Immotile Sperm Research Articles

346 The Association Between Toxic Life Stress and Semen Quality

About 15% of couples who are of childbearing age cannot achieve a wanted pregnancy and about 50% of infertility is related to male factor. Stress is one of the determinants of infertility, which leads to disruption of spermatogenesis. We undertook a retrospective study of men to elucidate whether early life stress might be a contributing factor that negatively affects semen quality in who are infertile. In this retrospective study we examine 468 idiopathic infertile men between the ages of 18 to 56 years to elucidate whether early life stress might be a contributing factor that negatively affects semen quality of them. A physical examination was performed by a physician-urologist for all eligible participants to insure they have no problems in their genitals system, followed by participants completed early life stress questionnaire and finally their semen were collected by masturbation. The results showed that the median of sperm counts and progressive motility in the participants who their parents were divorced in their childhood, were significantly lower than others. In the infertile male who one of their parents died in their childhood, the median of immotile sperm was significantly higher also people who had experienced physical and sexual abuse or physical and emotional neglect in their childhood or adolescence, had significantly lower sperm counts also lower normal morphology than others. Furthermore, participants who work in their childhood to help financial of their families had lower progressive motility of sperm.

CFAP43 modulates ciliary beating in mouse and Xenopus

Malfunctions of motile cilia cause a variety of developmental defects and diseases in humans and animal model organisms. Defects include impaired mucociliary clearance of the airways, sperm immotility, hydrocephalus and organ laterality. Here, we characterize the evolutionary conserved Cfap43 gene by loss-of-function experiments in the mouse and the frog Xenopus laevis. Cfap43 is expressed in tissues carrying motile cilia and acts as a target gene of the transcription factor FOXJ1, which is essential for the induction of motile ciliogenesis. We show that CFAP43, a protein of unknown biochemical function, localizes to the ciliary axoneme. CFAP43 is involved in the regulation of the beating frequency of tracheal cilia and loss of CFAP43 causes severe mucus accumulation in the nasal cavity. Likewise, morphant and crispant frog embryos revealed impaired function of motile cilia of the larval epidermis, a model for airway mucociliary epithelia. CFAP43 participates in the formation of flagellar axonemes during spermatogenesis as mice mutant for Cfap43 display male infertility, consistent with observations in male sterile patients. In addition, mice mutant for Cfap43 display early onset hydrocephalus. Together, these results confirm the role of CFAP43 in the male reproductive tract and pinpoint additional functions in airway epithelia mucus clearance and brain development.

Single-nucleotide polymorphism c.474G>A in the SEPT12 gene is a predisposing factor in male infertility.

SEPT12 is a testis-specific gene involved in the terminal differentiation of male germ cells. SEPT12 protein is required for sperm head-tail formation and acts as a fundamental constituent of sperm tail annulus. In this study, we screened genetic variations in exons 5, 6, 7 of the SEPT12 and assessed the annulus status in teratozoospermic, globozoospermic, and patients with immotile short tail sperm. DNA sequencing was performed for 90 teratozoospermic and 30 normozoospermic individuals. Immunocytochemistry, transmission electron microscopy and western blotting were conducted to evaluate annulus status and the expression level of SEPT12 in patients with a distinct exonic variation (c.474G>A), respectively. Five polymorphisms identified within the desired regions of the SEPT12, among them c.474G>A had the potential to induce aberrant splicing results in the expression of a truncated protein. The annulus was detected in most of the spermatozoa from teratozoospermic and normozoospermic men with c.474G>A. In contrast, in the patient with short tail sperm defect carrying c.474G>A, 99% of spermatozoa were devoid of the annulus. Based on our findings there would be no association between exons 5, 6, 7 polymorphisms of the SEPT12 gene and the occurrence of mentioned disease but c.474G>A would be a predisposing factor in male infertility.

Smartphone based sperm counting - an alternative way to the visual assessment technique in sperm concentration analysis

Sperm Counting is the first phase of the infertility diagnosis. Computer Aided Sperm Analysis (CASA) and Visual Assessment (VA) are two evaluation techniques employed in analyses. VA is carried out by observing sperm on counting chambers. Therefore, diagnosis strongly depends on the skills and experiences. CASA isolates the human factor by utilizing the computer based techniques. However, it is more costly than VA and requires exhausted parameter settings. In this study, we present a novel approach that uses smartphone and computer for sperm counting analysis. Smartphone is used to obtain images similar to VA technique. Then, sample videos are transferred to the computer side where we developed the Computerized Sperm Counting Software (CSCS) to count the sperm using counting chambers and eliminate human effects. CSCS consists of four modules: (1) Data Acquisition and Organization, (2) Regions of Interest (ROI) detection, (3) Motile/Immotile Sperm Detection, (4) Counting. Smartphone based data acquisition provided less costly design contrary to CASA systems. ROI extraction was realized by a combinational approach of line detection and segmentation methods. Background and Foreground extractions were employed in immotile and motile sperm detection process, respectively. Additionally, active contour was implemented to enhance the segmentation of immotile sperm. As the final step, detected sperms were counted by pixel based blob analysis. According to experimental results, the proposed smartphone based sperm concentration analysis can be adapted in laboratories due to its modularity, functionality, accuracy and cost when compare to CASA and VA based sperm counting analysis.

Bi-allelic Mutations in TTC29 Cause Male Subfertility with Asthenoteratospermia in Humans and Mice

As a type of severe asthenoteratospermia, multiple morphological abnormalities of the flagella (MMAF) are characterized by the presence of immotile spermatozoa with severe flagellar malformations. MMAF is a genetically heterogeneous disorder, and the known MMAF-associated genes can only account for approximately 60% of human MMAF cases. Here we conducted whole-exome sequencing and identified bi-allelic truncating mutations of the TTC29 (tetratricopeptide repeat domain 29) gene in three (3.8%) unrelated cases from a cohort of 80 MMAF-affected Han Chinese men. TTC29 is preferentially expressed in the testis, and TTC29 protein contains the tetratricopeptide repeat domains that play an important role in cilia- and flagella-associated functions. All of the men harboring TTC29 mutations presented a typical MMAF phenotype and dramatic disorganization in axonemal and/or other peri-axonemal structures. Immunofluorescence assays of spermatozoa from men harboring TTC29 mutations showed deficiency of TTC29 and remarkably reduced staining of intraflagellar-transport-complex-B-associated proteins (TTC30A and IFT52). We also generated a Ttc29-mutated mouse model through the use of CRISPR-Cas9 technology. Remarkably, Ttc29-mutated male mice also presented reduced sperm motility, abnormal flagellar ultrastructure, and male subfertility. Furthermore, intracytoplasmic sperm injections performed for Ttc29-mutated mice and men harboring TTC29 mutations consistently acquired satisfactory outcomes. Collectively, our experimental observations in humans and mice suggest that bi-allelic mutations in TTC29, as an important genetic pathogeny, can induce MMAF-related asthenoteratospermia. Our study also provided effective guidance for clinical diagnosis and assisted reproduction treatments.

Evaluation of sperm DNA fragmentation and chromatin structure in infertile men with immotile short‐tail sperm defect

Teratozoospermia is characterised by the presence of spermatozoa with abnormal morphology. One of the morphological disorders that lead to male infertility is immotile short-tail sperm (ISTS) defect. In this study, we evaluated the levels of chromatin packing and DNA fragmentation in patients with immotile short-tail sperm defect. Semen samples were obtained from 31 infertile men with ISTS as case group and 31 normozoospermic men as a control group. Protamine status was evaluated using chromomycin A3 (CMA3) staining and sperm DNA fragmentation assessed by sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labelling (TUNEL). The percentage of positive CMA3 spermatozoa was significantly higher in patients' samples (22.6±6.9) compared with controls (16.3±4.2) (p<.05) and also mean (±SD) of sperm DNA fragmentation was significantly higher in patients compared with controls, as measured by TUNEL assay (10.45±4.60 vs. 7.03±2.86, p<.05) and SCSA (24.80±13.1 vs. 15.2±7.2, p<.05). According to our study, sperm DNA fragmentation and chromatin packing abnormality are significantly higher in the ISTS samples compared with normal samples. A possible explanation for this relationship is that sperm chromatin condensation and sperm flagellum formation occur during the same phase of spermatogenesis.

The association of seminal leucocytes, interleukin-6 and interleukin-8 with sperm DNA fragmentation: A prospective study.

The effect of seminal leucocytes on sperm DNA integrity has been discussed controversially in literatures. Moreover, the studies investigating the in vivo effect of pro-inflammatory cytokines interleukin-6 and interleukin-8 on sperm DNA fragmentation are scarce and inconsistent. The association of standard sperm parameters with sperm DNA fragmentation is also a matter of ongoing discussion. Hence, the aims of this study were, first, to evaluate the effect of seminal leucocytes, interleukin-6 and interleukin-8 on sperm DNA integrity and, second, to examine whether standard semen parameters are associated with sperm DNA fragmentation. Seminal leucocytes, interleukin-6, interleukin-8 and standard semen parameters, including total sperm number, sperm concentration, progressive motility, nonprogressive motility, immotility and normal morphology, were determined in 134 consecutive men. The concentrations of seminal leucocytes, interleukin-6 and interleukin-8, did not correlate with sperm DNA fragmentation. In contrast, total sperm number, sperm concentration, progressive motility, nonprogressive motility and normal morphology exhibited significant inverse correlations with sperm DNA fragmentation. Immotile spermatozoa were directly correlated with sperm DNA fragmentation. In conclusion, seminal leucocytes, interleukin-6 and interleukin-8, are not associated with sperm DNA fragmentation. Poor standard semen parameters are significantly related to the high levels of sperm DNA fragmentation.

Comparative Evaluation of Motility and Kinematics of Fresh Versus Frozen-Thawed Spermatozoa of Cattle and Buffalo Bull by CASA

A study was undertaken on semen of three mature bulls each of Gir, Surti and Murrah breed to evaluate the comparative motion characteristics and kinematics of their fresh and frozen-thawed spermatozoa by Biovis CASA. The ejaculates (n= 24/breed) having &gt;75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed. Amongst motility traits, the total motile, rapid progressive motile and slow progressive motile spermatozoa percentage decreased significantly by 23.08 - 30.09, 43.57 - 55.18, 9.12 - 22.75 %, respectively, plessthan0.01), while non-progressive motile sperm (4.78 - 21.48%) and immotile sperm (164.38 - 178.38 %) percentage increased significantly ( plessthan0.01) in frozen-thawed semen compared to that of fresh semen. The post-thaw quality of semen of all three breeds was in acceptable range. The mean values of sperm velocity/kinematic parameters observed in frozen-thawed semen of Gir, Surti and Murrah bulls, based on total motile sperms, viz., average path velocity, curvilinear velocity, straight line velocity, linearity, straightness, beat-cross frequency, amplitude of lateral head displacement, dancing frequency and dancing mean decreased significantly by 13.70 - 17.79; 9.76 - 12.95; 13.28 - 21.90; 7.28 - 9.68; 4.36 - 7.79; 15.56 - 25.15; 8.78 - 10.50; 6.16 - 18.67 and 12.98 - 15.96 %, respectively, as compared to that of their fresh semen samples. However, wobbling index remained almost same for both fresh and frozen semen. All motility traits differed but none of kinematics/velocity traits differed significantly between breeds/species. The values of all velocity parameters for progressive motile sperms were higher than total motile sperms in all three breeds. The effect of freezing-thawing on velocity and kinematic attributes was much less compared to absolute sperm motility, and both the fresh and frozen-thawed sperms behaved identically with respect to their velocity and kinematics. The rapid progressive motile sperm in both fresh (r=0.41 to 0.92) and frozen-thawed (r=044 to 0.88) semen had significant correlations with most of their velocity traits, and the later were significantly and positively or negatively inter-related among each other in semen of all three breeds. It was therefore concluded that cryopreservation process significantly reduces the motility and kinematics attributes of bovine spermatozoa and, CASA analysis of fresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/ kinematics of spermatozoa.

Use of dimethylxanthine theophylline (SpermMobil®) does not affect clinical, obstetric or perinatal outcomes.

To evaluate whether the use of a commercially available dimethylxanthine theophylline compound (SpermMobil®) for artificial sperm activation would negatively affect clinical, obstetric and perinatal outcomes. Artificial sperm activation (ASA) was used when sperm motility after preparation was low or absent in our clinical standard procedure practice. ICSI cycles using either testicular or ejaculated sperm with concentration smaller than 5million/ml from August 2012 to January 2018 were retrospectively analyzed (n = 815) and divided into two groups, a control group where no ASA was needed and the SpermMobil® group with ASA. The fertilization rate was significantly higher in the control group, but pregnancy and implantation rates did not differ significantly. Number of embryos transferred, good quality embryos for ET and number of frozen blastocysts were similar in both groups. Clinical pregnancy loss was significantly reduced in the SpermMobil® group, which was reflected in slightly better live birth rates than in the control group. Furthermore, there were no significant differences regarding gestational age, weight, height and z score for singletons or multiples in the SpermMobil® (n = 27 and n = 10) or control (n = 144 and n = 67) groups. There were no reports of malformation, perinatal mortality or intensive therapy in the SpermMobil® group, whereas in the control group, 12 babies needed intensive care, besides one intrauterine death. The use of SpermMobil® in samples with mostly immotile sperm not only facilitates the embryologists work but also optimizes the treatment outcomes for those patients with a bad prognosis. This is the first report of obstetric and perinatal outcomes after applying a theophylline derivative in human clinical use.

What do we know about the genetic background of primary ciliary dyskinesia (PCD) and the reproductive approach?

Introduction: Primary ciliary dyskinesia (PCD; MIM 244400) is a heterogeneous autosomal recessive genetic disorder associated with infertility due to impaired sperm motility in men. We describe two such cases and give a literature review on the genetic background and reproductive outcome. Materials and methods: Two primary infertile couples were referred to our clinic. Native semen, analyzed according to the WHO manual (2010), showed that male partners have completely immotile sperm with a negative pentoxifylline test. Sample vitality was compared according to different semen preparation methods (gradient density and washing method only). Genetic testing was done by Whole Exome next-generation Sequencing (WES) analysis. For the IVF/ICSI procedure, semen was prepared using the gradient density method, and prior to the ICSI procedure itself, a hypo-osmotic swelling test (HOST) was done. Results: Semen analysis showed oligoasthenozoospermia in Patient 1 and oligoasthenoteratozoospermia in Patient 2. Blood draw for hormones and karyotype showed no irregularities in either case. Patient 1 was previously diagnosed with PCD, while Patient 2 was not. Parallel vitality testing did not show any differences between the two semen preparation methods in either of the two cases. Genetic testing in Patient 2 showed a pathogenic apparently homozygous CCDC40:c.2440C&amp;gt;T variant in exon 14 of the CCDC40 gene (MIM 613799). Fertilization rate after HOST/ICSI in both patients was 100%, and the final outcome for both patients was the birth of a healthy child. Conclusion: Although PCD has diverse etiology, assisted reproduction techniques such as HOST give these couples a good chance for parenthood. Advances in testing and strict adherence to advised procedures are to be credited for such outcome improvement. Additionally, our recommendation for PCD patients and patients with immotile sperm is to do genetic testing and counselling prior to the IVF/ICSI procedure.

Human Sperm Motility, Viability, and Morphology Decreased after Cryopreservation

The aim of this study was to analyze human sperm motility, viability, and morphology before and after cryopreservation. This true laboratory experimental study had pre and post randomized one group design. The study was conducted at the Embryology, Andrology, and Genetics Laboratory, Department of Medical Biology, Faculty of Medicine, Universitas Airlangga from August to November 2017. The eighteen samples of fresh semen were collected from male volunteers who agreed and signed the informed consent of the study. Samples were analyzed their motility, viability, and morphology before and after cryopreservation. Results of this study indicated differentiation between motility before and after cryopreservation. Cryopreservation process decreased progressive motility (42.22 + 9.46%; 17.83 + 6.24%; p&lt; 0.0001) and increased the number of immotile spermatozoa (35.44 + 10.15%; 60.11 + 12.53%; p&lt; 0.0001). Cryopreservation also decreased human sperm viability (73.78 + 8.91%; 40.83 + 12.89%; p&lt; 0.0001) and morphology (10.94 + 4.96%; 7.39 + 3.90%; p&lt; 0.0001). Cryopreservation of human spermatozoa caused the decreased of motility, viability, and morphology.

Real-Time Single Cell Monitoring: Measurement and Counting of Motile Sperm Using LCR Impedance-Integrated Microfluidic Device.

In this research, we aimed to count the ratio of the number of motile to immotile sperm for patients with infertility problems based on a low-sperm-concentration examination. The microfluidic system consists of two series of applications: The conventional separation of motile sperm and the proposed inductance (L), capacitance (C), and resistance (R) or LCR impedance sperm counter. In the experiment, 96% of motile sperm were isolated from nonmotile sperm in the first part and transported to the second part to count and calculate real-time sperm concentration. A pair of microelectrodes composed of thin metal film were integrated between microchannels, resulting in a peak signal for LCR single-cell detection, as well as the estimated total sperm concentration. A minimum of 10 µL of the sperm sample was completely analyzed with an accuracy of 94.8% compared with the standard computer-assisted semen analysis (CASA) method. This method could be applied for low-cost sperm separation and counting in the future.

Optimal analysis conditions for sperm motility parameters with a CASA system in a passerine bird, Passer montanus

BackgroundSperm motility parameters, which can be measured objectively and repeatedly by a computer-assisted sperm analysis (CASA) system, are important indicators of sperm quality. However, the sperm motility parameters assessed by a CASA system can be affected by various factors, including instrument components and settings, sperm preparation or analysis procedures. To date, no standardized protocol is available that would permit to assess sperm kinetic characteristics in passerine birds and this lack precludes any comparison of sperm swimming ability and sperm quality across species.MethodsIn this study, we chose the Tree Sparrow (Passer montanus) as the object to evaluate sperm motility parameters, including sperm motility, sperm velocity and sperm movement trajectory, at different analysis time, temperatures and pH using the WLJY-9000 CASA system.ResultsSperm motility parameters remained statistically unchanged at 1‒9 min. Progressive motility was similar at 38 °C and 40 °C, but a greater percentage of slow progressive sperm was detected at 38 °C compared to 40 °C and 42 °C. Additionally, progressive motility was lower and immotility was higher at 42 °C than 38 °C and/or 40 °C (close to the body temperature of the Tree Sparrow). The percentages of rapid progressive sperm, progressive sperm and immotile sperm were statistically similar at pH 7.0, 7.5 and 8.0 with the exception of lower percentage of progressive sperm at pH 7.0 compared to pH 7.5. In addition, slower sperm velocity and worse sperm movement trajectory were found at pH 6.0 and 9.0 than those at pH 8.0, 7.5 or 7.0.ConclusionsOur study indicates that the ideal conditions for sperm motility parameters assessment in Tree Sparrow are obtained between 1 and 9 min after dilution, an environment at body temperature (40 °C) and a pH around 7.5‒8.0. The results of this study provide a reference for the evaluation of sperm characteristics and sperm quality using a CASA system in passerine birds.

Beta-defensin126 is correlated with sperm motility in fertile and infertile men†.

A crucial function of the epididymis is providing a surface glycocalyx that is important for sperm maturation and capacitation. Defensins are antimicrobial peptides expressed in the epididymis. In the macaque epididymis, defensin beta 126 (DEFB126) is important for sperm motility, however, it is not known whether this is the case in humans. The objectives were to determine: (1) if DEFB126 on human ejaculated sperm was correlated with sperm motility in fertile and infertile men, (2) that recombinant DEFB126 could induce immature sperm motility in vitro. Immunofluorescence staining indicated that the proportion of DEFB126-positive sperm was significantly higher in motile sperm. Furthermore, the proportion of DEFB126-labeled sperm was positively correlated with sperm motility and normal morphology. Additional studies indicated that the proportion of DEFB126-positive spermatozoa in fertile volunteers was significantly higher than in volunteers with varicocele, and in infertile volunteers with semen deficiencies. To determine the role of DEFB126 on sperm motility, the DEFB126 gene was cloned and used to generate recombinant DEFB126 in H9C2 cells (rat embryonic heart myoblast cells). Deletion mutations were created into two regions of the protein, which have been linked to male infertility. Immotile testicular spermatozoa were incubated with cells expressing the different forms of DEFB126. Full-length DEFB126 significantly increased motility of co-cultured spermatozoa. However, no increase in sperm motility was observed with the mutated forms of DEFB126. In conclusion, these results support the notion that DEFB126 is important in human sperm maturation and the potential use of DEFB126 for in vitro sperm maturation.

Biallelic mutations in CFAP65 lead to severe asthenoteratospermia due to acrosome hypoplasia and flagellum malformations

BackgroundThe genetic causes for most male infertility due to severe asthenozoospermia remain unclear.ObjectiveOur objective was to identify unknown genetic factors in 47 patients with severe asthenozoospermia from 45 unrelated Chinese...

Mitochondrial OXPHOS is involved in the protective effects of L-arginine against heat-induced low sperm motility of boar.

The present study aimed to analyze the time- and temperature-responses of boar sperm and clarify the mechanism underlying the protective effects of L-arginine on heat-induced low sperm motility. Mature boar sperm was used to evaluate the effects of temperature, exposure time, L-arginine level and their interactions on sperm motility, respectively. Results showed increasing exposure time resulted in the decreased total motility and rate of rapid progressive sperm, and the increased rates of the immotile sperm and the sperm shaking in place at 38 and 39 °C, respectively (P < 0.05). L-arginine supplementation at the dose of 1.0 mM increased total motility and decreased rate of immotile sperm (P < 0.05). Heat at 39 °C decreased total motile and rate of rapid progressive sperm (P < 0.05), increased the level of sperm reactive oxygen species (ROS) (P < 0.05), reduced mitochondrial membrane potential (ΔΨm), ATP content and the activities of mitochondrial respiratory chain complexes (MRCC) ΙΙΙ and V (P < 0.05), which were attenuated by L-arginine supplementation. There were significant increases in the relative mRNA expression of nuclear respiratory factor 1 and peroxisome proliferator-activated receptor gamma coactivator-1 alpha in heat-exposed group without L-arginine supplementation. In conclusion, the rising temperatures impacted boar sperm motility in a time-dependent manner. In vitro addition of L-arginine to boar semen had a dose-dependent effect on sperm motility and sperm incubated with 1.0 mM L-arginine showed elevated motility. L-arginine supplementation can ameliorate heat-induced increase in ROS level and decreases in MRCC activities, which further maintain mitochondrial oxidative phosphorylation function, ATP synthesis and boar sperm motility.

SEMEN CHARACTERISTICS IN HIV-INFECTED MEN

The goal of the study was to evaluate the impact of antiretroviral therapy (ART) on the semen characteristics in HIV-infected men.Materials and methods. A prospective case-control study enrolled 66 HIV-infected male patients who presented for fertility assessment. Group 1 included 51 male patients treated with ART; Group 2 included 15 males who were not receiving ART. 97 and 25 semen samples were analyzed, respectively. HIV status was assessed based on the data regarding the stage and phase of the disease, viral load, CD3+, CD4+, CD8+ counts and the ART duration.Study results. At enrollment in the study, the duration of HIV infection in Group 1 was longer than that in Group 2 (the difference being statistically significant): 5 years (2–9 years) versus 2,5 years (0,4–4,5), respectively (р=0,0004). Median duration of treatment in Group 1 was 1,5 years. The most common type of abnormal semen morphology in ART-treated patients was teratozoospermia (40%), and the percentage of abnormally shaped spermatozoa in this group was higher compared to Group 2 (the difference being statistically significant): 97 (96–98) versus 96 (94,5–96,5); р=0,006). Group 1 demonstrated a negative relationship between the count of abnormally shaped spermatozoa and CD4+ cell count (r=–0,362; р=0,026), disease duration (r=0,173; р=0,173) and sperm count (r=–0,242; р=0,020), progressively motile (category B) sperm count (r=–0,241; р=0,024). The ART duration showed an inverse correlation with the progressively motile sperm count (category B) (r=–0,224; р=0,036). Group 2 demonstrated a statistically significant positive relationship between the CD4+ cell count, the semen volume (r=0,778; p=0,014), and the count of progressively motile (category B) spermatozoa (r=0,667; р=0,05). We also revealed a statistically significant inverse relationship between the viral load and the semen volume (r=–0,669; р=0,035). Sperm DNA fragmentation was found to be higher in HIV-infected patients treated with ART compared to HIV-infected men not receiving ART (15,8% (12,4–23,0) and 14% (10,9–20,5); р=0,533), respectively. Thus, HIV-infected patients treated with ART demonstrated abnormal sperm morphology (increased abnormal sperm count), decreased sperm motility and increased sperm DNA fragmentation level. The percentage of abnormally shaped sperm was higher in patients with lower CD4+ cell counts. In patients with a long history of HIV infection, the sperm count decreases and the count of immotile spermatozoa increases. HIV-infected individuals who are not receiving ART and have high viral loads show decreased semen volume. This value increases upon an increase in the CD4+ cell count.

Proteomic markers of low and high fertility bovine spermatozoa separated by Percoll gradient.

In the context of artificial insemination, male fertility is defined as the ability to produce functional spermatozoa able to withstand cryopreservation. We hypothesized that interindividual variations in fertility depend on the proportion of the fully functional sperm population contained in the insemination dose. The objective of this study was to identify protein markers of the fully functional sperm subpopulation. Insemination doses from four high-fertility (HF) and four low-fertility (LF) bulls with comparable post-thaw quality parameters were selected for proteomic analysis using iTRAQ technology. Thawed semen was centrifuged through a Percoll gradient to segregate the motile (high density [HD]) from the immotile (low density [LD]) sperm populations. Sperm proteins were extracted with sodium deoxycholate and four groups were compared: LD and HD spermatozoa from LF and HF bulls. A total of 498 unique proteins were identified and quantified. Comparison of HD spermatozoa from HF and LF bulls revealed that five proteins were significantly more abundant in the HF group (AK8, TPI1, TSPAN8, OAT, and DBIL5) whereas five proteins were more abundant in the LF group (RGS22, ATP5J, CLU, LOC616319, and CCT5). Comparison of LD spermatozoa from HF and LF bulls revealed that four proteins were significantly more abundant in the HF group (IL4I1, CYLC2, OAT, and ARMC3) whereas 15 proteins were significantly more abundant in the LF group (HADHA, HSP90AA1, DNASE1L3, SLC25A20, GPX5, TCP1, HIP1, CLU, G5E622, LOC616319, HSPA2, NUP155, DPY19L2, SPERT, and SERPINE2). DBIL5, TSPAN8, and TPI1 showed potential as putative markers of the fully functional sperm subpopulation.

PO-01-101 Epididymovasostomy: Patency, pregnancy rate and predictive factors for success in 109 patients over a decade

This study aims to assess, and identify predictive factors for, the patency and pregnancy rates in patients with obstructive azoospermia(OA) following microsurgical epididymovasostomy (EDV). We retrospectively analysed data on 129 infertile men diagnosed with OA, who underwent EDV over a ten year period at a tertiary referral centre. Complete follow up data including semen analyses, pregnancy and patency rate was analysed in 109 patients. The patient and partner’s age, and cause of azoospermia was also noted. EDV was performed using an end to side anastomosis, with or without intussusception. Data collection on intra¬ and post-operative parameters concentrated on microsurgical anastomotic technique, evidence of spermatogenesis in aspirated epididymal fluid (none, immotile, motile), and level of epididymal anastomosis (head, body, tail). Patency rate (sperm concentration at least 0.1x106 or natural conception), pregnancy and live births rate (total number of live births) and surgeon experience was also collected. The median ages of patients and their partners were 39 ± 8.3 years and 34 ± 5.4 years respectively. The aetiology of obstruction included idiopathic (37.6%), post vasectomy (26.6%), iatrogenic (3.7%), and trauma (3.7%).Microsurgical end to side anastomosis was performed in 64.2% of cases, with 28.4% of patients undergoing end to side intussusception. The most common level of anastomoses (77%)was at the level of the epididymal body, followed by 20.2% at the level of the tail and 13.8% at the head. Intraoperative aspiration of the epididymal fluid demonstrated motile and immotile sperm in 45.9% (n=50), and 40.4% (n=44) of cases respectively, with absence of sperm in 6.4%(n=7). Patients underwent unilateral (66%) or bilateral 36.7% EDV. The majority of procedures (57.8%) were carried out by a surgeon performing >10 EDV procedures per year. There were n=6 conservatively managed complications including haematoma and infection. The patency and birth rate was 56% (n=61) and 12.5% (n=13) respectively, over a 15 month follow up period.

Intraflagellar transport protein 74 is essential for spermatogenesis and male fertility in mice†.

Intraflagellar transport protein 74 (IFT74) is a component of the core intraflagellar transport complex, a bidirectional movement of large particles along the axoneme microtubules for cilia formation. In this study, we investigated its role in sperm flagella formation and discovered that mice deficiency in Ift74 gene in male germ cells were infertile with low sperm count and immotile sperm. The few developed spermatozoa displayed misshaped heads and short tails. Transmission electron microscopy revealed abnormal flagellar axonemes in the seminiferous tubules where sperm are made. Clusters of unassembled microtubules were present in the spermatids. Testicular expression levels of IFT27, IFT57, IFT81, IFT88, and IFT140 proteins were significantly reduced in the conditional Ift74 mutant mice, with the exception of IFT20 and IFT25. The levels of outer dense fiber 2 and sperm-associated antigen 16L proteins were also not changed. However, the processed A-Kinase anchor protein, a major component of the fibrous sheath, a unique structure of sperm tail, was significantly reduced. Our study demonstrates that IFT74 is essential for mouse sperm formation, probably through assembly of the core axoneme and fibrous sheath, and suggests that IFT74 may be a potential genetic factor affecting male reproduction in man.