Increased Sperm DNA Fragmentation Research Articles

Bleomycin in vitro exposure decreases markers of human male gamete competence

To investigate the in vitro impact of bleomycin on human sperm DNA integrity, functionality, and morphology, with the aim of elucidating the underlying mechanism and anticipating potential repercussions on patients' reproductive function. Controlled laboratory-based in vitro investigation. Surplus human ejaculate donated for research by 45 reproductive-age participants exhibiting normozoospermic sperm parameters following clinical semen analysis. None of the participants had received a cancer diagnosis or undergone radiotherapy, chemotherapy, or both. After clinical semen analysis, sperm samples were centrifuged, diluted in sperm preparation medium (SPM), and exposed to bleomycin (100 μg/mL) for two hours at 37ºC in a humidified incubator with 5% CO2. In vitro human sperm competence was evaluated by comparing raw sperm, sperm incubated with SPM, and sperm exposed to bleomycin. Competence indicators included sperm motility, vitality, DNA and acrosome integrity, and mitochondrial membrane potential. Transmisson electron microscopy was employed to correlate the ultrastructural morphological findings with functional assays. Exposure to bleomycin for two hours in vitro significantly (P < 0.001) decreased sperm vitality, motility, and chromatin condensation compared to raw and control sperm. It also significantly (P < 0.001) increased sperm DNA fragmentation and the proportion of sperm with low mitochondrial membrane potential. Additionally, bleomycin significantly (P < 0.05) retarded the acrosomal response compared to control but did not affect the formation of intracellular and extracellular reactive oxygen species. Bleomycin-induced ultrastructural morphological changes supported the detected functional alterations. Bleomycin negatively impacts male gamete competency in humans. Healthcare professionals should vigilantly monitor and further investigate the gonadotoxicity effects of bleomycin, in addition to its recognized lung toxicity. Meanwhile, it is recommended that cancer patients undergoing bleomycin-containing chemotherapy regimens receive guidance on fertility preservation strategies.

Влияние распространенных бактериальных инфекций на индекс фрагментации ДНК сперматозоидов. Обзор литературы

Sperm DNA fragmentation is an important factor in male infertility, miscarriage and unsuccessful ART cycles. The purpose of thisreview wasto summarize and briefly analyze the available data on the effect of common infections of the male genitourinary tract on sperm DNA fragmentation (SDF), the mechanisms of their influence, efficacy and outcomes of treatment. Based on the results of these studies, it can be said that Chlamydia trachomatis, Ureaplasma spp. and Mycoplasma genitalium have a significant effect and lead to a pronounced increase in SDF. The effect of opportunistic bacteria on SDF depends on the type of microorganisms, their diversity and quantity. The presence of certain bacteria is associated with higher levels of ,some are presumably associated with beneficial effects and lower levels of SDF. Candidiasis can presumably affect SDF, but the available data are insufficient for an accurate answer. Treatment ofsuch infectionsleads to a decrease in sperm DNA fragmentation index, an increase in the effectiveness of ART and spontaneous pregnancy rates. Keywords: sperm DNA fragmentation, infection, male infertility, oxidative stress, fertility, semen analysis, sperm, sperm chromatin.

The paternal clock: Uncovering the consequences of advanced paternal age on sperm DNA fragmentation

The objective of the study was to investigate the relationship between advanced paternal age and sperm DNA fragmentation (SDF) levels, specifically identifying the age at which a significant increase in SDF occurs. This is a retrospective cohort study involving 4250 consecutive semen samples from patients presenting for infertility evaluation. Patients were stratified into seven age groups: < 26 (n = 36; 0.8 %), 26–30 (n = 500; 11.8 %), 31–35 (n = 1269; 29.9 %), 36–40 (n = 1268; 29.8 %), 41–45 (n = 732; 17.2 %), 46–50 (n = 304; 7.2 %), > 50 (n = 141; 3.3 %). The main outcome measures included comparing mean SDF levels throughout different age groups and assessing the prevalence of normal, intermediate, and high SDF among the age groups. A positive correlation was observed between paternal age and SDF (r = 0.17, p < 0.001). SDF remained relatively constant until the age of 35 but increased significantly beyond age 35. Mean SDF levels in the older age groups (36–40, 41–45, 46–50, and >50 years) were significantly higher than in the younger age groups (<26, 26–30, and 31–35 years) (p < 0.001). The prevalence of normal SDF was highest among the younger age groups, whereas the prevalence of high SDF was highest among the older age groups. Interestingly, the prevalence of intermediate SDF was relatively constant throughout the age groups (ranging between 29.8 % to 37.2 %). The increase in SDF after the age of 35 highlights the importance of considering male age in infertility evaluations. Assessing SDF in men over the age of 35 is crucial in couples seeking to conceive.

P-015 Impaired spermatogenesis in colitis: the role of interferon-γ mediated ferroptosis in sertoli cells by colitic CD4+ T cells

Abstract Study question Does colitis have an impact on spermatogenesis and what could be the potential mechanism? Summary answer Colitis causes sertoli cell ferroptosis and disrupts spermatogenesis in mice due to the migration of colitic CD4+ T cells that produce interferon-γ. What is known already In recent years, there has been a significant increase in rheumatic and inflammatory diseases. Epidemiological data reveals that dysregulated immune conditions can directly or indirectly impact male fertility and sexual function in patients of reproductive age. Inflammatory bowel disease (IBD), which encompasses Crohn’s disease and ulcerative colitis, is a chronic, recurring gastrointestinal inflammatory disorder characterized by bloody stools and weight loss. A study from a Swedish national cohort showed that IBD patients had diminished sperm fertility and increased sperm DNA fragmentation compared to healthy individuals. However, the precise mechanisms underlying male fertility problems in IBD remain unclear. Study design, size, duration In this study, C57BL/6 mice were kept under specific pathogen-free conditions. A murine colitis model that simulates human colitis histological features and symptoms was created using Dextran Sulfate Sodium (DSS). To establish the model, 2-2.5% DSS was added to the mice’s drinking water from days 0 to 7, followed by a three-day recovery period. Participants/materials, setting, methods Testis, epididymis and colon sections were stained with hematoxylin and eosin for histological analysis. The localization and phenotype of infiltrated lymphocytes in epididymis and testis were studied by immunostaining and flow cytometry. Sperm functions including motility, viability, and DNA integrity were assessed by standard assays. Iron, malondialdehyde, and reactive oxygen species (ROS) levels were analysed by commercial kit. Main results and the role of chance Colitis was found to impair male fertility in DSS-induced colitic mice, causing testis and epididymis tissue damage along with inflammatory cell infiltration. Colitis altered the immune cell phenotypes in the testis and epididymis. The proportion and absolute cell count of CD4+ T cells and CD19+ B cells increased after colon inflammation. Testicular sertoli cells in colitic mice exhibited ferroptosis, characterized by increased levels of iron deposition, malondialdehyde, and ROS. We assessed sperm parameters from the cauda epididymis of mice. Compared to the control group, there were significant reduction in sperm concentration, motility, viability and DNA integrity in the group with murine colitis induction. Consistently, we observed lower numbers of active pups from the fathers of the colitis group compared to control mice. CD4+ T cells with interferon-γ producing capabilities accumulated in the testes of colitis mice. Leukocyte flow cytometric analysis and parabiotic mouse model further demonstrated that colitis promoted pathogenic CD4+ T cells migration to the testis through circulation. The treatment of anti-CD4 antibody, anti-interferon-γ antibody or ferroptosis inhibitor can attenuate the colitis-associated testis pathologies. Limitations, reasons for caution Our study was conducted exclusively in mice. The pathogenic effect and the corresponding mechanism of colitis on male fertility in humans still needs to be confirmed. Wider implications of the findings Our team identified the mechanism linking dysregulated gut immune cell activity to testicular dysfunction. By identifying specific molecules on colitic immune cells, which allow their infiltration into the testis, may pave the way for a new therapeutic approach potentially improving the prognosis of colitis and related fertility impairments. Trial registration number not applicable

P-053 Comparative analysis between microfluidic sperm sorting and Swim-up technique to select spermatozoa with better characteristics in hyper viscous samples

Abstract Study question What is the best technique to select spermatozoa in hyper viscous semen samples? Summary answer Microfluidics is the best technique that allows to select spermatozoa with better seminal characteristics and hyperactivated motility, and less sperm DNA fragmentation. What is known already Density gradient centrifugation and Swim-up (SU) are the two traditional selection methods used during assisted reproduction techniques (ART) to collect the highly motile sperm fraction. Many studies reported that centrifugation increase sperm DNA fragmentation and oxidative status which could affect reproductive outcomes. To overcome the limitations of centrifugation, an alternative approach by using microfluidic-based devices was recently designed. In microfluidic protocol, sample is applied to the device’s inlet port, connected to a lower chamber. This chamber consists of a microporous filter, where the most motile spermatozoa migrate upward through the filter. Study design, size, duration An observational study was conducted on 29 semen samples from patients undergoing routine semen analysis for couple infertility in the Andrology Laboratory of Careggi University Hospital of Florence from January 2023 to January 2024. In these samples we compare two different methods for sperm selection: indirect SU technique and microfluidic devices (MSS). Participants/materials, setting, methods We evaluated seminal parameters (sperm progressive and total motility, viability, and concentration), sperm kinematic parameters by Computer-Assisted Sperm Analysis (C.A.S.A. system), sperm DNA fragmentation by TUNEL/PI assay and sperm oxidative status by CellROX® Orange (which we have recently shown to identify a fraction of viable oxidized spermatozoa with better characteristics (Traini G., et al. 2022)) in 29 normozoospermic and 18 hyper viscous semen samples selected by SU and MSS. Main results and the role of chance In normozoospermic samples, sperm selection with both indirect SU technique and microfluidic devices, ameliorated sperm characteristics compared to not selected samples. Spermatozoa selected by MSS showed statistically significant higher progressive and total motility, viability and concentration compared to those selected by indirect SU. Moreover, an increase in most of kinematic semen parameters (like VAP, VCL and VSL) and hyperactivated motility was observed in MSS selected samples. Also, sperm DNA fragmentation was significantly reduced after MSS selection compared to indirect SU. We evaluated the percentage of viable oxidized spermatozoa by using CellROX® Orange. A significant increase of the percentage of CellROX® Orange positive spermatozoa was observed in most of MSS selected samples. In all 18 hyper viscous samples, the results obtained in normozoospermic samples were confirmed. Indeed, an overall improvement of sperm progressive and total motility, viability and concentration together with an increase in most of kinematic semen parameters and in hyperactivated motility were evidenced in MSS selected spermatozoa respect to those selected by indirect SU. Again, a significant decrease of sperm DNA fragmentation and an increase of the percentage of CellROX® Orange positive spermatozoa, were observed after MSS selection. Limitations, reasons for caution The number of semen samples included in the study is not very elevated. Results obtained in hyper viscous semen samples should be confirmed in a greater number of samples. Wider implications of the findings Our data indicate that MSS selects the sperm fraction with better characteristics and less DNA fragmentation. These results suggest that MSS could improve ART outcomes, particularly for specific categories of patients, including hyper viscous semen samples, for which the selection by SU does not always guarantee good yields. Trial registration number not applicable

Effects of Slow Freezing and Vitrification of Human Semen on Post-Thaw Semen Quality and miRNA Expression.

Semen cryopreservation has played an important role in medically assisted reproduction for decades. In addition to preserving male fertility, it is sometimes used for overcoming logistical issues. Despite its proven clinical usability and safety, there is a lack of knowledge of how it affects spermatozoa at the molecular level, especially in terms of non-coding RNAs. Therefore, we conducted this study, where we compared slow freezing and vitrification of good- and poor-quality human semen samples by analyzing conventional sperm quality parameters, performing functional tests and analyzing the expression of miRNAs. The results revealed that cryopreservation of normozoospermic samples does not alter the maturity of spermatozoa (protamine staining, hyaluronan binding), although cryopreservation can increase sperm DNA fragmentation and lower motility. On a molecular level, we revealed that in both types of cryopreservation, miRNAs from spermatozoa are significantly overexpressed compared to those in the native semen of normozoospermic patients, but in oligozoospermic samples, this effect is observed only after vitrification. Moreover, we show that expression of selected miRNAs is mostly overexpressed in native oligozoospermic samples compared to normozoospermic samples. Conversely, when vitrified normozoospermic and oligozoospermic samples were compared, we determined that only miR-99b-5p was significantly overexpressed in oligozoospermic sperm samples, and when comparing slow freezing, only miR-15b-5p and miR-34b-3p were significantly under-expressed in oligozoospermic sperm samples. Therefore, our results imply that cryopreservation of normozoospermic sperm samples can modulate miRNA expression profiles in spermatozoa to become comparable to those in oligozoospermic samples.

Advanced Paternal Age: A New Indicator for the Use of Microfluidic Devices for Sperm DNA Fragmentation Selection.

New social conditions and progress in ART have both contributed to the delay in parenthood in developed countries. While the effects of maternal age have been widely studied, paternal age is poorly understood, and there are no specific guides on ART techniques to treat its deleterious effects. It is known that there is an increase in sperm DNA fragmentation (SDF) in elderly men, and new sperm selection devices using microfluids have been developed. This study analyses 189 ICSI cycles with donor oocytes performed between January 2018 and February 2022. Spermatozoa were selected using an MSS device or density gradients, followed by ICSI fertilization and fresh/thawed embryo transfer. We assessed the association between the selection technique, paternal age (< or ≥45) and reproductive outcomes. Fertilization (FR), blastulation (BR), implantation (IR), live-birth (LBR) and miscarriage (MR) rates were calculated. The results showed significantly higher IR (57.7% vs. 42.5%) and LBR (42.9% vs. 30.3%) when applying MSS selection, and particularly higher BR, IR and LBR when the paternal age was equal to or over 45 years (BR: 64.4 ± 23% vs. 50.1 ± 25%, IR: 51.5% vs. 31.6% and LBR: 42.4% vs. 23.7%). We also found a negative correlation between BR and paternal age (r2 = 0.084). The findings show that MSS enhances success in assisted reproduction cycles with ICSI, especially in couples with advanced paternal age. We propose advanced paternal age as a new indicator for the application of sperm selection techniques that reduce fragmentation.

Age-Related COVID-19 Influence on Male Fertility.

The impact of coronavirus on the reproductive health of men attracts the special attention of many researchers. While studies suggest changes in sperm parameters and the possibility of testicular inflammation, further studies are needed to elucidate any potential age-related changes in these findings, which is the purpose of the present study. The semen quality parameters, cytokine concentration, and markers of the pro- and antioxidant system were assessed in 60 men five to seven months after the coronavirus infection and in 77 controls (without a history of coronavirus infection). Additionally, participants were divided into two age groups: less than 35 years and 35 years or older. Notably increased round cell count in ejaculate and reduced sperm hyaluronan binding ability were observed among post-infection patients younger than 35 years. In the same group, a decline in seminal plasma zinc levels and nitrotyrosine in the cell fraction was found. In men over 35 years of age, Coronavirus Disease 2019 (COVID-19) led to increased sperm DNA fragmentation, a decrease in the total antioxidant capacity, and an elevation in the levels of interleukin-1β and interleukin-10. The concentration of interleukin-1β decreased over time following recovery in all affected patients. The data obtained suggest the potential adverse impact of the coronavirus infection on male reproductive health; however, these effects appear to be age-dependent.

Effects of cigarette smoking on semen quality, reproductive hormone levels, metabolic profile, zinc and sperm DNA fragmentation in men: results from a population-based study.

Cigarette smoking seems to have a negative impact on men's reproductive health, but our knowledge of its effects on the reproductive function of Russian men is still very limited. The purpose of this study was to evaluate the effect of cigarette smoking on semen quality, including sperm DNA fragmentation, hormonal, zinc and metabolic status in young men from the general multi-ethnic Russian population (n=1,222, median age 23 years) and to find out the ethno-specific effects of smoking by comparing male groups of different ethnicity. Each participant filled out a standardized questionnaire, provided one blood and semen sample. Semen parameters, serum reproductive hormones, lipids, glucose, uric acid and seminal zinc were analyzed. Participants were classified as smokers (n=450) and non-smokers (n=772), and smokers were stratified into moderate (≤10 cigarettes/day) and heavy (>10 cigarettes/day) smokers. In the entire study population, heavy smokers were characterized by a decrease in semen volume, total sperm count, sperm concentration and motility, and an increase in sperm DNA fragmentation and teratozoospermia compared with non-smokers (p<0.05). There was also a reduction in the serum and seminal zinc level as well as an impairment in metabolic health in smokers compared with non-smokers (p<0.05). No significant differences between smokers and non-smokers were found for serum levels of LH, FSH, inhibin B, testosterone and estradiol. In the second part of our study, the most numerous ethnic groups of Slavs (n=654), Buryats (n=191), and Yakuts (n=125) were selected from the entire study population. Among three ethnic groups, the smoking intensity was higher in Slavs than in Buryats or Yakuts suggesting a greater tobacco addiction in Slavs than in Asians. A decrease in semen parameters and seminal zinc levels, and an increase in sperm DNA fragmentation and teratozoospermia was observed only in smoking Slavs (p<0.05); moderate decrease in testosterone and increase in triglyceride levels were revealed in smoking Yakuts (p<0.05), but no significant changes were detected in smoking Buryats. We concluded that cigarette smoking has an ethno-specific effect on male reproductive function, probably due to the different activity of the seminal antioxidant system, which is yet to be elucidated.

Omega 6/Omega 3 Ratio Is High in Individuals with Increased Sperm DNA fragmentation.

An imbalance between omega-6 and omega-3 fatty acids in sperm has been linked with lipid peroxidation and DNA damage in sperm, indicating a possible correlation to fertility potential. This cross-sectional study involved 56 infertile men (aged 25-45), and assessed the relationship between the omega-6 to omega-3 fatty acid ratio in sperm and seminal plasma with sperm DNA fragmentation. Individuals were categorized based on high or low levels of sperm DNA fragmentation according to two tests (TUNEL and SCSA assay less or greater than 10 and 30%, respectively), and their fatty acid composition, as well as sperm functional tests, were analyzed. Results showed that men with high DNA fragmentation exhibited higher percentages of total saturated, monounsaturated, and omega-6 to omega-3 fatty acid ratios in both sperm (P < 0.001) and seminal plasma (P < 0.001) compared to men with low DNA fragmentation. The percentage of sperm lipid peroxidation, and residual histone (P < 0.05) were higher, while the percentage of sperm motility (P < 0.001) was lower in the former compared to the latter group. Moreover, Pearson's correlation revealed positive associations between the omega-6 to omega-3 fatty acid ratio with sperm lipid peroxidation, DNA fragmentation, and residual histones in both sperm and seminal plasma. Overall, these observations suggest that consumption of omega-3 fatty acids may be related to male fertility potential, as it appears that individuals with a high percentage of omega-3 fatty acids have better sperm quality compared to men with a lower omega-3 fatty acid.

The prevalence of social risk factors for the development of male infertility: smoking, alcohol and narcotic use in men from infertile couples, the influence of the “northern type” of alcohol consumption on ejaculate indicators

Introduction. Lifestyle factors, including smoking, alcohol (AU) and drug use (DU), can affect male fertility. The aim of the study was to investigate the frequency and characteristics of smoking, AU and DU in men from infertile couples, to identify the most significant associations of disorders of spermatogenesis and social risk factors.Material and methods. At the 1st stage of the study, the case histories of 1198 men from infertile couples were analyzed to determine the frequency of smoking, AU and DU, at the 2nd stage, a more detailed questionnaire of 239 patients from the general sample was conducted for detailed characteristics of smoking and/or AU and/or DU, at the 3rd stage, a comprehensive analysis of ejaculate from men who consumed strong alcohol, but did not smoke or use DU (n = 46) was performed in comparison with men without bad habits taken into account (n = 60).Results. In the general sample, the frequency of AU was 73 %, smoking – 41 %, DU –17 % (mostly in anamnesis). 47.9 % of AU men consumed beer (1.5 (1–2.5) liters per week, 25.6 % – strong alcohol (250 (100–500) ml per week), 7.6 % – champagne/wine (500 (250–725) ml per week); 92.7 % of smokers used cigarettes, 7.3 % – electronic cigarettes (smoking experience was 15 (10–20) years, the number of cigarettes per day was 15 (10–20) pieces); more than 90 % of DU men have a history of non-injection DU. Men who consumed only strong alcohol, but did not smoke and did not use drugs, increased sperm DNA fragmentation compared to men without bad habits taken into account: 16.0 (13.5–19.6) and 12.8 (8.8–19.4) %, respectively, p = 0.018.Conclusions. Among men from infertile couples, a high frequency of AU was revealed with the predominant use of beer or strong alcohol, the use of the latter increases sperm DNA fragmentation; smoking is characterized by a long experience; active DU is rare. Given the specifics of the requirements for the state of reproductive health of men applying to reproductive medicine centers, it is important to have knowledge about the frequency and severity of smoking, AU and DU among them, as well as about the relationship of risk factors of infertility with the parameters of ejaculate.

P-465 Effect of male age on the outcome of frozen embryo trnasfer cycle in women over 40 years

Abstract Study question Does male age affect the outcome of in vitro fertilization in women over 40? Summary answer Advanced male partner's age is associated with decreased pregnancy rate, implantation rate and ongoing pregnancy/live birth rate. What is known already Fertility is known to have the greatest correlation with female age. Older women have a relatively lower pregnancy rate and higher miscarriage rate. It is also widely known that male partner's age is associated to pregnancy rate with increased sperm DNA fragmentation and decreased sperm motility. Age of marriage is delayed both male and female partners, assisted reproductive techniques are often conducted in women over 40 years. The effect of male age on IVF/ICSI outcomes on women over 40 is poorly explored. Study design, size, duration This is a retrospective cohort study analysed 1046 cycles of couples with an age of 40 or more of female partners and their male partners, performed at a single IVF center of university hospital between 2019 and 2022. The primary outcome was ongoing live birth and secondary outcomes were clinical pregnancy and miscarriage. Participants/materials, setting, methods Couples of primary/secondary infertility with female partner age over 40 years who underwent frozen-embryo transfer cycles were selected. couples who collected their sperm from TESE/TESA were excluded. Main results and the role of chance We decided the male partner's age in three groups, age less than 40, 41-45, and age over 45. There was a negative effect of male age on ongoing live birth and clinical pregnancy rate.(20.0%, 10.3%, 10.9%, p &amp;lt; 0.001, 33.7%, 28.4%, 19.9%, p = 0.017). And we sub-analyzed ongoing live birth rate and clinical pregnancy rate into female age group(female age of 40, 41-42, and over 43). The ongoing pregnancy/live birth rate and clinical pregnancy rate were negatively associated with male partner's age especially in female age group of over 43 years. Limitations, reasons for caution This study is limited to the information on confounding factors. The study may also be limited due to its selection criteria. Wider implications of the findings This study provides information regarding IVF/ICSI outcomes on women over 40 years of age Trial registration number not applicable

Negative effect of varicocele on sperm mitochondrial dysfunction: A cross-sectional study.

Varicocele is an abnormal dilation and enlargement of the scrotal venous pampiniform plexus that impairs normal blood drainage and finally leads to infertility if not treated. This study aimed to figure out the impact of mitochondria status through the mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) assessment and its correlation with semen parameters to illuminate the impact of sperm mitochondria healthiness on normal sperm functionality. This analytical cross-sectional study was conducted with 100 men including 50 cases in the normozoospermic group (normal) and 50 in an infertile group with the non-varicocelectomy operation (varicocele) referring to Infertility Research and Treatment Center, ACECR Khuzestan, Iran. Routine semen analysis was performed according to World Health Organization guidelines, DNA fragmentation index, the MMP assay, ATP content, and apoptosis were carried out for all samples. The results showed that the concentration, progressive motility, normal morphology, MMP, and ATP contents of sperm in varicocele were significantly lower than the normal group. In addition, the sperm DNA fragmentation index was significantly higher in the varicocele group in comparison with the normal group. Reduction in MMP and ATP contents, besides the loss of sperm parameters quality and increase in sperm DNA fragmentation, were seriously implicating sperm mitochondria dysfunctionality in varicocele men.

How to select healthy sperm for intracytoplasmic sperm injection in samples with high sperm DNA fragmentation?

The body of evidence supports the negative impact of increased sperm DNA fragmentation (SDF) on natural fertility as well as assisted reproduction conditions. High SDF has been correlated with low pregnancy and delivery rates following intrauterine insemination. Also, high SDF is accused of reducing the rates of fertilization, implantation, pregnancy, and live birth following in-vitro fertilization (IVF). Despite no impact of high SDF on fertilization or pregnancy rates following intracytoplasmic sperm injection (ICSI), it has been correlated with poor embryo quality and a higher risk of miscarriage. Several methods have been introduced to help select sperm with the best DNA quality to be used in assisted reproductive technology procedures. These include magnetic-activated cell sorting, intracytoplasmic morphologically selected sperm injection, physiologic ICSI, and microfluidic sperm sorters, among others. This article aimed to discuss the impact of high SDF in infertile men on the reproductive outcome of couples undergoing IVF/ICSI. Additionally, this review highlights the principles, advantages, and limitations of different techniques that are currently used for the selection of sperm with intact DNA to be utilized for ICSI.

Integridad del ADN espermático y toxicidad reproductiva del nitrato de plomo en ratones machos adultos

Introduction: Lead toxicity has been linked to different diseases in humans and several evidences suggest a strong relationship with the observed damage on reproductive function in humans and rodents. Methods: we provide to mice a single dose of lead nitrate (LN) of 50mg/kg body weight. Mice were euthanized seven days post-injection with the aim of evaluating sperm to come out from the seminiferous tubules and are in transit through the epididymis. Also the TUNEL test was done to evaluate the sperm DNA fragmentation. Results: The decrease in body weight in mice treated with LN (p &lt;0.05), suggest a toxic systemic effect. However, we didn´t find an effect on the reproductive system because the weights of the testes, epididymis, prostate and seminal vesicles were not altered (p&gt;0.05), in the same way physiological values such as sperm concentration and motility didn´t decrease with the treatment (p&gt; 0.05). Transit and sperm maturation in the epididymis would not be affected by the LN, and because we did not observe increased sperm DNA fragmentation in the treated group (p&gt;0.05), sperm protamination would be fulfilling its protective role on murine genetic material avoiding genotoxic damage by LN. Conclusion: The intraperitoneal administration of 50mg/kg/pc of lead nitrate (LN) for seven days does not cause systemic toxicity or effect on spermatogenesis in mice.

The Effects of Glutathione on Clinically Essential Fertility Parameters in a Bleomycin Etoposide Cisplatin Chemotherapy Model.

Chemotherapeutic agents used in the treatment of testicular cancer cause damage to healthy tissues, including the testis. We investigated the effects of glutathione on sperm DNA integrity and testicular histomorphology in bleomycin etoposide cisplatin (BEP) treated rats. Twelve-week-old male rats of reproductive age (n = 24) were randomly divided into three groups, the (i) control group, (ii) BEP group, and (iii) BEP+ glutathione group. Weight gain increase and testes indices of the control group were found to be higher than that of the BEP group and BEP+ glutathione group. While the BEP treatment increased sperm DNA fragmentation and morphological abnormalities when compared to the control group, GSH treatment resulted in a marked decrease for both parameters. Moreover, BEP treatment significantly decreased serum testosterone levels and sperm counts in comparison to the control group, yet this reduction was recovered in the BEP+ glutathione treated group. Similarly, seminiferous tubule epithelial thicknesses and Johnsen scores in testicles were higher in the control and BEP+ glutathione groups than in the BEP-treated group. In conclusion, exogenous glutathione might prevent the deterioration of male reproductive functions by alleviating the detrimental effects of BEP treatment on sperm quality and testicular histomorphology.

The impact of leptin on sperm.

Despite its important role in numerous physiological functions, including regulation of appetite and body weight, immune function and normal sexual maturation, raised leptin levels could result in significant damaging effects on sperm. The adverse effects of leptin on the male reproductive system result from its direct actions on the reproductive organs and cells instead of the hypothalamus-pituitary-gonadal axis. Binding of leptin to the receptors in the seminiferous tubular cells of the testes increases free radical production and decreases the gene expression and activity of endogenous enzymatic antioxidants. These effects are mediated via the PI3K pathway. The resultant oxidative stress causes significant damage to the seminiferous tubular cells, germ cells and sperm DNA leading to apoptosis, increased sperm DNA fragmentation, decreased sperm count, increased fraction of sperm with abnormal morphology, and decreased seminiferous tubular height and diameter. This review summarises the evidence in the literature on the adverse effects of leptin on sperm, which could underlie the often-reported sperm abnormalities in obese hyperleptinaemic infertile males. Although leptin is necessary for normal reproductive function, its raised levels could be pathologic. There is, therefore, a need to identify the cut-off level in the serum and seminal fluid above which leptin becomes pathological for better management of leptin associated adverse effects on male reproductive function.

Bacterial DNase activity as a putative inductor of sperm DNA fragmentation in infected bull frozen-thawed semen samples

The aim of this study was to investigate the relationship between DNase activity associated with bacterial contamination of incubated bovine frozen-thawed spermatozoa and elevated sperm DNA fragmentation. Electrophoresis analysis of plasmid PBR322 incubated for 30 min at 37 °C with the supernatant of the diluent of frozen-thawed centrifuged bovine semen straws infected with bacteria showed clear evidence of DNase activity when compared to plasmid incubated in similarly prepared non-infected bovine diluent supernatant (Experiment 1). This DNase activity was subsequently found to be time dependent (0–60 min) and its activity prevented in the presence of EDTA (10 and 20 mM; Experiment 2). Semen straws infected (n = 10) and not infected (n = 10) with bacteria where incubated at 37 °C for up to 48h post-thaw. Semen infected with bacteria showed an exponential increase in bacterial growth and a corresponding increase in sperm DNA fragmentation. Non-infected semen samples showed no change in the incidence of sperm DNA fragmentation over the same period of incubation (Experiment 3). Our experiments reinforce the idea that exogenous DNases present in the semen should be considered as one of the primary contributing causes of sperm DNA fragmentation post ejaculation. In the case of the bull, post-thaw incubation of commercial straws contaminated with bacteria, resulted in increased levels of sperm DNA fragmentation, most likely associated with DNase activity (potentially restriction endonucleases) derived from the bacteria. Such adverse changes in sperm DNA fragmentation, as described here in vitro, may be also operative after insemination in the female reproductive tract (in vivo) and highlight the importance of implementing high levels of hygiene practice during semen processing, especially in light of future trends of bacterial resistance to the common antibiotics used in semen diluents.

The effect of human papilloma virus on women reproductive system, pregnancy outcome of mothers: a review

Human papillomavirus is a type of sexually transmitted virus that is common in people with factors such as multiple sexual partners, a weakened immune system caused by anxiety, or alcohol and tobacco users. This virus (human papilloma) causes negative effects on vaginal and cervical epithelial cells, sperm quality and other fertility parameters. Human papilloma virus can exist in the seminal fluid caused by ejaculation and play a role in the fertility quality of men. Different viruses may directly or indirectly infect male gametes, sperms. Decreased fertility quality is caused by changes in sperm structure, increased sperm DNA fragmentation, and increased sperm aneuploidy. As a result, it causes a decrease in the quality of fertility in women and men and failure of assisted reproductive methods, and pregnancy complications such as frequent miscarriages, premature births and weight loss at birth. HPV causes precancerous cells, low-grade and high-grade cancer (LSIL-HSIL).

Temporal decline of sperm concentration: role of endocrine disruptors.

Male infertility is a widespread disease with an etiology that is not always clear. A number of studies have reported a decrease in sperm production in the last forty years. Although the reasons are still undefined, the change in environmental conditions and the higher exposure to endocrine-disrupting chemicals (EDCs), namely bisphenol A, phthalates, polychlorinated biphenyls, polybrominated diphenyl esters, dichlorodiphenyl-dichloroethylene, pesticides, and herbicides, organophosphates, and heavy metals, starting from prenatal life may represent a possible factor justifying the temporal decline in sperm count. The aim of this study is to provide a comprehensive description of the effects of the exposure to EDCs on testicular development, spermatogenesis, the prevalence of malformations of the male genital tract (cryptorchidism, testicular dysgenesis, and hypospadias), testicular tumor, and the mechanisms of testicular EDC-mediated damage. Animal studies confirm the deleterious impact of EDCs on the male reproductive apparatus. EDCs can compromise male fertility by binding to hormone receptors, dysregulating the expression of receptors, disrupting steroidogenesis and hormonal metabolism, and altering the epigenetic mechanisms. In humans, exposure to EDCs has been associated with poor semen quality, increased sperm DNA fragmentation, increased gonadotropin levels, a slightly increased risk of structural abnormalities of the genital apparatus, such as cryptorchidism and hypospadias, and development of testicular tumor. Finally, maternal exposure to EDCs seems to predispose to the risk of developing testicular tumors. EDCs negatively impact the testicular function, as suggested by evidence in both experimental animals and humans. A prenatal and postnatal increase to EDC exposure compared to the past may likely represent one of the factors leading to the temporal decline in sperm counts.