OBJECTIVE:To study the emotional responses and family-building decisions made by individuals and couples following unsuccessful microsurgical testicular sperm extraction (micro-TESE), as described in publicly accessible online narratives. METHODS:Public posts describing personal or partner experiences after unsuccessful micro-TESE for nonobstructive azoospermia were systematically identified across online forums. Two independent investigators conducted a qualitative theme analysis using grounded theory methodology. RESULTS:Fifty-two posts were analyzed, authored by patients (46%) and partners (54%), with most published in 2025. The most common choice was pursuing sperm donation (46%), followed by additional medical interventions (19.2%) and ongoing decision-making (21%). Others reported embryo donation (3.8%), adoption/fostering (6.7%), or remaining child-free (3.8%). Five major thematic categories emerged: (1) Emotional processing, with many users expressing grief over the loss of biological parenthood; (2) Family planning, highlighting financial limitations and medical decision-making; (3) Therapy and counseling, including reflections on emotional and genetic support; (4) Timeline of decision-making, influenced by partner alignment and emotional readiness; and (5) Role of online communities, which offered validation, peer advice, and emotional support in lieu of formal follow-up care. Three broader concepts were identified: emotional recovery precedes decision-making; partner consensus influences chosen pathways; and online platforms serve as informal support systems. CONCLUSION:Online narratives illustrate the complex journey couples face after a failed micro-TESE. Emotional readiness, partner consensus, financial barriers, and peer support influence reproductive decisions. These findings highlight gaps in structured post-procedural guidance and underscore the need for earlier, integrated, patient-centered emotional and clinical counseling.

OBJECTIVE:This study aims to investigate the impact of alcohol on blood-testis barrier (BTB) integrity using a novel in vitro model and to elucidate potential nonhormonal mechanisms underlying alcohologenic reversible azoospermia. DESIGN:Primary rhesus macaque Sertoli cells were exposed to ethanol to evaluate dose-dependent effects on BTB integrity. Barrier function was assessed through electrical resistance and permeability assays, with recovery evaluated after a 48-hour withdrawal period. Gene expression of Sertoli cells and tight junction markers and cytokine levels associated with barrier degradation were analyzed. SUBJECTS:Primary rhesus macaque Sertoli cells were used. EXPOSURE:Clinically relevant concentrations (10, 60, and 100 mM), equivalent to 1-2 drinks to requiring hospitalization, of ethanol were used. MAIN OUTCOME MEASURES:The main outcome measures were BTB integrity and permeability, assessed via transepithelial electrical resistance and Dye Flux assays, recovery of barrier function after ethanol withdrawal, gene expression changes in Sertoli cells and tight junction markers, and cytokine levels associated with barrier impairment. These metrics evaluated the functional and molecular impacts of in vitro ethanol exposure on the BTB. RESULTS:Ethanol exposure was associated with a dose-dependent decrease in BTB integrity, with partial recovery observed at lower concentrations (10 and 60 mM) after 48 hours of withdrawal, but not at 100 mM. Additionally, ethanol increased the expression of key Sertoli cells and tight junction genes such as SOX9 and CLDN3, and elevated cytokines associated with barrier degradation at higher concentrations. CONCLUSION:Clinically relevant ethanol concentrations reversibly disrupt BTB function through a nonhormonal mechanism, with partial recovery at lower concentrations. These findings provide novel insights into the role of BTB dysfunction in alcohologenic reversible azoospermia.

OBJECTIVE:To determine if state populations have equitable access to vasectomy under Medicaid and evaluate the relationship between accessibility of Medicaid-covered vasectomy and state abortion restrictions. METHODS:Individual state-based Medicaid rules were assessed by publicly available data for all 50 states and Washington, D.C. A novel scoring system quantifying restrictions to Medicaid coverage for vasectomy was developed utilizing state-specific Medicaid income eligibility criteria, presence or absence of extended family planning benefits, and any additional prohibitive vasectomy coverage policies (0-4 points; 0 no restrictions-4 most restrictions). Abortion access was quantified using the Guttmacher Institute's ranking of state abortion policy restrictions (1 most protective, 7 most restrictive). Fisher's exact test (α <.05) was used to evaluate the relationship between vasectomy access score and abortion access. RESULTS:Fifty-one Medicaid regions (50 states and Washington, D.C.) were analyzed. For vasectomy access, each had a point assigned for a mandatory 30-day waiting period. Eighteen states (35%) had only 1 point assigned, 25 (49%) had 2, and 7 (14%) had 3 points. Texas was the only state to receive 4 points, correlating with the greatest number of barriers to vasectomy, due an additional consent form beyond the conventional paperwork. There was a statistically significant relationship between state Medicaid vasectomy coverage and restrictions on abortion access (P = .01). States with more barriers to Medicaid vasectomy coverage were more likely to have laws restricting abortion. CONCLUSION:Access to vasectomy under Medicaid varies greatly by state. States with the most limited access to coverage also had the most restrictive abortion laws.

Male fertility is complex and influenced by genetic, hormonal, environmental and lifestyle factors. However, limitations to human studies necessitate the use of reliable preclinical models to better understand the underlying mechanisms of male fertility. Rhesus macaques (Macaca mulatta), with their close genetic and physiological similarities to humans, offer an invaluable model for male reproductive health studies. The suitability of rhesus macaques for studying male infertility is based on similarities in spermatogenesis, hormonal cycles and the way in which assisted reproductive technologies can be applied, and key differences and similarities between human and rhesus macaque sperm structure, function and cryopreservation techniques highlight the translational potential of findings derived from macaque models. Furthermore, insights into the epigenetic and proteomic characteristics of sperm in both species improve understanding of how these findings can help to advance clinical diagnostics, male contraception and fertility preservation and illuminate the regulatory omics of normal reproduction. Thus, the rhesus macaque model offers critical insights into male fertility and studies in this species could contribute to advances in therapies for male infertility. Non-human primates, especially the rhesus macaque, provide a good preclinical model for research into male fertility, owing to their physiological and genetic similarities to humans. In this article, the authors examine the value and limitations of using non-human primates in studies to improve understanding of spermatogenesis, reproductive endocrinology and innovations in assisted reproductive technologies.

The prevalence of substance use globally is rising and is highest among men of reproductive age. In Africa, and South and Central America, cannabis use disorder is most prevalent and in Eastern and South-Eastern Europe, Central America, Canada and the USA, opioid use disorder predominates. Substance use might be contributing to the ongoing global decline in male fertility, and emerging evidence has linked paternal substance use with short-term and long-term adverse effects on offspring development and outcomes. This trend is concerning given that substance use is increasing, including during the COVID-19 pandemic. Preclinical studies have shown that male preconception substance use can influence offspring brain development and neurobehaviour through epigenetic mechanisms. Additionally, human studies investigating paternal health behaviours during the prenatal period suggest that paternal tobacco, opioid, cannabis and alcohol use is associated with reduced offspring mental health, in particular hyperactivity and attention-deficit hyperactivity disorder. The potential effects of paternal substance use are areas in which to focus public health efforts and health-care provider counselling of couples or individuals interested in conceiving.

You have accessJournal of UrologyCME1 Apr 2023MP01-02 PROTEOMIC ANALYSIS OF SEMINAL FLUID IN RHESUS MACAQUES EXPOSED TO DELTA-9-TETRAHYDROCANNABINOL Jasper C. Bash, Lyndsey E. Shorey-Kendrick, Carol B. Hannah, Charles A. Easley, Eliot Spindel, Jamie O. Lo, and Jason C. Hedges Jasper C. BashJasper C. Bash More articles by this author , Lyndsey E. Shorey-KendrickLyndsey E. Shorey-Kendrick More articles by this author , Carol B. HannahCarol B. Hannah More articles by this author , Charles A. EasleyCharles A. Easley More articles by this author , Eliot SpindelEliot Spindel More articles by this author , Jamie O. LoJamie O. Lo More articles by this author , and Jason C. HedgesJason C. Hedges More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003212.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Cannabis use has been associated with male infertility, but the etiology is not well understood. Seminal fluid is comprised of multiple factors (e.g. proteins), that influence fertility. Our objective was to determine the impact of chronic delta-9-tetrahydrocannabinol (THC) use on the seminal fluid proteome in a rhesus macaque (RM) model. METHODS: 6 adult RMs were given a daily THC edible over 280 days (∼4 spermatogenesis cycles) with serial non-sedated semen collection. Differential expression of proteins in the seminal fluid at baseline (pre-THC), high-THC dose (2.5 mg/7 kg/day), and post-THC discontinuation for 140 days (∼2 spermatogenesis cycles) was evaluated. Proteomic analysis was performed by liquid chromatography tandem mass spectrometry followed by statistical analysis to identify differentially expressed proteins. RESULTS: Proteomic analysis resulted in 1,395 quantifiable proteins (Figure 1A). The average percent identity was 88.6% (SD=14.9%). All RM proteins were matched to human proteins with 99.4% of the RM proteins (1,386) exceeding an identity cutoff of 44.3% (average minus 3 SDs). 7 proteins reached FDR significance in any contrast, thus we focused downstream analyses on nominally significant candidate proteins. Ingenuity Pathway Analysis (Figure 1B) revealed that high-THC exposure was associated with dysregulation of pathways related to “Remodeling of Epithelial Adherens Junctions”, “Axonal Guidance Signaling”, and “Glucocorticoid Receptor Signaling”. THC discontinuation for 140 days was associated with decreased “LXR/RXR Activation”, “Acute Phase Response Signaling”, and “Production of Nitric Oxide and ROS in Macrophages”. 5 proteins dysregulated with THC and significantly restored to baseline included products of CHIT1, FGG, MMP9, TGFBI, and CFB genes (Figure 1D). CHIT1 has been associated with oligozoospermia and MMP9 protein with sperm motility. CONCLUSIONS: This study demonstrated the translational strength of the RM model to study male reproductive health; there was significant seminal fluid proteome homology between humans and RMs. THC-exposed RMs are likely to exhibit proteins in their seminal ejaculate that are involved in regulatory processes that could potentially contribute to male infertility. Source of Funding: OHSU Faculty SEED Award and ASRM Pilot and Exploratory Grant © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jasper C. Bash More articles by this author Lyndsey E. Shorey-Kendrick More articles by this author Carol B. Hannah More articles by this author Charles A. Easley More articles by this author Eliot Spindel More articles by this author Jamie O. Lo More articles by this author Jason C. Hedges More articles by this author Expand All Advertisement PDF downloadLoading ...

Cannabis is the most commonly used illicit drug by reproductive age males and emerging evidence suggests it may impact fertility. Our objective was to determine the effect of chronic delta-9-tetrahydrocannabinol (THC) use on the sperm epigenome and whether these changes are permanent with THC cessation in a rhesus macaque (RM) model. Prior studies have linked sperm DNA methylation alterations with impaired spermatogenesis and infertility. We performed whole genome bisulfite sequencing (WGBS) of DNA extracted from sperm collected from 6 adult RM at baseline (pre-THC), at a high-THC dose (2.5mg/7kg/day), and post-THC discontinuation for 140 days (∼2 spermatogenesis cycles). Overlap of genes annotated to differentially methylatedregions (DMRs) in our RM following THC with published gene lists of human cannabis users and non-users was more than expected by chance (p= 1.7e-13). Pre-THC vs post-THC DMRs were similarly enriched for genes annotated to methylation differences between pre-cannabis exposure vs. post-cannabis discontinuation in humans. We identified 1,613 DMRs between pre-THC and heavy-THC that intersected DMRs in heavy-THC versus post-THC sperm samples and demonstrated a reversal in methylation post-THC toward the level of methylation pre-THC. Genes annotated to a smaller number of DMRs (1,601) between pre-THC and heavy-THC did not return to baseline levels after the THC washout period. Both the restored and non-restored DMRs were highly enriched for genes in the "synaptogenesis signaling pathway" (e.g., BDNF, several integrin, glutamate receptor, and synapsin genes), as well as for genes in the "sperm motility" canonical pathway (e.g., ADCY10, CNG, and PLA2). From the literature we identified 217 genes reported to play a key role in male infertility, of which 55 were differentially methylated with THC. We highlight a subset of these THC-DMRs in Table 1. Chronic THC use in RM alters genes related to several key regulatory pathways including fertility. Discontinuation of THC restored methylation in only a subset of DMRs.

Background With the growing availability of cannabis and the popularization of additional routes of cannabis use beyond smoking, including edibles, the prevalence of cannabis use in pregnancy is rapidly increasing. However, the potential effects of prenatal cannabis use on fetal developmental programming remain unknown. Results We designed this study to determine whether the use of edible cannabis during pregnancy is deleterious to the fetal and placental epigenome. Pregnant rhesus macaques consumed a daily edible containing either delta-9-tetrahydrocannabinol (THC) (2.5 mg/7 kg/day) or placebo. DNA methylation was measured in 5 tissues collected at cesarean delivery (placenta, lung, cerebellum, prefrontal cortex, and right ventricle of the heart) using the Illumina MethylationEPIC platform and filtering for probes previously validated in rhesus macaque. In utero exposure to THC was associated with differential methylation at 581 CpGs, with 573 (98%) identified in placenta. Loci differentially methylated with THC were enriched for candidate autism spectrum disorder (ASD) genes from the Simons Foundation Autism Research Initiative (SFARI) database in all tissues. The placenta demonstrated greatest SFARI gene enrichment, including genes differentially methylated in placentas from a prospective ASD study. Conclusions Overall, our findings reveal that prenatal THC exposure alters placental and fetal DNA methylation at genes involved in neurobehavioral development that may influence longer-term offspring outcomes. The data from this study add to the limited existing literature to help guide patient counseling and public health polices focused on prenatal cannabis use in the future.

Objective: To determine whether discontinuation of delta-9-tetrahydrocannabinol (THC) use mitigates THC-associated changes in male reproductive health using a rhesus macaque model of daily THC edible consumption. Design: Research animal study. Setting: Research institute environment. Patient(s): Adult male rhesus macaques (age, 8-10 years; n 1/4 6).Intervention(s): Chronic daily THC edible administration at medically and recreationally relevant contemporary doses followed by cessation of THC use. Main Outcome Measure(s): Testicular volume, serum male hormones, semen parameters, sperm deoxyribonucleic acid (DNA) frag-mentation, seminal fluid proteomics, and whole genome bisulfite sequencing of sperm DNA. Result(s): Chronic THC use resulted in significant testicular atrophy, increased gonadotropin levels, decreased serum sex steroid levels, changes in seminal fluid proteome, and increased DNA fragmentation with partial recovery after discontinuation of THC use. For every increase of 1 mg/7 kg/day in THC dosing, there was a significant decrease in the total testicular volume bilaterally by 12.6 cm(3) (95% confidence interval [CI], 10.6-14.5), resulting in a 59% decrease in volume. With THC abstinence, the total testicular volume increased to 73% of its original volume. Similarly, with THC exposure, there were significant decreases in the mean total testosterone and estradiol levels and a significant increase in the follicle-stimulating hormone level. With increasing THC dose, there was a significant decrease in the liquid semen ejaculate volume and weight of coagulum; however, no other significant changes in the other semen parameters were noted. After discontinuing THC use, there was a significant increase in the total serum testosterone level by 1.3 ng/mL (95% CI, 0.1-2.4) and estradiol level by 2.9 pg/mL (95% CI, 0.4-5.4), and the follicle-stimulating hormone level significantly decreased by 0.06 ng/mL (95% CI, 0.01-0.11). Seminal fluid proteome analysis revealed differential expression of proteins enriched for processes related to cellular secretion, immune response, and fibrinolysis. Whole genome bisulfite sequencing identified 23,558 CpGs differentially methylated in heavy-THC vs. pre-THC sperm, with partial restoration of methylation after discontinuation of THC use. Genes associated with altered differentially methylated regions were enriched for those involved in the development and function of the nervous system. Conclusion(s): This is the first study demonstrating that discontinuation of chronic THC use in rhesus macaques partially restores adverse impacts to male reproductive health, THC-associated sperm differentially methylated regions in genes important for development, and expression of proteins important for male fertility. (Fertil Sterile 2023;120:163-74. (c) 2023 by American Society for Reproductive Medicine.)