Complete Azoospermia Research Articles

Independent factors associated with intracytoplasmic sperm injection outcomes in patients with complete azoospermia factor c microdeletions

Independent factors associated with intracytoplasmic sperm injection outcomes in patients with complete azoospermia factor c microdeletions

Autologous Bone Marrow-Derived Mesenchymal Stem Cells in the Reversal of Unobstructed Azoospermia in Rats.

Non-obstructive azoospermia (NOA) is an important cause of male infertility. This study is being proposed to assess the efficacy of autologous bone marrow-derived mesenchymal stem cells (MSCs) in the reversal of busulfan-induced NOA in rats. Twenty adult 3-month-old male rats were divided into two groups: a control group and a study group. In the study group, bone marrow was aspirated to culture MSCs. NOA was created by stopping endogenous spermatogenesis in all the animals by injecting two doses of busulfan 10 mg/kg body weight with a 3week interval. Four weeks after the last dose of busulfan, two animals were euthanized and the testes were studied histologically to confirm complete azoospermia. In the study group, five million MSCs in 1 mL normal saline were injected into seminiferous tubules; and in the control group, 1 mL of normal saline was injected. After 4weeks of MSC injection, all the rats were euthanized and epididymis tails and testes were harvested and sent for measurement of serological indices, including luminal, cellular, and total diameters, luminal, cellular, and cross-sectional areas, number of tubules per unit area of testis, numerical density of the tubules, and spermatogenesis index, pre- and post-MSC transplantation. The effect of busulfan on the testicular tissue was universally devastating. In the control group, there was variable length and width of markedly necrotic seminiferous tubules, whereas in the group treated with autologous bone marrow-derived MSCs there was variable height of germinal epithelium in seminiferous tubules, with active spermatogenesis, showing spermatogonia, spermatocytes, and sperm. MSC injection in the testis has the potential to reverse the testicular function of spermatogenesis after cytotoxic therapy. Human trials should be undertaken to confirm our findings and bring the results into clinical practice.

Способ создания модели варикоцеле в хроническом эксперименте.

Introduction. Varicocele is diagnosed in every seventh teenager over 15 years of age, and in total up to 15% of all men suffer. The connection between varicocele and male infertility has been poorly studied, however, spermogram abnormalities in infertile men with varicocele can vary from normozoospermia to complete azoospermia. This determines the relevance of further study and creation of experimental animal models of this disease in order to study the mechanism of its occurrence, the pathophysiology of the conditions associated with it, as well as addressing the issue of prevention and treatment. Purpose of the study: to create a model of persistent varicocele in a laboratory animal in a chronic experiment. Materials and methods. In a chronic experiment, 20 male rabbits were used, randomized into 2 groups: group 1 (control group, n=10) – animals in which a varicocele model was created by narrowing the lumen of the left renal vein by 1/2-2/3 of the original diameter. Group 2 (n=10) – modeling of varicocele by nar- rowing the lumen of the left renal vein by 1/2-2/3 of the original diameter and daily intramuscular injection of gonadotropin 300 units/kg body weight and 0,2 ml of 1% progesterone solution, starting from the first preoperative day. The duration of the course of drug treatment was 10 days. The criterion for creating a varicocele model was considered to be registration during duplex angioscanning of an increase in the diameter of the veins of the spermatic cord by 2 times from the original/the very fact of their visualization/registration of a reflux wave of blood lasting more than 1 second. Results. In all cases, by the 14th day, dilated spermatic veins were detected in rabbits and reflux was recorded from them. The maximum dilation of the veins was recorded in group 2 after 30 days (up to 2,1 mm), while in the first group, in 2 individuals, according to Doppler echo data, on the contrary, dilation of the veins and reflux were no longer detectable. The general trend of the effect is consolidated at the 8th week of observation – with continued dilation of the veins and the presence of reflux in the second group, the diameter of the spermatic veins returns to their original diameter in the control group – in 7 individuals the diameter of the veins is not determined, and in all 10 animals venous reflux is not recorded. At 12 and 24 weeks of the experiment, in all animals of the control group (group 1), the blood flow was compensated and measurement of the diameter of the spermatic veins and registration of reflux again became unavailable, while in the second group the effect of gonadotropin and progesterone is confirmed by relatively stable figures for measuring the diameter of the veins and the presence venous reflux in 10 cases. Conclusions. The proposed model is effective and can be used in a chronic experiment for up to 6 months or more to study the influence of venous congestion of the spermatic veins on the occurrence and course of pathological processes in the testis. And also to study the effectiveness of potential therapeutic effects both on the varicocele itself and on the pathological processes occurring against its background and in connection with it.

MP30-08 EVALUATING THE CHANGES IN SEMEN PARAMETERS OVER TIME FOR MEN WITH AZOOSPERMIA FACTOR C (AZFc) MICRODELETIONS

You have accessJournal of UrologyCME1 Apr 2023MP30-08 EVALUATING THE CHANGES IN SEMEN PARAMETERS OVER TIME FOR MEN WITH AZOOSPERMIA FACTOR C (AZFc) MICRODELETIONS Jessica Marinaro, Jonathan Gal, Anna Mielnik, and Peter Schlegel Jessica MarinaroJessica Marinaro More articles by this author , Jonathan GalJonathan Gal More articles by this author , Anna MielnikAnna Mielnik More articles by this author , and Peter SchlegelPeter Schlegel More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Men with complete azoospermia factor C (AZFc) microdeletions may have variable degrees of spermatogenic failure. While some are persistently azoospermic, most intermittently or consistently have sperm in the ejaculate. It has been proposed that these men may experience a temporal decline in semen quality with age. The objective of this study was to assess changes in semen parameters over time among men with complete AZFc deletions. METHODS: We retrospectively reviewed 163 men with complete AZFc deletions who were tested at our center from 4/1996 to 7/2022. Of these, 41 had ≥2 semen analyses performed (SA). ANOVA tests were used to compare mean demographic values between azoospermic, intermittently azoospermic, and non-azoospermic men. Wilcoxon matched-pairs signed rank tests were used to compare initial and final semen analyses. RESULTS: In total, 179 SAs were reviewed. The mean number of SAs per patient was 4.4 ± 2.2. The mean time between each SA was 189.6 ± 420.0 days, and the mean time between the first and last SA was 638.9 ± 850.9 days. Of 41 men, 13 (32%) were azoospermic, 9 (22%) intermittently azoospermic, and 19 (46%) were non-azoospermic. Of the non-azoospermic men, most (n=14, 73.7%) had severe oligospermia (<1 million/mL).Of 9 men with intermittent azoospermia, most had an increase in sperm concentration (n=8, 89%), total sperm count (TSC) (n=8, 89%), and total motile sperm count (TMSC) (n=6, 67%) between the first and last SA. Similarly, most non-azoospermic men also had an increase in sperm concentration (n=12, 63%), TSC (n=11, 58%), and TMSC (n=11, 58%). In comparing semen parameters, men with intermittent azoospermia had an increase in sperm concentration (p=0.0078) and TSC (p=0.0078) between the first and last SA, but no change in TMSC. There was no difference in concentration, TSC, or TMSC over time for non-azoospermic men.While non-azoospermic men were older (mean: 37.0 +/- 5.6 years; p=0.004) versus intermittently azoospermic (mean: 33.4 +/- 3.3 years) or azoospermic men (mean: 30.3 +/- 5.6 years), there were no differences in baseline testosterone (p=0.24), baseline FSH (p=0.31), or testis size (p=0.17 right, p=0.50 left). CONCLUSIONS: In this study, there was no significant decline in sperm concentration, TSC, or TMSC over time for men with complete AZFc deletions; rather, sperm number tended to increase over time. These data suggest that men with AZFc deletions are not at substantive risk of decline in sperm production and do not need to take aggressive early measures for fertility preservation. Source of Funding: Authors JM and JG are supported in part by the Frederick J. and Theresa Dow Wallace Fund of the New York Community Trust. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e393 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jessica Marinaro More articles by this author Jonathan Gal More articles by this author Anna Mielnik More articles by this author Peter Schlegel More articles by this author Expand All Advertisement PDF downloadLoading ...

Untoward Azoospermia by absurd testosterone therapy

The established role of the male partner in overall infertility is almost 50%.1 The most significant cause is degraded number as well as the quality of sperms.2 Spermatogenesis is governed by gonadotropin-releasing hormone (GnRH) then follicle-stimulating hormone (FSH) and finally by testosterone, which is linked with puberty, and essentially required well controlled hypothalamic-pituitary axis leading the Leydig cells along with Sertoli cells.3 In Pakistan incidence of azoospermia is 12.32%, and another study reported it to be 16%, this incidence is comparable to infertility in the USA at 10% while 11.35% in Kenya.4The non-obstructive azoospermia (NOA) is mostly treated with testicular sperm extraction and then intracytoplasmic sperm injection (ICSI), but only a small percentage can afford it, and mostly it requires multiple attempts. So, most couples end up with adaptation or sperm donation.5,6 One percent of all men and 10% of infertile are azoospermic. In non-obstructive azoospermia primary testicular failure occur, testosterone therapy (TTh) negative feedback mechanism can cause suppression of luteinizing hormone (LH).7 In 30% of infertile men, the cause cannot be established, but genetics, chronic infection, prostatitis, anti-sperm antibodies, and persistent obesity can lead to it.8Unfortunately, in Pakistan, most of our physicians are fond of prescribing a high dosage of testosterone for the long term, to infertile male patients. Patients become satisfied due to the positive characteristics of testosterone, but the most painful aspect is that high TTh leads to complete azoospermia. Moreover, in most such cases, it is permanent due to damage to the germinal layer. More than six-month high TTh is contraceptive.9 In 2018 the Endocrine Society and the American Urological Association recommend testosterone therapy to treat infertility and preserve fertility.10 The first management of such cases is the cessation of TTh. An integrated analysis of 1549 men on high TTh was reported as the median time to recover was 3.4 months, to a sperm concentration of 20 million/ mL and the median time to recover to baseline sperm concentration was 5.4 months.10 According to this analysis, 90% of men recovered in 12-month time after stopping TTh with 20 million/ml sperms, and 100% recovered after 24 months, all of them were on average 31.8 years of age. They were on TTh for 9.5 months.11 Hypothalamic-pituitary-gonadal (HPG) axis is almost terminated by testosterone replacement therapy (TRT) and anabolic androgenic steroids (AAS) resulting in very much suppressed or even the complete arrest of the spermatogenesis. Spontaneous cessation of such overwhelming male infertility treatments can recover spermatogenesis depending upon the length and strength of such treatments.12,13 The Prime concern is the weightage of the decision by a physician to start such therapies, which have an abusive role in treating infertility. Most physicians are unfortunately causing such azoospermia.Many very reputed international guidelines and recommendations are against such TTh, TRT, and even AAS for the treatment of male infertility. Clomiphene citrate (CC) and human chorionic gonadotropin (HCG) along with the revival of the hypothalamic-pituitary-gonadal axis can help to restore spermatogenesis in these men, after 1-2 years of the cessation of the TTh.14

The Etiology of Primary infertility in the Population of Kashmir Valley, India

Objective: To study the etiology of primary infertility in the population of Kashmir Valley, India. Design: A prospective study was undertaken of the couples who attended the clinic of the author over a period of 4 years, 2012 to 2016 for Consultation of Primary Infertility. Setting: This was study done in a community hospital. All patients who attended our hospital for infertility over a period of 4 years. A NICE protocol was followed for the evaluation. Ethical committee permission was obtained for this study. Patient Population: Couples married for at least 1 year and those who had regular unprotected sexual intercourse. Process of Study: A longitudinal study protocol was followed to identify the cause of infertility. Results and outcome: A predominant female factor was identified in 74% of the couples. Predominant male factor was identified in 7.8 % males and in all others, there was either a male as well as female factor or no male factor at all. Complete azoospermia was seen in 7.8% of the screened men and 18% showed some abnormality. Among females. 37.2% % showed evidence of anovulation (mostly Polycystic Ovary disease), 14.89% had hypothyroidism, 7% showed some sort of uterine abnormalities like malformed uterus, uterine septa or massive fibroids in uterus, and 6.3% of the patients showed tubal abnormalities. Conclusion: Polycystic ovary disease, either unrecognized or untreated is most common cause of infertility followed by hypothyroidism and uterine issues. Tubal blockage is not the common etiology as seen in most of the western world. Male factor like azoospermia does occur and sperm abnormalities are a common cause in our setting but not the sole factor for subfertility.

Sperm impairing microbial factor: potential candidate for male contraception

BackgroundDespite significant advances in contraceptive options for women, vasectomy and condoms are the only options available for male contraception. Due to this limitation, the burden of contraception resides on the shoulders of females only. Therefore, there is an urgent need to develop a safe, effective and reversible method of contraception for men. Amongst the alternative approaches, microbial derived products are gaining attention of the scientific world to combat unintended pregnancies. Earlier in our laboratory, sperm impairing microbial factor (Sperm immobilization factor) isolated from Staphylococcus aureus has shown excellent contraceptive efficacy in female mice. Keeping this in mind, the present study was carried out to exploit the sperm immobilization factor (SIF) as potential male contraceptive using vas deferens for administration in mouse model.MethodsSIF (10, 50, 100 or 200 μg) was inoculated in the lumen of right vas deferens whereas the left vas deferens served as control. The mice were sacrificed at Day 3, 7, 14, 21, 30, 45, 60 and 90 after inoculation and the results in terms of change in body weight, seminal parameters, Tissue somatic indices (TSI), haematological parameters, serum level of testosterone, lipid peroxidation and histology were studied. In order to ratify the SIF induced azoospermia SIF (200 μg) was administered with different doses viz. 100, 200, 300, 400 or 500 μg of SIF binding receptor extracted from mouse spermatozoa.ResultsThe weight profile studies of all the experimental groups showed no significant change in the initial and final body weight. In case of seminal parameters, the results revealed that right vas deferens treated with SIF showed azoospermia and with 200 μg of SIF it persisted up to 90 days. TSI of reproductive organs and non-reproductive organs showed no significant change in all the experimental groups. The haematological indices were found to be unaltered throughout the course of investigation however significant decrease in testosterone level was observed in the treated mice. The treatment also affected the oxidative status of the testis. Further, histological studies revealed hypospermatogenesis and late maturation arrest on treated side whereas the left side which served as control showed normal tissue histology. SIF induced azoospermia was ameliorated when administered with 400 μg of SIF binding receptor from mouse spermatozoa.ConclusionSIF, when administered via intra vas deferens route, could lead to complete azoospermia. Therefore, it could be considered as a potential male contraceptive.

Detection of low-grade mosaicism and its correlation with hormonal profile, testicular volume, and semen quality in a cohort of Egyptian Klinefelter and Klinefelter-like patients

Klinefelter syndrome (KS) is the most common chromosomal syndrome, causing infertility in men and leading to non-obstructive azoospermia. Previous studies on mosaicism have shown contradictory results on its correlation with both serum hormone levels and the presence of spermatozoa in the ejaculate of KS, KS-like, and non-KS-like infertile patients. So, the present study was designed to detect low-grade mosaicism in the peripheral blood lymphocytes and buccal mucosa cells of 14 KS and 8 KS-like patients by using fluorescence in situ hybridization (FISH) and to investigate its correlation with luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T) levels, testicular volume, and semen analysis compared with 10 normal healthy fertile men. Our results indicated that mosaicism was only found in 42.9 % of the KS patients and completely absent in all KS-like patients. Moreover, mosaicism has led to complete azoospermia and non-significant differences in both hormone levels and testicular volume between mosaic and non-mosaic KS patients. All KS patients demonstrated significant differences in both hormone levels and testicular volume compared with normal men. Conversely, they revealed non-significant differences in hormone levels and significant differences in testicular volume compared with KS-like patients. Additionally, the KS-like patients exhibited non-significant variations in both LH and FSH levels and significant variations in T level and testicular volume compared with normal men. Moreover, all KS-like patients had azoospermia, except for one patient who showed oligozoospermia. Therefore, no correlations were found either between mosaicism and serum hormone levels or with testicular volume and semen analysis.

Polymorphisms in the human PRDM9 gene may lead to meiotic arrest and azoospermia

PRDM9 is responsible for directing the location of programmed double strand breaks and subsequent crossover events between homologous chromosomes during meiosis in human gametes. Prdm9 is also essential for meiosis and fertility in mice, with PRDM9 knockout males displaying complete azoospermia. PRDM9 contains a rapidly evolving DNA binding zinc finger array that is coded by a ∼84 nucleotide repeating unit mini-satellite sequence. The human A-type allele, which accounts for ∼90% of the alleles in the human population, contains 13 repeating units. We sought to develop a s strategy to effectively genotype this mini-satellite with PacBio sequencing and to determine whether PRDM9 variation, including mini-satellite length polymorphisms, is associated with infertility in the human male. Observational study. Using a normospermic human control, a two-step polymerase chain reaction (PCR) protocol was established to successfully amplify the PRDM9 zinc finger array mini-satellite and sequence it using both Sanger and PacBio next generation sequencing, which has the advantage of circular consensus sequencing and long reads. We next amplified PRDM9 from the genomic DNA of 48 azoospermic men and 5 controls. The samples were visually analyzed with gel electrophoresis, and potential mutant alleles that had an atypical band appearance were compared to the wild type. PacBio sequencing results from the normospermic control were found to be an identical match to the known human A allele, confirming our ability to effectively genotype the mini-satellite array in a single PacBio run. Gel electrophoresis of PCR amplified PRDM9 alleles from azoospermic men identified 6 potential mutant variants of distinct mini-satellite lengths differing from the A-allele. We have developed a protocol to effectively genotype the human PRDM9 zinc finger array mini-satellite to evaluate a potential etiology of azoospermia in the infertile human male. We found 6 potential PRDM9 alleles differing from the known A-allele in a small (n=48) azoospermia cohort. Current/ongoing research includes applying our PacBio sequencing protocol to genotype all 48 azoospermic men in comparison to a control group and evaluate if there is an association between PRDM9 mini-satellite repeat length, polymorphisms, or de novo mutations and azoospermia.

Genomic integrity in the male germ line: evidence in support of the disposable soma hypothesis.

The Big Blue λSelect-cII selection system has been employed along with whole-exome sequencing to examine the susceptibility of the male germ line to mutation in two challenging situations (i) exposure to a chemotherapeutic regime including bleomycin, etoposide and cis-platinum (BEP) and (ii) the ageing process. A 3-week exposure to BEP induced complete azoospermia associated with a loss of developing germ cells and extensive vacuolization of Sertoli cell cytoplasm. Following cessation of treatment, spermatozoa first appeared in the caput epididymis after 6 weeks and by 12 weeks motile spermatozoa could be recovered from the cauda, although the count (P < 0.001) and motility (P < 0.01) of these cells were significantly reduced and superoxide generation was significantly elevated (P < 0.001). Despite this increase in free radical generation, no evidence of chromatin instability was detected in these spermatozoa. Furthermore, embryos obtained from females mated at this 12-week time point showed no evidence of an increased mutational load. Similarly, progressive ageing of Big Blue mice had no impact on the quality of the spermatozoa, fertility or mutation frequency in the offspring despite a significant increase in the mutational load carried by somatic tissues such as the liver (P < 0.05). We conclude that the male germ line is highly resistant to mutation in keeping with the disposable soma hypothesis, which posits that genetic integrity in the germ cells will be maintained at the expense of the soma, in light of the former's sentinel position in safeguarding the stability of the genome.

Contraception with RISUG® and functional reversal through DMSO and NaHCO3 in male rabbits.

The study aimed to evaluate reversal of short- and long-term vas occlusion with reversible inhibition of sperm under guidance (RISUG) using dimethyl sulfoxide (DMSO) and sodium bicarbonate (NaHCO3) in male rabbits (Oryctolagus cuniculus). Animals were divided into seven groups containing five animals each. Fortnightly, semen analysis revealed that sperm concentration and output steadily declined after vas occlusion and complete azoospermia was attained at 30–60 days postinjection. Spermatozoa reappeared at 60–75 days of reversal and normozoospermia was noticed between 135 days and 150 days in the reversal groups. All spermatozoa were found nonmotile prior to azoospermia and a gradual recovery in sperm motility was observed between 105 days and 135 days of reversal. A significant decline in viability of sperms was noticed during vas occlusion up to 30–60 days which recovered at 60–75 days postreversal and normalized by 75–105 days in the reversal groups. A significant enhancement in the sperm abnormalities was recorded in all vas occluded animals as well as those in initial periods of reversal. Other parameters, namely, semen volume, ejaculation time, pH, color, and consistency, remained unaltered during all phases of the study. Fertility test, at the intervals of 15 days, demonstrated that animals exhibited complete sterility during the entire period of vas occlusion. A gradual recovery in fertility was observed with the appearance of spermatozoa following vas occlusion reversal and 100% fertility was observed following 135–150 days of reversal. F1 progeny of reversed animals was found normal. The results suggest that reversal with DMSO or NaHCO3 is feasible, with normal progeny, following short- and long-term contraception.

Complete Azoospermia Factor b Deletion of Y Chromosome in an Infertile Male With Severe Oligoasthenozoospermia: Case Report and Literature Review

To report on a male patient with complete deletion of azoospermia factor b (AZFb) who presented with severe oligoasthenozoospermia, but who successfully fathered a child via intracytoplasmic sperm injection (ICSI). Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding. Y chromosome microdeletions were detected by multiplex polymerase chain reaction amplification using AZF-specific, sequence-tagged site markers. The ICSI procedure was performed using ejaculated motile spermatozoa. Cytogenetic analysis of the patient revealed a normal male karyotype, 46,XY. Multiplex polymerase chain reaction screening showed complete deletion of AZFb demonstrated by the absence of specific sequence-tagged site markers sY121, sY127, sY134, and sY143. Following successful ICSI, an ultrasound scan of the patient's partner revealed a single pregnancy with cardiac activity. A healthy boy was born by cesarean section at 38 weeks of gestation. Genetic testing 2 years later revealed that the infant had inherited his father's AZFb deletion. Evidence from this case supports the fact that carriers of AZFb deletions can sometimes produce spermatozoa and father a son with the same AZFb deletion. This possibility reinforces the need for genetic counseling in patients with Y chromosome microdeletions.

Azoospermia with variable testicular histology after 7 months of treatment with a deslorelin implant in toms

The main aim of the study was to assess whether the longer use of a GnRH-agonist implant (deslorelin 4.7 mg, Suprelorin) in toms would lead to the suppression of spermatogenesis comparable with histologic appearance in juvenile animals as was previously described in dogs. The other aims were to monitor the progression of the testes size decrease and development of azoospermia 5 to 7 months after treatment with a GnRH-agonist implant. In animals, 5, 6, and 7 months after GnRH-agonist implant insertion, variable histological appearance of germinal epithelium was found, when tubules with elongating spermatids, round spermatids, spermatocytes, and spermatogonia as the most developed germinal cells were found in each group of toms. In all male cats, 5, 6, and 7 months after implant insertion, testosterone concentrations and testes size significantly differed between the first and the last visit. All animals, except one tom castrated 5 months after implant insertion, developed complete azoospermia. However, in this tom, all spermatozoa were immotile. Treatment with the subcutaneous GnRH-agonist implant was well tolerated, and no treatment-related adverse effects were noted. These results reported the efficacy of 4.7-mg deslorelin implant (Suprelorin) during its 7 months of use. The complete azoospermia confirms its contraceptive effect. However, the histologic evaluation revealed a great individual variability in the degree of spermatogenic suppression. The question as to whether spermatogenesis in toms can be suppressed in all males to the level of spermatogonia/primary spermatocytes after prolonged exposure to deslorelin has yet to be answered.

CHD5 is required for spermiogenesis and chromatin condensation

Haploid spermatids undergo extensive cellular, molecular and morphological changes to form spermatozoa during spermiogenesis. Abnormalities in these steps can lead to serious male fertility problems, from oligospermia to complete azoospermia. CHD5 is a chromatin-remodeling nuclear protein expressed almost exclusively in the brain and testis. Male Chd5 knockout (KO) mice have deregulated spermatogenesis, characterized by immature sloughing of spermatids, spermiation failure, disorganization of the spermatogenic cycle and abnormal head morphology in elongating spermatids. This results in the inappropriate placement and juxtaposition of germ cell types within the epithelium. Sperm that did enter the epididymis displayed irregular shaped sperm heads, and retained cytoplasmic components. These sperm also stained positively for acidic aniline, indicating improper removal of histones and lack of proper chromatin condensation. Electron microscopy showed that spermatids in the seminiferous tubules of Chd5 KO mice have extensive nuclear deformation, with irregular shaped heads of elongated spermatids, and lack the progression of chromatin condensation in an anterior-to-posterior direction. However, the mRNA expression levels of other important genes controlling spermatogenesis were not affected. Chd5 KO mice also showed decreased H4 hyperacetylation beginning at stage IX, step 9, which is vital for the histone-transition protein replacement in spermiogenesis. Our data indicate that CHD5 is required for normal spermiogenesis, especially for spermatid chromatin condensation.

Abnormal Reproductive Measures and Seminal Plasma Findings in Men With Thalassemia Major (TM) and Iron Overload

Continuous improvement in the life span of TM patients allows patients to address critical issues of specific organ function. Among these hypogonadotrophic-hypogonadism (HH) and infertility are common complications, having a significant adverse impact on quality of life (QOL). While pregnancies are reported in TM women, paternity is less common in TM men, with significant gaps in our understanding of the underlying pathogenesis of male reduced fertility. Growing evidence indicates that oxidative stress negatively affects sperm quality and function, playing a central role in certain types of male infertility. In addition, abnormal low trace elements, in particular zinc, are important causes for infertility. We hypothesized that deleterious effects of iron overload impair sperm quality beyond that of iron deposition in the pituitary and the resultant HH state, and thus aimed to evaluate potential mechanisms for infertility in TM men: Systemic and pituitary iron burden (liver iron concentration; LIC, pituitary iron by MRI, ferritin), reproductive hormones (LH, FSH) and semen fluid were measured in 7 TM men (median age 26, range 21-30 years). Semen fluid parameters included volume, sperm count, motility, DNA damage determined by DNA Fragmentation Index (DFI), concentration of glutathione (GSH), a major cellular antioxidant which plays a central role in the defense against oxidative stress, and that of pertinent metal/trace elements (iron, zinc, copper). Results were compared to normal reference and Pearson correlation analysis was performed to identify markers that best predict fertility potential and highlight pathogenesis.Sperm and semen plasma analysisLab resultsImagingPtVol. mlCount Mill/mlMotility %DFI %Zn mg/LFe mg/LGSH µmol/LLH/FSH mIU/mLFerritin ng/mLLICMRI R213.51787513141.41.189.54.9/7.96685.413.5± 0.222.500-54.42.732.20.3/0.9369625.216.4± 0.731.400-6.72.530.880.0/0.0428822.816.8± 1.442.314659.7163.02.218.661.6/2.710926.614.5± 0.951.51137---7.52.8/3.419248.4-64.594758.6120.407.54-37752016.7± 0.671.800-80.22.341.060.07/0.330002019.5± 1.2nl&gt;2&gt;20&gt;50&lt;151400.3±0.12030-60/ 0.9-15&lt;300&lt;39.5-13 Semen analysis showed that 5/7 (70%) had either low sperm count or complete azoospermia, and 4/7 exhibited reduced sperm motility (57%). The three patients with a measurable sperm count did not exhibit increased DNA fragmentation (normal DFI). Overall, patients had low seminal plasma GSH concentration, indicating increased oxidative stress. Seminal plasma had elevated iron concentration in 5/6 patients; zinc levels were low in 4/6, while copper was undetectable in all patients (nl 0.064 mg/L). Patients with highest pituitary iron (2,3,7) had the lowest LH/FSH levels and azoospermia, but pituitary iron (R2) did not correlate with LH/FSH, with sperm or systemic iron measures. Seminal fluid GSH (p=0.001), iron (p=0.06), and zinc (p=0.02) inversely correlated with sperm motility. Systemic iron status, as measured by LIC and ferritin, was inversely correlated with GSH (p=0.02; 0.04, respectively) and with zinc (p=0.04; 0.04, respectively). Our data suggest that increased free iron, iron-induced oxidative damage and low trace elements are associated with impaired semen integrity and may explain low fertility potential in TM men, beyond the expected effect of pituitary iron toxicity and HH. These abnormalities, however, did not result in sperm DNA damage, in a small analyzed sample size. Oxidative and metal profiles in seminal fluid may be a valuable tool to improve the reproductive evaluation of TM men. Anti-oxidant treatment will likely have an important role in increasing their reproductive potential. Disclosures: No relevant conflicts of interest to declare.

The Effect of Microsurgical Varicocelectomy on Semen Parameters in Men with Non-Obstructive Azoospermia

Introduction: The aim of this study was to evaluate the effect of microsurgical subinguinal varicocelectomy on semen parameters in azoospermic men with clinical varicocele and to determine the predictive parameters of postoperative improvement. Methods: Twenty-three men with non-obstructive complete azoospermia and varicocele underwent subinguinal open microsurgical varicocele repair. The outcome was assessed in terms of improvement in semen parameters after surgical repair for varicocelectomy. Results: Bilateral varicocelectomy was performed on 15 patients and unilateral (left) varicocelectomy was performed on 8 patients. In the post-operative period, of the 23 patients, 7 (30.4%) had motile sperm in the ejaculate. The mean sperm concentration of these patients was 1.34 ± 2.6 × 10⁶/ml and the mean total sperm motility was 37.5 ± 15.5%. Conclusion: Infertile men with non-obstructive azospermia can have improvement in semen analysis after subinguinal microsurgical repair of varicoceles. Motile sperm in ejaculate were detected after microsurgical varicocele repair.

Ongoing graft-versus-host disease is a risk factor for azoospermia after allogeneic hematopoietic stem cell transplantation: a survey of the Late Effects Working Party of the European Group for Blood and Marrow Transplantation

The aim of this study was to assess the degree of spermatogenesis defects in sperm analysis in long-term male survivors after allogeneic hematopoietic stem cell transplantation in order to identify the risk factors related to potential infertility after hematopoietic stem cell transplantation and to provide data on longitudinal sperm recovery after hematopoietic stem cell transplantation. Here, the Late Effects Working Party of the European Group for Blood and Marrow Transplantation reports data of sperm analysis from 224 males who underwent hematopoietic stem cell transplantation. Median time between transplantation and sperm analysis was 63 months (8-275 months). At last sperm analysis, presence of any degree of spermatozoa was reported in 70 (31%) and complete azoospermia in 154 (69%) patients. In multivariate analysis, being conditioned with total body irradiation (RR 7.1; 95% CI: 3.4-14.8) and age over 25 years at transplantation (RR 2.4; 95% CI: 1.09-5.2) were significantly associated with higher risk for azoospermia. In patients not conditioned with total body irradiation, ongoing chronic graft-versus-host disease is the main adverse factor for sperm recovery (RR of 3.11; 95% CI: 1.02-9.47; P=0.045). Already established risk factors, such as total body irradiation and age older than 25 years at hematopoietic stem cell transplantation, were seen to be the most relevant adverse risk factor for sperm production after hematopoietic stem cell transplantation. Furthermore, for the first time, ongoing graft-versus-host disease has been shown to be the most relevant adverse factor for sperm recovery, particularly in patients conditioned without total body irradiation. We also introduce a useful scoring system to predict the probability of male long-term survivors' azoospermia.

SRY-negative 46,XX infertile male with Leydig cell hyperplasia: clinical, cytogenetic, and molecular analysis and review of the literature

To describe a 46,XX male whose infertility is not accounted for by a translocation of the SRY gene to the X chromosome or to the autosomes. Case report. Fertility Center of CHA Gangnam Medical Center, Seoul, South Korea. A 29-year-old male with normal male phenotype, in whom seminal analysis showed complete azoospermia. Laboratory evaluations, radiologic studies, testicular biopsy, G-banding karyotype, in situ fluorescence hybridization, and polymerase chain reaction. Clinical and laboratory findings. Peripheral blood culture for chromosome studies revealed 46,XX chromosome complement. Cytogenetic and molecular analyses excluded the presence of SRY gene. Radiologic studies displayed male structures without Müllerian ducts. Gonadal biopsy showed testicular Leydig cell hyperplasia. This is a very rare case of testicular differentiation in a 46,XX chromosomal constitution without SRY. This finding suggests that some unknown genes downstream participate in sex determination.

Microtese avoidance program?

OBJECTIVE: For men with non-obstructive azoospermia (NOA) or crypto-spermia (CS - rare non-motile sperm), microdissection testicular sperm extraction (mTESE) can obtain sperm for in-vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). In patients with NOA, occasional sperm may be noted on one or more semen analyses (S/A) despite complete azoospermia on others. We investigated the rate of appearance of sperm on serial S/As in men with NOA or CS, and the suitability of these sperm for use with IVF/ICSI.

Reversible inhibition of spermatogenesis by danazol with combination of testosterone enanthate in rabbit.

Danazol (15 mg/kg b.w./day; oral) with combination of testosterone enanthate (5 mg/kg b.w./15 days; S.C.) was tested in male rabbits for reversible suppression of spermatogenesis. Semen analyses for efficacy and reversibility were performed biweekly. The regimen resulted in complete azoospermia or severe oligozoospermia in all the animals within 75 days of treatment accompanied with decreased motility, vitality and increased sperm abnormalities. Semen volume and pH did not alter significantly. Libido was unaffected. All seminal characteristics were within normal range after 115 days of recovery. In conclusion, the drug combination in given dose regimen resulted in reversible inhibition of spermatogenesis in rabbits.