Male Subfertility Research Articles

P-061 What is the optimal number of IUIH cycles before changing treatment ?

Abstract Study question Is there an optimal number of intrauterine inseminations before offering another treatment for unexplained infertility, taking into account cumulative pregnancy rate and drop out? Summary answer Up to 3-4 intrauterine inseminations are advisable for unexplained and mild male infertility, as this will provide the optimal chances for the patient. What is known already Intrauterine insemination has been used as a first line treatment for unexplained infertility and mild male subfertility. However, during the years great debate and only few randomised controlled studies has been published supporting this. It has been shown that up to 3 inseminations are comparable to an IVF cycle. Further, multiple gestations can be managed and reduced in inseminations as well. Thus, up to 3 cycles of intrauterine inseminations can increase the chances of a healthy offspring and in controlled trials are comparable to an IVF cycle. Study design, size, duration The study is a retrospective cohort study including all patients in treatment for unexplained infertility, anovulation disorders or mild male factor from 2005 to 2022 in a single fertility clinic. Inclusion criteria: more than 1 year of infertility, normal or moderate sperm parameters. A total of 3729 women were included and all followed until pregnancy or drop out. A total of 9459 cycles were included. Participants/materials, setting, methods Women between 18 and 40 years were included. All had more than 1 year of infertility and suffered from unexplained infertility, mild male factor or ovulation disorders. All women received a mild oral tamoxifen or clomiphene stimulation for 5 days and when the leading follicle was 17 mm in diameter an ovulation induction with Fyrmadel 0,25 mg was used. All inseminations after Percoll centrifugation. Pregnancy in 12 weeks were recorded including number of gestations. Main results and the role of chance We found a cumulative pregnancy rate for 3 cycles to be 29%, for 4 cycles 32%, for 5 cycles 35%, for 6 cycles 37%, and a corresponding cumulative drop out of; for 3 cycles; 33%, for 4 cycles;43% for 5 cycles; 50%, for 6 cycles 55%. Multiple gestations were found to be 4%. Using this large data material, we found that 3-4 intrauterine inseminations were acceptable due to the cumulative pregnancy rate and drop numbers. Increasing the number to more cycles did not benefit the patient’s chance significantly and should not be recommended. The study was able to identify all cycles from any individual person starting the treatment and therefore the drop out form a main fertility clinic could be estimated properly. No hyperstimulation’s syndrome were identified and a multiple gestational risk of 4 % were identified. Limitations, reasons for caution A retrospective study from 2005 to 2021 with the limitations that a retrospective carries. The lab procedure/stimulation protocol were the same which makes the results robust. This study involved a large number of patients (3729) every cycle were followed until they got pregnant and dropped out until the 6th cycle. Wider implications of the findings Intrauterine insemination is a safe and efficient treatment option for a group of infertile women. This study describes the relative high drops out after 4-5 cycles and a cumulative pregnancy rate of 30-33% following 3-4 cycles. hereafter the cumulative pregnancy rate does not rise significantly and drop out increases. Trial registration number na

P-026 Lack of association between single polymorphic variants of the mitochondrial nicotinamide adenine dinucleotide dehydrogenase 3, and 4L (MT-ND3 and MT-ND4L) and male infertility

Abstract Study question Is there any association between male infertility and the polymorphic variants of Mitochondrial Nicotinamide Adenine Dinucleotide Hydride dehydrogenase (NADH) Subunit 3 and 4L (MT-ND3 and MT-ND4L)? Summary answer The genotypes and allele frequencies of the study population have shown a lack of statistically significant association between MT-ND3 and MT-ND4L SNPs and male infertility. What is known already Male infertility is a multifactorial condition associated with different genetic abnormalities in at least 15%–30% of cases. The rate of mutations in the mtDNA is high due to the lack of histones and DNA repair mechanisms. Moreover, mtDNA is more vulnerable compared to nuclear DNA due to the lack of an efficient repairing system and high exposure to oxidative species produced by the mitochondria. Eventually, this leads to abnormal sperm function, structure and even infertility. Study design, size, duration Prospective study was carried out between 2018 and 2019. 112 semen samples were collected in this study. Participants/materials, setting, methods The present study was carried out at the department of Obstetrics and Gynecology, University of Saarland,Germany. After semen analysis, samples were purified. Mitochondrial DNA was extracted using a QIAamp DNA Mini Kit, then, the mitochondrial DNA was amplified by using the REPLI-g Mitochondrial DNA Kit. Polymerase chain reaction (PCR) was used to amplify MT-ND3 and MT-ND4L genes. After that samples were purified and sequenced using the Sanger method in the Microsynth Seq Lab in Germany. Main results and the role of chance Eight single nucleotide polymorphisms (SNPs) in MT-ND3 (rs2853826, rs28435660, rs193302927, rs28358278, rs41467651, rs3899188, rs28358277 and rs28673954) and seven SNPs in MT-ND4L (rs28358280, rs28358281, rs28358279, rs2853487, rs2853488, rs193302933 and rs28532881) were detected and genotyped. The genotypes and allele frequencies of the study population have shown a lack of statistically significant association between MT-ND3 and MT-ND4L SNPs and male infertility. Besides, no statistically significant difference was found between the asthenozoospermia, oligozoospermia, teratozoospermia, asthenoteratozoospermia, oligoasthenoteratozoospermia and oligoteratozoospermia subgroups of subfertile males. However, rs28358278 genotype of the MT-ND3 gene was reported in the subfertile group but not in the fertile group, which implies a possible role of this SNP in male infertility. Limitations, reasons for caution The size number of the study samples. Wider implications of the findings Further studies are required to evaluate these findings. Moreover, the subfertile individuals who exhibit a polymorphism at rs28358278 require further monitoring and evaluation. Trial registration number Basic Science

O-135 Sperm-borne small ribonucleic acid profile significantly impacts embryo development

Abstract Study question Do normozoospermic males with reduced pre-implantation embryo development have aberrant sperm small RNA profiles? Summary answer Small RNA sequencing suggests the small RNA profile may differ in normozoospermic males with low blastocyst development rates, compared to males with higher blastocyst progression. What is known already Current male factor infertility diagnostics are insufficient, with 30-50% of subfertile males having unknown etiology. Spermatozoa contain a complex, epigenetically-marked genome and a collection of RNAs and proteins, which are not adequately assessed by current diagnostic methods. The sperm small RNA payload is reportedly modified during epididymal transit and in response to paternal exposures, influencing which sperm small RNA species are delivered to the oocyte. Mechanistic animal studies and correlative human and animal studies have suggested that sperm small RNAs may be important for early embryonic development and health of offspring, though their diagnostic and therapeutic value are still unclear. Study design, size, duration Human semen samples were collected between April 2017 and August 2020 from a total of 56 male patients presenting to CReATe Fertility Centre for fertility evaluation. Clinical data was accessed retrospectively. All patients were normozoospermic, according to standard semen analysis and were using donor oocytes. Samples were divided into high (n = 20), average (n = 16), and low (n = 20) fertility groups based on their deviation (1 standard deviation) from the mean blastocyst rate. Participants/materials, setting, methods Semen analysis was undertaken immediately following sample collection and spermatozoa were isolated by centrifugation. Sperm small RNA was purified and eluted using the RNeasy and MiRNeasy Kits (Qiagen). Barcoded and amplified cDNA libraries were prepared from small RNA using the NEXTFLEX Small RNA-Seq Kit v3 (Bioo Scientific). Resulting libraries were pooled, size-selected to a range of 140-190 base pairs, denatured and diluted for sequencing. Single-end, 75 bp sequencing was performed using the NextSeq 550 (Illumina). Main results and the role of chance Sequencing generated approximately 300 million raw reads, with 30 samples exceeding 2 million reads included in the differential expression analysis. Most reads were mapped to rRNAs (69%), miRNAs (11%), and piRNAs (12%). However, transfer RNA fragments from tRNA-Gly-GCC and tRNA-Val-CAC were the most abundant sequences. Top annotated miRNAs include: miR-12136-5p; miR-21-5p; and miR-122-5p. Principal component analysis revealed 222 genes that were differentially expressed between the high (n = 14) and low (n = 11) fertility groups (p < 0.05). Interestingly, the top 50 differentially expressed sRNAs are sufficient to effectively cluster sperm with poor blastocyst development rates. Limitations, reasons for caution The results are limited by a relatively low sequencing depth (mean of 4.1 million reads per sample) and sample size. Fertility groups were determined by blastocyst rates, which can be confounded by non-sperm-derived variables, including technical skill, embryo culturing conditions, and maternal factors (though donor oocytes were used). Wider implications of the findings With additional validation, a clinically-useful panel of differentially expressed sperm small RNAs could be used to predict IVF success and evaluate therapies aimed at improving male reproductive health. Augmenting traditional semen analytics with diagnostic sperm small RNA analysis could reduce time to pregnancy and the psychosocial impacts of fertility treatment. Trial registration number Not applicable

Loss of perinuclear theca ACTRT1 causes acrosome detachment and severe male subfertility in mice.

The perinuclear theca (PT) is a cytoskeletal element encapsulating the sperm nucleus; however, the physiological roles of the PT in sperm are largely uncertain. Here, we reveal that ACTRT1, ACTRT2, ACTL7A and ACTL9 proteins interact to form a multimeric complex and localize to the subacrosomal region of spermatids. Furthermore, we engineered Actrt1-knockout (KO) mice to define the functions of ACTRT1. Despite normal sperm count and motility, Actrt1-KO males were severely subfertile owing to a deficiency in fertilization. Loss of ACTRT1 caused a high incidence of malformed heads and detachment of acrosomes from sperm nuclei, caused by loosened acroplaxome structure during spermiogenesis. Furthermore, Actrt1-KO sperm showed reduced ACTL7A and PLCζ protein content as a potential cause of fertilization defects. Moreover, we reveal that ACTRT1 anchors developing acrosomes to the nucleus, likely by interacting with the inner acrosomal membrane protein SPACA1 and the nuclear envelope proteins PARP11 and SPATA46. Loss of ACTRT1 weakened the interaction between ACTL7A and SPACA1. Our study and recent findings of ACTL7A/ACTL9-deficient sperm together reveal that the sperm PT-specific ARP complex mediates the acrosome-nucleus connection.

A Role of the TEX101 Interactome in the Common Aetiology Behind Male Subfertility and Testicular Germ Cell Tumor.

Patients who develop testicular germ cell tumours (TGCT) are at higher risk to be subfertile than the general population. The conditions are believed to originate during foetal life, however, the mechanisms behind a common aetiology of TGCT and male subfertility remains unknown. Testis-expressed 101 (TEX101) is a glycoprotein that is related to male fertility, and downregulation of the TEX101 gene was shown in pre-diagnostic TGCT patients. In this review, we summarize the current knowledge of TEX101 and its interactome related to fertility and TGCT development. We searched literature and compilation of data from curated databases. There are studies from both human and animals showing that disruption of TEX101 result in abnormal semen parameters and sperm function. Members of the TEX101 interactome, like SPATA19, Ly6k, PICK1, and ODF genes are important for normal sperm function. We found only two studies of TEX101 related to TGCT, however, several genes in its interactome may be associated with TGCT development, such as PLAUR, PRSS21, CD109, and ALP1. Some of the interactome members are related to both fertility and cancer. Of special interest is the presence of the glycosylphosphatidylinositol anchored proteins TEX101 and PRSS21 in basophils that may be coupled to the immune response preventing further development of TGCT precursor cells. The findings of this review indicate that members of the TEX101 interactome could be a part of the link between TGCT and male subfertility.

‘Fertility awareness methods’ als eerste stap voor koppels met fertiliteitsproblemen

Fertility awareness methods as the first step in subfertility management An unfulfilled child wish is due to unexplained or mild male subfertility in more than half of the consulting subfertile couples. Since many of these couples can still conceive naturally within 1 year without treatment, expectant management (6-12 months) is proposed. However, in practice it often proves to be difficult to adhere to this approach, resulting in a quick shift to expensive assisted reproductive therapy (ART). Recent fertility awareness methods (FAMs) train couples to distinguish fertile from infertile days of the menstrual cycle, allowing them to target sexual intercourse on the most fertile days. By incorporating FAMs into expectant management, the latter is made more effective, increasing the chances of pregnancy. Furthermore, the health risks associated with ART are reduced. The symptothermal method is a combination of the temperature-based and cervical mucus secretion method. By applying this FAM and having fertility-focused intercourse, 92% of the women had become pregnant after 1 year, compared to 82% in studies without FAMs. For a subfertile subgroup, the cumulative spontaneous pregnancy rate after 8 months was less (38%), but still significantly above the spontaneous pregnancy rate without fertility awareness training (21.6%). Via educational materials and a network of trained teachers, FAMs are proposed. In order to fulfil this role properly, FAMs need to be recognised as the first step in fertility care in Belgium. Integrating FAMs can cause a judicious reduction in healthcare costs by reducing the number of multiple pregnancies and selecting only couples who genuinely need ART.

Fabrication on the microscale: a two-photon polymerized device for oocyte microinjection

PurposeIntracytoplasmic sperm injection (ICSI) addresses male sub-fertility by injecting a spermatozoon into the oocyte. This challenging procedure requires the use of dual micromanipulators, with success influenced by inter-operator expertise. We hypothesized that minimizing oocyte handling during ICSI will simplify the procedure. To address this, we designed and fabricated a micrometer scale device that houses the oocyte and requires only one micromanipulator for microinjection.MethodsThe device consisted of 2 components, each of sub-cubic millimeter volume: a Pod and a Garage. These were fabricated using 2-photon polymerization. Toxicity was evaluated by culturing single-mouse presumptive zygotes (PZs) to the blastocyst stage within a Pod, with several Pods (and embryos) docked in a Garage. The development was compared to standard culture. The level of DNA damage/repair in resultant blastocysts was quantified (γH2A.X immunohistochemistry). To demonstrate the capability to carry out ICSI within the device, PZs were microinjected with 4-μm fluorescent microspheres and cultured to the blastocyst stage. Finally, the device was assessed for oocyte traceability and high-throughput microinjection capabilities and compared to standard microinjection practice using key parameters (pipette setup, holding then injecting oocytes).ResultsCompared to standard culture, embryo culture within Pods and a Garage showed no differences in development to the blastocyst stage or levels of DNA damage in resultant blastocysts. Furthermore, microinjection within our device removes the need for a holding pipette, improves traceability, and facilitates high-throughput microinjection.ConclusionThis novel device could improve embryo production following ICSI by simplifying the procedure and thus decreasing inter-operator variability.

Citrylglutamate synthase deficient male mice are subfertile with impaired histone and transition protein 2 removal in late spermatids.

Chromatin remodelling in spermatids is an essential step in spermiogenesis and involves the exchange of most histones by protamines, which drives chromatin condensation in late spermatids. The gene Rimklb encodes a citrylglutamate synthase highly expressed in testes of vertebrates and the increase of its reaction product, β-citrylglutamate, correlates in time with the appearance of spermatids. Here we show that deficiency in a functional Rimklb gene leads to male subfertility, which could be partially rescued by in vitro fertilization. Rimklb-deficient mice are impaired in a late step of spermiogenesis and produce spermatozoa with abnormally shaped heads and nuclei. Sperm chromatin in Rimklb-deficient mice was less condensed and showed impaired histone to protamine exchange and retained transition protein 2. These observations suggest that citrylglutamate synthase, probably via its reaction product β-citrylglutamate, is essential for efficient chromatin remodelling during spermiogenesis and may be a possible candidate gene for male subfertility or infertility in humans.

Antioxidants for male subfertility.

Antioxidants for male subfertility.

Resveratrol reverses male reproductive damage in rats exposed to nicotine during the intrauterine phase and breastfeeding.

Nicotine leads to reproductive changes culminating in male infertility and subfertility. Resveratrol, a polyphenol, is a biological modulator. Sirtuin 1 (SIRT1) protein can positively act on male reproduction, and its expression can be affected by nicotine and modulated by resveratrol. The capability of resveratrol to reverse the reproductive damage in adult male offspring, which was nicotine-exposed during the intrauterine phase and breastfeeding, was investigated. Four groups were established with male offspring born from nicotine-exposed and non-exposed rat dams during pregnancy and lactation. Forty-eight male Wistar rats were distributed into four groups: sham control (SC), resveratrol (R), nicotine (N), and nicotine + resveratrol (NR). Rat dams of the N and NR offspring were exposed to nicotine (2mg/kg/day) during pregnancy and lactation using a subcutaneously implanted minipump. The offspring of the R and NR groups received resveratrol (300mg/kg of body weight, gavage) for 63 days from puberty. At 114 days of age, the male rats were euthanized. Nicotine did not alter the body weight, biometry of reproductive organs, or quantitative sperm parameters of adult offspring but caused an evident worsening of all sperm qualitative parameters studied. Daily treatment with resveratrol from puberty up to adulthood improved all qualitative sperm parameters significantly, leading some of them close to the control values. Resveratrol also improved the morphological integrity and expression of SIRT1 in the seminiferous epithelium of nicotine-exposed offspring. Resveratrol reversed the male reproductive damage caused by nicotine. Nicotine crosses the blood-placental membrane and is present in the breast milk of mothers who smoke. Resveratrol restored the altered reproductive parameters in the male adult offspring that were nicotine-exposed during intrauterine life and breastfeeding. The epigenetic modulating action of resveratrol can be involved in this nicotine damage reversion. Resveratrol may be a promising candidate to be investigated regarding the adjuvant strategies in the treatment of male infertility.

Animal models of male subfertility targeted on LanCL1-regulated spermatogenic redox homeostasis.

Oxidative stress in spermatozoa is a major contributor to male subfertility, which makes it an informed choice to generate animal models of male subfertility with targeted modifications of the antioxidant systems. However, the critical male germ cell-specific antioxidant mechanisms have not been well defined yet. Here we identify LanCL1 as a major male germ cell-specific antioxidant gene, reduced expression of which is related to human male infertility. Mice deficient in LanCL1 display spermatozoal oxidative damage and impaired male fertility. Histopathological studies reveal that LanCL1-mediated antioxidant response is required for mouse testicular homeostasis, from the initiation of spermatogenesis to the maintenance of viability and functionality of male germ cells. Conversely, a mouse model expressing LanCL1 transgene is protected against high-fat-diet/obesity-induced oxidative damage and subfertility. We further show that germ cell-expressed LanCL1, in response to spermatogenic reactive oxygen species, is regulated by transcription factor specific protein 1 (SP1) during spermatogenesis. This study demonstrates a critical role for the SP1-LanCL1 axis in regulating testicular homeostasis and male fertility mediated by redox balance, and provides evidence that LanCL1 genetically modified mice have attractive applications as animal models of male subfertility.

Insights into ovarian response with a fixed low dose FSH stimulation in an IUI programme: the PRORAILS study.

Are patients' characteristics, such as anti-Müllerian hormone (AMH) and BMI, reliable factors to predict ovarian response in couples with unexplained subfertility undergoing IUI with ovarian hyperstimulation (IUI-OH)? We observed no solid relationship between serum AMH and ovarian response. Ovarian stimulation during IUI treatment could lead to a higher chance of pregnancy, but also a higher incidence of multiple pregnancies, unless strict cancellation criteria are being used. Several factors could influence the result of the stimulation, such as age, BMI and hormonal status of the female. In IVF treatment, AMH has shown to be a useful predictor of ovarian stimulation to optimize the outcome; however, in a milder stimulation protocol, such as IUI, this has not been investigated. We performed a prospective cohort study and evaluated the first IUI stimulation cycle of 492 patients. The study was conducted between 2012 and 2017. Follow-up ended if patients were not pregnant after the first cycle. If pregnancy did occur, follow-up lasted until delivery. PRORAILS is a large multicentre nationwide cohort study executed in the Netherlands. Eligible women aged 18-43 years who were diagnosed with unexplained subfertility or mild male subfertility according to the Dutch guideline, with a regular indication for IUI-OH, were asked to participate. Ovarian response was assessed using a transvaginal ultrasound 5-7 days after initiation of the stimulation and was repeated according to the size of the leading follicles. Ovarian response was defined as optimal or suboptimal based on the total number of dominant follicles >15 mm. A successful stimulation was defined as the presence of two to three follicles >15 mm on the day of hCG administration. Serum AMH (µg/l) was measured by ELISA, and samples were taken on day 2, 3 or 4 of the menstrual cycle. Poisson regression was used to estimate the risk of a suboptimal ovarian response. Of the 492 participants, the mean age was 33 years and the mean subfertility duration was 2.5 years. The median serum AMH was 2.1 (µg/l). The majority of patients had a suboptimal response: 326 women (66%), of whom 224 (45%) had a hypo response (defined as <two follicles sized > 15 mm) and 102 (21%) had a hyper response (defined as more than three follicles sized >15 mm). The lowest AMH category showed a trend towards a smaller risk of a suboptimal response (relative risk ratio 0.76 (95% CI 0.54, 1.06)), but this effect did not reach statistical significance. In the prediction models, BMI and serum basal FSH were significant predictors of a hypo response, while for hyper response the factors age, BMI and serum FSH were significant. A higher BMI showed a higher risk for hypo response, as did a higher FSH whereas a lower BMI and lower FSH showed a higher risk for hyper response. The addition of AMH to the models did not improve the predictive abilities. Although the study was prospective, the main analyses were cross-sectional with characteristics measured at one time-point. The study was not powered to provide insight into predictors of pregnancy and live births and, therefore, the result for pregnancy should be interpreted with caution. This was the first large multicentre study that investigated the characteristics of ovarian response categories using standardized methods and centrally analysed laboratory measures. PRORAILS is a nationwide study with 15 hospitals and, therefore, these results are generalizable to other hospitals in the Netherlands. This study provides high-quality outcomes advancing the subfertility research field. Future studies would benefit from a randomized design investigating the effectiveness of an individualized approach versus a fixed dose. Also, the relation between a good ovarian stimulation and pregnancy rate could be further investigated. The PRORAILS study is sponsored by Merck B.V., Schiphol-Rijk, the Netherlands, an affiliation of Merck KGaA, Darmstadt, Germany (EMR700623_612). Merck KGaA, Darmstadt, Germany, reviewed the manuscript prior to submission. The opinions remain those of the authors. Merck KGaA, Darmstadt, Germany, had no influence on the use of medication in this study. The recombinant FSH was mostly provided by Merck B.V. or MSD. F.B. is a member of the external advisory board for Merck B.V., Schiphol-Rijk, the Netherlands, and has received a research grant from Merck B.V., Schiphol-Rijk. H.v.B. is an employee from IQVIA, which is a commercial data-analysing company, and received payment for her part in the article. NCT01662180.

The role of impaired acrosomal exocytosis (IAE) in stallion subfertility: A retrospective analysis of the clinical condition, and an update on its diagnosis by high throughput technologies

Acrosomal dysfunction has been considered as a cause of subfertility in males of different species, including stallions. A subset of subfertile stallions with acrosomal dysfunction is unique because they have normal sperm quality (motility, morphology, viability, and DNA quality). The current work aims to describe the clinical characteristics of subfertile stallions that were diagnosed with Impaired Acrosomal Exocytosis (IAE) by using two high throughput methods: flow cytometry and molecular genetic analysis, and to identify the prevalence of subfertility due to IAE in stallions evaluated at Texas A&M University. Clinical data from 1,128 stallions evaluated during 17 years at a Veterinary Teaching Hospital was retrospectively analyzed. Only stallions with a history of subfertility not explained following a breeding soundness examination and/or conventional semen analysis, were included. For those stallions, the acrosomal exocytosis test (AE test), in which sperm is incubated at 37 °C for up to 2 h in the presence of the calcium ionophore A23187, was used to determine IAE. The difference in AE-Rate (AE-Diff) between each pair of fertile control stallion and subfertile stallion was categorized as: Normal: AE-Diff < 14%; Questionable: AE-Diff 15–29%; Abnormal: AE-Diff > 30%. In selected cases, blood or hair was procured for identification of the susceptibility genotype for IAE, A/A-A/A, in the FKBP6 gene, exon 5. Twenty-one (21) stallions (1.86% total population analyzed) had reduced fertility despite having acceptable sperm quality. Sperm from these stallions were subjected to the AE Test. Of these, 8 stallions had reduced sperm AE-rate, based on the AE Test (8/21; 38.1%). Subsequently, blood or hair samples from these 8 stallions which had either questionable (AE-Diff 15 – 29%; n = 5) or abnormal (AE-Diff > 30%; n = 3) responses to the AE Test were analyzed for the susceptibility genotype for IAE, A/A-A/A (FKBP6 gene, exon 5). Seven out of the eight (7/8) stallions carried this susceptibility genotype. All of these were Thoroughbreds. After 2 h of incubation, the viability in fertile stallion sperm was lower than in A/A-A/A stallions (4% vs. 25%, respectively; P < 0.05), while the AE-rate was higher for fertile than for A/A-A/A stallions (85% vs. 56%, respectively; P < 0.05). The use of two high throughput tests (i.e., flow cytometry and molecular genetic analysis) may complement each other in the diagnosis of IAE in breeding stallions. In this study, 5/7 subfertile stallions diagnosed with the IAE susceptibility genotype would have been diagnosed as normal with the AE Test. This study introduces a subset of stallions with the IAE genotype with fertility higher than has been previously reported (i.e., <15% per-cycle pregnancy rate), suggesting that IAE manifests as a broader range of subfertility.

Racial Disparities in Sexual Health Care Received by Kidney Transplant Recipients in a Metropolitan Area

ABSTRACT Introduction Issues regarding disparities in urologic health care delivery and access among different racial and ethnic groups in the United States is well described in oncologic care. Such issues also exist in male sexual health care and kidney transplantation, but are less studied when these areas overlap. As a result of improved outcomes with kidney transplants (KT), quality of life (QOL) metrics become increasingly important to address. Sexual function is one such QOL domain in which KT recipients are particularly affected. Objective This study aims to elucidate the racial composition of male KT recipients with and without a sexual health issue, how that compares to the racial breakdown of the metropolitan area, and whether a particular racial group is more likely to be diagnosed with sexual dysfunction before versus after their transplant surgery. Methods After obtaining IRB approval we retrospectively reviewed the charts of men age 18 and older who received a KT at a single institution in a metropolitan city between January 2016 and February 2020. We sought to identify those with a diagnosis of ED, low or decreased libido, hypogonadism, ejaculatory dysfunction, Peyronie's disease (PD), or male factor subfertility. Demographic information, time of initial diagnosis of ED, and treatment profile were documented. The causes of end stage renal disease (ESRD) were obtained, but not controlled for. Results A total of 833 men received KT's during this time, 43% of which identified as Hispanic or Latino, 26% white (non-Hispanic or Latino), 13% Asian, and 12% Black. Of the men with a sexual dysfunction diagnosis, 26% identified as Hispanic or Latino, 31% white (non-Hispanic or Latino), 14% Asian, and 23% Black (Figure 1). No men were found to have a diagnosis of ejaculatory dysfunction, PD, or male factor subfertility. Latino men were more likely to have a diagnosis of sexual dysfunction after receiving a KT when compared to their non-Latino counterparts (OR: 2.91, 95% CI: [0.41 to 20.90]). Finally, there was no statistically significant difference in treatment patterns for ED between racial groups. Conclusions In this study we show that the overall racial composition of men age 18 and over who received a KT is representative of the demographics in its corresponding metropolitan city. However, we do note that Latino men are underdiagnosed for sexual dysfunction issues when compared to their non-Latino counterparts. Latino men are also more likely to have a diagnosis of sexual dysfunction after their transplant, likely implying they suffer from these conditions longer than men of other races. These findings highlight the need for continued research to reduce and eliminate health care disparities in the sexual health arena, specifically for patients with ESRD. Disclosure No

Critical Underdiagnosis and Undertreatment of Sexual Health Issues in Male Kidney Transplant Recipients

ABSTRACT Introduction Patients with end stage renal disease (ESRD) often endure a worse quality of life (QOL) than the general population. As a result of improved outcomes with kidney transplants, it has become increasingly important to address QOL. Sexual function is one such QOL domain in which kidney transplant (KT) patients are particularly affected by. Objective This study aims to elucidate the underdiagnosis and undertreatment of sexual health issues in male KT recipients, as well as the treatment profile for erectile dysfunction (ED) in these patients. Methods With IRB approval we retrospectively reviewed the charts of men age 18 and older who received a KT between January 2016 and February 2020 at an academic institute. We sought to identify those with a diagnosis of ED, low or decreased libido, hypogonadism, ejaculatory dysfunction, Peyronie's disease (PD), or male factor subfertility. Next, for those with a diagnosis of ED, information regarding time of initial diagnosis of ED, time when treatment was started, treatment profile and adverse events were documented. Causes of ESRD were obtained, but not controlled for. Results A total of 35 men were diagnosed with some form of sexual dysfunction, which represents 4.2% of the 833 men aged 18+ years who received a KT during this time. Seventy-one percent of men had a diagnosis of ED, 17.1% had ED and hypogonadism, 2.9% had ED and low libido, 8.6% had hypogonadism alone, and there were no diagnosis of ejaculatory dysfunction, PD, or infertility. Seventy-five percent of men with a diagnosis of ED were treated with phosphodiesterase-5 inhibitors (PDE5i), 25% with intracavernosal injections (ICI), 3.1% with intraurethral (IU) suppositories, and 9.4% with a penile prosthesis (PP). No severe medical or surgical adverse events were noted. Of those with ED, 87.5% were diagnosed prior to their transplant surgery and only 62.5% received any treatment prior to it. Conclusions In this study we show that a small fraction of men who received a KT had a diagnosis of a sexual health issue. This is in spite of the known high prevalence of ED in ESRD patients that can be as high as 75%, with up to a 10-20% improvement of ED with transplant alone. Furthermore, we note that KT recipients with ED are treated similarly to the general population and have a similar side effect profile, however, a significant number of patients do not receive treatment until after their transplant. Disclosure No

Syndromic male subfertility: A network view of genome-phenome associations.

BackgroundMale infertility is a disorder of the reproductive system with a highly complex genetic landscape. In most cases, the reason for male infertility remains unknown; however, the importance of genetic abnormalities in the diagnosis of subfertility/infertility is becoming increasingly recognized. Several syndromes include impaired male fertility in the clinical picture, although a comprehensive analysis of genetic causes of the syndromology perspective of male reproduction is not yet available.Objectives(1) To develop a catalog of syndromes and corresponding genes associated with impaired male fertility and (2) to visualize an up‐to‐date genome–phenome network of syndromic male subfertility.Materials and methodsPublished literature was retrieved from the Online Mendelian Inheritance in Man, Orphanet, Human Phenotype Ontology and PubMed databases using keywords “male infertility,” “syndrome,” “gene,” and “case report”; time period from 1980 to September, 2021. Retrieved data were organized as a catalog and complemented with identification numbers of syndromes (MIM ID) and genes (Gene ID). The genome–phenome network and the phenome network were visualized using Cytoscape and Gephi software platforms. Protein–protein interaction analysis was performed using STRING tool.ResultsRetrieved syndromes were presented as (1) a catalog containing 63 syndromes and 93 associated genes, (2) a genome–phenome network including CHD7 and WT1 genes and Noonan and Kartagener syndromes, and (3) a phenome network including 63 syndromes, and 25 categories of clinical features.DiscussionThe developed catalog will contribute to the advances and translational impact toward understanding the factors of syndromic male infertility. Visualized networks provide simple, flexible tools for clinicians and researchers to quickly generate hypotheses and gain a deeper understanding of underlying mechanisms affecting male reproduction.ConclusionRecognition of the significance of genome–phenome visualization as part of network medicine can help expedite efforts toward unravelling molecular mechanisms and enable advances personal/precision medicine of male reproduction and other complex traits.

Systematic Review of Literature Documenting the Link between BMI, Sperm Parameters, and Leptin

Background: Leptin administration to rats causes adverse effects on sperm that resemble those reported in obese males. The purpose of this review is, therefore, to examine whether human and animal studies have taken this information on board and have attempted to link the poor sperm quality in obese or overweight individuals to leptin in their methodology, data analysis, and discussion. The review also documents the link between low sperm quality in the obese and leptin. Method: A search was made for articles reporting body mass index (BMI), sperm parameters, and leptin, published until September 2020 in PubMed, Science Direct, Google Scholar, and Springer Link using a number of search terms. Results: Forty-four of the 53 human and animal studies on BMI and sperm parameters reported abnormal sperm parameters in the obese but only 9 of those had measured serum leptin. None, however, attempted to link sperm abnormalities to leptin. Nine animal studies report of adverse effects of leptin on sperm in normal weight rats that were found to be similar to those reported in obese males. Conclusion: Although none of the human studies had actually linked the raised leptin to sperm abnormalities in the obese, there is nevertheless, convincing evidence linking leptin to obesity-related poor sperm quality and male infertility. There is a need, therefore, to include measures that will help reduce the impact of raised leptin levels on male reproductive function, particularly, in obese infertile or subfertile males.

In vitro and in vivo parameters for identification of landrace pigs with low reproductive performance

In pig farming, measurements of production parameters play a fundamental role in the success of the activity. Minimal differences in fertility between breeders can lead to less reproductive efficiency and, less productivity. However, assessing the fertility of each male and the early identification of subfertile males is a difficult task to be performed. Thus, the aim of this study was to evaluate the use of in vitro and in vivo parameters in the identification of subfertile males of the Landrace breed, aiming to collaborate with genetic improvement programs, routine optimization in the Genetic Diffusion Units (GDUs) and the results of performance. In experiment 1, an approach to identify males with subfertility was evaluated based on retrospective data. For this, the results (averages of birth rates, number of total births and average percentages of female and male piglets per litter) were evaluated for a total of 996 matings and 847 parturitions. The inseminations came from ejaculates of 32 males, who had at least 19 females inseminated with homospermic doses in the concentration of 2.5 x 109 total sperm from the same male. As for the birth rate (BR), an average of 85.47% ± 6.05 was observed with a group of median males, seven males that stood out and one individual (M32) with a performance of 58.06% ± 9.0. For the total number of piglets born (PB) the average was 13.41 ± 0.56, with three males with better performance and one (M32) with very poor performance (8.62 ± 0.59). In experiment 2, it was verified whether evaluations of inseminating doses (ID) of semen in vitro (motility and sperm morphology) after 96 hours of storage had correlations with fertility in vivo, which can be used to identify subfertile males. The evaluations were performed on 30 ejaculates regarding the means of BR and PB, considering only those who had at least 7 females inseminated. There were no correlations between the motility assessments and semen morphological changes and the reproductive parameters evaluated. The results obtained in vivo, referring to BR and PB, demonstrated that it was possible to identify differences between males, the individual (M32) had the worst results for the percentages of BR and PB. It is concluded that there are males of high and low fertility and that only the in vitro analyzes carried out in this study are not enough to categorize them, however, the evaluation of retrospective data was efficient for this purpose.

Women with combined adenomyosis and endometriosis on MRI have worse IVF/ICSI outcomes compared to adenomyosis and endometriosis alone: A matched retrospective cohort study

Study objectivesTo assess the effect of adenomyosis, endometriosis and combined adenomyosis and endometriosis, diagnosed on MRI, on IVF/ICSI outcomes versus male subfertility controls. Study DesignThis single-centre matched retrospective cohort study was carried out at Catharina Hospital in Eindhoven, The Netherlands. The study group consisted of infertile women undergoing their first, fresh embryo transfer during IVF/ICSI, with adenomyosis only (N = 36), endometriosis only (N = 61), and combined adenomyosis and endometriosis (N = 93) based on MRI. The control group consisted of IVF/ICSI patients undergoing treatment due to male subfertility (N = 889). 1:2 case-control matching based on age during IVF/ICSI, parity and number of embryos transferred was performed. Odds ratios were calculated for biochemical pregnancy, ongoing pregnancy and live birth rate versus matched male subfertility controls, and were corrected for embryo quality. ResultsOnly the combined adenomyosis and endometriosis group showed a significantly reduced OR for biochemical pregnancy (p = 0.004, OR 0.453 (95% CI :(0.284–0.791)), ongoing pregnancy (p = 0.001, OR 0.302 (95% CI: (0.167–0.608)) and live birth (p = 0.001, OR 0.309 (95% CI: (0.168–0.644)) compared to matched male subfertility controls. ConclusionsThe lower (ongoing) pregnancy and live birth rates in the combined adenomyosis and endometriosis women can be attributed to more severe disease in these women, ultimately resulting in increased chances for failed implantation and miscarriage. This highlights the importance of screening for adenomyosis in endometriosis patients, and identifies these women target for additional (hormonal) treatment prior to IVF/ICSI.

Seed extract of Thai Mucuna pruriens reduced male reproductive damage in rats induced by chronic stress

Context Thai Mucuna pruriens (L.) DC. var. pruriens (Fabaceae) (TMP) is known to enrich reproduction but preventive effects on stress related adverse reproductive parameters are not documented. Objective This study investigates the protective property of TMP seed extract on reproductive damage under chronic stress (CS). Materials and methods Male Sprague-Dawley rats were divided into four groups. The control and CS groups received distilled water, whereas the pre-treated rats received the aqueous TMP seed extract at doses of 150 and 300 mg/kg BW for 20 days before co-treatments with CS induction (immobilization and forced swimming) for 81 days. Serum was used to determine the cortisol and testosterone levels. Histology of testis and epididymis was observed with localization of androgen receptor (AR). Sperm parameters and the expression of steroidogenic acute regulatory (StAR), cytochrome P450 family 11 subfamily a member 1 (CYP11A1), AR, HSP70, caspases (3 and 9) and tyrosine phosphorylation (TyrPho) proteins were investigated. Results TMP extract improved cortisol level (0.84 ± 0.02 µg/dL) and protected against the damage of reproductive tissues and sperm parameters (count [49.78 ± 3.74 million sperm/mL], viability [90.01 ± 1.17%] and precocious acrosome reaction [1.38 ± 0.48%]). Expression of testicular StAR, CYP11A1, AR and HSP70 proteins was improved. Caspase expression was decreased in treated rats. TMP increased AR expression in CS sperm. Moreover, TyrPho protein expression was corrected after TMP administration. Conclusions TMP seed protected against adverse reproductive parameters in CS via improvements of functionally testicular markers and reductions of apoptotic proteins. It is possible to develop the TMP beans as an alternative medicine in treating of male subfertility caused by CS.