• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.34-41
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• 2020

Objective: The strong antioxidant activity of Commiphora mukul prompted us to conduct the present study to explore whether treatment with C. mukul extract (CME) would have any protective influence on sperm parameters, testosterone levels, and plasma glucose levels in streptozotocin (STZ)-induced diabetic rats. Methods: Male Wistar rats were randomly divided into four groups: control, control animals treated with CME, diabetic animals, and diabetic animals treated with CME. CME extract (300 mg/kg) was administered for 60 days by daily gavage. Diabetes was induced by an intraperitoneal injection of 50 mg/kg STZ. The epididymal sperm count, weight, motility, morphology, viability, and serum testosterone and glucose levels were determined. Results: In the diabetic animals, CME decreased blood glucose levels (p< 0.05), increased the total sperm count (p< 0.05), and decreased the proportion of sperm with abnormal morphology (p< 0.05). Diabetes reduced sperm motility (p< 0.001), and CME supplementation partially reversed this effect of diabetes (p= 0.003). Furthermore, in diabetic animals, CME decreased the proportion of immotile sperm (p< 0.001). In rats, diabetes caused a significant decrease (p< 0.05) in serum testosterone levels (F[3, 28] = 3.283, p= 0.035), but treatment of diabetic animals with CME increased serum testosterone levels. Conclusion: The present study demonstrated that C. mukul possesses proandrogenic activity and exerts a beneficial effect on sperm parameters in diabetic rats.

• Journal of Embryo Transfer
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• v.10 no.1
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• pp.15-22
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• 1995

Since the turn of the century, scientists have earnestly sought to develop a single laboratory assay or combination of laboratory assays which accurately predict the fertility of a semen sample. Most of these assays have focused on evaluating physical characteristics of sperm such as motility, viability, acrosomal integrity and morphology. In recent years new approches have been used to assess the functional aspects of a sperm that are needed to reach the oocyte, fertilize it and contribute to successful embryo development. Among these techniques are the ability of sperm to undergo a heparin induced acrosome reaction and in vitro fertilization, and the affinity of sperm to bind heparin binding protein. Intensification of research efforts in the area of control of sperm fertilizing ability should be a high priority, in view of undoubted benifits both to our basic understanding of sperm fertilizing ability and to our ability to modify it for Al industry.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.2
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• pp.73-78
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• 2017

Objective: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. Methods: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. Results: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. Conclusion: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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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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• v.47 no.1
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• pp.54-60
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• 2020

Objective: Oxidative stress plays a key role in the pathogenesis of male infertility. But, the adverse effects of oxidative biomarkers on sperm quality remain unclear. This study aimed to investigate the levels of nitric oxide (NO), 8-hydroxydesoxyguanosine (8-OHdG), and total antioxidant capacity (TAC) oxidative biomarkers in seminal plasma and their relationship with sperm parameters. Methods: A total of 77 volunteers participated in the study, including fertile (n = 40) and infertile men (n = 37). NO, 8-OHdG, and TAC levels were measured using the ferric reducing ability of plasma, Griess reagent method and an enzyme-linked immunosorbent assay kit, respectively. Results: The mean values of sperm parameters in the infertile group were significantly lower than those in the fertile group (p< 0.001). The mean 8-OHdG in the seminal plasma of infertile men was significantly higher (p= 0.013) than those of controls, while the mean TAC was significantly lower (p= 0.046). There was no significant difference in NO level between the two groups. The elevated seminal 8-OHdG levels were negatively correlated with semen volume, total sperm counts and morphology (p< 0.001, p= 0.001 and p= 0.052, respectively). NO levels were negatively correlated with semen volume, total sperm counts and morphology (p= 0.014, p= 0.020 and p= 0.060, respectively). Positive correlations between TAC and both sperm count and morphology (p= 0.043 and p= 0.025, respectively) were also found. Conclusion: These results suggested that increased levels of NO and 8-OHdG in seminal plasma could have a negative effect on sperm function by inducing damage to the sperm DNA hence their fertility potentials. Therefore, these biomarkers can be useful in the diagnosis and treatment of male infertility.

• Korean Journal of Veterinary Research
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• v.24 no.2
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• pp.245-266
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• 1984

As a fundamental study to increase the reproductive efficiency in cattle, highly motile sperm were separated and collected from raw semen, extended semen and frozen semen by different methods using various concentrations of bovine serum albumin or Tyrode's solution. Various characteristics and light microscopic and electron-microscopic morphology of sperm separated by different methods were compared. The results obtained were as follows; 1. The sperm separated from raw semen using bovine serum albumin showed significantly high value in motility, motile sperm count, percent of normal sperm and progressive motility, as compared with control sperm and revealed the highest sperm recovery rate when separated with 6% bovine serum albumin. 2. The sperm motility, percent of normal sperm and progressive motility of the highly motile sperm frozen after being separated from raw semen with bovine serum albumin, showed significantly high value than those of a control sperm and especially found the highest value when separated with 20% bovine serum albumin. 3. Light-microscopic percent of abnormality was significantly low in the prefrozen and postfrozen highly motile sperm separated with bovine serum albumin, as compared with control sperm. 4. Electron-microscopic finding of the highly motile sperm separated with bovine serum albumin showed low percent of deformity in the dilatation and vesiculation of cell membrane, in dilatation and density loss of acrosome than in those of control sperm. 5. It was impossible to separate the highly motile sperm from frozen semen with bovine serum albumin, but it was possible with Tyrode's solution. 6. Recovery rate of highly motile sperm from raw semen extended semen and frozen semen was the highest when the sperm pellet stood in Tyrde's solution for 80 minutes. 7. The highly motile sperm separated from raw semen, extended semen and frozen semen with Tyrode's solution showed significantly high value in motility, progressive motility and percent of normal sperm, as compared with control sperm. 8. Highly motile sperm, when separated from raw semen, extended semen and frozen semen with Tyrode's solution, showed significantly low percent of microscopic abnormality as compared with control sperm.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.3
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• pp.132-140
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• 2017

Objective: Correlations between semen parameters and sperm DNA fragmentation index (DFI) were investigated to identify characteristics of sperm without DNA damage that could be used in selecting sperm for intracytoplasmic sperm injection (ICSI). Pregnancy outcomes were compared to determine whether in vitro fertilization (IVF) or ICSI is a better choice for patients who have sperm with a high-DFI. Methods: Semen analysis was carried out in 388 patients who visited our IVF center for the first time to investigate correlations between sperm DFI and semen parameters. In addition, 1,102 IVF cycles in 867 patients were carried out in the present study; 921 cycles in the low-DFI group (DFI < 30%) and 181 cycles in the high-DFI group ($DFI{\geq}30%$). Both the low- and high-DFI groups were subdivided into IVF and ICSI cycle groups. Results: Sperm DFI showed significant inverse correlations with sperm motility (r = -0.435, p< 0.001) and morphology (r = -0.153, p< 0.05). Sperm DFI also showed significant correlations with rapid motility (r = -0.436, p< 0.001), and the kinetic parameters of average-path velocity (r = -0.403) and linearity (r = -0.412). Although there was no significant difference in the pregnancy rates between IVF (48.6%) and ICSI (44.8%) in the low-DFI group, the pregnancy rate of ICSI cycles (44.8%, p< 0.05) was significantly higher than IVF cycles (25.0%) in the high-DFI group. No significant difference was observed in the abortion rates between the low-DFI (52 of 921, 5.6%) and high-DFI groups (7 of 181, 3.8%). Conclusion: ICSI is a better choice than IVF for improving the pregnancy outcomes of patients who have sperm with a high DFI.

• Applied Microscopy
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• v.35 no.4
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• pp.32-41
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• 2005

The sperm morphology of two Korean frogs, Rana catesbeiana and R. nigromaculata (Ranidae) was studied with the light microscope, scanning and transmission electron microscopes. Sperm heads of the R. catesbeiana and R. nigromaculata were long-cylinder shapes. In the total length of R. nigromaculata sperm ($51.2{\mu}m$) was longer than those of R. catesbeiana ($44.6{\mu}m$). Both R. catesbeiana and R. nigromaculata had the long head and principal piece, and short middle piece, but the neck was undeveloped. The axonemal structures contain a 9+9+2 arrangement of microtubules. The number of mitochondria in middle piece have less than mammalian sperm, which irregular surrounded the axone of the middle piece. In conclusion, R. catesbeiana and R. nigromaculata shared many similar features in sperm heads, neck, middle piece, and a site of mitochondria and axial structure, and it suggests that these species should be closely related.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.237-245
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• 2010

Only an optimum number of viable spermatozoa in a frozen-thawed insemination dose can ensure conception at artificial insemination (AI). We report here the percentages of normal, abnormal and viable spermatozoa present in the frozen-thawed semen of 20 Black Bengal bucks used for commercial AI. Bucks in this experiment were of 19.3~46.1 months old and 25~42 kg body weight. Four semen straws (0.25 ml) from each buck were collected for evaluation of their kinetic parameters. Scrotal circumference was measured by using a scrotal tape, sperm motility was estimated on eye estimation and sperm concentration was determined by using a haemocytometer. Sperm morphology was studied in paraformaldehyde fixed spermatozoa under differential interference contrast (DIC) microscope. To determine the proportion of live (plasma membrane intact) spermatozoa, semen was stained with SYBR-14 and propidium iodide and examined under fluorescent microscope. Scrotal circumference, post-thaw sperm motility, sperm concentration per insemination dose and proportion of normal spermatozoa were $21.5{\pm}0.7\;cm$, $43.5 {\pm}5.4%$, $83.5{\pm}6.7$ million and $88.3{\pm}4.1%$, respectively. The percentages of spermatozoa with head shape and acrosome abnormalities were lower ($2.7{\pm}1.1$ and $1.4{\pm}1.3$, respectively), whereas higher percentages of abnormalities ($7.0{\pm}1.8$) were observed in mid piece and tail portion. The proportion of live spermatozoa was $28.5{\pm}5.4$. It is concluded that although a good number of morphologically normal spermatozoa are present in the insemination dose, the proportion of live spermatozoa is low, which warrants further improvements of buck semen freezing procedures to ensure good quality at AI.

• Clinical and Experimental Reproductive Medicine
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• v.13 no.2
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• pp.175-180
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• 1986

It has been reported that oral kallikrein therapy exerts a favourable effect on sperm motility in asthenozoospermic patients. In order to evaluate the efficacy of kallikrein on asthenozoospermia, a total of 20 subfertile male patients with varicocele, whose sperm counts were less than $40{\times}10^6/ml$ and sperm motility was less than 30%, was subjected to this clinical study (Table 1). They were divided into 2 study groups: 1) Varicocelectomy group consisted of 10 patients with varicocele (grade II-III) who underwent varicocelectomy. 2) Kallikrein group was composed of 10 patients with varicocele (grade I) who were given kallikrein orally 600 KU (kallikrein unit) daily divided 3 times after meal for 3 to 9 months. Semen analyses were repeated twice before the study, once a month during the study and twice after the study. Effective results designate that sperm parameters improved more than 30% from the basical levels after varicocelectomy or kallikrein exposure. Sperm counts increased from $32.5{\times}10^5/ml$ to $45.5{\times}10^6/ml$ after varicocelectomy in 3 patients and sperm motility increased from 25% to 38.5% after varicocelectomy in 3 patients. Pregnancy occurred in 2 patients of 3 responders and 1 patient of 7 non-responders 3 to 6 months after varicocelectomy in Varicocelectomy group. Sperm motility increased from 28% to 40.2% after kallikrein treatment in 3 patients. Pregnancy occurred in 2 patients of the 3 responders in Kallikrein group (Tables 2-3). There were no significant changes in volume and morphology in Varicocelectomy group before after varicocelectomy and no significant changes in volume, counts, and morphology before and after kallikrein exposure. No remarkable side effects were noted with kallikrein treatment.