Human Sperm Parameters Research Articles

Supplementation of sperm freezing medium with myoinositol improve human sperm parameters and protects it against DNA fragmentation and apoptosis.

The aim of this study is to evaluate the beneficial effect of Myoinositol (MYO) supplement in freezing media on the post thaw sperm quality. Semen samples from 40 normozoospermic men were divided into two aliquots and frozen with simple or 2mg/mL MYO supplemented freezing medium. Post thaw process including, computer-assissted sperm analysis was used to analyze sperm motility and morphology. Reactive oxygen species was evaluated by the fluorometry of DCFH-DA, as well as total antioxidant capacity and lipid peroxidation were measured based on colorimetric assay by ELISA reader. Eventually, DNA fragmentation was assessed using TUNEL staining. MYO significantly improved progressive motility and normal morphology in treated samples (p < 0.05). Lipid peroxidation (malondialdehyde level) can be diminished in samples were frozen by MYO supplemented freezing media (p < 0.05). While MYO did not affect the amount of ROS (p > 0.05), it was associated with high values of total antioxidant capacity (p < 0.05). DNA integrity was significantly affected by MYO, as in MYO treated samples, DNA fragmentation was decreased compared to control ones (p < 0.001). The findings support the use of 2mg/mL myoinositol supplemented freezing media in sperm cryopreservation to increase sperm quality after freezing-thawing procedures.

Alterations in human sperm parameters as a corollary of incubation with various uropathogenic microorganisms

Purpose: To analyze the impact of different uropathogenic microorganisms on various sperm parameters those are generally used to determine male fertilizing potential. Method: The standard strains viz. Serratia marcescens, Klebsiella pneumoniae, Enterobacter aerogenes, Candida albicans, Proteus mirabilis, Streptococcus pyogenes, Klebsiella oxytoca were checked for their impact on sperm motility, viability, morphology, Mg++ ATPase activity and acrosome status of spermatozoa. Results: S. marcescens, K. pneumoniae, E. aerogenes compromised sperm motility via agglutination and C. albicans impaired motility without agglutination. Further, these strains could also reduce viability resulting in sperm death. These strains induced morphological defects in spermatozoa as evident by electron microscopic studies. Incubation of spermatozoa with these uropathogenic strains led to reduction in Mg2+ dependent ATPase activity and also resulted in premature acrosomal loss. On the other hand, P. mirabilis, S. pyogenes, K. oxytoca did not alter any of these parameters. Conclusion: It can be concluded from the present study that certain uropathogenic strains can induce detrimental effects on spermatozoon thereby reducing its fertilizing ability.

Alterations in human sperm parameters as a corollary of incubation with various uropathogenic microorganisms

Alterations in human sperm parameters as a corollary of incubation with various uropathogenic microorganisms

Relationships between bacteriospermia, DNA integrity, nuclear protamine alteration, sperm quality and ICSI outcome

The Aim of this study was to evaluate the effects of bacteriospermia on human sperm parameters, nuclear protamines, DNA integrity and ICSI outcome in patients enrolled for ICSI treatment. 84 unselected couples consulting in infertility and obstetrics clinic and enrolled for ICSI treatment were included in this study. The semen specimens were screened bacteriologically; semen and sperm parameters were also evaluated according to WHO guidelines. DNA integrity, protamines concentration and protamine deficiency were estimated by TUNEL assay, AU-PAGE and Chromomycin (CMA3) respectively. The results of this study revealed that 34.52% of studied semen samples were infected with bacteria. The isolated bacteria were identified as Staphylococcus aureus, Staph. epidermidis, Staph. haemolyticus, Escherichia coli, Enterococcus faecalis and Streptococcus agalactiae. Bacteriospermia had a significant (p < .010) negative effect on sperm parameters; concentration, motility, progressive motility and chromatin condensation. Moreover, high DNA fragmentation with low P1 and P2 concentrations were noticed in infected patients in comparison to non-infected patients but non-significant. Also, the fertilization rate decreased significantly (p < .05) with infected patients. In conclusion: bacteriospermia has significant negative effect on sperm quality and fertilization rate in patients who underwent ICSI treatment.

Abnormal Human Sperm Parameters Contribute to Sperm DNA Fragmentation in Men with Varicocele.

PurposeThis study was performed to evaluate and compare threshold sperm parameters and sperm DNA fragmentation index (DFI), and further analyzed whether sperm DFI could be predicted from sperm parameters in men with varicocele.Materials and MethodsA total of 157 semen samples underwent both semen analysis and sperm DNA fragmentation (SDF) testing in men with varicocele. Sperm parameters were assessed using the World Health Organization guidelines. SDF testing was performed using the Halosperm kit. Sperm parameters and sperm DFI results were compared.ResultsThe overall sperm parameter results and sperm DFI showed normal values; however, the morphology value was at the lower limit of normal. High sperm DFI was associated with significantly lower motility and viability (p<0.001, respectively). Sperm motility and morphology were significantly higher in the higher sperm count group compared to the lower sperm count group (p<0.05), while sperm DFI was higher in the lower sperm count group (p<0.05). Sperm count and viability and sperm DFI were significantly associated with the quality of sperm motility (p<0.001). Sperm motility and sperm DFI were significantly different (p<0.001) between normal and abnormal sperm viability groups. Between normal and abnormal sperm morphology groups, sperm count, motility, and sperm DFI showed significant differences (p<0.001).ConclusionsIn this study, a correlation between SDF and sperm parameters was confirmed in men with varicocele. SDF may be contributing factors to sperm motility, viability, and morphology. Abnormal sperm count, motility, and viability showed high sperm DFI. Therefore, lower sperm parameters were indicative of increasing SDF in men with varicocele.

Melatonin affects membrane integrity, intracellular reactive oxygen species, caspase3 activity and AKT phosphorylation in frozen thawed human sperm.

Cryopreservation is known to induce oxidative stress in spermatozoa. Although melatonin has powerful antioxidant properties, little is known about its effects on human sperm quality during cryopreservation. The present study was undertaken to investigate the effects of melatonin treatment on human sperm parameters essential for fertilization. We first evaluated the effects of various concentrations of melatonin (0-15mM) on human sperm parameters such as motility, viability and levels of intracellular reactive oxygen species during cryopreservation in order to identify an optimal dose with the greatest effects for further studies. Liquefied semen samples were then divided into three aliquots: cryopreserved without melatonin (control), cryopreserved with 3mM melatonin and fresh groups. After being thawed, samples were evaluated for motility, viability, membrane integrity, intracellular reactive oxygen species levels, caspase-3 activity and AKT phosphorylation. Treatment of spermatozoa with the various concentrations of melatonin significantly increased their motility and viability and decreased their intracellular reactive oxygen species levels compared with the control group. The optimal melatonin concentration (3mM) significantly decreased the intracellular reactive oxygen species levels, caspase-3 activity and the percentage of both dead and apoptotic-like sperm cells and increased the vitality, progressive motility and total motility and AKT phosphorylation compared with the control group. Thus, melatonin exerts protective effects against cryodamage during human spermatozoa cryopreservation and may exert its effects via the PI3K/AKT signaling pathway.

The influence of semen quality on male pronucleus demethylation process during ICSI cycle

ABSTRACTThere is growing evidence that the spermatozoon’s epigenetic structure is of the utmost importance in the health of the future embryo. Following fertilization, sperm chromatin undergoes epigenetic reprogramming including DNA demethylation and remethylation, which resets gene expression. In some infertile patients, it is inevitable that sperm cells that are not within the range of normal human sperm parameters will be used for intracytoplasmic sperm injection. Understanding the relationship between the human sperm parameters and male pronucleus DNA demethylation seems necessary. We hypothesized that demethylation of the male pronucleus might be altered in zygotes conceived from a spermatozoa obtained from a sample exhibiting an abnormal semen analysis profile. To test the hypothesis, sperm cells from normal and abnormal human semen samples were injected into mouse oocytes. A group of cultured zygotes was fixed before the onset of DNA demethylation and the other group was fixed after DNA demethylation. Both groups were then labeled with a 5 methylcytosine antibody and the level of pronuclei methylation was detected as a function of fluorescent intensity. The level of demethylation was then determined as the difference between 5 methylcytosine fluorescent intensity before and after DNA demethylation. A negative correlation (p<0.05) was observed between sperm motility, morphology, percentage of head defects, protamine deficiency, and DNA demethylation level. However, no correlation was found between the demethylation level and sperm count. In conclusion, these observations suggest that demethylation is altered in the male pronucleus when low quality sperm samples are used.

Relationship between Semenogelins bound to human sperm and other semen parameters and pregnancy outcomes

BackgroundSemenogelins (SEMGs) are major components of human seminal vesicle secretions. Due to SEMG’s sperm-motility inhibitor, a significant negative correlation between sperm motility and the proportion of SEMG-bound spermatozoa (SEMG+) was found in asthenozoospermic patients. SEMGs also show intrinsic inhibitory capability for sperm capacitation; however, studies on actual clinical specimens have not been conducted.MethodsTo reveal the relationship between SEMGs and the fertilizing capacity of sperm from male infertile patients who are not restricted to asthenozoospermia, we measured the proportion of SEMG+ in the spermatozoa of 142 male infertile patients. The pregnancy outcomes in partners of these patients were retrospectively analyzed using questionnaires.ResultsAmong examined semen parameters, only the total SEMG-unbound sperm count showed a tendency to be different between the spontaneous pregnancy or intra-uterine-insemination-pregnancy groups and in-vitro-fertilization- or intracytoplasmic-sperm-injection-pregnancy groups. It was elevated in the former group, which includes patients who used in vivo fertilization.ConclusionsThe total SEMG-unbound sperm count would be a relevant parameter for in vivo fertilization. This result suggests that SEMGs inhibit ectopic capacitation before sperm reach the fertilization site and that the number of total SEMG-unbound sperm is a parameter directly linked to the possibility of in vivo fertilization.

Effect of selenium on human sperm parameters after freezing and thawing procedures

ObjectiveTo evaluate the effects of pre-freezing treatment of human semen samples with selenium on semen parameters after thawing procedure. MethodsEach sample was divided into four groups: two groups were washed and other two remained unwashed. 5 μg/mL of selenium was added to one of the washed and unwashed group. ResultsThe results showed that sperm motility in unwashed selenium-treated samples was higher compared to washed untreated samples (P < 0.05). DNA damage percentage in unwashed treated samples compared to both untreated samples showed no difference (P > 0.05) but in washed sperm samples, DNA damaged percentage decreased in treated samples compared to untreated ones (P < 0.05). Moreover, the effect of selenium on morphology of washed treated sperms was higher than unwashed untreated samples after thawing procedure. Normal morphology percentage in unwashed treated samples was significantly higher than both unwashed samples after thawing (P < 0.001). In the washed treated samples a higher number of sperms with normal morphology compared to washed untreated samples (P < 0.001) were observed. ConclusionsBased on the results, we recommend 5 μg/mL of selenium should be used in infertility clinics to increase sperm quality after freezing and thawing procedures.

Tribulus terrestris Extract Improves Human Sperm Parameters In Vitro.

Objective.The object of present study was to investigate the effects of direct addition of Tribulus terrestris extract on human sperm parameters.Design.Semen specimens from 40 healthy men volunteers were divided into 4 groups: one group received no treatment (control group) while the others were incubated with 20, 40, and 50 µg/mL of T terrestris extract (experimental groups). Motility, viability, and DNA fragmentation were assessed in all groups.Results.The incubation of human semen with 40 and 50 μg/mL of T terrestris extract significantly enhanced total sperm motility, number of progressive motile spermatozoa, and curvilinear velocity over 60 to 120 minutes’ holding time (P < .05 or P < < .01). Furthermore, viability was significantly enhanced by using T terrestris extract (P < .01).Conclusions.In vitro addition of the T terrestris extract to human sperm could affect male fertility capacity.

The human RHOX gene cluster: target genes and functional analysis of gene variants in infertile men.

The X-linked reproductive homeobox (RHOX) gene cluster encodes transcription factors preferentially expressed in reproductive tissues. This gene cluster has important roles in male fertility based on phenotypic defects of Rhox-mutant mice and the finding that aberrant RHOX promoter methylation is strongly associated with abnormal human sperm parameters. However, little is known about the molecular mechanism of RHOX function in humans. Using gene expression profiling, we identified genes regulated by members of the human RHOX gene cluster. Some genes were uniquely regulated by RHOXF1 or RHOXF2/2B, while others were regulated by both of these transcription factors. Several of these regulated genes encode proteins involved in processes relevant to spermatogenesis; e.g. stress protection and cell survival. One of the target genes of RHOXF2/2B is RHOXF1, suggesting cross-regulation to enhance transcriptional responses. The potential role of RHOX in human infertility was addressed by sequencing all RHOX exons in a group of 250 patients with severe oligozoospermia. This revealed two mutations in RHOXF1 (c.515G > A and c.522C > T) and four in RHOXF2/2B (-73C > G, c.202G > A, c.411C > T and c.679G > A), of which only one (c.202G > A) was found in a control group of men with normal sperm concentration. Functional analysis demonstrated that c.202G > A and c.679G > A significantly impaired the ability of RHOXF2/2B to regulate downstream genes. Molecular modelling suggested that these mutations alter RHOXF2/F2B protein conformation. By combining clinical data with in vitro functional analysis, we demonstrate how the X-linked RHOX gene cluster may function in normal human spermatogenesis and we provide evidence that it is impaired in human male fertility.

Toxic effects of polychlorinated biphenyls (Aroclor 1254) on human sperm motility.

Polychlorinated biphenyls (PCBs) are common environmental contaminants that represent a considerable risk to reproductive toxicity in exposed human populations. Although some experimental studies have suggested an association between the levels of PCBs and semen quality, the direct effects of PCBs on human sperm parameters remain largely unexplored. To this aim, a short-term in vitro incubation experiment that better imitated the putative exposure of sperm to Aroclor 1254 (a commercial PCB mixture) in male reproduction tissue was conducted. Human sperm were incubated with various concentrations (0, 1, 5, or 25 mg l−1) of Aroclor 1254 for different amounts of time (3 and 6 h) in vitro. Sperm motility parameters were analyzed with computer-assisted sperm analysis (CASA). The proportion of sperm with high mitochondrial membrane potential (ΔΨm) and the levels of intracellular reactive oxygen species (ROS) were detected to explore the probable cause of sperm impairment. Human sperm exposed to continuous Aroclor 1254 exhibited: (i) reduced sperm motility and kinematic parameters, (ii) a proportion of sperm with high ΔΨm that decreased in a dose-dependent manner (P < 0.05), and (iii) increased levels of ROS compared with controls (P < 0.05). In conclusion, Aroclor 1254 can decrease sperm motility, which may culminate in increased ROS and general mitochondrial dysfunction, thus affecting the fertilization potential of sperm. Our findings suggest a broader understanding of the effect of Aroclor 1254 on human sperm.

Azoospermia and testicular biopsy before intra-cytoplasmic sperm injection: Does the type of anesthesia make a difference?

Aim:Azoospermia is the most common form of male factor infertility, due to which sperms are retrieved for intra-cytoplasmic sperm injection (ICSI) under general or local anesthesia. The aim of the present study was to compare the efficacy of general and local anesthesia in an attempt to extract a sufficient quality of sperm for men with azoospermia, who intend to undergo ICSI.Materials and Methods:A total number of 50 patients with azoospermia, who were within the age range of 20-40, were randomly scheduled for testicular sperm extraction with either general or local anesthesia before ICSI. The hormonal abnormalities and severe varicocele were evaluated and treated before the testicular sperm extraction. The inclusion criteria obliged the researchers to focus on men with azoospermic, those who were diagnosed by two semen analyses after a 3-day abstinence from coitus according to the modified WHO criteria, and 19< body mass index ≤28. The concentration, motility, and morphology of the spermatozoa of the two groups were also compared.Results:A total number of 76 men, with a mean age of 35.1 ± 6.0, were selected, 26 were excluded from the study, and the remaining participants were randomly divided into two groups such as general anesthesia and lidocaine group. According to the results, the differences between the values of sperm parameters within various hours after the testicle biopsy were not statistically significant. Also, there was no significant difference between the two groups in terms of sperm motility and sperm morphology during various timing after the processing.Conclusion:The present study demonstrated that there was no evidence found for values of sperm parameters between the two groups, undergoing local and general anesthesia, within various hours after the testicular biopsy. Further investigations with more focus on concentration-dependent lidocaine on human sperm parameters need to be carried out.

The Role of Honey Supplementation to Cryopreservation Solution on Human Sperm Parameters and DNA Integrity during Cryopreservation

The Role of Honey Supplementation to Cryopreservation Solution on Human Sperm Parameters and DNA Integrity during Cryopreservation

Effect of L- carnitine and Co Enzyme Q10 Addition to SMART Pro- Medium on Human Sperm Parameters During In Vitro Sperm Activation

Effect of L- carnitine and Co Enzyme Q10 Addition to SMART Pro- Medium on Human Sperm Parameters During In Vitro Sperm Activation

The potential adverse effects of khat chewing and cigarette smoking on human sperm parameters

The combined effect of khat chewing and cigarette smoking is confusing and no standard general guideline documented. The current study aims to evaluate the potential adverse effects of khat chewing and cigarette smoking on human sperm parameters of patients. Semen samples were collected from 507 patients of couples consulting for infertility recruited in this study and divided into three groups; (i) non-smokers and non-khat chewers group (n=77), (ii) smokers group (n=142) and (iii) smokers and khat chewers group (n=288). Seminal volume, sperm count, motility, vitality, and normal morphology were evaluated according to guidelines of World Health Organization (WHO). A highly significant differences (p<0.0001) were recognized between the three groups in volume (ml) of semen and the percentage of count, motility, vitality and normal morphology of the sperms. Moreover, semen volume (ml) was significantly (p<0.010) connected with sperm motility (%) (r=0.100) and normal morphology (%) (r=0.168), also, sperm count (mill/ml) was significantly (p<0.010) associated with sperm motility (%) (r=0.379), normal morphology (%) (r=0.386) and vitality (%) (r=0.244). In addition, sperm motility (%) was highly and significantly (p=0.0001) correlated with sperm normal morphology (%) and vitality (%) (r=0.446; r= 0.284, respectively). A significant (p=0.000) correlation also conducted between sperms with normal morphology (%) and sperm vitality (%) (r=0.225). These findings suggest that chewing khat and cigarette smoking may be a source of high oxidative stress and accordingly deteriorate semen quality.

Effect of Gonadotropins Addition to SMART Medium on Human Sperm Parameters and Chromatin Structure during in vitro Sperm Activation

A study was conducted to investigate the effect Gonadotropins supplied to culture medium on human sperm parameters and sperm DNA structure during in vitro sperm activation. Thirty infertile patients were included. Semen sample was divided into 3 aliquots. One mL of SMART medium either alone (control group) or supplied with one concentration of Gn (0.25 IU or 0.5 IU) was over layered the pellet, and the three tubes were incubated at 37 o C for 30 min in air incubator. Sperm concentration, motility, grades activity, pro gressive motility, normal morphology and agglutination were assessed pre- and post-activation in vitro. Results of the present study revealed that, percentage, sperm motility, progressive motility, normal sperm morphology and sperm DNA fragmentation index (DFI) were significantly enhanced (P<0.05) for both treated groups as compared to the control group post - sperm activation in vitro. Using 0.5IU hormones Gn within SMART medium (G3) was significantly improved (P<0.05) sperm parameters post- sperm activation in vitro as compared to G2 group (0.25IU hormones). Post-sperm activation in vitro significant increment (P<0.05) in the sperm DNA fragmentation index (DFI) was assessed when using SMART medium enrich with 0.5IU hormones as compared to 0.25IU. The result showed that addition of hormones (GN) to the culture medium enhance sperm motility (%), percentage of progressive sperm activity and normal sperm morphology (%) using centrifugation techniques.

Distinct isolates of uropathogenicEscherichia colidifferentially affect human sperm parametersin vitro

Sperm motility and vitality are decreased in male genital tract infection. Uropathogenic Escherichia coli (UPEC) are frequently associated with sperm parameter loss, but there are no reports to date regarding the effects of different E. coli isolates on human spermatozoa. The aim of this work was to compare the effect in vitro of different E. coli isolates on human sperm parameters. Normal spermatozoa were incubated with E. coli isolated from nine men with urinary tract infection. After 1 h of incubation, sperm motility, vitality and mitochondrial membrane potential (ΔΨm) were measured. The E. coli isolates were serotyped with specific antisera. Sperm motility was decreased with five of nine E. coli isolates. Two UPEC were typed as O6 strains, and they did not decrease sperm motility in the same experimental conditions as the other five isolates, despite the described high pathogenicity of the O6 strain in urogenital infections. Neither UPEC analysed affected vitality or ΔΨm. UPEC isolates were shown to be heterogeneous in their effects, suggesting the need to characterise the pattern defining the pathogenicity of E. coli on human spermatozoa.

Environmental and occupational pesticide exposure and human sperm parameters: A systematic review

Of continuing concern are the associations between environmental or occupational exposures to pesticides and semen quality parameters. Prior research has indicated that there may be associations between exposure to pesticides of a variety of classes and decreased sperm health. The intent of this review was to summarize the most recent evidence related to pesticide exposures and commonly used semen quality parameters, including concentration, motility and morphology. The recent literature was searched for studies published between January 2007 and August 2012 that focused on environmental or occupational pesticide exposures. Included in the review are 17 studies, 15 of which reported significant associations between exposure to pesticides and semen quality indicators. Two studies also investigated the roles genetic polymorphisms may play in the strength or directions of these associations. Specific pesticides targeted for study included dichlorodiphenyltrichloroethane (DDT), hexachlorocyclohexane (HCH), and abamectin. Pyrethroids and organophosphates were analyzed as classes of pesticides rather than as individual compounds, primarily due to the limitations of exposure assessment techniques. Overall, a majority of the studies reported significant associations between pesticide exposure and sperm parameters. A decrease in sperm concentration was the most commonly reported finding among all of the pesticide classes investigated. Decreased motility was also associated with exposures to each of the pesticide classes, although these findings were less frequent across studies. An association between pesticide exposure and sperm morphology was less clear, with only two studies reporting an association. The evidence presented in this review continues to support the hypothesis that exposures to pesticides at environmentally or occupationally relevant levels may be associated with decreased sperm health. Future work in this area should focus on associations between specific pesticides or metabolic products and sperm quality parameters. Analysis of effects of varying genetic characteristics, especially in genes related to pesticide metabolism, also needs further attention.

In vitro effects of cationic compounds on functional human sperm parameters

To determine the mechanisms of spermicidal activity by cationic compounds. Invitro study with human sperm. Academic research institute in collaboration with a university hospital. Normozoospermic men providing samples for routine analysis or assisted reproductive technologies. Invitro incubation of sperm with commercially available cationic surfactants. The effects of benzalkonium bromide (C12Bzk), pyridinium bromide (C12Pyr), and dodecyl trimethylammonium bromide (C12TAB) on human sperm viability, motility, mitochondrial status, capacitation, acrosomal status, and calcium movements were monitored. There was a universal decrease in mitochondrial membrane potential after 180 minutes. Incubation with C12Bzk led to less capacitated sperm at LD50 after 180 minutes (46.0%) as well as to 20.2% of cells significantly increasing their intracellular calcium. C12Pyr was the stronger compound, affecting capacitation and acrosomal status at LD50 and promoting a higher calcium influx (81.7%). C12TAB increased the number of acrosome-reacted sperm at LD50 after 180 minutes and led to a significant increase in calcium signaling (61.2% and 67.9%, respectively). Cationic surfactants seem to impair sperm function at various levels, although C12Pyr was shown to be more effective. Given the concentrations used, these compounds are not necessarily acting as surfactants. To our knowledge, this is the first study that evaluates contraceptive compounds by monitoring sperm calcium signaling, and this seems to be a sensitive parameter in terms of determining the action of putative spermicides.