Evaluation of the relation between preapoptotic sperm rate in ejaculate and intrauterine insemination success rate in patients with unexplained infertility.

The parameters measured in the standard semen analysis may be insufficient for exact differentiation between fertile and infertile men. Therefore, we assume that the high rate of apoptotic sperm in ejaculate may play a role on the aetiology of unexplained infertility. Couples with unexplained infertility treated by ovulation induction and intrauterine insemination were consecutively enrolled (n = 94). To determine the proapoptotic sperm rate, the ejaculate from patients was stained with annexin V. Thirteen of the 94 couples (13.8%) conceived after intrauterine insemination. The annexin V-positive sperm rate was found to be 20.0% in the whole group. In women failing to conceive, the annexin V-positive sperm rate was 20.8% compared to 15.7% in patients who achieved pregnancy. Although there is a trend towards higher preapoptotic sperm rate in couples failing to get pregnant with insemination, the difference did not reach statistical significance.

Obesity is associated with increased seminal insulin and leptin alongside reduced fertility parameters in a controlled male cohort.

BACKGROUND: Obesity appears to be associated with male reproductive dysfunction and infertility, although this has been inconsistent and inconclusive. Insulin and leptin are known mediators and modulators of the hypothalamus-pituitary-testes axis, contributing to the regulation of male reproductive potential and overall wellbeing. These hormones are also present in semen influencing sperm functions. Although abdominal obesity is closely associated with insulin resistance (hyperinsulinaemia), hyperleptinaemia and glucose dysfunction, changes in seminal plasma concentrations of insulin, leptin and glucose in obese males has not previously been investigated. METHODS: This small case controlled study assessed serum and seminal concentrations of insulin, leptin and glucose in obese (BMI > =30; n = 23) and non-obese (BMI < 30; n = 19) males. Following a detailed medical history and examination, participants meeting the inclusion criteria were entered for data analysis. Body parameters such as BMI, waist and hip circumference and the waist hip ratio were measured. Serum and semen samples were collected and assayed for insulin, leptin and glucose. Semen samples also underwent a standard semen analysis, with sperm mitochondrial membrane potential (MMP) and DNA fragmentation (DF). RESULTS: Obesity was associated with increased serum and seminal insulin and leptin, with no significant difference in seminal glucose. Serum and seminal concentrations of insulin and leptin were positively correlated. Furthermore, obesity was associated with decreased sperm concentration, sperm vitality and increased MMP and DF, with a non-significant impact on motility and morphology. CONCLUSIONS: Hyperinsulinaemia and hyperleptinaemia are associated with increased seminal insulin and leptin concentrations, which may negatively impact male reproductive function in obesity. Insulin was also found to be highly concentrated in the seminal plasma of both groups. This data will contribute to the contradictive information available in the literature on the impact of obesity and male reproduction.

Male reproductive health statement (XIIIth international symposium on Spermatology, may 9th-12th 2018, Stockholm, Sweden.

On the occasion of the XIIIth International Symposium on Spermatology held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest speakers and audience) collectively felt the need to make a public statement on the general issue of male reproductive health. Our intention is to raise awareness of what we believe is a neglected area of research despite alarming situations around the world. The disclosure strategy desired by the co-authors is to bring it to the attention of the greatest number partly by considering co-publication in the various periodicals dealing with Reproductive Biology and Andrology. BaCA's editorial office accepted this mission and found it natural that our periodical, the official journal of the French Andrology Society (SALF), should carry this message.

A l'occasion du XIII eme Symposium international sur la Spermatologie qui s'est. tenu du 9 au 13 Mai 2018 à Stockholm (Suède), les participants (orateurs invités et l'auditoire) ont ressenti collectivement le besoin de faire une déclaration publique sur la question générale de la santé reproductive masculine. Notre intention est. de mieux faire connaître ce que nous pensons être un domaine de recherche négligé malgré des situations alarmantes dans le monde entier. La stratégie de divulgation souhaitée par les co-auteurs est. de le porter à l'attention du plus grand nombre en envisageant pour partie une co-publication dans les différents périodiques traitant de Reproduction et d'Andrologie. Le bureau éditorial de BaCA, a accepté cette mission et a trouvé naturel que notre périodique, journal officiel de la Société d'Andrologie en Langue Française (SALF) porte ce message.

Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients.

OBJECTIVE: To investigate whether there is an impact of different sources of reactive oxygen species (ROS) on sperm functions. DESIGN: Prospective study. SETTING: Patients at the Center for Dermatology and Andrology, Giessen, Germany. PATIENT(S): Semen collected from 63 randomly collected patients attending the IVF unit of the University of Giessen, Germany. INTERVENTION(S): Only patients with nonleukocytospermia were included in this study. MAIN OUTCOME MEASURE(S): Sperm count and motility before and after sperm separation by swim-up, morphology, DNA fragmentation, and extrinsic (by leukocytes) and intrinsic ROS production (by spermatozoa) were evaluated. RESULT(S): Leukocytes correlated significantly with extrinsic ROS production (r = 0.576), but markedly less with intrinsic ROS production (r = 0.296). Sperm count, morphology, and motility in the ejaculate were markedly more affected by extrinsic than by intrinsic ROS. The DNA fragmentation was strongly positively correlated with intrinsic ROS production, whereas this correlation was weaker for extrinsic ROS production. No correlation was found between DNA fragmentation and the number of leukocytes, whereas the correlations with motility in the ejaculate and the motile sperm count after swim-up were highly significant. Moreover, significant differences were observed for extrinsic and intrinsic ROS production between groups of patients having a high (> or = 1 x 10(6)/mL) and a low number (<1 x 10(6)/mL) of leukocytes in the ejaculate. CONCLUSION(S): The origin of ROS seems to have an influence on the site of the damage. Because leukocyte counts <1 x 10(6)/mL caused a significant decrease of motility and DNA integrity, the threshold given by the World Health Organization (WHO) should be re-evaluated.

Relationship between zona pellucida-induced acrosome reaction, sperm morphology, sperm-zona pellucida binding, and in vitro fertilization.

OBJECTIVE: To evaluate the possible relationships between sperm morphology, acrosome responsiveness to solubilized human zona pellucida, and sperm-zona binding potential among [1] consecutive andrology referrals and [2] randomly selected in vitro fertilization (IVF) cases. DESIGN: Prospective analytical study. SETTING: Academic training hospital.Randomly selected couples consulting for infertility. INTERVENTION(S): Acrosome reaction response to solubilized human zona pellucida was recorded. MAIN OUTCOME MEASURE(S): We determined the difference in the percentage of sperm that acrosome reacted after exposure to solubilized zona pellucida and spontaneous acrosome reaction. The results were expressed as percentage zona induced acrosome reaction (ZIAR). RESULT(S): Data were analyzed using correlation coefficients (r) and receiver operator characteristics (ROC curve analyses). The ROC curve analyses indicated ZIAR to be a sensitive indicator for fertilization failure during IVF therapy, with sensitivity and specificity of 81% and 75%, respectively. For andrology referrals, a positive and statistically significant correlation existed between ZIAR data and sperm morphology (r = 0.65) and sperm-zona binding (r = 0.57). CONCLUSION(S): ZIAR results provide further information regarding dysfunctional sperm and can be used as an additional diagnostic test. Our results predicted fertilization failure during IVF treatment.

Monitoring technologist reading skills in a sperm morphology quality control program.

OBJECTIVE: The value of sperm morphology as a predictor of a man's fertilizing potential has often been challenged because of different classification systems used to distinguish between normal and abnormal spermatozoa. The study aimed to monitor the reading skills of 53 laboratory technologists who are enrolled in a continuous quality control program for sperm morphology assessment. DESIGN: Prospective analytical study. SETTINGS: Academic hospital and academic institution setting. PATIENT(S): Sperm samples from the sperm donor program and andrology technicians from sub-Saharan Africa. INTERVENTION(S): Papanicolaou-stained sperm slides were prepared and shipped on a quarterly basis to participating laboratories. Papanicolaou-prestained sperm morphology slides were used as test material for 21 months. MAIN OUTCOME MEASURE(S): Percentage normal spermatozoa.A new statistical model was developed to record reading skills of the participating technicians. Reading skills were classified as marginal (5.7% of cases), good (11.3% of cases), and excellent (83% of cases). RESULT(S): Participants maintained their morphological reading skills and agreed with the reference laboratory by not exceeding a SD limit of 0.2 to set stringent standards for the program. CONCLUSION(S): Technician proficiency can be monitored using the results of a quality control program. A continuous quality control program can be initiated only after intensive training, because baseline values at the onset of the quality control program serves as an internal reference value.

Influence of deoxyribonucleic acid damage on fertilization and pregnancy.

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

Effects of folic acid and zinc sulfate on male factor subfertility: a double-blind, randomized, placebo-controlled trial.

OBJECTIVE: To study the effects of folic acid and zinc sulfate treatment on semen variables in fertile and subfertile men. DESIGN: Double-blind, placebo-controlled interventional study. SETTING: Two outpatient fertility clinics and nine midwifery practices in The Netherlands. PARTICIPANT(S): One hundred eight fertile and 103 subfertile men. INTERVENTION(S): Both groups were randomly assigned to receive one of four treatments for 26 weeks: folic acid and placebo, zinc sulfate and placebo, zinc sulfate and folic acid, and two placebos. Folic acid was given at a daily dose of 5 mg, and zinc sulfate was given at a daily dose of 66 mg. MAIN OUTCOME MEASURE(S): Before and after treatment, standardized semen and blood samples were obtained for determinations of sperm concentration, motility, and morphology according to World Health Organization guidelines; semen morphology according to strict criteria; and blood folate and zinc concentrations. Effects of the four interventions were evaluated separately in subfertile and fertile men. RESULT(S): Subfertile men demonstrated a significant 74% increase in total normal sperm count and a minor increase of 4% abnormal spermatozoa. A similar trend was observed in fertile men. Pre-intervention concentrations of folate and zinc in blood and seminal plasma did not significantly differ between fertile and subfertile men. CONCLUSION(S): Total normal sperm count increases after combined zinc sulfate and folic acid treatment in both subfertile and fertile men. Although the beneficial effect on fertility remains to be established, this finding opens avenues of future fertility research and treatment and may affect public health.

Sperm morphology assessment using strict (tygerberg) criteria.

Although sperm morphology evaluation is one of the most important aspects of the semen analysis if done correctly and accurately, a trend is developing in which many laboratories or clinicians no longer regard sperm morphology as relevant due to the very low normal reference value of only 4% morphological normal spermatozoa given in the newest (2010) WHO-5 semen analysis manual. However, to maintain its relevance, sperm morphology evaluation, like the rest of the standard semen analysis, should be performed according to well-defined procedures. If performed correctly and according to high standards, morphology data are of high predictive value for male fertility potential and are critical in selecting the assisted reproductive techniques (ART) treatment. With the new low normal value it is becoming even more important that not only percentage normal and abnormal be reported but also that an in-depth report be given on the types of abnormalities present which will be important in selecting the type of clinical procedure to be adopted in ART, being, IUI, IVF, or ICSI. The methods to properly evaluate human sperm morphology are described in this chapter.

Measurement and significance of sperm morphology.

The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male's fertility potential. In this review the background of the development of the evaluation criteria for sperm morphology will be discussed. Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen. One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns. It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual, sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.

Clinical significance of the low normal sperm morphology value as proposed in the fifth edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen.

The very low cut-off value for sperm morphology of 4% morphologically normal spermatozoa, as proposed in the new edition of the World Health Organization (WHO) manual on semen analysis, is in agreement with recently published values and reflects the trend of a decline in reported mean values for normal sperm morphology. The reduced value for morphologically normal spermatozoa over the years may be due to several factors. The first is the introduction of strict criteria for the evaluation of sperm morphology. Other reasons may include the introduction of additional criteria for sperm morphology abnormalities and the suggested decrease in semen parameters because of increasing negative environmental influences. Although on its own the newly proposed very low normal value may not provide the strong predictive value for a males' fertility potential, as originally reported for sperm morphology evaluated according to strict criteria, a good predictive value can still be obtained if the holistic, strict approach for sperm morphology evaluation is followed together with additional sperm morphology parameters now available, because certain morphology patterns and sperm abnormalities are now known to be of strong prognostic value. In addition, better international standardization of the technical methodology, consensus on the interpretation of sperm morphology evaluation criteria and standardized international external quality control (EQC) schemes, are of utmost importance to maintain the good predictive value of sperm morphology.

Comparison of three staining methods for the morphological evaluation of human spermatozoa.

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

Relationship between human sperm morphology and acrosomal function.

PURPOSE: In this study, we investigated the relationship between functionality of the acrosome and sperm morphology. METHODS: Acrosome reaction (AR) was separately determined in live and dead sperm and in those with normal, small, and large sized acrosomes by means of the triple stain. Morphology was analyzed according to strict criteria after Papanicolaou stain. RESULTS: AR and morphology correlated regarding detection of large and small sized acrosomes, but not for normal sized acrosomes. Spontaneous AR was significantly influenced by acrosomal size. Sperm with large (11.4%) and normal (9.2%) acrosomes exhibited a significantly higher percentage of life spontaneously acrosome-reacted sperm than those with small acrosomes (4.5%). Sperm with small acrosomes were associated with a higher percentage of cell death. CONCLUSION: The results indicate that sperm with small acrosomes are more susceptible to cell death and nonphysiological acrosomal loss. Acrosome size reflects the physiological capability of sperm function and therefore male fertility potential.

DNA fragmentation of spermatozoa and assisted reproduction technology.

Despite the ever-increasing knowledge of the fertilization process, there is still a need for better understanding of the causes of sperm DNA fragmentation and its impact on fertilization and pregnancy. For this reason, human sperm DNA fragmentation was investigated by means of the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) assay and the production of reactive oxygen species (ROS) in the ejaculate and in the spermatozoa themselves. These data were correlated with fertilization and pregnancy data from IVF and intracytoplasmic sperm injection (ICSI) patients. Sperm DNA fragmentation did not correlate with fertilization rate, but there was a significantly reduced pregnancy rate in IVF patients inseminated with TUNEL-positive spermatozoa. ICSI patients exhibited the same tendency. This implies that spermatozoa with damaged DNA are able to fertilize an oocyte, but at the time the paternal genome is switched on, further development stops. The determination of ROS in the ejaculate and the percentage of ROS-producing spermatozoa revealed markedly stronger correlations between sperm functions (i.e. motility) and the percentage of ROS-producing spermatozoa. The influence of seminal leukocytes, known to produce large amounts of oxidants, on sperm DNA fragmentation should not be neglected.