Search

Is seminal oxidation-reduction potential (ORP) clinically relevant to reproductive outcome? Prospective observational study including a total of 144 couples who had an intracytoplasmic sperm injection (ICSI) cycle between June 2018 and December 2020. The study included patients undergoing fresh ICSI cycles with autologous gametes. Cycles that had day 3 embryo transfers and cryopreservation cycles were excluded. There was no restriction on patients with severe male infertility; couples with unexplained infertility and unexplained male infertility were included, those with azoospermia were excluded. Semen analysis, seminal ORP as determined by means of the MiOXSYS system, sperm DNA fragmentation (SDF) and reproductive outcomes (fertilization, blastocyst development, clinical pregnancy and live birth) were determined. Seminal ORP was significantly negatively correlated with fertilization rate (r = –0.267; P = 0.0012), blastocyst development rate (r = –0.432; P < 0.0001), implantation/clinical pregnancy (r = –0.305; P = 0.0003) and live birth (r = –0.366; P < 0.0001). Receiver operating characteristic curve analysis showed significant predictive power for ORP for fertilization (≥80%; area under the curve [AUC] 0.652; P = 0.0012), blastocyst development rate (≥60%; AUC 0.794; P < 0.0001), implantation/clinical pregnancy (AUC 0.680; P = 0.0002) and live birth (AUC 0.728; P < 0.0001). Comparable results were obtained for SDF (fertilization: AUC 0.678; blastocyst development: AUC 0.777; implantation/clinical pregnancy: AUC 0.665; live birth: AUC 0.723). Normal sperm morphology showed the lowest predictive power for all reproductive outcome parameters. With male age as confounding factor, ORP (cut-off value of 0.51 mV/106 sperm/ml) has significant (P < 0.04667) effects on odds ratios for all reproductive outcome parameters. Multivariate logistic regression to investigate potential seminal and female confounding factors revealed that seminal ORP significantly (P < 0.0039; P < 0.0130) affects reproductive outcome. Seminal ORP is relevant for good fertilization, blastocyst development, implantation, clinical pregnancy and live birth. [ABSTRACT FROM AUTHOR]

Objective: To compare different staining methods to evaluate human sperm morphology. Design: Prospective study. Setting: Patients at the Departments of Dermatology and Urology, University of Jena, Germany. Patient(s): A total of 94 randomly collected patients attending the andrological outpatient clinics of the Departments of Dermatology and Urology, University of Jena, Germany. Intervention(s): None. Main Outcome Measure(s): Statistical comparison of resultant standard morphological parameters (mean percentages) after staining according to Papanicolaou and Shorr methods and with Testsimplets® prestained slides. Result(s): All morphological parameters investigated (percent normal morphology, percent head, midpiece, and flagellar abnormalities) correlated statistically significantly positively, however with markedly lower correlation coefficients for the Testsimplets®results. As compared with the mean Papanicolaou (4.78% ± 2.54%) and Shorr staining (4.75% ± 2.64%) results, a statistically significantly lower percentage of morphologically normal spermatozoa was determined after using the Testsimplets® slides (3.89% ± 2.53%). In general, the mean values of all parameters differed for all comparisons with the Testsimplets® slides and especially for the percentage of flagellar defects but not between the Papanicolaou and the Shorr staining results. Conclusion(s): The results show an extensive agreement between the Papanicolaou- and Shorr-stained smears, whereas Testsimplets® staining exhibited statistically significant deviations. Because the correct evaluation of sperm morphology is of essence within the scope of assisted reproduction and in andrological diagnostics, the use of rapid staining methods cannot be recommended. [Copyright &y& Elsevier]

Objective: To investigate the molecular mechanism for declining sperm motility in older men. Design: Retrospective and prospective study. Setting: University-based andrology unit. Patient(s): Semen was collected from 2,111 patients and sperm donors. Intervention(s): None. Main Outcome Measure(s): Age, sperm concentration, ejaculate volume, sperm motility (including velocity average path, velocity straight line, velocity curvilinear, linearity, and lateral head displacement), normal sperm morphology, percentage of abnormally blue-stained flagella, and total and free T, FSH, and LH concentrations were analyzed. In addition, the zinc concentration in the seminal fluid, flagella, and the whole spermatozoa was measured by means of atomic absorption spectrometry. Result(s): Patient age correlated negatively with T concentration and sperm motility, including velocity, but positively with the percentage of abnormally stained flagella and the flagellar zinc. Whereas the percentage of abnormally stained flagella correlated negatively with motility and sperm velocity, there was a positive relationship with the flagellar zinc content. Flagellar zinc content was negatively correlated with motility. Testosterone showed a positive relationship with motility and sperm concentration. Conclusion(s): Because the epididymis is functionally T-dependent, our data suggest that the removal of zinc from the outer dense fibers during epididymal sperm maturation is affected in aging men, which in turn will result in decreased sperm motility. [Copyright &y& Elsevier]

Objective: To investigate whether there is an impact of different sources of reactive oxygen species (ROS) on sperm functions. Design: Prospective study. Setting: Patients at the Center for Dermatology and Andrology, Giessen, Germany. Patient(s): Semen collected from 63 randomly collected patients attending the IVF unit of the University of Giessen, Germany. Intervention(s): Only patients with nonleukocytospermia were included in this study. Main outcome measure(s): Sperm count and motility before and after sperm separation by swim-up, morphology, DNA fragmentation, and extrinsic (by leukocytes) and intrinsic ROS production (by spermatozoa) were evaluated. Result(s): Leukocytes correlated significantly with extrinsic ROS production (r = 0.576), but markedly less with intrinsic ROS production (r = 0.296). Sperm count, morphology, and motility in the ejaculate were markedly more affected by extrinsic than by intrinsic ROS. The DNA fragmentation was strongly positively correlated with intrinsic ROS production, whereas this correlation was weaker for extrinsic ROS production. No correlation was found between DNA fragmentation and the number of leukocytes, whereas the correlations with motility in the ejaculate and the motile sperm count after swim-up were highly significant. Moreover, significant differences were observed for extrinsic and intrinsic ROS production between groups of patients having a high (≥1 × 106/mL) and a low number (<1 × 106/mL) of leukocytes in the ejaculate. Conclusion(s): The origin of ROS seems to have an influence on the site of the damage. Because leukocyte counts <1 × 106/mL caused a significant decrease of motility and DNA integrity, the threshold given by the World Health Organization (WHO) should be re-evaluated. [Copyright &y& Elsevier]

: ObjectiveTo investigate sperm DNA damage in relation to fertilization and pregnancy.: DesignProspective study.: SettingThe Institute of Reproductive Medicine, Giessen, Germany.: Patient(s)Semen collected from 249 patients attending the IVF program.: Main outcome measure(s)The percentage of terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling- (TUNEL-), Fas-, and annexin-V-positive sperm and the proportion of green-fluorescing sperm in the acridine orange stain was determined and correlated with sperm concentration, motility, fertilization, and pregnancy.: Result(s)Significant correlations with the concentration of motile sperm were only found for the acridine orange stain (before and after sperm separation) and for the TUNEL assay (after sperm separation). Moreover, patients whose sperm had a high percentage of DNA fragmentations showed significantly lower pregnancy rates (TUNEL assay: 19.05% vs. 34.65%; acridine orange stain: 24.58% vs. 37.93%). The apoptosis parameters (annexin V binding and Fas expression) showed no statistically significant differences.: ConclusionsOur data clearly demonstrate that DNA fragmentation, as determined by the TUNEL assay, is predictive for pregnancy in IVF. This implies that spermatozoa with DNA fragmentation can still fertilize an oocyte but that when paternal genes are “switched on,” further embryonic development stops, resulting in failed pregnancy. It seems that, at least in the patients we analyzed, apoptosis in the sperm does not play a role for fertilization. This would imply that DNA fragmentation in human spermatozoa is caused by external factors, such as reactive oxygen species, rather than by apoptosis. [Copyright &y& Elsevier]

Both oxidative stress (OS) and reductive stress (RS) are the two extreme facets of redox imbalance that can have deleterious effects on sperm function. However, there is a lack of information on the physiological range of oxidation-reduction potential (ORP). The aim of this study was to investigate the effect of OS and RS on functions and associated molecular changes in normal spermatozoa in order to establish the physiological range of ORP. In the current study, total and progressive motility remained unchanged in spermatozoa exposed to ORP values 0.33 and 0.72 mV/106 sperm/mL. However, a significant (P < 0.05) decline in total and progressive motility were observed at ORP values 1.48, 2.75, −11.24, −9.76 and −9.35 mV/106 sperm/mL. Sperm vitality also decreased significantly (P < 0.0001) at 2.75, −11.24 and −9.76 mV/106 sperm/mL. Spermatozoa exposed to ORP levels 2.75 and −11.24 mV/106 sperm/mL showed a significant (P < 0.01) decrease in mitochondrial membrane potential. Intracellular reactive oxygen species (iROS) production increased (P < 0.05) in spermatozoa exposed to ORP levels of 1.48 and 2.75 mV/106 sperm/mL, while iROS decreased (P < 0.05) at ORP levels −9.76 and −11.24 mV/106 sperm/mL. No significant change in sperm DNA fragmentation was noted in sperm exposed to OS/RS and the values were below the reference range (<19.25%). Western blot analysis revealed decreased expression of CV-ATPA, CIII-UQCRC2 and CIV-MTCO1 proteins at 60 and 120 min (P < 0.05) in both OS and RS conditions. This is the first study to report physiological range of ORP (between −9.76 and 1.48 mV/106 sperm/mL) and to elucidate the role of altered expression of oxidative phosphorylation (OXPHOS) complexes proteins in mediating detrimental effects of oxidative and reductive conditions on sperm functions. A decreased expression of OXPHOS proteins and associated mitochondrial dysfunction contributes to decreased sperm motility and vitality under oxidative and reductive stress. Image 1 • This novel study defines the physiological range of ORP and demonstrates that both OS and RS damages sperm equally. • Semen samples with ORP values ≤ −9.76 mV/106 sperm/mL and ≥1.48 mV/106 sperm/mL can be characterized as pathological RS and OS, respectively. • The proposed ORP will help in the identification of OS and RS induced male infertility. [ABSTRACT FROM AUTHOR]

: ObjectiveTo investigate the influence of zinc on the generation of motility.: DesignProspective study.: SettingOutpatient clinic of the Center of Dermatology and Andrology.: Patient(s)Seventy-three patients and 10 sperm donors.: Intervention(s)Motile spermatozoa were isolated by swim-up and incubated for 1, 2, 4 or 6 hours with DL-penicillamine, 2,3-dimercaptopropan-1-sulfonate, and meso-2,3-dimercapto-succinimic acid at concentrations of 1, 10, and 100 μmol/L.: Main outcome measure(s)Motility was analyzed by means of computer-assisted semen analysis (CASA).: Result(s)Significant dose-dependent changes in nonlinear motility, progressive motility, and velocity straight line (VSL) were observed. After only 1 hour of incubation, nonlinear motility decreased, and progressive motility and VSL increased. The percentage of immotile spermatozoa was not affected. Time-dependent changes in motility were observed on a significantly higher or lower level as compared to the control. Comparing the chelators after 2 hours revealed that DL-penicillamine showed the strongest effect on the sperm.: Conclusion(s)The results imply that chelators can eliminate at least some part of the zinc from the flagellum. This zinc elimination appears to lead to comparable changes of the outer dense fibers as seen in vivo during epididymal maturation, finally resulting in improved motility. [Copyright &y& Elsevier]

• A high sugar diet (HSD) compared to normal chow induces obesity and decreases sperm concentration and progressive motility in male Wister rats. • Metformin and high dose Nigella sativa oil improves obesity, sperm concentration, sperm MMP and serum testosterone in HSD induced obesity. • Low dose Nigella sativa oil improves obesity and serum testosterone compared in HSD induced obesity. • Metformin and Nigella sativa seed oil may provide novel therapeutic options for males with obesity induced infertility. Obesity is a significant global health and socio-economic challenge, and considered an important risk factor for poor health outcomes including male reproductive dysfunction and infertility. As excess adiposity causes testicular dysfunction and infertility, novel therapeutic strategies require investigation. Nigella sativa (Ns) seed oil and metformin have both demonstrated a potential positive effect on obesity, although both remain poorly investigated in male fertility. Therefore, this study aimed to determine the effect of Ns oil and metformin on total body weight (TBW), mitochondrial membrane potential (MMP), serum testosterone and semen parameters in an obese animal model. Wistar rats (n = 54) were divided into six groups: normal chow (NC), high sugar diet (HSD) only, HSD and saline, HSD and metformin (75 mg/Kg/day), HSD and Ns (200 mg/Kg/day) (NS200), HSD and Ns (400 mg/Kg/day) (NS400). Intervention was force fed for the last 8 weeks of the 14 weeks dietary exposures. Results showed that the HSD increased TBW (P = 0.001) and reduced sperm concentration (P = 0.013) and progressive motility (P = 0.009) compared to the NC group. Metformin, NS200 and NS400 improved TBW (P = 0.035, P = 0.006 and P = 0.005, respectively) and testosterone (P < 0.001) compared to the HSD saline group, where metformin and NS400 improved sperm concentration (P < 0.001 and P = 0.049, respectively) and MMP (P < 0.001). There were no changes in sperm motility and viability for all experimental exposures, although NS400 (P = 0.047) negatively affected sperm viability. Metformin and Ns may be novel treatment options in obesity-induced infertility, although a potential negative impact on viability is cautioned for high dose Ns. These results warrant further investigation of Ns and Metformin for the management of obese infertile males. [ABSTRACT FROM AUTHOR]

Unraveling the role of reactive oxygen species and associated oxidative stress (OS) in male reproduction is one of the key areas of male reproductive research. This article illustrates the scientific landscape of OS in male reproductive research over the past several decades (1941–2018) using a scientometric approach. Scientometric data (articles per year, authors, affiliations, journals, and countries) on OS related to male reproduction were retrieved from the Scopus database and analyzed for each decade. Our analysis revealed an increasing trend in OS-based male reproductive research from 1941 to 2018 with a steep raise in publications and research collaborations starting from the period 1991–2000 (R2 = 0.81). Semen abnormalities and varicocele were the major areas investigated in relation to OS with the highest positive trend in publications from the time interval 1981–1990 to 2011–2018. Analysis of publications based on OS assessment techniques revealed chemiluminescence (n = 180) and evaluation of antioxidants (n = 300) as the most widely used direct and indirect tests, respectively. Furthermore, prognostic/diagnostic studies on OS evaluation increased significantly over the time. Our analysis highlights the evolution of OS in male reproductive research and its emergence as an important prognostic and diagnostic tool in the evaluation of male infertility. Image 1 • Scientometric analysis revealed an increasing trend in OS-based male reproductive research from 1941 to 2018. • Chemiluminescence was the most widely used direct technique to assess OS. • Showcase the evolution of OS in male reproductive research. [ABSTRACT FROM AUTHOR]

Objective: To test the ability of the smartphone-based YO Home Sperm Test to accurately and precisely measure motile sperm concentration (MSC) versus the SQA-Vision, an automated laboratory semen analyzer.Design: MSC compared for the YO device on Galaxy and iPhone smartphones versus the SQA-Vision in a double-blind manner.Setting: Academic medical center.Patient(s): Donor semen samples from 24 men in 144 aliquots.Intervention(s): None.Main Outcome Measure(s): Accuracy, precision, and agreement assessed between the YO device and the SQA-Vision for MSC results.Result(s): The YO device demonstrated good correlation and good to moderate agreement with the SQA-Vision for MSC results up to a range of 94 × 106/mL with Pearson and concordance correlation coefficient above 0.92. The YO also showed a very high level of accuracy (97.8%) with positive and negative percent agreement above 94%. The difference in coefficient of variation between the YO and the SQA-Vision was low (between 9.4% and 11.2%) and not statistically significant. The precision among the YO phone devices was lower (16.0%) than the manufacturer's claim of ≤20%.Conclusion(s): The smartphone-based device has a high level of accuracy and precision when compared with the SQA-Vision. It can detect samples with abnormally "low" MSC (below 6 × 106/mL cutoff), which supports its use as an effective home sperm test for screening "low" and "moderate/normal" MSC cases. In addition, the device effectively identifies varying levels of normal MSC in a precise manner over a wide range of normal MSC. Thus, the YO Score can improve patient satisfaction and empowerment. [ABSTRACT FROM AUTHOR]

Abstract The cryopreservation of spermatozoa was introduced in the 1960s as a route to fertility preservation. Despite the extensive progress that has been made in this field, the biological and biochemical mechanisms involved in cryopreservation have not been thoroughly elucidated to date. Various factors during the freezing process, including sudden temperature changes, ice formation and osmotic stress, have been proposed as reasons for poor sperm quality post-thaw. Little is known regarding the new aspects of sperm cryobiology, such as epigenetic and proteomic modulation of sperm and trans-generational effects of sperm freezing. This article reviews recent reports on molecular and cellular modifications of spermatozoa during cryopreservation in order to collate the existing understanding in this field. The aim is to discuss current freezing techniques and novel strategies that have been developed for sperm protection against cryo-damage, as well as evaluating the probable effects of sperm freezing on offspring health. [ABSTRACT FROM AUTHOR]

To manage male infertility caused by hormonal imbalance, infections and other predicaments, multifarious treatment strategies are emerging worldwide. Contemporary treatments, such as assisted reproductive techniques, are costly with low success rates of only 10–30%; however, herbal remedies are gaining more attention as an alternative or supplementary therapeutic modality for male infertility. The beneficial effects induced by oral intake of the roots of a small evergreen shrub, Withania sominifera (Ashwagandha) on semen quality of infertile men have previously been studied. Oral intake of Ashwagandha roots has been found to inhibit lipid peroxidation, improve sperm count and motility, and regulate reproductive hormone levels. The molecular mechanisms of these effects, however, are yet to be unveiled. In this review, we will discuss the role of herbal medicines in male infertility; provide a detailed analysis of various human and animal studies involving Withania somnifera; describe a proposed direct oxidative mechanism involving mitigation of oxidative stress as well as an indirect mechanism consisting of a gamma - aminobutyric acid-like-mimetic pathway ameliorating hormonal balance through crosstalk among different endocrine glands to improve male fertility; and how Withania somnifera supplementation mitigates risk factor-induced male infertility as well as ameliorates male fertility. [ABSTRACT FROM AUTHOR]

Objective: To develop a method whereby sperm with phosphatidylserine externalization can be separated from those without this feature. Because annexin V binds phosphatidylserine, this study is using this feature to select functional spermatozoa. In addition, the relationship between annexin V binding in human spermatozoa and normal sperm morphology according to strict criteria was to be assessed. Design: Prospective study. Setting: Department of Obstetrics and Gynaecology of Stellenbosch University at Tygerberg Academic Hospital, Tygerberg, South Africa. Patient(s): Semen from 14 healthy sperm donors. Exclusion criterion was the presence of less than 20 × 106/mL total motile spermatozoa in the original sample. Main Outcome Measure(s): Annexin V-negative sperm, annexin V-positive sperm, normal sperm morphology. Intervention(s): An aliquot of a semen sample after double density gradient centrifugation was incubated with annexin V fluorescein isothiocyanate conjugate (FITC). Cell fluorescence signals were determined using a FACScalibur flow cytometer equipped with a FACSSort fluidic sorting module. The sorting procedure delivered two sperm subpopulations: annexin V-negative and annexin V-positive. Morphology slides were made and the sperm morphology was assessed according to strict criteria. Result(s): There was a significant enrichment of annexin V-negative sperm as well as morphologically normal sperm in the annexin V-negative subgroup after separation with flow cytometry. The percentage of morphologically normal sperm increased from 8.3% in the control to 11.9% in the annexin V-negative fraction, whereas the percentage of annexin V-positive sperm decreased to 5.7%. Conclusion(s): The annexin V-negative sperm subpopulation had morphologically superior quality sperm compared to annexin V-positive sperm. It is important to select morphologically normal sperm during intracytoplasmic sperm injection (ICSI) as it may contribute to increased implantation and pregnancy rates (PR). [Copyright &y& Elsevier]