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RESEARCH QUESTION: Is seminal oxidation-reduction potential (ORP) clinically relevant to reproductive outcome? DESIGN: 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. RESULTS: 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. CONCLUSION: Seminal ORP is relevant for good fertilization, blastocyst development, implantation, clinical pregnancy and live birth.

Cancer impairs spermatogenesis, whereas results on sperm DNA integrity are controversial and no data are available about sperm oxidative stress. In cancer patients, we detected sperm DNA fragmentation (sDF) and both viable (ROS production in viable sperm fraction/viable spermatozoa) and total (ROS production in viable sperm fraction/total spermatozoa) oxidative stress. We found that cancer (22.50 (17.00–26.75)%, n = 85) increased sDF with respect to the control groups in both normozoospermic subfertile patients (NSP) (12.75 (8.63–14.88)%, n = 52, p < 0.001) and in healthy donors (HD) (8.50 (7.00–14.00)%, n = 19, p < 0.001). The induction of viable oxidative stress (n = 96) with cancer was even higher: 36.60 (24.05–58.65)% versus 11.10 (8.63–14.90)% in NSP (p < 0.001) and 9.60 (8.00–14.03)% in HD (p < 0.001). Similar, albeit lower, differences were found for total oxidative stress. SDF sharply correlated to viable oxidative stress when we considered all subjects (cancer patients and controls) (r = 0.591, p < 0.001, n = 134), but no correlation was found when only cancer patients were studied (r = 0.200; p > 0.05, n = 63). In conclusion, cancer significantly increases sDF and sperm oxidative stress levels. Additional mechanisms to oxidative attack might be responsible for increased sDF in cancer patients. Because sperm oxidative stress might affect the outcomes of sperm cryopreservation, of cancer treatments and of sperm epigenoma, the detection of oxidative stress could be of help in managing the reproductive issues of cancer patients.

Chromosomal polymorphisms are structural variations in chromosomes that define the genomic variance of a species. These alterations are recurrent in the general population, and some of them appear to be more recurrent in the infertile population. Human chromosome 9 is highly heteromorphic, and how its rearrangement affects male fertility remains to be fully investigated. In this study, we aimed to investigate the association between the polymorphic rearrangements of chromosome 9 and male infertility via an Italian cohort of male infertile patients. Cytogenetic analysis was carried out, along with Y microdeletion screening, semen analysis, fluorescence in situ hybridization, and TUNEL assays using spermatic cells. Chromosome 9 rearrangements were observed in six patients: three of them showed a pericentric inversion, while the others showed a polymorphic heterochromatin variant 9qh. Of these, four patients exhibited oligozoospermia associated with teratozoospermia, along with a percentage of aneuploidy in the sperm of above 9%, in particular, an increase in XY disomy. Additionally, high values for sperm DNA fragmentation (≥30%) were observed in two patients. None of them had microdeletions to the AZF loci on chromosome Y. Our results suggest that polymorphic rearrangements of chromosome 9 might be associated with abnormalities in sperm quality due to incorrect spermatogenesis regulation.

Sperm separation plays a critical role in assisted reproductive technology. Based on migration, density gradient centrifugation and filtration, a properly selected sperm could help in increasing assisted reproductive outcomes in teratozoospermia (TZs). The current study aimed to assess the prognostic value of four sperm selection techniques: density gradient centrifugation (DGC), swim-up (SU), DGC-SU and DGC followed by magnetic-activated cell sorting (DGC-MACS). These were evaluated using spermatozoa functional parameters. A total of 385 infertile couples underwent the procedure of intracytoplasmic sperm injection (ICSI), with an isolated teratozoospermia in the male partner. Semen samples were prepared by using one of the mentioned sperm preparation techniques. The improvements in the percentage of normal mature spermatozoa, rate of fertilization, cleavage, pregnancy and the number of live births were assessed. The normal morphology, spermatozoa DNA fragmentation (SDF) and chromatin maturity checked by using chromomycin A3 (CMA3) with DGC-MACS preparation were better compared to the other three methods. Embryo cleavage, clinical pregnancy and implantation were better improved in the DGC-MACS than in the other tested techniques. The DGC-MACS technique helped in the selection of an increased percentage of normal viable and mature sperm with intact chromatin integrity in patients with teratozoospermia.

Background: Sperm DNA fragmentation has been proposed as a candidate test for the assessment of sperm function on the premise that damage to the sperm chromatin is associated with a detrimental reproductive outcome. The objective of our study was to investigate whether sperm DNA fragmentation testing has a prognostic value, and thus can play a pivotal role in selecting future patients for intra-uterine insemination (IUI) therapy. Methods: This was a prospective cohort study conducted in a University Hospital setting. SDF was measured through TUNEL assay on the fresh semen sample presented at diagnosis and at insemination in couples with idiopathic/mild male infertility undergoing natural cycle IUI treatment. The generalized estimating equation (GEE)-model and multivariable model were used to analyze the probability of live birth and clinical pregnancy, respectively. ROC analysis was carried out to determine an SDF cut-off. Results: There was an inverse relationship between SDF in the ejaculate of the diagnostic semen sample and CP (p = 0.02; OR 0.94 95% CI (0.90, 0.989)) as well as LB (p = 0.04; OR 0.95 95% CI (0.90, 0.9985)). No significant association was found between SDF after gradient and IUI outcome in the diagnostic sample nor between SDF (ejaculate/after gradient) in the IUI samples. The ROC analysis proposed a cutoff of 17.5% as the best compromise between sensitivity and specificity in the diagnostic SDF for live birth; however, the test diagnostics are low, with an AUC of 0.576. Conclusions: Overall, this study strengthens the hypothesis of an inverse relationship between SDF and CP/LB. Furthermore, SDF taken together with other clinical characteristics might provide more insight into male reproductive potential and predicting IUI outcome. Couples with SDF ≥ 17.5% in the diagnostic semen sample did not reach live birth. Further research is necessary to establish the diagnostic and prognostic potential of SDF as an add-on test.

Currently, traumatic spinal cord injury disease is a common problem in men of sexual and reproductive active age. The result may be disturbance of erectile and ejaculatory functions, which often leads to inability of natural conception. Vibrostimulation, electrical stimulation or surgical sperm extraction, intrauterine insemination procedures and assisted reproductive technologies are the standard procedures in the treatment of fertility disorders in men with spinal cord injury and dysejaculation. The effectiveness of the techniques directly depends on the quality of sperm and the female reproductive health. Currently, the potential mechanisms of the disorders of spermatogenesis in patients with spinal cord injury are not clearly learned. The literature review of sexual dysfunction and pathogenetic aspects of pathospermia in men with spinal cord injury is provided.

Purpose The objective of this scientometric analysis was to recognize the top 100 cited articles on 'Male infertility and Antioxidants' and analyze its publication characteristics. Materials and methods The Scopus database was used to retrieve related articles and the top 100 identified based on citation rate. Results The articles were published in 56 journals between 1995 and 2019 with a median (interquartile range) citation score of 17 (5-62). Among the top 100 articles, 69 were clinical studies, which included controlled and blinded (33.33%), prospective (27.54%), randomized-controlled trials (26.09%), uncontrolled (11.59%), and retrospective (1.45%) studies. In addition to conventional semen parameters, advanced sperm function tests such as oxidative stress (51%) and sperm DNA damage (23%) were reported. Pregnancy rate (33%) was found to be the most reported reproductive outcome. Antioxidant therapy was mostly investigated in male cohorts with sperm abnormalities such as asthenozoospermia (28%) and clinical conditions such as idiopathic male infertility (20%), varicocele/varicocelectomy (17%) and general male infertility (16%). Conclusions The most influential publications on antioxidants and male infertility were identified for the first time in the literature. This will serve as a reliable source of information for researchers and clinicians alike.

Currently, recurrent pregnancy loss (RPL) examinations focus on the woman, although paternal factors are also involved. Men in couples with RPL have higher sperm DNA fragmentation levels than fertile men, but the effect of sperm DNA damage on couple's later prognosis is unknown. Advanced maternal age and obesity are associated with RPL, but paternal lifestyle factors are less studied. Therefore, we aimed to study the associations of couples' lifestyle factors, causes of RPL, and sperm DNA fragmentation with their prognosis of future live birth.This descriptive cohort study comprised 506 couples investigated for RPL at Helsinki University Hospital, Finland, between 2007 and 2016, linked with national health and population registers. The primary outcome was couple's live birth after RPL investigations. Data on couple's background factors, including age, body mass index, smoking, and alcohol use, were collected from medical records. Sperm DNA fragmentation index was analyzed from 211 men using the sperm chromatin dispersion test. The associations between background factors, sperm DNA fragmentation, and cumulative probability of live birth over time were analyzed using cross-tabulations and age-adjusted Cox regression.In all, 352 of 506 couples (69.6%) achieved live birth. Maternal age, unexplained RPL, prolonged pregnancy attempts before investigations, paternal obesity, and maternal smoking were associated with prognosis: unadjusted hazard ratio for couple's live birth for women aged 35-39 vs younger than 30 years was 0.63 (95% confidence interval [CI] 0.47-0.84), and for 40 years or older was 0.36 (95% CI 0.22-0.58). Age-adjusted hazard ratio for unexplained vs explained RPL was 1.39 (95% CI 1.12-1.72), for couple's pregnancy attempt at least 4 years vs less than 2 years was 0.50 (95% CI 0.33-0.76), for paternal body mass index at least 30 kg/mIn couples with RPL, prolonged pregnancy attempts, a cause found in RPL examinations, lifestyle factors, and maternal age are negatively associated with their prognosis of future live birth. Sperm DNA fragmentation was not associated, but the number of men with damaged spermatozoa was small. We suggest that clinicians include women and men in RPL counseling because couple's joint lifestyle seems to determine their later prognosis.

The origin and quality of gametes are likely to influence the kinetics of embryonic development. The purpose of the study was to assess the impact of sperm nuclear quality, and in particular sperm chromatin condensation, on the kinetics of early embryo development after intracytoplasmic sperm injection (ICSI). Our study included 157 couples who benefitted from ICSI for male factor infertility. Chromatin condensation and DNA fragmentation were assessed in spermatozoa prior to ICSI. Above the 20% threshold of sperm condensation defect, patients were included in the abnormal sperm chromatin condensation (ASCC) group; below the 20% threshold, patients were included in the normal sperm chromatin condensation (NSCC) group. After ICSI, the oocytes were placed in the time-lapse incubator. The kinetics of the cohort’s embryonic development have been modeled. The fading times of pronuclei and the time to two blastomeres (t2, first cleavage) and four blastomeres (t4, third cleavage) differed significantly between the NSCC and ASCC groups, with earlier events occurring in the ASCC group. On the other hand, the state of sperm chromatin condensation did not seem to have an impact on live birth rates or the occurrence of miscarriages. The kinetics of early embryonic development was accelerated in males with a sperm chromatin condensation defect without compromising the chances of pregnancy or promoting miscarriage. However, our study highlights the paternal contribution to early embryonic events and potentially to the future health of the conceptus.

Simple Summary There is quite some variability observed in both the semen and sperm DNA fragmentation (SDF) parameters in a longitudinal intrauterine insemination (IUI) trial over multiple IUI cycles. Using fertile threshold values, three patient categories were distinguished: those with a high SDF in all samples, those with low SDF in all samples and those who fluctuated between high and low during the whole IUI trial. Semen preparation via density gradient centrifugation increases SDF. However, the three patient categories react differently after semen processing. A large percentage of those with high SDF retain their high SDF even after gradient centrifugation. The SDF fluctuaters react with a high SDF after gradient centrifugation. The low SDF category, on the contrary, distributes itself evenly between the three categories after gradient centrifugation. SDF testing after semen processing might be indispensable for therapeutic purposes, probably influencing medical decision-making. In order to isolate the fluctuaters, a second SDF testing might be advocated in certain cases. The biological variability of semen and sperm DNA fragmentation (SDF) parameters in a longitudinal intrauterine insemination (IUI) trial over multiple IUI cycles was investigated. A TUNEL assay was used for SDF testing, both before and after density gradient centrifugation. A significant age effect was observed: while semen parameters deteriorated with advancing age, on average, higher SDF values were observed for older males. There was quite some variability observed for both semen and SDF variables. Using fertile threshold values, three patient categories were distinguished: those with a high SDF in all samples, those with low SDF in all samples and those who fluctuated between high and low during the whole IUI trial. Density gradient centrifugation increases SDF. However, the three patient categories react differently after semen processing. A large percentage of those with high SDF retain their high SDF even after gradient centrifugation. The SDF fluctuaters react with a high SDF after gradient centrifugation. The low SDF category, on the contrary, distributes itself evenly between the three categories after gradient centrifugation. SDF testing after semen processing might be indispensable for therapeutic purposes, probably influencing medical decision-making. In order to isolate fluctuaters, a second SDF testing might be advocated in certain cases. SDF after semen processing is indispensable for therapeutic management.

Introduction. According to Russian Association of Urology a number of infertile couples ranges from 8 to 17.2 % in various regions of Russia. One of the causes of which is the male factor. Raising the age of the father reduces the chance of conception, increases the risk of early termination of pregnancy and the risk of certain diseases of the child. In this regard the existing knowledge of age-related changes in ejaculate parameters needs to be de veloped. This is particularly true for men of infertile couples.The study objective – to investigate the age patterns of change in the spermological status of infertile men and identify the most age-related variations.Materials and methods. Research design – observation, retrospective, crossection. Analysis of the medical records of 517 Novosibirsk male residents from couples with infertility in marriage was made. The sampling is divided into 5 age groups: 1st group (n = 108) – men of ≤29.9 years old; 2nd group (n = 171) – 30.0–34.9 years old; 3rd group (n = 130) – 35.0–39.9 years old; 4th group (n = 68) – 40.0–44.9 years old; 5th group (n = 40) – ≥45.0 years old.Results. Median of male age in total sample – 34 years, frequency of persons over 40 years – 21 %. Overall sample shows a decline in reference values of proportion of morphologically normal forms – 3.0 (1.5–5.0) % and increase in sperm DNA fragmentation index (SDFI) – 16.0 (11.0–22.2) %. In groups from 1st to 5th there is an age-specific decrease in ejaculate volume (3.5 (2.5–4.8) ml; 3.3 (2.4–4.2) ml; 3.2 (2.4–4.5) ml; 3.0 (1.7–4.9) ml; 2.7 (1.9–3.5) ml, р1–5 1–5, 2–5 1–4 Conclusion. It was first time shown for the male population of Novosibirsk of Siberian Federal District that a reduction in the proportion of morphologically normal sperm below the reference values observed in the total sample and it did not depend on the age of the men examined. Only three of the examined indicators of sperm status studied infertile males had a significant age dependency: ejaculate volume decrease, reduction of total sperm, increased DNA fragmentation of sperm. The lack of a reliable influence of age on HBA-test indicators was first shown. However the trends identified require further study. The results show that age-associated increase in DNA fragmentation of sperm starts with young men under 30, and underscores the need to define SDFI in actual clinical practice. And negative processes of age influence on the quantitative, morphological and ultrastructural characteristics of sperm occur most frequently in men after 40 years.

This systematic review aims to compare and evaluate the outcome of using either testicular sperm (Testi-ICSI) or ejaculated sperm (Ejac-ICSI) in intracytoplasmic sperm injections in patients with recurrent ICSI failure. The Cochrane Library, MEDLINE, EMBASE, and PubMed were used to search for relevant papers up till October 2020. Four cohort studies and two case series studies were included. Four studies investigated males with high sperm DNA fragmentation (SDF) and were classified as “high SDF”, which included 247 couples and 2712 injected oocytes. The other three studies provided paired data to an unselected population of infertile men with either untested SDF or when anomalous SDF was not used as the basis for deciding to use Testi-ICSI, and were classified as “noclassify” in this study. This subgroup consisted of a total of 290 couples and 1061 injected oocytes. There was a higher level of clinical pregnancy rates (CPRs) in the “high SDF” subgroup when Testi-ICSI was used as compared to Ejac-ICSI, at 43.4% and 20.8% respectively, with a pooled odds ratio (OR) of 2.87 (95% confidence interval (CI) 1.44–5.71; P = 0.003). Furthermore, in the “high SDF” subgroup, Testi-ICSI use was associated with better take home baby rates (38%) as compared to Ejac-ICSI (16%), with a pooled OR of 3.24 (95% CI 1.20–8.76; P = 0.02). In the “noclassify” group, there was no statistically significant difference in the CPRs and take home baby rates of Testi-ICSI and Ejac-ICSI, although there was a trend of better CPRs and take home baby rates with Testi-ICSI use. Utilization of Testi-ICSI in recurrent ICSI failure couples, where males were confirmed to have high SDF in their ejaculated sperm, were correlated with greater CPRs and take home baby rates. However, Testi-ICSI may not result in better ICSI outcomes among men with untested SDF or when anomalous SDF was not the main factor influencing the decision to utilize Testi-ICSI.

The article provides an overview of the most significant publications on the topic of male infertility. The main selection criteria were the practical significance of the article, as well as the impact factor of the journal in which it was published, according to the SCImago Journal Rank (SJR). As a result, a list of 10 works published in the first quarter (January - March) of 2021 was formed. The review included articles related to the following issues: sperm DNA fragmentation, the use of testicular spermatozoa in ART programs (assisted reproductive technologies), advanced paternal age, the role of the human papillomavirus for male fertility, hormonal modulation by clomiphene, the risk of birth defects in children in ART programs, andrological examination of patients with CFTR gene mutations. And also, an analysis of the new clinical guidelines of the American Urological Association (AUA) was carried out.

Introduction. According to the WHO data, depression is a common disease among women and men of reproductive age. One line of the correction of depressive disorders is selective serotonin reuptake inhibitors (SSRIs). The ingestions have shown that using SSRIs harms sperm quality. The literature date of evaluation of male fertility after discontinuation of antidepressants is quite limited.Purpose of the study. To evaluate the influence of Fluoxetine intake on semen parameters, sperm DNA fragmentation and hormonal status.Materials and methods. Twenty-five men (mean age - 35.2 ± 4.5 yo) with depression were included in the study. Fluoxetine (20 mg per day) was prescribed to all the patients for 12 wk. Semen parameters, sperm DNA fragmentation, sex hormones levels were measured before-after treatment and 3 mo behind discontinuation.Results. After 12 weeks of the treatment the mean semen volume decreased from 3.1 ± 0.7 to 2.9 ± 0.7 ml (p = 0.638), sperm concentration - from 39.4 ± 18.5 to 34.3 ± 16.8 mln/ml (p = 0.384), sperm motility decreased from 41.7 ± 7.6 to 35.5 ± 7.8% (p < 0.05), the mean percent of normal morphology form - from с 12.7 ± 2.8 to 10.7 ± 2.2% (p < 0.001). Sperm DNA fragmentation increased 16.2 ± 4.9 to 22.2 ± 4.3% (p < 0.001). The mean semen volume, sperm concentration, motility, percentage of normal morphology increased and reverted to the normal levels after 3 mounts of drug discontinuation. Sperm DNA fragmentation index decreased, and it had the values less than before the treatment that positively correlated with the reduction of depression's symptoms. It was not significant dynamics in hormonal parameters before and after the therapy.Conclusion. Using fluoxetine has a reversible negative effect on male fertility. It is important to inform the patients about the temporary side effects of SSRIs in fatherhood planning cases.

Recently, sperm quality and the presence of double-stranded breaks (DSB) has been pointed out as a possible cause of recurrent miscarriage, and the use of antioxidants has expanded as a treatment for male infertility. The aim of the present study was to analyze the proteomic effects of antioxidants on sperm from RM patients with high incidence of DSB. Proteomic analysis was performed using a tandem mass tag labeling technique, and subsequently compared with the PANTHER database for DEPs, and the STRING database for protein–protein interactions (PPI). Differentially expressed proteins (DEPs) both before and after antioxidant oral treatment were identified. PPI involving DEPs clustered into networks related to cell metabolism, cytoskeleton, and DNA damage. Results show that the sperm proteomic profiles before and after antioxidant treatment do not significantly differ from each other. However, some DEPs found after the antioxidant treatment shifted towards a DEPs profile typical of fertile donors. This indirect measurement suggests an improvement caused by antioxidants on the expression of several proteins. Among them were proteins involved in sperm DNA remodeling (LMO7, MMP28, BNC2, H2B, and PRDM2). The results presented here represent the first approach in the analysis and repair of the proteomic change caused by antioxidants in recurrent miscarriage patients, elucidating biomarkers that may be useful for the diagnosis and further sperm selection in this type of patient. Further studies should be conducted to validate the usefulness of these biomarkers in larger study groups.

Endogenous and exogenous factors can severely affect the integrity of genetic information by inducing DNA damage and impairing genome stability. The extent to which men with and without subfertility are exposed to several adverse lifestyle factors and the impact on sperm DNA fragmentation (SDF), sperm chromatin maturity (condensation and decondensation), stability (hypo- and hypercondensation) and sperm aneuploidy are assessed in this study. Standardized assays employing flow cytometry were used to detect genome instability in 556 samples. Semen parameters deteriorated with age, BMI, increased physical activity and smoking. Age and BMI were associated with increased SDF. Increased BMI was associated with increased hypocondensed chromatin and decreased decondensed chromatin. Increase in age also caused an increase in sex chromosome aneuploidy in sperms. Surprisingly, alcohol abuse reduced chromatin hypercondensation and drug abuse reduced SDF. Although genome instability was more pronounced in the subfertile population as compared to the fertile group, the proportion of men with at least one lifestyle risk factor was the same in both the fertile and subfertile groups. While one in three benefited from nutritional supplementation, one in five showed an increase in SDF after supplementation. Whilst the message of ‘no smoking, no alcohol, no drugs, but a healthy diet’ should be offered as good health advice, we are a long way from concluding that nutritional supplementation would be beneficial for male fertility.

Bu çalışma, 10-13 Mart 2022 tarihlerinde Bursa düzenlenen 16. Uludağ Jinekoloji ve Obstetrik Kış Kongresi‘nde bildiri olarak sunulmuştur. Üremeye yardımcı tedavi uygulamalarında tercih edilen tedavi yaklaşımına göre semen parametrelerinin embriyoloji laboratuvarı sonuçlarına ve klinik başarıya etkisi değişmektedir. Semen parametreleri abstinens süresine ve androloji laboratuvarında uygulanan yıkama protokollerine göre değişmekte ve insemine edilecek sperm materyalinin kalitesini etkilemektedir. Bu çalışmada normozoospermik erkeklerde kısa abstinens süresinin rutin semen parametrelerine, sperm kromatin ve DNA bütünlüğüne, oksidatif strese karşı gelişen antioksidan kapasiteye etkisinin değerlendirilmesi amaçlandı. Aynı hastadan ardışık ejakülasyonla 2-5 günlük abstinens süresi sonrası (n=36) ve 1 saat abstinens süresi sonrası (n=36) alınan numuneler yıkama öncesi ve yıkama sonrası değerlendirildi. Yıkama öncesinde sperm volümünün ve total motil sperm sayısının kısa abstinens grubunda anlamlı olarak azaldığı bulundu. Yıkama sonrasında gruplar arasında motilitenin değişmediği, konsantrasyonun kısa abstinens grubunda anlamlı olarak azaldığı görüldü. Abstinens süresi kısa tutulduğunda sperm kromatin hasarının ve DNA fragmantasyon oranının azaldığı, antioksidan kapasitede bir değişiklik oluşturmadığı saptandı. Sonuç olarak normozoospermik olgularda, abstinens süresinin kısa tutulması sperm konsantrasyonunu ve total progressif motil sperm sayısını azaltmakla birlikte, uygulanacak üremeye yardımcı tedavi yaklaşımına göre inseminasyonda kromatin ve DNA bütünlüğü açısından daha kaliteli sperm kullanılmasına imkan sağlayacaktır. Ardışık ejakülasyon ve abstinens süresindeki kısalma aktioksidan kapasitede olumlu ya da olumsuz bir etki oluşturmamaktadır. The effects of semen parameters on embryology laboratory results and clinical success vary according to the preferred treatment approach in assisted reproductive technology. Semen parameters vary according to the duration of abstinence and the washing protocols applied in the andrology laboratory and effect the quality of the sperm material to be inseminated. This study aimed to evaluate the effect of a short abstinence period on routine semen parameters, sperm chromatin, DNA integrity, and antioxidant capacity against oxidative stress in normozoospermic men. Samples taken from the same patient after 2-5 days of abstinence with consecutive ejaculation (n=36) and after 1 hour of abstinence (n=36) were evaluated before and after washing. It was found that sperm volume and total motile sperm count were significantly decreased in the short abstinence group before washing. It was observed that motility did not change between the groups after washing, and the concentration decreased significantly in the short abstinence group. It was determined that sperm chromatin damage and DNA fragmentation rate decreased when the abstinence period was kept short, and there was no change in antioxidant capacity. As a result, in normozoospermic cases, keeping the abstinence period short will decrease sperm concentration and total progressive motile sperm count and allow the use of higher quality sperm in terms of chromatin and DNA integrity in insemination compared to the assisted approach reproductive treatment. Sequential ejaculation and shortening of the abstinence time do not have a positive or negative effect on the antioxidant capacity.

Seminal oxidative stress and sperm DNA damage are potential etiologies of male factor infertility. The present study aims to evaluate the relationship between oxidation-reduction potential (ORP), a measure of oxidative stress, and sperm DNA fragmentation (SDF) by conducting a systematic review and meta-analysis of relevant clinical data. A literature search was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The COVIDENCE tool was used to screen and identify studies evaluating seminal ORP and SDF. Studies (n = 7) that measured seminal ORP and SDF of 3491 semen samples were included in the analysis. The fixed-effects model revealed a significant pooled correlation coefficient (r = 0.24; p < 0.001) between seminal ORP and SDF. Furthermore, subgroup analyses indicated that the pooled correlation coefficient between ORP and sperm chromatin dispersion (SCD) assay was less than other SDF assays (0.23 vs. 0.29). There was a moderate level of heterogeneity (I2 = 42.27%) among the studies, indicating a lack of publication bias. This is the first meta-analysis to reveal a positive correlation between seminal ORP and SDF. Furthermore, this study indicates the role of oxidative stress in the development of sperm DNA damage and thus warrants prospectively exploring the clinical value of these sperm function tests.

Purpose While SARS-CoV-2 infection appears not to be clinically evident in the testes, indirect inflammatory effects and fever may impair testicular function. To date, few long-term data of semen parameters impairment after recovery and comprehensive andrological evaluation of recovered patients has been published. The purpose of this study was to investigate whether SARS-CoV-2 infection affect male reproductive health. Methods Eighty patients were recruited three months after COVID-19 recovery. They performed physical examination, testicular ultrasound, semen analysis, sperm DNA integrity evaluation (TUNEL), anti-sperm antibodies (ASA) testing, sex hormone profile evaluation (Total testosterone, LH, FSH). In addition, all patients were administered International Index of Erectile Function questionnaire (IIEF-15). Sperm parameters were compared with two age-matched healthy pre-COVID-19 control groups of normozoospermic (CTR1) and primary infertile (CTR2) subjects. Results Median values of semen parameters from recovered SARS-CoV-2 subjects were within WHO 2010 fifth percentile. Mean percentage of sperm DNA fragmentation (%SDF) was 14.1 ± 7.0%. Gelatin Agglutination Test (GAT) was positive in 3.9% of blood serum samples, but no positive semen plasma sample was found. Only five subjects (6.2%) had total testosterone levels below the laboratory reference range. Mean bilateral testicular volume was 31.5 ± 9.6 ml. Erectile dysfunction was detected in 30% of subjects. Conclusion Our data remark that COVID-19 does not seem to cause direct damage to the testicular function, while indirect damage appears to be transient. It is possible to counsel infertile couples to postpone the research of parenthood or ART procedures around three months after recovery from the infection.

Objective: Oxidative stress (OS) plays a key role in the etiology of unexplained male infertility. Coenzyme Q10 (CoQ10) is a potent antioxidant that may improve semen quality and OS in infertile men with idiopathic oligoasthenoteratospermia (OAT), but the underlying mechanism is unknown. Therefore, the present study was undertaken to investigate the effect of CoQ10 on OS markers and sperm DNA damage in infertile patients with idiopathic OAT. Methods: This prospective controlled study included 50 patients with idiopathic OAT and 50 fertile men who served as controls. All patients underwent a comprehensive medical assessment. Patients and controls received 200 mg of oral CoQ10 once daily for 3 months. Semen and blood were collected and analyzed for sperm parameters, seminal CoQ10 levels, reactive oxygen species (ROS) levels, total antioxidant capacity, catalase, sperm DNA fragmentation (SDF), and serum hormonal profile. Results: The administration of CoQ10 to patients with idiopathic OAT significantly improved sperm quality and seminal antioxidant status and significantly reduced total ROS and SDF levels compared to pre-treatment values. Conclusion: CoQ10, at a dose of 200 mg/day for 3 months, may be a potential therapy for infertile patients with idiopathic OAT, as it improved sperm parameters and reduced OS and SDF in these patients.

The article provides an overview of the most significant publications on the topic of male infertility. The main selection criteria were considered the practical significance of the article, as well as the impact factor of the journal in which it was published, according to the SCImago Journal Rank (SJR). As a result, a list of 10 papers was formed that published from October - December 2020. The review included articles on the following issues: sperm DNA fragmentation and oxidative-reductive balance in the male reproductive system, advanced paternal age, hormonal stimulation in patients with nonobstructive azoospermia, the relationship of cancer with infertility, as well as embryological methods to increase the effectiveness of assisted reproductive technologies procedures.

Objective: To evaluate the outcome of microscopic subinguinal varicocelectomy on sperm DNA fragmentation (SDF) and pregnancy rate in men with normal semen parameters. Patients and methods: A pilot study that included male patients with a minimum of a 1-year history of male subfertility, normal semen parameters, a high percentage of SDF, and clinically palpable varicoceles. Microscopic subinguinal varicocelectomy was carried out for 45 patients (study group), while 40 patients had no intervention (control group). Semen analysis and SDF were measured before and at 6 months after the varicocelectomy. The pregnancy rate was assessed at the 6- and 12-month follow-ups. Results: Between July 2014 and January 2019, 85 subfertile men were included in the study and completed 12 months of follow-up. The two groups were comparable in terms of their age, body mass index, infertility duration, infertility type, varicoceles laterality, and varicoceles grade (P values = 0.84, 0.34. 0.35, 1, 0.39, and 0.46, respectively). At 6 months after varicocelectomy, the mean SDF was reduced in both groups, and this reduction was statistically higher in the varicocelectomy group (P

Cryopreservation impairs sperm quality and functions, including motility and DNA integrity. Antioxidant additives in sperm freezing media have previously brought improvements in postthawed sperm quality. Green tea extract (GTE) is widely considered as an excellent antioxidant, and its beneficial role has been proven in other human cells. This study aims to evaluate the GTE as a potential additive in cryopreservation media of human spermatozoa. In part one, the semen of 20 normozoospermic men was used to optimize the concentration of GTE that maintains sperm motility and DNA integrity against oxidative stress, induced by hydrogen peroxide (H2O2). Spermatozoa were treated with GTE at different concentrations before incubation with H2O2. In part two, the semen of 45 patients was cryopreserved with or without 1.0 ng ml−1 GTE. After 2 weeks, the semen was thawed, and the effect on sperm motility and DNA fragmentation was observed. Our data showed that GTE significantly protected sperm motility and DNA integrity against oxidative stress induced by H2O2 when added at a final concentration of 1.0 ng ml−1. We found that the addition of 1.0 ng ml−1 GTE to cryopreservation media significantly increased sperm motility and DNA integrity (both P < 0.05). More interestingly, patients with high sperm DNA damage benefited similarly from the GTE supplementation. However, there was no significant change in the reactive oxygen species (ROS) level. In conclusion, supplementing sperm freezing media with GTE has a significant protective effect on human sperm motility and DNA integrity, which may be of clinical interest.

Introduction. In recent years, the effects of obesity on male fertility have been extensively investigated. The results of existing studies are extremely contradictory.The study objective was to determine the effect of obesity on the male reproductive system using the biological model of laboratory rats as an example.Materials and methods. In vivo modeling of diet-induced obesity. The study was conducted on 22 laboratory sexually mature white rats weighing 140–160 g. The animals were divided into two groups: 1 control (10 animals) and 2 rats with diet-induced obesity (12 animals). After 12 weeks, the animals were removed from the experiment. All rats underwent: calculation of the Lee index (body mass index in rats), determination of the concentration and viability of spermatozoa in a suspension of sperm from the epididymis, determination of glucose level of total cholesterol and triglycerides in the blood, study of sperm DNA fragmentation, histological examination testis: calculating the crosssectional area of the seminiferous tubule; determination of the number of non-functioning tubules and tubules with desquamated spermiogenic epithelium; determination of the average spermatogenesis index.Results. In the study groups there were no differences in glucose and total cholesterol levels. However, a statistically significant, significant difference in the level of triglycerides in the blood was revealed. The concentration of sperm and their viability in the studied groups did not differ. The level of sperm DNA fragmentation in the experimental group is significantly higher than in the control group (31.5 ± 10.1 and ± 1.4 %, respectively, p Conclusion. Diet-induced obesity causes impaired spermatogenesis, and damage to the sperm genetic material in male white rats.

The article provides an overview of the most influential papers on the topic of “male infertility”. The selection criteria were the practical significance of the article and the impact factor of the journal in which it was published, according to SCImago Journal Rank (SJR). As a result, we created the list of 10 papers published in the I quarter (January – March) of 2020. The review included articles on the following issues: the effectiveness and safety of antioxidants, zinc and folic acid in the treatment of male infertility; the effect of obesity on sperm DNA-fragmentation; the possibility of obtaining of spermatogonia in children and adolescents with Kleinfelter syndrome; comparison of intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected spermatozoa injection (IMSI); the reaction of the female reproductive system to the seminal fluid; viral profile in seminal fluid of men receiving the antiretroviral therapy, the effect of sleep duration on fertility and the risk of diabetes mellitus in children, conceived using assisted reproductive technologies.

Background This study investigates the effect of letrozole on hormone profiles, semen parameters, body mass index (BMI), degree of oxidative stress and sperm chromatin integrity in men with idiopathic oligo/astheno/teratozoospermia (iOAT) and T:E2 ratio ≤ 10. Materials and methods This study is a longitudinal, prospective, interventional and open-labelled clinical trial. Semen samples were collected from 20 iOAT men with low serum testosterone (T) to estradiol (E2) ratio (T:E2 ratio ≤ 10). The participants were treated with 2.5 mg letrozole orally per day for 3 months. Then, sperm parameters, hormone profiles, BMI, chromatin integrity and intracellular reactive oxygen species (ROS) level were assessed pre- and post- treatment. The chromatin integrity was evaluated by assessment of DNA fragmentation (with TUNEL assay) and protamine deficiency (with Chromomycin A3, CMA3). Also, the intracellular ROS levels were investigated by 2′, 7′-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. Finally, the differences between the parameters evaluated before and after letrozole treatment were analyzed with the t-test and the Wilcoxon signed-rank test. Results Sperm concentration, percentage of sperm motility and its normal morphology increased significantly after letrozole treatment. Moreover, serum testosterone level increased but estradiol level decreased significantly following treatment. The mean of T:E2 ratio improved 1600%. Also, letrozole treatment significantly reduced the percentage of sperm TUNEL positivity and sperm CMA3 positivity. While no significant difference was observed between intracellular ROS levels and BMI before and after treatment. Finally, as a notable result, four spontaneous pregnancies (20%) were achieved after treatment. Conclusions Letrozole treatment can effectively increase spontaneous pregnancies by improving sperm parameters and sperm chromatin integrity in men with iOAT and T:E2 ratio ≤ 10. Trial registration Trial registration: IRCT, IRCT20191030045283N1. Registered 16 November 2019 - Retrospectively registered, https://fa.irct.ir/user/trial/43484/view

The article provides an overview of the most influential papers on the topic of «male infertility». The main selection criteria were the practical significance of the article and the impact factor of the journal in which it was published, according to SCImago Journal Rank (SJR). As a result, we created the list of 10 papers published in the IV quarter (October − December) of 2019. The review included articles on the following issues: the effectiveness and prognostic factors of the effectiveness of microsurgical testicular sperm extraction (micro-TESE), the significance of sperm DNA fragmentation and «advanced paternal age» in the effectiveness of assisted reproduction technologies (ART) procedures, the role of latent chlamydial infection in azoospermia, management of adolescents with Kleinfelter syndrome, the effectiveness and safety of aromatase inhibitors in treatment of male infertility, sperm microbiome, the association between ART procedures and the cancer risk in children, follicle-stimulating hormone (FSH) treatment of male idiopathic infertility.

According to world statistics, 15 % of married couples do not reach pregnancy within one year and seek medical help. At the same time, an important component of the complex problem of infertility, in addition to not getting pregnant, are also pregnancy losses. According to modern ideas, one of the reasons for termination of pregnancy is sperm DNA fragmentation (SDF). According to the European Association of Urology guidelines on male infertility, SDF reduces the chances of natural conception and increases the risk of pregnancy loss. In the European Society of Human Reproduction and Embryology guideline on the management of recurrent pregnancy loss, the definition of SDF is recommended in the algorithm for examining a couple after a single episode of miscarriage. The causes, mechanisms and approaches to correcting increased SDF are the subject of active research; according to PubMed, more than 2,500 papers have been published on this problem to date. Given the huge amount of factual material, the purpose of this review is to highlight the key points of etiology, pathogenesis and approaches to the correction of increased levels of SDF

Background: Opioids have been implicated to induce infertility. Although codeine remains the most used opioid for recreational purpose, no study has documented its effect on sperm quality. Elucidating the effect of codeine on sperm cells and the associated mechanisms may provide an insight into preventing drug-induced sperm damage. Twenty-one New Zealand white rabbits were randomized into three groups; control and codeine-treated. The codeine-treated groups received either 4 or 10mg/kg b.w of codeine for six weeks. Results: Codeine treatment led to significant decrease in sperm count, motility, viability, normal morphology, and sperm membrane integrity. This was associated with significant rise in sperm DNA fragmentation, oxidative damage, and caspase 3 activity. The percentage of sperm DNA fragmentation correlates positively with 8-hydroxy-2'-deoxyguanosine, a biomarker of oxidative DNA damage, and caspase 3 activity, a biomarker of apoptosis. The observed correlation was stronger between sperm DNA fragmentation and oxidative DNA damage than sperm DNA fragmentation and caspase 3 activity. Conclusion: This study revealed that chronic codeine exposure causes sperm DNA fragmentation and poor sperm quality primarily via oxidative stress rather than activation of caspase 3-dependent apoptosis. Findings of the present study may explain drug-induced male factor infertility, particularly, those associated with opioid use.

OBJECTIVE: To better study the effect of sperm deoxyribonucleic acid fragmentation (SDF) on intracytoplasmic sperm injection (ICSI) outcomes from an ovum donation program by assessing the cumulative live birth rates (CLBRs) per number of embryo transfers (ETs), embryos replaced (EmbR), and metaphase II (MII) oocytes required in consecutive treatments to achieve the first newborn. DESIGN: A multicenter retrospective cohort study was conducted, and the Kaplan-Meier survival curves were generated to calculate the CLBR with regard to the SDF degree. SETTING: Private university-affiliated in vitro fertilization centers. PATIENT(S): Data from 864 couples using donated eggs and undergoing ICSI from 2000 to 2019 were analyzed. Sperm deoxyribonucleic acid fragmentation was measured using terminal deoxynucleotidyl transferase biotin dUTP nick end labeling assay on their ejaculated sperm. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Live birth rate (LBR) per first ET and per all consecutive ETs within the same patient and CLBR per ET, per EmbR, and per MII oocyte used considering the SDF level. RESULT(S): A total of 1,903 ICSI cycles were considered, encompassing 6,340 donated oocytes, 2,543 embryos, and 1,145 ETs. Comparing =15% SDF (low) with >15% SDF (high) or by 10% SDF ranges, the LBRs per first ET and per all ETs did not significantly differ. The Kaplan-Meier curves of the CLBR per ET, per EmbR, and per donor oocyte consumed were similar between the SDF groups evaluated. CONCLUSION(S): Elevated SDF does not reduce the LBR or cumulative probability to obtain a child when calculated per ET, per EmbR, and per donated MII oocyte used in couples undergoing ICSI cycles.

RESEARCH QUESTION: Does sperm DNA fragmentation (SDF) affect reproductive success of IVF and intracytoplasmic sperm injection (ICSI) cycles measured as cumulative live birth rates (CLBR) in unselected couples? DESIGN: Clinical data from 1339 couples undergoing 2759 IVF/ICSI cycles using autologous oocytes with a SDF test by TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling (TUNEL) assay on their ejaculated spermatozoa were retrospectively evaluated. Main outcomes were calculated according to two different analyses: using 15% SDF as cut-off point (low =15% and high >15%); and categorizing participants based on four SDF ranges (30%). Live birth rate and CLBR per number of embryo transfers, per number of embryos replaced and consumed oocytes required to achieve the first live birth according to level of SDF were the main outcomes assessed. RESULTS: No significant difference was found in clinical pregnancy rate and miscarriage rate between both groups. No differences in LBR per embryo transfer were found for the first or for all embryo transfers when comparing =15% and >15% sperm DNA fragmentation or by SDF ranges. The CLBR according to the number of embryo transfers and the number of embryos replaced showed no statistically significant differences between different SDF groups. When the same number of oocytes were inseminated, similar CLBR were obtained regardless of the degree of male sperm DNA fragmentation. CONCLUSIONS: High SDF did not impair live birth rates of unselected males undergoing IVF/ICSI cycles with autologous oocytes per transfer or the cumulative probability of a live birth.

L’esterilitat, una malaltia que presenta increments anuals d’incidència, afecta actualment entre el 7% i el 15% de les parelles en edat fèrtil. És conegut que en la meitat dels casos hi contribueix un factor masculí, pel que la qualitat seminal és un dels paràmetres importants a estudiar, ja que existeixen múltiples factors que la poden alterar. Alguns d’aquests factors, com l’exposició a tòxics o diferències en l’estil de vida poden variar segons l’àmbit geogràfic. Davant l’increment de parelles estèrils, les tècniques de reproducció han suposat una important solució en molts casos, però encara presenten una disponibilitat limitada a nivell públic, motiu pel qual s’ha suggerit que una millor direcció dels pacients al tractament més adequat podria ajudar a incrementar les taxes d’èxit. Els objectius principals de la present tesi doctoral han estat elucidar si ha existit una reducció de la qualitat seminal en els últims 20 anys en els pacients que acudeixen a l’Hospital Parc Taulí, i determinar quins dels paràmetres masculins i femenins tenen un efecte en la consecució d’un embaràs clínic en tractaments d’inseminació artificial conjugal, per a construir un model amb capacitat predictiva sobre aquesta tècnica. Els resultats del primer objectiu han mostrat que el volum, el pH, la motilitat progressiva, el percentatge d’espermatozoides immòbils i d’espermatozoides amb morfologia normal van disminuir al llarg del les darreres dues dècades, el que tradueix una reducció de la qualitat seminal en els homes que acudeixen per esterilitat a l’Hospital Parc Taulí de Sabadell. El segon objectiu va consistir en un estudi prospectiu que va incloure 197 parelles consecutives candidates a cicles d’inseminació intrauterina conjugal, dividits en dos anys. Al final del primer any es van identificar els paràmetres tant femenins com masculins que s’associaven a la falta d’embaràs clínic. El segon any va servir per a validar aquests paràmetres i per a generar un model multivariable amb possibilitats d’aplicar-se a la clínica. El model va donar lloc a una sensibilitat del 38,4% (IC 95% 31,9% a 45,3%) i una especificitat del 96,8% (IC 95% 83,8% a 99,8%). L’odds ratio obtinguda va ser d’11,9; el valor predictiu positiu va ser 65,26% i el valor predictiu negatiu de 90,94%. Així doncs, l’aplicació retrospectiva d’aquest model hauria augmentat un 30% la taxa d’embaràs clínic, en cas d’haver estat aplicat en els mateixos pacients inclosos en l’estudi. La estertilidad es una enfermedad que presenta aumentos anuales de incidencia, afecta actualmente entre el 7% y el 15% de las parejas en edad fértil. Se sabe que en la mitad de los casos contribuye un factor masculino, por lo que la calidad seminal es uno de los parámetros importantes a estudiar, ya que existen múltiples factores que pueden alterarla. Algunos de estos factores, como la exposición a sustancias tóxicas o las diferencias en el estilo de vida, pueden variar dependiendo de la zona geográfica. Ante el aumento de parejas estériles, las técnicas de reproducción han sido una solución importante en muchos casos, pero aún así tienen una disponibilidad limitada a nivel público, por lo que se ha sugerido que una mejor orientación de los pacientes al tratamiento más adecuado podría ayudar a aumentar las tasas de éxito. Los principales objetivos de esta tesis doctoral han sido determinar si se ha producido una reducción de la calidad seminal en los últimos 20 años en los pacientes que acudieran al Hospital Parc Taulí, y determinar cuáles de los parámetros masculinos y femeninos tienen un efecto en la consecución de un embarazo clínico en tratamientos de inseminación artificial conyugal, para construir un modelo con capacidad predictiva sobre esta técnica. Los resultados del primer objetivo han demostrado que el volumen, el pH, la motilidad progresiva, el porcentaje de espermatozoides inmóviles y espermatozoides con morfología normal disminuyeron en las últimas dos décadas, dando lugar a una reducción de la calidad seminal en los hombres que acuden por infertilidad al Hospital Parc Taulí de Sabadell. El segundo objetivo consistió en un estudio prospectivo que incluyó 197 parejas consecutivas candidatas a ciclos de inseminación intrauterina conyugal, divididos en dos años. Al final del primer año, se identificaron parámetros femeninos y masculinos asociados con la falta de embarazo clínico. El segundo año sirvió para validar estos parámetros y generar un modelo multivariante con posibilidades de ser aplicado a la clínica. El modelo resultó en una sensibilidad de 38,4% (IC 95% 31,9% a 45,3%) y una especificidad de 96,8% (IC 95% 83,8% a 99,8%). El odds ratio obtenido fue de 11,9; el valor predictivo positivo fue del 65,26% y el valor predictivo negativo del 90,94%. Por lo tanto, la aplicación retrospectiva de este modelo habría aumentado la tasa de embarazo clínico en un 30%, si se aplicara en las mismas pacientes incluidas en el estudio. Infertility, a disorder with increasing annual incidence, currently affects between 7% and 15% of couples at reproductive age. The contribution of a male factor in half of cases is well accepted, making seminal quality one of the main parameters to study, since there are multiple factors that can affect it. Among these factors, such as exposure to toxic substances or lifestyle differences, may vary depending on the geographical area. In face of the increasing amount of sterile couples, assisted reproductive techniques have been an important contribute in many cases, though often with limited availability, suggesting that a better guidance of patients towards the most appropriate treatment could help increasing treatment success rates. The main objectives of this doctoral thesis have been to assess whether there has been a reduction in seminal quality in the last 20 years in patients who attend Hospital Universitari Parc Taulí, and determine which male and female parameters have an effect on achieving a clinical pregnancy through conjugal artificial insemination treatments, in order to build a model with predictive capability for this technique. The results of the first objective showed a decrease in volume, pH, progressive motility, percentage of non-motile sperm and of normal morphology sperm over the past two decades, suggesting a reduction in seminal quality in men who consult for infertility at the Hospital Universitari Parc Taulí in Sabadell. The second objective consisted of a prospective study that included 197 consecutive couples candidates for cycles of intrauterine insemination, divided into two years. At the end of the first year, both female and male parameters associated with absence of clinical pregnancy were identified. The second year served to validate these parameters and to generate a multivariate model that could be applied in clinic practice. The model presented a sensitivity of 38.4% (CI 95% 31.9% to 45.3%) and a specificity of 96.8% (CI 95% 83.8% to 99.8%). The odds were 11.9; the positive predictive value was 65.26% and the negative predictive value of 90.94%. Therefore, the retrospective application of this model would have increased the rate of clinical pregnancy by 30%, if applied in the same patients included in the study. Universitat Autònoma de Barcelona. Programa de Doctorat en Pediatria, Obstetrícia i Ginecologia

Male infertility, a condition that has during the last decade raised significant concern, is a diagnostically demanding and socially sensitive topic. The number of unsolved issues on infertility etiology, especially potential environmental causes, in couples demonstrates the need for further investigations into infertility biomarkers. Semen parameters are often insufficient for reliable profiling of male infertility. Thus, this study aims to evaluate for the first time seminal plasma N-glycosylation as a biomarker of environmental exposure in semen samples from 82 normozoospermic men and 84 men with abnormal semen parameters and compare it with genome damage measured by DNA fragmentation. We obtained information about chronic exposure to environmental factors from the self-reported questionnaire, and determined sperm DNA fragmentation by sperm chromatin dispersion, while N-glycans were characterized with liquid chromatography-mass spectrometry (LC-MS). Based on previously published results, ten N-glycans were selected. Results show that the selected seminal plasma N-glycans were significantly associated with smoking, exposure to pesticides, air pollution, agents emitted during photocopying, alcohol consumption, and obesity. Some N-glycans showed a simultaneous association with DNA fragmentation, semen parameters, and environmental stressors. These subgroups of N- glycans are new potential candidates for biomonitoring of exposure to different environmental factors in men with semen abnormalities.

El estudio de la fragmentación del ADN espermático para la evaluación de varones infértiles ha sido de gran interés en los últimos años. Este daño puede ser de cadena sencilla (SSB), mayoritariamente debido a la presencia de estrés oxidativo y relacionado con la capacidad de conseguir un embarazo y la tasa de fecundación; o de cadena doble (DSB), con un origen mayoritariamente enzimático y relacionado con una cinética embrionaria lenta, una menor tasa de implantación y un riesgo elevado de aborto. La fragmentación en el ADN de los espermatozoides se puede estudiar mediante diferentes técnicas como el test Comet (versiones alcalina y neutra), el test de dispersión de la cromatina o el test TUNEL. El primer objetivo de la presente Tesis Doctoral ha sido encontrar una posible explicación a la falta de asociación entre los valores de fragmentación y los resultados reproductivos en tratamientos de Inyección intracitoplasmática de espermatozoides (ICSI). Para ello, se ha realizado un estudio multimetodológico de la fragmentación con protocolos adaptados para el estudio de espermatozoides microseleccionados bajo criterios de ICSI (ICSI-S). También se han estudiado espermatozoides descartables para ICSI por presentar alteraciones en su movilidad o morfología (ICSI-N). Los resultados muestran la existencia de un sesgo diagnóstico al analizar los valores de SSB y DSB en la muestra de eyaculado en comparación con la subpoblación de espermatozoides candidatos para ICSI. Mientras que los valores de SSB se reducen significativamente en los ICSI-S en comparación con el eyaculado y la muestra tras realizar un Swim-up, los valores de DSB se mantienen en el mismo nivel. Por su parte, los espermatozoides ICSI-N muestran valores incrementados de SSB y DSB, posiblemente relacionado con la presencia de un nivel elevado de estrés oxidativo y de apoptosis abortiva. El segundo objetivo ha sido estudiar el efecto de las roturas de cadena doble en ciclos de ICSI. Se han comparado las tasas de fecundación y euploidía, así como su efecto clínico, tras el uso de un método convencional de preparación seminal (Gradientes de densidad) y tras el uso de un 2 nuevo método de selección microfluídico (dispositivo ZyMōt™ICSI). Previamente, el método convencional ha mostrado una reducción eficiente de los valores de SSB, pero no de DSB. Por el contrario, el dispositivo microfluídico sí ha mostrado resultados prometedores para reducir significativamente los niveles de ambos tipos de roturas. Los resultados muestran que el uso del dispositivo microfluídico permite aumentar la tasa de fecundación en comparación a los Gradientes de densidad, posiblemente debido a la mayor reducción en los valores de SSB. El dispositivo microfluídico también ha permitido aumentar la tasa de euploidía de los embriones obtenidos, especialmente cuando la pareja femenina es menor de 35 años en comparación con el uso del sistema convencional, posiblemente debido a la reducción de los valores de DSB. En cuanto a resultados clínicos, los datos muestran que el uso del dispositivo ZyMōt™ICSI permite obtener mejores tasas de embarazo bioquímico y embarazo clínico y una menor tasa de aborto en comparación con métodos convencionales. The study of sperm DNA fragmentation to evaluate infertile men has been an emerging field during the last years. DNA breaks can affect one or both strands, resulting in Single-Strand Breaks (SSB) or Double-Strand Breaks (DSB). SSB in sperm DNA have an oxidative origin and showed to reduce the chances of achieving a natural pregnancy. DSB have an enzymatic origin and showed to reduce embryo’s kinetics, implantation rates and to increase the miscarriage risk. DNA breaks can be studied using different techniques such as the Comet assay (alkaline and neutral versions), the Sperm chromatin dispersion test or the TUNEL test. The main objective of this Doctoral Thesis was to study the effect of sperm DNA damage on Intracytoplasmic sperm injection (ICSI) treatments. For this purpose, DNA breaks were analysed on microselected sperm candidates for ICSI (ICSI-S). Sperm presenting motility or morphological alterations were also studied (ICSI-N). The results revealed a diagnostic bias when SSB and DSB are measured on the ejaculate sample compared to those values found on ICSI-S. SSB were significantly reduced after semen preparation through a Swim-up procedure and after ICSI selection compared to the ejaculate. On the contrary, DSB were present in sperm after the Swim-up and after ICSI sperm selection in the same proportion than in the ejaculate sample. On the other hand, ICSI-N sperm showed significantly increased values of SSB and DSB compared to the ejaculate, the Swim-up sample and ICSI-S, probably related to high oxidative stress conditions and to the abortive apoptosis process. 3 After showing that conventional methods based on sperm motility may reduce SSB values but do not affect DSB, the second objective of the present Doctoral Thesis was to study the effect of these breaks on reproductive outcomes after ICSI. Fertilization and euploidy rates were compared after using a conventional method for semen preparation (Density gradients) and using a novel microfluidic sperm sorting device (ZyMōt™ICSI). The conventional method has previously shown to reduce SSB values, but not DSB values. On the contrary, the microfluidic device has recently shown to reduce both types of damage. The use of the microfluidic device resulted in better fertilization rates compared to the use of Density gradients. These results might be attributed to a better reduction in SSB values after using the ZyMōt™ICSI. The use of this device also increased the euploidy rate of the obtained embryos after ICSI, especially when the female partner was under 35 years of age. This increase might be attributed to the reduction in DSB values. The use of the microfluidic device also increased the euploidy rate when women were older than 35 years, but the presence of aneuploidies related to women’s age persisted. In addition to the second objective, the effect of the microfluidic sorting device on clinical outcomes after ICSI was studied. The use of this device when patients presented high DSB values resulted in better biochemical and clinical pregnancy rates and lower miscarriage rates compared to conventional methods. Universitat Autònoma de Barcelona. Programa de Doctorat en Biologia Cel·lular

El cos humà està colonitzat per bacteris comensals que habiten superfícies i fluids de l'organisme, una comunitat coneguda com a microbiota, amb implicacions en la salut humana. La presència o absència de determinades espècies bacterianes altera el funcionament fisiològic, fet que ha impulsat estudis dirigits a conèixer el seu paper en algunes malalties, com per exemple la infertilitat. Sota la hipòtesi que la microbiota pot alterar paràmetres seminals i espermàtics associats a la infertilitat masculina, s'han proposat un seguit d'objectius per a estudiar-ho. S'han valorat paràmetres relacionats amb la integritat de l'ADN espermàtic com l'estat de compactació de la cromatina, la fragmentació de l'ADN o l'estrès oxidatiu, juntament amb els paràmetres clínics d'un seminograma. En primer lloc, s'ha valorat la relació entre aquests paràmetres per, a continuació, caracteritzar la microbiota seminal a través de la seqüenciació del gen ribosomal 16S complet mitjançant les plataformes MiSeq d'Illumina i MinION d'Oxford Nanopore Technologies, i buscar-hi associacions amb els paràmetres valorats. S'han pogut establir relacions entre un biomarcador recentment descrit, el potencial oxidant-reductor, i altres paràmetres més coneguts com el pH, l'estat de compactació de la cromatina o la fragmentació d'ADN. S'ha descrit la composició bacteriana seminal, integrada principalment per Firmicutes, Bacteroidetes, Proteobacteria i Actinobacteria, i s'han establert relacions entre alguns gèneres i famílies amb els paràmetres de qualitat seminal. El cuerpo humano está colonizado por bacterias comensales que habitan superficies y fluidos del organismo, una comunidad conocida como microbiota, con implicaciones en la salud humana. La presencia o ausencia de determinadas especies bacterianas altera el funcionamiento fisiológico, lo que ha impulsado estudios dirigidos a conocer su papel en algunas enfermedades, como por ejemplo la infertilidad. Bajo la hipótesis de que la microbiota puede alterar parámetros seminales y espermáticos asociados a la infertilidad masculina, se han propuesto una serie de objetivos para estudiarlo. Se han valorado parámetros relacionados con la integridad de l'ADN espermático como el estado de compactación de la cromatina, la fragmentación de l'ADN o el estrés oxidativo, junto con los parámetros clínicos de un seminograma. En primer lugar, se ha valorado la relación entre estos parámetros para, a continuación, caracterizar la microbiota seminal a través de la secuenciación del gen ribosomal 16S completo mediante las plataformas MiSeq de Illumina y MinION de Oxford Nanopore Technologies, buscando asociaciones con los parámetros valorados. Se han podido establecer relaciones entre un biomarcador recientemente descrito, el potencial oxidante-reductor, y otros parámetros más conocidos como el pH, el estado de compactación de la cromatina o la fragmentación d'ADN. Se ha descrito la composición bacteriana seminal, integrada principalmente por Firmicutes, Bacteroidetes, Proteobacteria y Actinobacteria, y se han establecido relaciones entre algunos géneros y familias con los parámetros de calidad seminal. The human body is colonized by commensal bacteria that inhabit the body's surfaces and fluids, a community known as microbiota, with implications for human health. The presence or absence of certain bacterial species alters physiological functioning, which has led to studies aimed at understanding their role in some diseases, such as infertility. Under the hypothesis that the microbiota may alter seminal and sperm parameters associated with male infertility, some objectives have been proposed. Parameters related to sperm DNA integrity such as chromatin compaction, DNA fragmentation, or oxidative stress have been assessed, along with the clinical parameters of a seminogram. The relationship between these parameters was first assessed, and then seminal microbiota was characterized through full-length 16S ribosomal gene sequencing using Illumina MiSeq and Oxford Nanopore Technologies MinION platforms, and relationship was assessed with the rated parameters. Relationships have been established between a recently described biomarker, oxidation-reduction potential, and other better-known parameters such as pH, chromatin compaction, or DNA fragmentation. The seminal bacterial composition, composed mainly of Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria, has been described, and relationships have been established between some genus and families with the seminal quality parameters.

Purpose: Seminal oxidative stress (OS) is a recognized factor potentially associated with male infertility, but the efficacy of antioxidant (AOX) therapy is controversial and there is no consensus on its utility. Primary outcomes of this study were to investigate the effect of AOX on spontaneous clinical pregnancy, live birth and miscarriage rates in male infertile patients. Secondary outcomes were conventional semen parameters, sperm DNA fragmentation (SDF) and seminal OS. Materials and Methods: Literature search was performed using Scopus, PubMed, Ovid, Embase, and Cochrane databases. Only randomized controlled trials (RCTs) were included and the meta-analysis was conducted according to PRISMA guidelines. Results: We assessed for eligibility 1,307 abstracts, and 45 RCTs were finally included, for a total of 4,332 infertile patients. We found a significantly higher pregnancy rate in patients treated with AOX compared to placebo-treated or untreated controls, without significant inter-study heterogeneity. No effects on live-birth or miscarriage rates were observed in four studies. A significantly higher sperm concentration, sperm progressive motility, sperm total motility, and normal sperm morphology was found in patients compared to controls. We found no effect on SDF in analysis of three eligible studies. Seminal levels of total antioxidant capacity were significantly higher, while seminal malondialdehyde acid was significantly lower in patients than controls. These results did not change after exclusion of studies performed following varicocele repair. Conclusions: The present analysis upgrades the level of evidence favoring a recommendation for using AOX in male infertility to improve the spontaneous pregnancy rate and the conventional sperm parameters. The failure to demonstrate an increase in live-birth rate, despite an increase in pregnancy rates, is due to the very few RCTs specifically assessing the impact of AOX on live-birth rate. Therefore, further RCTs assessing the impact of AOX on live-birth rate and miscarriage rate, and SDF will be helpful.

Simple Summary Freezing dog semen is not always possible due to low quality sperm or poor survival during freezing. In order to make this assisted reproductive technique available to a larger number of dogs, this study investigated the benefit of selecting the best spermatozoa before freezing using single layer centrifugation (SLC). The results indicated that this technique was effective in separating spermatozoa according to their quality, although this resulted in losing some good quality spermatozoa. After thawing, spermatozoa centrifuged by SLC were of better quality than after standard centrifugation. However, spermatozoa from suboptimal quality semen did not survive freezing as well as spermatozoa from semen of optimal quality, even after SLC. Single layer centrifugation, therefore, makes it possible to obtain better quality spermatozoa after thawing but is not sufficient on its own to improve the inferior freezing ability of spermatozoa from suboptimal quality semen. So far, eighteen pups were born after insemination with SLC-selected frozen-thawed semen, proving that these selected spermatozoa remain fertile. This study evaluated how semen selection by single layer centrifugation (SLC) with Canicoll affects semen freezability in dogs. A total of eighteen ejaculates, collected from dogs with optimal and suboptimal semen quality (optimal: normal morphology (NM) >= 80%, n = 9; suboptimal: NM between 60 and 79%, n = 9), were divided into two aliquots and subjected to standard centrifugation or SLC before cryopreservation. Motility, NM, membrane integrity, mitochondrial membrane potential (MMP), and DNA integrity were improved in fresh samples after SLC, regardless of semen quality, but at the expense of some good quality spermatozoa. After thawing, NM and membrane integrity were improved in SLC-selected semen in both semen qualities. Interestingly, MMP was also higher but only in optimal quality semen. Still, spermatozoa from suboptimal quality semen did not survive freezing to the same extent as spermatozoa from optimal quality semen, even after selecting superior spermatozoa. Semen selection with Canicoll is, therefore, an effective technique to isolate a subpopulation of high-quality spermatozoa and obtain sperm samples of better quality after thawing, but is not sufficient to improve the intrinsic inferior freezability of suboptimal quality semen. So far, eighteen pups were born after insemination with SLC-selected frozen-thawed semen, proving that these selected spermatozoa remain fertile.

El cos humà està colonitzat per bacteris comensals que habiten superfícies i fluids de l’organisme, una comunitat coneguda com a microbiota, amb implicacions en la salut humana. La presència o absència de determinades espècies bacterianes altera el funcionament fisiològic, fet que ha impulsat estudis dirigits a conèixer el seu paper en algunes malalties, com per exemple la infertilitat. Sota la hipòtesi que la microbiota pot alterar paràmetres seminals i espermàtics associats a la infertilitat masculina, s’han proposat un seguit d’objectius per a estudiar-ho. S’han valorat paràmetres relacionats amb la integritat de l’ADN espermàtic com l’estat de compactació de la cromatina, la fragmentació de l’ADN o l’estrès oxidatiu, juntament amb els paràmetres clínics d’un seminograma. En primer lloc, s’ha valorat la relació entre aquests paràmetres per, a continuació, caracteritzar la microbiota seminal a través de la seqüenciació del gen ribosomal 16S complet mitjançant les plataformes MiSeq d’Illumina i MinION d’Oxford Nanopore Technologies, i buscar-hi associacions amb els paràmetres valorats. S’han pogut establir relacions entre un biomarcador recentment descrit, el potencial oxidant-reductor, i altres paràmetres més coneguts com el pH, l’estat de compactació de la cromatina o la fragmentació d’ADN. S’ha descrit la composició bacteriana seminal, integrada principalment per Firmicutes, Bacteroidetes, Proteobacteria i Actinobacteria, i s’han establert relacions entre alguns gèneres i famílies amb els paràmetres de qualitat seminal. El cuerpo humano está colonizado por bacterias comensales que habitan superficies y fluidos del organismo, una comunidad conocida como microbiota, con implicaciones en la salud humana. La presencia o ausencia de determinadas especies bacterianas altera el funcionamiento fisiológico, lo que ha impulsado estudios dirigidos a conocer su papel en algunas enfermedades, como por ejemplo la infertilidad. Bajo la hipótesis de que la microbiota puede alterar parámetros seminales y espermáticos asociados a la infertilidad masculina, se han propuesto una serie de objetivos para estudiarlo. Se han valorado parámetros relacionados con la integridad de l’ADN espermático como el estado de compactación de la cromatina, la fragmentación de l’ADN o el estrés oxidativo, junto con los parámetros clínicos de un seminograma. En primer lugar, se ha valorado la relación entre estos parámetros para, a continuación, caracterizar la microbiota seminal a través de la secuenciación del gen ribosomal 16S completo mediante las plataformas MiSeq de Illumina y MinION de Oxford Nanopore Technologies, buscando asociaciones con los parámetros valorados. Se han podido establecer relaciones entre un biomarcador recientemente descrito, el potencial oxidante-reductor, y otros parámetros más conocidos como el pH, el estado de compactación de la cromatina o la fragmentación d’ADN. Se ha descrito la composición bacteriana seminal, integrada principalmente por Firmicutes, Bacteroidetes, Proteobacteria y Actinobacteria, y se han establecido relaciones entre algunos géneros y familias con los parámetros de calidad seminal. The human body is colonized by commensal bacteria that inhabit the body's surfaces and fluids, a community known as microbiota, with implications for human health. The presence or absence of certain bacterial species alters physiological functioning, which has led to studies aimed at understanding their role in some diseases, such as infertility. Under the hypothesis that the microbiota may alter seminal and sperm parameters associated with male infertility, some objectives have been proposed. Parameters related to sperm DNA integrity such as chromatin compaction, DNA fragmentation, or oxidative stress have been assessed, along with the clinical parameters of a seminogram. The relationship between these parameters was first assessed, and then seminal microbiota was characterized through full-length 16S ribosomal gene sequencing using Illumina MiSeq and Oxford Nanopore Technologies MinION platforms, and relationship was assessed with the rated parameters. Relationships have been established between a recently described biomarker, oxidation-reduction potential, and other better-known parameters such as pH, chromatin compaction, or DNA fragmentation. The seminal bacterial composition, composed mainly of Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria, has been described, and relationships have been established between some genus and families with the seminal quality parameters. Universitat Autònoma de Barcelona. Programa de Doctorat en Biologia Cel·lular

Clinical outcome in assisted reproduction techniques (ARTs) is mainly influenced by the quality of gametes used. It is known that a high percentage of sperm DNA fragmentation (DNAf) decreases the success of ART clinical results. Therefore, techniques such as magnetic-activated cell sorting (MACS) help to improve results in cases of patients with a high percentage of DNAf. Cryopreservation of sperm in donor intrauterine insemination (D-IUI) treatments increases sperm DNAf, so patients using these sperm samples can benefit from using this technique. This prospective randomized national multicenter study analyzed clinical outcomes of 181 D-IUI treatments. MACS was performed after density gradient centrifugation (DGC) in 90 thawed semen donor samples (MACSG), whereas only DGC was performed in 91 thawed semen donor samples (CG). To our knowledge, this is the first study analyzing the effect of MACS on D-IUI cycles. Our results show no significant differences in gestation, live birth, or miscarriage rates between the two groups. We believe that further studies with a larger sample size are needed to evaluate the application of MACS in combination with standard IUI donor sperm preparations in fertility clinics.

Purpose: In recent years, a growing number of studies have supported the beneficial effects of a very short abstinence period on sperm parameters, especially in patients with oligoasthenozoospermia. However, the results are controversial and no consensus exists regarding whether to request a second semen collection in clinical practice. Therefore, this systematic review and meta-analysis aimed to evaluate the influence of a very short abstinence period (within 4 h) on conventional sperm parameters and sperm DNA fragmentation (SDF) rate. Materials and Methods: The literature search was performed using Scopus and PubMed databases. The meta-analysis was conducted according to the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocol (PRISMA-P) guidelines. All eligible studies were selected according to the Population, Intervention, Comparison/Comparator, Outcomes, and Study design (PICOS) model. The quality of evidence of the included studies was analyzed through the Cambridge Quality Checklists. The standardized mean difference (SMD) was used to analyze the outcomes. Cochran-Q and I2 statistics were used to evaluate statistical heterogeneity. Results: We assessed for eligibility 1334 abstracts, and 19 studies were finally included. All 19 articles evaluated the effects of a very short abstinence period on sperm parameters and, among these, 5 articles also evaluated the effects on SDF rate. The quantitative analysis showed a significant reduction in semen volume after a very short abstinence period in both normozoospermic men and patients with oligozoospermia, asthenozoospermia, and/or teratozoospermia (OAT) patients. We found a statistically significant increase in sperm concentration and total and progressive motility in the second ejaculation of patients with OAT. In contrast, the SDF rate decreased significantly in the second ejaculate of OAT patients. Conclusions: This is the first systematic review and meta-analysis investigating the impact of a very short abstinence period on sperm parameters and SDF rate. The results suggest that collecting a second consecutive ejaculation after a very short time from the first could represent a simple and useful strategy for obtaining better-quality spermatozoa, especially in patients with abnormal sperm parameters.

Semen parameters are unable to inform on the function or fertilizing capacity of the male gamete. Standardized methods are provided by the WHO but, the lower reference limits have reduced sensitivity to predict chances of conception. Subfertile men may be falsely classified as “normal” and a male factor contributing to genome instability may be overlooked. Semen parameters, sperm DNA fragmentation (SDF), sperm chromatin maturity and stability, and sperm aneuploidy were assessed in fertile (F), subfertile normozoospermic (SN) and subfertile non-normozoospermic males (SN-N). Standardized assays employing flow cytometry were used to detect genome instability. Sperm DNA fragmentation did not differ significantly whether the semen samples were from a fertile (F), subfertile normozoospermic (SN) or subfertile non-normozoospermic male (SN-N). Chromatin decondensation was significantly reduced and hyperstability significantly increased in the SN group as compared to the F group. The frequency of diploidy was significantly different in the three study groups with significance between F and SN and between F and SN-N groups. Subfertile men with normal semen parameters are often excluded from extensive genetic testing. Genome instability might be an independent attribute of semen quality detecting problems not seen with semen analysis alone.

The evaluation of morpho-functional sperm characteristics alone is not enough to explain infertility or to predict the outcome of Assisted Reproductive Technologies (ART): more sensitive diagnostic tools are needed in clinical practice. The aim of the present study was to analyze Sperm DNA Fragmentation (SDF) and sperm-borne miR-34c-5p and miR-449b-5p levels in men of couples undergoing ART, in order to investigate any correlations with fertilization rate, embryo quality and development. Male partners (n = 106) were recruited. Semen analysis, SDF evaluation and molecular profiling analysis of miR-34c-5p and miR-449b-5p (in 38 subjects) were performed. Sperm DNA Fragmentation evaluation- a positive correlation between SDF post sperm selection and the percentage of low-quality embryos and a negative correlation with viable embryo were found. SDF > 2.9% increased the risk of obtaining a non-viable embryo by almost 4-fold. Sperm miRNAs profile—we found an association with both miRNAs and sperm concentration, while miR-449b-5p is positively associated with SDF. Moreover, the two miRNAs are positively correlated. Higher levels of miR-34c-5p compared to miR-449b-5p increases by 14-fold the probability of obtaining viable embryos. This study shows that SDF, sperm miR-34c-5p, and miR-449b-5p have a promising role as biomarkers of semen quality and ART outcome.

International audience; Semen analysis is the cornerstone of male fertility evaluation with WHO guidelines providing the basis for procedural standardization and reference values worldwide. The first WHO manual was published in 1980, and five editions have been subsequently released over the last four decades. The 6th Edition was published in July 2021. In this review, we identify the key changes of this 6th Edition. Additionally, we evaluate the utility of this 6th Edition in clinical practice using SWOT (strengths, weaknesses, opportunities, and threats) analysis. This new Edition has made the analysis of basic semen parameters more robust, taking into account the criticisms and grey areas of the previous editions. The tests assessing sperm DNA fragmentation and seminal oxidative stress are well-described. The main novelty is that this latest edition abandons the notion of reference thresholds, suggesting instead to replace them with “decision limits”. While this seems attractive, no decision limits are proposed for either basic semen parameters, or for extended or advanced parameters. This critical review of the 6th Edition of the WHO laboratory manual combined with a SWOT analysis summarizes the changes and novelties present in this new Edition and provides an in-depth analysis that could help its global use in the coming years.

Objective: Our study aims to retrospectively examine the relationship between two different sperm preparation methods used in IUI among eight years in terms of pregnancy and live birth rates. Methods: We evaluated the data of semen samples between December 2012 and March 2020. Three hundred eighty-four samples prepared with Conventional Swim-up (CSW) and 361 samples prepared with Density Gradient-Swim up (DGC-SW) obtained from men applying for IUI were analyzed. Spermiogram results of the semen samples given by men applying for IUI were examined. Data about sperm preparation method, post washed sperm parameters, pregnancy, and live birth rate were collected. Statistical analysis was performed. Results: Basal progressive sperm count was significantly higher in pregnant couples in both CSW and DGCSW groups (p = 0,032, p = 0,035, respectively). In each group, the post washed total progressive motile sperm count obtained by CSW and DGC-SW methods were significantly higher in pregnant patients (p < 0.05). There was no significant difference between CSW and DGC-SW methods in pregnancy achievement (p = 0,399, X-2 = 0,712). Live birth and miscarriage rates were not different between the groups (p = 0,243, X-2 = 2.827). Conclusion: In conclusion, there is no significant difference between CSW and DGC-SW for pregnancy and live birth rates. Our results suggest that both sperm preparation techniques used in IUI are not superior to each other. In other words, the choice of sperm preparation method does not affect the pregnancy rate in couples undergoing IUI.(c) 2022 Elsevier Masson SAS. All rights reserved.

To date 15% of couples are suffering from infertility with 45–50% of males being responsible. With an increase in paternal age as well as various environmental and lifestyle factors worsening these figures are expected to increase. As the so-called free radical theory of infertility suggests, free radicals or reactive oxygen species (ROS) play an essential role in this process. However, ROS also fulfill important functions for instance in sperm maturation. The aim of this review article is to discuss the role reactive oxygen species play in male fertility and how these are influenced by lifestyle, age or disease. We will further discuss how these ROS are measured and how they can be avoided during in-vitro fertilization., Graphical abstract Here we review how reactive oxygen species are created in sperm cells and how they influence fertility. We include a discussion on physiology and pathophysiology. We further review lifestyle factors and issues related to in-vitro fertilization.Image 1

Background. Antioxidants are commonly used for the treatment of idiopathic male infertility. Previous studies have shown that antioxidants are able to improve sperm quality, but little is known about their impact on sperm DNA fragmentation (SDF). Preliminary findings showed that superoxide-dismutase (SOD)-based antioxidant plus hydroxytyrosol and carnosol (FertiPlus® SOD) therapy was associated with SDF improvement in a small cohort of infertile men. Therefore, we aimed to assess rates of and predictors of semen parameters and SDF improvements in infertile men treated with FertiPlus® SOD therapy (SOD+) or with other antioxidants without SOD (SOD−) in the real-life setting. Methods. Data from 60 consecutive infertile men with baseline SDF ≥ 30% and treated with SOD+ or SOD− for at least three months were analyzed. Clinical parameters and serum hormones were collected. Sperm parameters and SDF were requested at baseline and after SOD+ or SOD− treatment. Clinically meaningful SDF change after treatment was defined as SDF improvement >20% compared to baseline. Propensity score matching was performed to adjust for baseline differences between groups. Descriptive statistics were used to compare clinical and hormonal characteristics between SOD+ and SOD− groups. Semen characteristics were compared before and after treatment. Logistic regression models investigated the association between clinical variables and SDF improvement. Results. Groups were similar in terms of clinical, serum hormones and semen parameters at baseline after matching. Compared to baseline, sperm progressive motility (17 (10–36)% vs. 27 (12–41)%) and normal morphology (2 (1–6)% vs. 4 (2–6)%) significantly improved after SOD+ treatment (all p < 0.01), but not after SOD−. SDF values significantly improved after treatment in both groups, compared to the baseline evaluation (all p < 0.01). However, SDF values were lower after SOD+ than SOD− treatment (30 (22–36)% vs. 37 (31–42)%, p = 0.01). Similarly, a clinically meaningful improvement in SDF at follow-up was more frequently found after SOD+ than SOD− treatment (76.7% vs. 20.0%, p = 0.001). Multivariable logistic regression analysis showed that SOD+ treatment (OR 5.4, p < 0.001) was an independent predictor of clinically meaningful SDF improvement, after accounting for age and baseline FSH values. Conclusions. This cross-sectional study showed that, in a cohort of primary infertile men with SDF ≥ 30%, SOD-based treatment was significantly effective in improving SDF compared to antioxidants without SOD. Approximately 80% of men treated with SOD+ achieved clinically meaningful improvement in SDF after three months of treatment. Sperm progressive motility and normal morphology also improved after SOD+ therapy but not after SOD−. These results suggest that SOD+ treatment could be considered an effective option for the management of idiopathic infertile men with elevated SDF.

Objective To evaluate the effect of varicocelectomy on sperm deoxyribonucleic acid fragmentation (SDF) rates in infertile men with clinical varicocele. Design Systematic review and meta-analysis. Setting Not applicable. Patient(s) Infertile men with clinical varicocele subjected to varicocelectomy. Intervention(s) Systematic search using PubMed/Medline, EMBASE, Cochrane’s central database, Scielo, and Google Scholar to identify relevant studies published from inception until January 2021. We included studies comparing SDF rates before and after varicocelectomy in infertile men with clinical varicocele. Main Outcome Measure(s) The primary outcome was the difference between the SDF rates before and after varicocelectomy. A meta-analysis of weighted data using random-effects models was performed. Results were reported as weighted mean differences (WMD) with 95% confidence intervals (CIs). Subgroup analyses were performed on the basis of the SDF assay, varicocelectomy technique, preoperative SDF levels, varicocele grade, follow-up time, and study design. Result(s) Nineteen studies involving 1,070 patients provided SDF data. Varicocelectomy was associated with reduced postoperative SDF rates (WMD −7.23%; 95% CI: −8.86 to −5.59; I2 = 91%). The treatment effect size was moderate (Cohen’s d = 0.68; 95% CI: 0.77 to 0.60). The pooled results were consistent for studies using sperm chromatin structure assay, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, sperm chromatin dispersion test, and microsurgical varicocele repair. Subgroup analyses showed that the treatment effect was more pronounced in men with elevated vs. normal preoperative SDF levels, but the impact of varicocele grade remained equivocal. Meta-regression analysis demonstrated that SDF decreased after varicocelectomy as a function of preoperative SDF levels (coefficient: 0.23; 95% CI: 0.07 to 0.39). Conclusion(s) We concluded that pooled results from studies including infertile men with clinical varicocele indicated that varicocelectomy reduced the SDF rates. The treatment effect was greater in men with elevated (vs. normal) preoperative SDF levels. Further research is required to determine the full clinical implications of SDF reduction for these men.

Since the publication of the first edition of the WHO (World Health Organization) Laboratory Manual for the examination of Human Semen in 1980, the reference values of sperm parameters have been updated on four occasions. Currently and globally, most of the laboratories analyzing semen samples use the latest, 5th edition of the manual that recommends ejaculatory abstinence from two to seven days before producing the sample for examination. While this standardized interval of time facilitates the interpretation of the results and research, no solid evidence exists to support the WHO-recommended abstinence time for a semen analysis in order to optimize clinical outcomes after assisted reproduction. Most of the studies refer to different clinical outcomes, different groups of patients and different editions of the WHO Laboratory Manual, including heterogeneous intervals of abstinence or sperm parameters. The aim of the current systematic review was to evaluate available evidence correlating ejaculatory abstinence time with clinical outcomes and sperm parameters analyzed according to the last edition of the World Health Organization Laboratory Manual reference values in different male populations. The results from the included studies indicate that WHO abstinence recommendations may need revision, given that a shorter ejaculatory abstinence interval appears to be associated with improved sperm parameters, such as sperm DNA fragmentation, progressive motility or morphology, while evidence suggests a potential increase in embryo euploidy rates and pregnancy outcomes.