Objective: To reveal the trend in alterations of sperm counts in Asian men over the past 50 years. Methods: This study reviewed all the published reports to unveil the specific pattern of alterations of sperm concentrations in Asian men from 1965 till 2015. The time-related changes in sperm concentration were studied using linear regression analyses. Results: The present study elucidated the trend using the reports from Carlsen et al (1965-1990) and non-Carlsen studies published until 2015, on fertile Asian men. In the reports of Carlsen et al, no overall declining trend in Asian men (r = 0.509, P = 0.760) was observed during this tenure, but non- Carsen reports showed a significant time-dependent decline of sperm concentration (r = -0.754, P = 0.005) in Asian men. This present review also showed a mild time-dependant decline in sperm concentration (-0.44×10⁶/mL/year, 95% CI: -0.65 to -0.23; r = -0.473, P = 0.040) which accounted for an overall 22.17% decrease in past 50 years. Conclusions: This study brings to the forefront that sperm concentration among Asian men follows a mild declining trend over the period of 50 years, and further studies addressing the causes of this decline are required.

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Objective: To optimize the cryopreservation of buffalo bull semen by altering freezing rates within critical temperature range (4 °C to -60 °C). Methods: A total of 20 ejaculates each from 5 Murrah buffalo bulls were cryopreserved using programmable biofreezer in 2 phases. In the 1st phase, 9 freezing rates were applied at -2, -5, -10, -20, -30, -40, -50, -60 or -4 °C/min (control) from 4 °C to -15 °C ; at -40 °C/min from -15 °C to -60 °C. In the 2nd phase, a fixed freezing rate was applied at -30 °C /min from 4 °C to -15 °C. Six freezing rates were applied at -10, -20, -30, -40 (control), -50 or -60 °C/min from -15 °C to -60 °C. The freezing from -60 °C to -140 °C were fixed at -50 °C/min in both the phases. Post thaw semen quality was assessed in terms of motility, viability, membrane integrity (hypo-osmotic swelling test), sperm abnormalities, and active mitochondria. Data were arc sine transformed and analyzed through one-way analysis of variance using SPSS software. Results: In the 1st phase, percent individual motility, progressive motility and viability were similar among various protocols. Percent hypo-osmotic swelling reactive sperm was higher with freezing at -30 °C/min. In the 2nd phase, percent individual motility, viability and hypo-osmotic swelling reactive sperm was higher with freezing at -50 °C /min. Sperm head abnormalities were lower at -30 °C /min in the 1st phase, but were similar among the protocols of the 2nd phase. Percent active mitochondria were higher at -30 °C /min in the 1st phase and at -50 °C/min in the 2nd phase. Conclusions: The optimum post thaw semen quality of buffalo bull could be obtained by applying freezing rate at -30 °C/min (4 °C to -15 °C) and at -50 °C/min (-15 °C to -140 °C ), followed by plunging of straws in into liquid nitrogen for storage.

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Objective: To evaluate correlation between the levels of vitamin D and male infertility as well as to determine the efficacy of vitamin D in improving the male fertility by up-regulating the levels of testosterone and spermatogenesis. Methods: In the present study, 130 male patients (aged 25-70 years) having fertility defects were screened and 145 healthy individuals were taken as control. All human subjects were screened for 4-hydroxynonenal, isoprostane-F2α, 8-hydroxy-2'-deoxyguanosine, vitamin D, luteinizing hormone, follicle stimulating hormone, testosterones, malondialdehyde, superoxide dismutase, catalase, glutathione peroxidase, and nitric oxide. Results: The screening analysis revealed that the levels of luteinizing hormone, follicle stimulating hormone, and testosterone were lower in male infertile subjects compared to healthy subjects. Similarly, the levels of vitamin D [(17.17 ± 2.30) ng/mL] and calcium[(6.29 ± 0.31) mg/dL] were significantly lower in infertile groups compared to the normal healthy groups. Moreover, the study revealed that the levels of superoxide dismutase, catalase, and glutathione peroxidase were significantly higher in healthy subjects compared to the infertile subjects. Conclusions: Vitamin D exhibits strong relevance to male fertility by maintaining the levels of sex hormones (luteinizing hormone, follicle stimulating hormone, and testosterone), up-regulating the antioxidant defense (superoxide dismutase, catalase, and glutathione peroxidase), and down-regulating the oxidative stress (malondialdehyde, nitric oxide, and inducible nitric oxide synthase species).

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Objective: To evaluate effect of tris-extender supplemented with various concentrations of strawberry (Fragaria spp.) on bull semen preservability. Methods: Pooled bull semen were extended with tris-citrate-fructose egg yolk diluent (control, 0% strawberry) and various concentrations of tris strawberry (TSB) (1%-6%) to achieve 60 million motile spermatozoa per milliliter. Extended semen were subjected to semen freezing protocol. Semen assessment including motility, alive%, abnormality%, intact sperm membrane (hypo-osmotic swelling test) and conception rate were carried out for both chilled and frozen semen. Results: Results showed that sperm motility after chilling was enhanced in groups treated with various concentrations of TSB from 1% to 5% and exhibited higher significance (P<0.000 1) at 6-day post-chilling. In frozen semen, 3%, 4%, 5% and 6% concentrations gave the best significance (P<0.000 1) on sperm motility in comparison with the control. Concentration 1% revealed the highest significance (P<0.000 1) on alive% as compared to the control. Hypo- osmotic swelling test was maintained as the control. Concentration 3% gave the lowest significance (P<0.000 1) considering abnormality%. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 5% and 6% in cattle. Conclusions: It is concluded that 1%-5% concentrations of TSB ameliorate bull semen characteristics after chilling, and 3%-6% concentrations of TSB improve bull semen characteristics after freezing. Higher conception rate exists in 5% and 6% concentration of TSB.

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Objective: To test the effect of voltage-gated sodium channels (VGSCs) blockers on the motility and viability of human sperm in-vitro and to evaluate the tested compounds as potential contact spermicidal. Methods: Sperm samples were obtained from healthy nonsmoking volunteers of age 25-30 years who had not taken any drug 3 months before and during the course of the study. The effect of VGSCs blockers evaluated from two pharmacological classes including antiarrhythmic (amiodarone, procainamide and disopyramide) and antiepileptic (carbamazepine, oxcarbazepine, phenytoin, and lamotrigine) drugs. They were tested on the in-vitro motility and viability of human sperm using Computer Assisted Semen Analyzer. Results: All tested drugs except oxcarbazepine showed dose dependent inhibition of total motility with significant reduction (P<0.05) at the maximum concentration of 200 μΜ when compared with the control. The concentrations of drugs that reduced total sperm motility to 50% of control (half maximal inhibitory concentration) were 2.76, 14.16 and 20.29 μΜ for phenytoin, lamotrigine and carbamazepine, respectively; and 2.53, 5.32 and 0.37 μΜ for amiodarone, procainamide and disopyramide, respectively. The anti-motility effects were reversible to various degrees. There was statistically insignificant difference in the inhibition of sperm viability among amiodarone, procainamide and disopyramide. Phenytoin demonstrated the most potent spermicidal action. Conclusions: VGSCs blockers have significant adverse effects on in-vitro motility of human spermatozoa. So in-vivo studies are required to determine their potential toxicological effects on human semen quality, which is an important factor regarding fertility. Moreover, these drugs have the potential to be developed into contact spermicidal.

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Objective: To understand the level of progesterone (P4) in different quality of buffalo cumulus oocyte complexes (COCs) and further to evaluate the effect of exogenous P4 supplementation on maturation and subsequent developmental ability of poor quality brilliant cresyl blue (BCB^- ) COCs. Methods: Progesterone secreted by different quality of buffalo oocytes was estimated by enzyme linked immunosorbent assay and the concentration differences were translated into P4 doses to be incorporated in the maturation medium of BCB^-ve COCs followed by expression analysis of genes involved in the cumulus expansion, extracellular matrix disintegration and progesterone receptor signalling. In addition, the study also evaluated the effect of exogenous P4 on sperm-cumulus interaction. Results: More than 10-fold up- regulated expression of progesterone receptor in P4 supplemented oocytes signified that P4 might be acting predominantly through this receptor. Also, exogenous P4 supplementation had significant effect on transcatheter arterial chemoembolization protease regulated by P4- progesterone receptor pathway which in turn had an important role in extracellular matrix disintegration. On the contrary, cumulus expansion genes HAS2, TNFAIP6, AREG were not altered upon P4 supplementation. Also, it was observed that P4 addition did facilitate passage of significantly more number of spermatozoa through P4 treated cumulus cells. Further, incorporation of different doses of P4 did not improve significantly the cleavage and blastocyst rates of BCB^-ve COCs. Conclusions: Different qualities of buffalo COCs secrete substantially diverse levels of P4, and its supplementation has a role in oocyte maturation via modulation of cumulus characteristics but perhaps not fertilization.

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Previous attempts have indicated that mesenchymal stem cells (MSCs) are a valuable source and candidate and new approach for tissue engineering and reproductive medicine. MSCs have this potential to be induced and differentiated in an appropriate in vivo and in vitro condition toward various cell lineages and then they can be applied in cell therapies and clinical applications. During recent two decades, various sources have demonstrated they are a great source for MSCs, including bone marrow, the human umbilical cord as well as Wharton's jelly. Due to discarding after birth, easily accessible cells and less ethical concerns, these cells have attracted more and more scientists' attention. Infertility and reproduction diseases have provided special opportunity to examine the efficiency of MSCs in this kind of application. Based on recent investigations, MSCs embedded in Wharton's jelly tissue are more appealing for cell therapies, especially in infertility treatment purposes. So, differentiation of MSCs embedded in Wharton's jelly tissue into germ layer cells for cell-based therapy purposes is now under intensive study.

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