays a crucial role in male fertility through its correlation with sperm kinematic characteristics, making it a potential marker for sperm motility-related assessments.