Normal Testis Research Articles

Impact of acetyl tributyl citrate on gonadal sex differentiation and expression of biomarker genes for endocrine disruption in Japanese medaka

Plasticizers are broadly classified as phthalate or nonphthalate. Recently, acetyl tributyl citrate (ATBC), an environmentally friendly nonphthalate plasticizer, was revealed to have the ability to disrupt thyroid hormone activity in fish species. Therefore, we aimed to assess whether ATBC exhibits any sex hormone (i.e., androgenic or estrogenic) activities. First, we examined the effects of ATBC on gonadal sex differentiation. Subsequently, we analyzed the different expression of biomarker genes that respond to endocrine-disrupting chemicals (EDCs) with sexual hormone activity in the liver. We observed normal testes and ovaries after both XX and XY medakas were exposed to ATBC, indicating that ATBC is not an EDCs with strong sex hormone activity and that it does not induce intersex (testis-to-ova or ovo-to-testis) or sex changes in Japanese medaka. The vitellogenin 1 (vtg1) and vitellogenin 2 (vtg2) mRNA expression levels in the liver of XX medakas were significantly reduced compared with those in the control group, whereas the expression levels of these genes in the liver of XY medakas remained unchanged. Finally, we examined the changes in the expression of biomarker genes that respond to EDCs with sex hormone activity in the gonads. The expression levels of biomarker genes did not differ significantly from that of the control group, although the expression levels of gsdf mRNA tended to increase while that of aromatase mRNA tended to decrease in the ovary of XX medakas following ATBC exposure. Conversely, the expression levels of gsdf and aromatase mRNAs in the testis of XY medakas remained unchanged. These results suggest that ATBC does not exhibit estrogenic activity, although it may have weak androgenic activity or no sexual hormone activity.

Comparison of conventional Doppler imaging techniques and superb microvascular imaging in determination of vascularization in undescended testes.

Our aim was to gain an idea about testicular injury by comparing the reduced volume, which is one of the indirect indicators of testicular damage in undescended testes, and by evaluating the reduced microvascular blood flow by superb microvascular imaging, and also to determine whether superb microvascular imaging modes could detect microvascular blood flow in more detail in the decreased volume of undescended testes. We compared testicular blood flow in undescended testes via conventional Doppler imaging, color superb microvascular imaging, and monochrome superb microvascular imaging techniques with contralateral normally located testis and normal control group. Each sample of testicular tissue was evaluated using a qualitative method. Spot color encoding and linear flow color encoding counts determined in testicular parenchyma were counted separately and expressed as numerical data. The localization of the examined testes in the grayscale was noted (proximal inguinal canal, medial inguinal canal, distal inguinal canal, and scrotal). The volume of undescended testes was calculated automatically via a formula for volume. Monochrome superb microvascular imaging is significantly superior in visualizing the vascularity of undescended testes compared with color Doppler, power Doppler and color superb microvascular imaging (p = 0.001). Also, undescended testes have a significantly lower blood flow compared with contralateral normal testes (p = 0.001). The volume of undescended testes was significantly lower than the contralateral normal testes. The volume, structure and blood flow are indirect signs of testicular damage in undescended testes. Monochrome superb microvascular imaging can detect vascularity in undescended testes better than the conventional Doppler imaging technique and color superb microvascular imaging. Based on our findings, we can report that monochrome superb microvascular imaging can be used to evaluate testicular injury and vascularity of undescended testes.

MP50-12 CLINICOPATHOLOGIC ANALYSIS AND DIGITAL EVALUATION OF ORCHIECTOMY SPECIMENS IN PATIENTS WITH GENDER DYSPHORIA

You have accessJournal of UrologyCME1 Apr 2023MP50-12 CLINICOPATHOLOGIC ANALYSIS AND DIGITAL EVALUATION OF ORCHIECTOMY SPECIMENS IN PATIENTS WITH GENDER DYSPHORIA Ivan De La Riva, Ruoji Zhou, Alcino Gama, Bonnie Choy, Bogdan Isaila, Diana Bowen, Robert Brannigan, and Ximing Yang Ivan De La RivaIvan De La Riva More articles by this author , Ruoji ZhouRuoji Zhou More articles by this author , Alcino GamaAlcino Gama More articles by this author , Bonnie ChoyBonnie Choy More articles by this author , Bogdan IsailaBogdan Isaila More articles by this author , Diana BowenDiana Bowen More articles by this author , Robert BranniganRobert Brannigan More articles by this author , and Ximing YangXiming Yang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003298.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Gender-affirmation therapy for transgender individuals is an area of increasing importance in genitourinary pathology. These patients undergo various forms of hormonal replacement therapy (HRT) followed by bilateral orchiectomy. To date, there are very limited studies on the histological and biochemical alterations in these testes. METHODS: We conducted a retrospective search of the database for gender-affirming orchiectomies performed at our institution from 2018 to 2022. Clinical features, including age, length and types of hormonal treatment, and pathologic features, were reviewed and summarized. 19 cases were subjected to whole slide imaging scanning for digital analysis using the Leica system. Immunohistochemistry for OCT4 and CD117 was performed on selected cases. RESULTS: Atrophic changes and decreased spermatogenesis were found in all the gender-affirming orchiectomy specimens examined, with decreased mean tubule diameter (102.9 μm, 202 tubules examined) compared to the tubular size of germ cell neoplasia in situ (GCNIS) and normal testis (139.3 μm and 143.7 μm, 326 and 185 tubules examined, respectively). All cases showed significant maturation arrest, with 55 testes showing aspermatogenesis and the remaining 14 testes showing hypospermatogenesis. Various degrees of basement membrane thickening were observed in all testes, and 55.1% of the testes were found to have focal tubule hyalinization/obliteration. The number of Leydig cells was reduced, with marked cytoplasmic vacuolization and degenerative changes. In a subset of specimens, scattered germ cells displayed prominent nucleomegaly. Using digital analysis, the average nuclear size of these abnormal germ cells was 14.4 μm in diameter (35 cells counted), approaching or exceeding the size of the neoplastic nucleus in GCNIS (mean diameter=12.1 μm, 33 cells counted). However, the enlarged nuclei of these germ cells demonstrated a smooth nuclear membrane, small nucleoli, and fine granular and smudged chromatin, significantly different from the nuclei in GCNIS. Furthermore, immunohistochemistry staining of the germ cells with nucleomegaly in tested cases demonstrated negative staining for both OCT4 and CD117, militating again the possibility of GCNIS. CONCLUSIONS: All the testes from gender-affirming orchiectomies in our study exhibit significantly reduced spermatogenesis, including maturation arrest and atrophic changes. In addition, the presence of scattered germ cells with nucleomegaly in these specimens can mimic GCNIS. Source of Funding: NA © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e689 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ivan De La Riva More articles by this author Ruoji Zhou More articles by this author Alcino Gama More articles by this author Bonnie Choy More articles by this author Bogdan Isaila More articles by this author Diana Bowen More articles by this author Robert Brannigan More articles by this author Ximing Yang More articles by this author Expand All Advertisement PDF downloadLoading ...

Toxic Effects of Nanographene Oxide on Testes of Rats

The current study aimed to examine the effects of nanographene oxide on the testes. A total of 48 male albino rats were randomly divided into 6 groups. The first, second, third, fourth, and sixth groups were treated with graphene oxide nanopowder at 20, 30, 40, 50, and 60 mg/kg concentrations, respectively. The sixth group was considered the control group. The results indicated a significant decrease in the average testis weight of rats treated with different nanographene oxide dosages, compared to the control group. There was also a significant decrease in the level of FSH and testosterone of treated rats with nanographene oxide, while there was no significant difference in the level of LH hormone when compared to the control group. The histological examination of the testes in the treated rats indicated hemorrhage, decreased sperm count, decreased thickness of the tubular epithelium, dissociation of connective tissue between the seminiferous tubules, in addition to hematological congestion, necrosis of the tubular epithelium, divergence of the seminal tubules, absence of sperm, shattering of the seminal tubule wall and degeneration sperm-forming cells and edema formation. Using the transmission electron microscope, the findings revealed a range of cellular changes, such as the presence of two-headed spermatids, the destruction of the nucleus membrane, spermatoblasts, the destruction of the cell membrane, and the denting of the nucleus membrane. It can be concluded that the nanographene oxide at 20-60 mg/kg concentrations can have harmful effects on spermatogenesis and normal function testis in rats.

Mapping of referral patterns for undescended testes – Risk factors for referral of children with normal testes

Accurate referral of boys with suspected undescended testes (UDT) is of importance to preserve fertility and reduce risk of future testicular cancer. While late referral is well studied, there is less knowledge about incorrect referrals, hence, referral of boys with normal testes. To evaluate the proportion of UDT referrals that did not lead to surgery or follow-up, and to assess risk factors for referral of boys with normal testes. All UDT referrals to a tertiary center of pediatric surgery during 2019-2020 were retrospectively assessed. Only children with suspected UDT in the referral (not suspected retractile testicles) were included. Primary outcome was normal testes at examination by a pediatric urologist. Independent variables were age, season, region of residence, referring care unit, referrer's educational level, referrer's findings, and ultrasound result. Risk factors for not needing surgery/follow-up were assessed with logistic regression and presented as adjusted odds ratios with a 95% confidence interval (aOR, [95% CI]). A total of 378 out of 740 included boys (51.1%) had normal testes. Patients >4 years (aOR 0,53, 95% CI [0,30-0,94]), referrals from pediatric clinics (aOR 0.27, 95% CI [0.14-0.51]) or surgery clinics (aOR 0.06, 95% CI [0.01-0.38]) had lower risk of normal testes. Boys referred during spring (aOR 1.80, 95% CI [1.06-3.05]), by a non-specialist physician (aOR 1.58, 95% CI [1.01-2.48]) or referrer's description of bilateral UDT (aOR 2.34, 95% CI [1.58-3.45]), or retractile testes (aOR 6.99, 95% CI [3.61-13.55]) had higher risk of not needing surgery/follow-up. None of the referred boys that had normal testes had been re-admitted at the end of this study (October 2022). Over 50% of boys referred for UDT had normal testes. This is higher or equal to previous reports. Efforts to reduce this rate should in our setting probably be directed towards well-child centers and training in examination of testicles. The main limitation of this study is the retrospective design and the rather short follow-up time, which however should have very modest effect on the main findings. Over 50% of boys referred for UDT have normal testes. A national survey regarding the management and examination of boys testicles has been launched and directed at well-child centers to further evaluate the findings of the current study.

Somatic FGFR2 is Required for Germ Cell Maintenance in the Mouse Ovary.

During sex determination in the mouse, fibroblast growth factor 9 signals through the fibroblast growth factor receptor 2c isoform (FGFR2c) to trigger Sertoli cell and testis development from 11.5 days post coitum (dpc). In the XX gonad, the FOXL2 and WNT4/RSPO1 pathways drive granulosa cell and ovarian development. The function of FGFR2 in the developing ovary, and whether FGFR2 is required in the testis after sex determination, is not clear. In fetal mouse gonads from 12.5 dpc, FGFR2 shows sexually dimorphic expression. In XX gonads, FGFR2c is coexpressed with FOXL2 in pregranulosa cells, whereas XY gonads show FGFR2b expression in germ cells. Deletion of Fgfr2c in XX mice led to a marked decrease/absence of germ cells by 13.5 dpc in the ovary. This indicates that FGFR2c in the somatic pregranulosa cells is required for the maintenance of germ cells. Surprisingly, on the Fgfr2c-/- background, the germ cell phenotype could be rescued by ablation of Foxl2, suggesting a novel mechanism whereby FGFR2 and FOXL2 act antagonistically during germ cell development. Consistent with low/absent FGFR2 expression in the Sertoli cells of 12.5 and 13.5 dpc XY gonads, XY AMH:Cre; Fgfr2flox/flox mice showed normal testis morphology and structures during fetal development and in adulthood. Thus, FGFR2 is not essential for maintaining Sertoli cell fate after sex determination. Combined, these data show that FGFR2 is not necessary for Sertoli cell function after sex determination but does play an important role in the ovary.

Identification of active spermatogenesis using a multiphoton microscope.

The sperm retrieval rate of microdissection testicular sperm extraction varies from 25% to 60%. Therefore, it is necessary to establish objective selection criteria for identifying seminiferous tubules with spermatozoa. Our aim was to develop a method for identifying spermatogenesis without sectioning testicular tissues. Testicular tissues of 10-week-old normal rats were fixed with 4% paraformaldehyde. Fluorescent labeling of seminiferous tubule nuclei and F-actin was performed, and the specimens were observed without sectioning using a multiphoton microscope. Cryptorchid rats were used as a model lacking elongated spermatids. Multiphoton images were compared with images of normal seminiferous tubules. In addition, seminiferous tubules of 10-week-old normal rats were labeled by testicular interstitial injection of fluorescent probes and observed by a multiphoton microscope without fixation. Terminal deoxynucleotidyl transferase dUTP nick end labeling-stained images of normal and probe-injected testes were compared. In fixed seminiferous tubules, elongated spermatids were identified. In addition, F-actin of apical ectoplasmic specialization was observed around elongated spermatids. Furthermore, spermatogenic stages were identified by an array of nuclei or F-actin. In cryptorchid testes, elongated spermatids and F-actin of the apical ectoplasmic specialization were not observed. In testes injected with fluorescent probes, F-actin of the apical ectoplasmic specialization was observed, and spermatogenic stages were identified without fixation. There was no significant difference in the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells per seminiferous tubule between normal and probe-injected testes. Seminiferous epithelium could be observed without sectioning of tissues by fluorescent probes and a multiphoton microscope. Active spermatogenesis was observed by labeling F-actin with and without fixation. Moreover, the toxicity of fluorescent probes was limited. Our method has a potential for live imaging of testicular tissue.

Diagnosis of Testicular Torsion and Differentiation From Other Pathologies Using Near-Infrared Spectroscopy

ObjectiveTo develop a near-infrared (NIR) spectroscopy device to diagnose testicular torsion with high sensitivity and specificity. Specifically, we aim to investigate the differentiation between testicular torsion from other pathologies such as orchidoepididymitis, varicocele, and hydrocele. MethodsTwo LEDs with wavelengths of 660 nm and 940 nm were used as light sources in the device. Each wavelength was sent to the testicle successively, and a photodiode detected back-reflected diffuse light. The ratio of the light intensities of 660 nm and 940 nm was used as a diagnostic parameter. A multi-center clinical trial was performed in 5 different hospitals. ResultsIn total, 62 patients in urology clinics with acute testicular pain have been recruited for the study. The developed NIR spectroscopy correctly defined all 8 testicular torsion cases. Besides, 3 orchidoepididymitis, 1 varicocele, and 3 hydrocele cases were correctly distinguished from testicular torsion. Only 1 hydrocele case was misdiagnosed as torsion. The range of the ratio was between 0.14 and 1.16 overall measurements. The ratio varied between 0.14 and 0.3 for the testicle with torsion. The ratio was between 0.49 and 1.16 for the normal testicle and testicle with other pathologies mentioned above. ConclusionWe have chosen the threshold ratio of 0.4 to differentiate between the normal and torsion testis and diagnosed all the torsion cases among all normal and other pathologies. The developed optical device to diagnose testicular torsion is inexpensive, user-friendly, and works based on objective criteria with high sensitivity and specificity in real time.

Unraveling the effect of the inflammatory microenvironment in spermatogenesis progression.

Experimental autoimmune orchitis (EAO) is a chronic inflammatory disorder that causes progressive spermatogenic impairment. EAO is characterized by high intratesticular levels of nitric oxide (NO) and tumor necrosis factor alpha (TNFα) causing germ cell apoptosis and Sertoli cell dysfunction. However, the impact of this inflammatory milieu on the spermatogenic wave is unknown. Therefore, we studied the effect of inflammation on spermatogonia and preleptotene spermatocyte cell cycle progression in an EAO context and through the intratesticular DETA-NO and TNFα injection in the normal rat testes. In EAO, premeiotic germ cell proliferation is limited as a consequence of the undifferentiated spermatogonia (CD9+) cell cycle arrest in G2/M and the reduced number of differentiated spermatogonia (c-kit+) and preleptotene spermatocytes that enter in the meiotic S-phase. Although inflammation disrupts spermatogenesis in EAO, it is maintained in some seminiferous tubules at XIV and VII-VIII stages of the epithelial cell cycle, thereby guaranteeing sperm production. We found that DETA-NO (2mM) injected in normal testes arrests spermatogonia and preleptotene spermatocyte cell cycle; this effect reduces the number of proliferative spermatogonia and the number of preleptotene spermatocytes in meiosis S-phase (36h after). The temporal inhibition of spermatogonia clonal amplification delayed progression of the spermatogenic wave (5days after) finally altering spermatogenesis. TNFα (0.5 and 1µg) exposure did not affect premeiotic germ cell cycle or spermatogenic wave. Our results show that in EAO the inflammatory microenvironment altered spermatogenesis kinetics through premeiotic germ cell cycle arrest and that NO is a sufficient factor contributing to this phenomenon.

Testicular abnormalities of rams in two slaughterhouses in Sokoto, Nigeria

This study was designed to determine the testicular abnormalities in rams slaughtered in Sokoto, Nigeria. The testicles of rams presented for slaughter were examined, and the age, breed, position, and sites of testicular abnormalities were noted. Out of 638 rams examined, 3 had testicular abnormalities, representing a prevalence of 0.47%, comprising 2 (0.31%) unilateral cryptorchidism and 1 (0.16%) unilateral hypoplasia. The prevalence of (0.58% (2/342) and 0.34% (1/296) were obtained at the Sokoto metropolitan abattoir and Batta Shuni slaughter slab, respectively. Two (0.31%) of the rams with abnormality were 1 < - ≤ 2 years old, comprising 1(0.16%) cryptorchid ram and testicular hypoplasia. There was 1 (0.16%) ram 2 < - ≤ 3 years with testicular hypoplasia. Based on breed, there was 1(0.16%) each of Yankasa, crosses (cryptorchidism) and Uda (testicular hypoplasia). The abnormalities were all on the right testis, while one cryptorchid testis each was in the subcutis and abdomen. There was a significant (p < 0.05) decrease in the testicular weight and mid-testicular circumference of the abnormal testis compared to the normal. The longitudinal length of the abnormal testis was smaller than the normal, although this was not significantly (p > 0.05) different. There was atrophy of the seminiferous tubules with a poorly developed tubular lumen in the cryptorchid ram. In addition, the epididymis was devoid of spermatogenic cells. The hypoplastic testis had reduced spermatogenic activity and slight testicular degeneration which were absent in the normal testis. The study shows that rams with testicular abnormalities are rare among rams slaughtered in Sokoto although those with abnormalities may be infertile, rendering them unfit for breeding.

Super Selective Angioembolization for Posttraumatic ArterioCavernosal Fistula Presenting with Nonischemic Priapism

Arteriocavernosal fistula generally occurs from penetrating injury and is rarely secondary to blunt trauma. Patients typically present with a painless partial erection following some type of genitoperineal trauma; the resultant arterial damage leads to an arterial-lacunar fistula and occasionally, to an associated pseudoaneurysm. The case reported in this article is unique in a way that the patient presented after 4 months yet super selective angioembolization of the feeder artery yielded promising results. A 24-year-old man was reported to the outpatient department of our center with a history of road traffic accidents and subsequent perineal injury 4 months back. The patient had complaint of inability to get full rigid erection and persistent partial erection of penis for 4 months since the trauma. On clinical examination, the patient had a partial detumescent penis and normal testes. Doppler ultrasonography revealed extravasation of blood from the lacerated cavernosal artery as a characteristic color blush that extended into the erectile tissue. Magnetic resonance imaging of the perineum suggested the possibility of left-sided posttraumatic arteriocavernous fistula. Diagnostic angiogram of left iliac artery demonstrated arteriocavernous fistula, and selective arterial embolization of feeding artery left cavernosal artery was performed with metallic coils. Procedure was uneventful. Complete detumescence was achieved in the immediate postoperative period. Super selective angioembolization of the feeder’s vessel remains the treatment of choice even in patients with long-standing fistula and priapism secondary to it.

Application of single-cell RNA sequencing on human testicular samples: a comprehensive review.

So far there has been no comprehensive review using systematic literature search strategies to show the application of single-cell RNA sequencing (scRNA-seq) in the human testis of the whole life cycle (from embryos to aging males). Here, we summarized the application of scRNA-seq analyses on various human testicular biological samples. A systematic search was conducted in PubMed and Gene Expression Omnibus (GEO), focusing on English researches published after 2009. Articles related to GEO data-series were also retrieved in PubMed or BioRxiv. 81 full-length studies were finally included in the review. ScRNA-seq has been widely used on different human testicular samples with various library strategies, and new cell subtypes such as State 0 spermatogonial stem cells (SSC) and stage_a/b/c Sertoli cells (SC) were identified. For the development of normal testes, scRNA-seq-based evidence showed dynamic transcriptional changes of both germ cells and somatic cells from embryos to adults. And dysregulated metabolic signaling or hedgehog signaling were revealed by scRNA-seq in aged SC or Leydig cells (LC), respectively. For infertile males, scRNA-seq studies revealed profound changes of testes, such as the increased proportion of immature SC/LC of Klinefelter syndrome, the somatic immaturity and altered germline autophagy of patients with non-obstructive azoospermia, and the repressed differentiation of SSC in trans-females receiving testosterone inhibition therapy. Besides, the re-analyzing of public scRNA-seq data made further discoveries such as the potential vulnerability of testicular SARS-CoV-2 infection, and both evolutionary conservatism and divergence among species. ScRNA-seq analyses would unveil mechanisms of testes' development and changes so as to help developing novel treatments for male infertility.

CSNK1G2-AS1 promotes metastasis, colony formation and serves as a biomarker in testicular germ cell tumor cells.

Background/Aim: Some long non-coding RNAs (lncRNAs) have been found to significantly participate in the progression of TGCTs. In comparison to the normal testis, the TGCT tissues showed significantly decreased CSNK1G2-AS1 expression, however, its effect on TGCTs and its mechanism are still unclear. The aim of this study is to investigate the effect of CSNK1G2-AS1 on TGCTs and explore the mechanism underlying its effect on TGCTs. Materials and Methods: In this study, to evaluate the expression of CSNK1G2-AS1 in tissue samples from TGCTs, the UCSC and GEPIA databases were applied and qRT-PCR was conducted. The Kaplan-Meier Plotter was applied to analyze the correlation between CSNK1G2-AS1 methylation levels and the prognosis of TGCTs patients. The assays of MTS, clone formation, transwell, and flow cytometry were performed to investigate the effect of CSNK1G2-AS1 overexpression on the proliferation, metastasis, and apoptosis of TGCT cells, respectively. Finally, western blotting was conducted to determine the expressions of the proteins associated with EMT and AKT. Results: Our study first found that, compared to the normal testis, TGCTs tissue showed significantly decreased CSNK1G2-AS1 expression, and hypomethylation of CSNK1G2-AS1 was significantly correlated with a better prognosis with TGCTs patients. In vitro, we found that overexpression of CSNK1G2-AS1 dramatically promoted the clone formation, invasion, and migration of TGCT cells, but inhibited apoptosis. And CSNK1G2-AS1 overexpression significantly decreased the expression of EMT-associated proteins ZO-1 but increased the expression and phosphorylation of AKT. Conclusions: CSNK1G2-AS1 may play an essential role in the pathogenesis and metastasis of TGCTs through the EMT- and AKT-mediated signal pathways.

Preserving the Normal Healthy Testis: Role of Beta-carotene

Background: Many drugs can have significant deleterious side effects on the different organs of the body which can change normal physiology to different pathologies. Recent research have been focusing on valuable methods to preserve the normal healthy testis from the medications’ adverse effects. The study aims to investigate the role of beta-carotene in the protection against potential amikacin’s testicular toxicity. Materials and methods: Male rats were involved in the study. Rats have divided into four groups: the first group was control, the second group has been given amikacin as an inducer of testicular toxicity; the third group has been given both amikacin and beta-carotene, and the fourth group has been given beta-carotene as prophylaxis before induction of the toxicity. The histological architecture of the testis was investigated for all the rats. Results: Amikacin caused a significant deleterious effect which included degeneration and necrosis of cells of seminiferous tubules as well as atrophy and congestion of blood vessels. Co-administration of both beta-carotene and amikacin resulted in a partial improvement in the testicular tissue while using beta-carotene as prophylaxis succeeded in protecting and preserving the normal histological features of the testis completely. Conclusion: The study concluded that beta-carotene can preserve the normal physiological testis and protect it against the amikacin’s deleterious effect on the testis.

A rare cause of scrotal abscess: xanthogranulomatous epididymitis

Xanthogranulomatous epididymitis is a rare benign process with destruction of tissue and replacement by cellular infiltration of foamy macrophages, dense lymphocytes and plasma cells. Literature has very few case reports. We reported here a case of a 72 years old male who presented as scrotal abscess with bilateral hydrocele on ultrasound. On exploration he had normal right testis with abscess localized to epididymis underwent right epididymectomy with orchidopexy. Histopathology reported necrosis and abundant foamy macrophages and plasma cells findings suggestive of xanthogranulomatous epididymitis.

Dynamic contrast-enhanced MRI in the assessment of testicular perfusion in infertile men with clinical varicocele.

Varicocele represents the most common correctable cause of male infertility. The presence of non-invasive imaging parameters providing evidence as to which patients with varicocele are at risk for infertility would be important. To explore the role of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) using semi-quantitative parameters in the assessment of testicular perfusion in infertile men with clinical varicocele. The study cohort included 11 infertile men with clinical varicocele and six controls, with prior paternity. Subtraction DCE-MRI was performed after gadolinium administration, using a three-dimensional fast field-echo sequence. Time-signal intensity curves were created and semi-quantitative parameters were calculated. The independent samples t-test was used to compare basic T1 perfusion parameters between infertile testes with clinical varicocele and normal testes. Logistic regression analysis was performed to assess the most significant predictor of the diagnosis of clinical varicocele. Both testes with clinical varicocele and normal testes enhanced moderately and homogeneously, with a linear increase of enhancement throughout the examination. Higher mean values of maximum enhancement (P = 0.026), maximum relative enhancement (P = 0.024), and wash-in rate (P = 0.013) were detected in the testes of infertile men with clinical varicocele, compared to the normal population. The wash-in rate proved the most significant predictor of the diagnosis of clinical varicocele (P = 0.013). DCE-MRI may provide a valuable insight into the testicular perfusion of infertile men with clinical varicocele. The wash-in rate proved a strong and independent predictor of the diagnosis of clinical varicocele.

New Insights into Testosterone Biosynthesis: Novel Observations from HSD17B3 Deficient Mice

Androgens such as testosterone and dihydrotestosterone (DHT) are essential for male sexual development, masculinisation, and fertility. Testosterone is produced via the canonical androgen production pathway and is essential for normal masculinisation and testis function. Disruption to androgen production can result in disorders of sexual development (DSD). In the canonical pathway, 17β-hydroxysteroid dehydrogenase type 3 (HSD17B3) is viewed as a critical enzyme in the production of testosterone, performing the final conversion required. HSD17B3 deficiency in humans is associated with DSD due to low testosterone concentration during development. Individuals with HSD17B3 mutations have poorly masculinised external genitalia that can appear as ambiguous or female, whilst having internal Wolffian structures and testes. Recent studies in mice deficient in HSD17B3 have made the surprising finding that testosterone production is maintained, male mice are masculinised and remain fertile, suggesting differences between mice and human testosterone production exist. We discuss the phenotypic differences observed and the possible other pathways and enzymes that could be contributing to testosterone production and male development. The identification of alternative testosterone synthesising enzymes could inform the development of novel therapies to endogenously regulate testosterone production in individuals with testosterone deficiency.

Prospective Randomized Controlled Trial Comparing Laparoscopic Palomo Surgery versus Scrotal Antegrade Sclerotherapy in Adolescent Varicocele.

Varicocele is a common condition in adolescence and the most common correctable cause of infertility. This study aimed to analyze and compare the outcomes of scrotal antegrade sclerotherapy (SAS) and laparoscopic Palomo surgery (LPS) in a tertiary referral centre. Patients with left grade 3 varicocele indicated for surgery were prospectively enrolled and randomly allocated to the SAS and LPS groups, with their respective contralateral normal testes taken as controls. The primary outcome measures were clinical varicocele recurrence, testicular catch-up growth, and post-operative hydrocele. All patients were evaluated clinically and using Doppler ultrasound by radiologists. From 2015 to 2020, 113 patients completed the study and were statistically analyzed (SAS, n = 57; LPS, n = 56). All patients had significantly smaller testes pre-operatively that the testicular volume differences with control testes were -23% in SAS and -19% in LPS. At 12-month follow-up, there were no statistical significant difference in clinical recurrences between the 2 groups (SAS = 5.3% vs LPS = 5.4%, P>.05, non-inferiority test). Testicular catch-up growths were observed in both groups, the mean testicular volume difference between the treatment and control testes reduced from -23% to -8.1% in SAS (P<.001) and -19% to -9.3% in LPS (P<.001) at 12-month follow-up. There was no post-operative hydrocele in SAS compared to 7 in LPS (0% vs 13%, P = .006). Both SAS and LPS are safe and effective procedures for treatment of adolescent varicocele with significant positive effect on testicular catch-up growth. SAS is not inferior to LPS in terms of clinical recurrence rate and with significantly less post-operative hydrocele.

Melatonin through blockade of Hif-1α signaling mediates the anti-fibrosis under hypoxia in canine Sertoli cells

The hypoxic microenvironment of cryptorchidism is an important factor in the impairment and fibrosis of Sertoli cells which result in blood-testis barrier (BTB) destruction and spermatogenesis loss. Recent studies have shown that melatonin, a well-known pineal hormone exerts beneficial effects against pathological fibrosis in a various of organs. However, it is still unknown whether melatonin can regulate hypoxia-induced fibrosis of Sertoli cells. In this study we evaluate melatonin levels, and its synthesizing enzymes, AANAT and HIOMT expression patterns in canine cryptorchidism and contralateral normal testis. Results show abdominal testes presented low melatonin levels and AANAT and HIOMT expression compared with testes located in the scrotum. Moreover, we established a hypoxia-induced fibrosis model in canine Sertoli cells induced by cobalt chloride (CoCl2) and found that melatonin inhibited the EMT markers expression and ECM production as well as Hif-1α expression of Sertoli cells in a dose-dependent manner. Furthermore, use of Lificiguat (synonyms YC-1, Hif-1α inhibitor) to interfere with the Hif-1α pathway showed a similar effect with melatonin suppression of the fibrosis in Sertoli cells. The results indicate that melatonin supplementation can alleviate the fibrosis process of Sertoli cells caused by hypoxia, which is associated with regulating the inhibition of Hif-1α signaling.

Testis-specific serine kinase 3 is required for sperm morphogenesis and male fertility.

The importance of phosphorylation in sperm during spermatogenesis has not been pursued extensively. Testis-specific serine kinase 3 (Tssk3) is a conserved gene, but TSSK3 kinase functions and phosphorylation substrates of TSSK3 are not known. The goals of our studies were to understand the mechanism of action of TSSK3. We analyzed the localization of TSSK3 in sperm, used CRISPR/Cas9 to generate Tssk3 knockout (KO) mice in which nearly all of the Tssk3 open reading frame was deleted (ensuring it is a null mutation), analyzed the fertility of Tssk3 KO mice by breeding mice for 4 months, and conducted phosphoproteomics analysis of male testicular germ cells. TSSK3 is expressed in elongating sperm and localizes to the sperm tail. To define the essential roles of TSSK3 in vivo, heterozygous (HET) or homozygous KO male mice were mated with wild-type females, and fertility was assessed over 4 months; Tssk3 KO males are sterile, whereas HET males produced normal litter sizes. The absence of TSSK3 results in disorganization of all stages of testicular seminiferous epithelium and significantly increased vacuolization of germ cells, leading to dramatically reduced sperm counts and abnormal sperm morphology; despite these histologic changes, Tssk3 null mice have normal testis size. To elucidate the mechanisms causing the KO phenotype, we conducted phosphoproteomics using purified germ cells from Tssk3 HET and KO testes. We found that proteins implicated in male infertility, such as GAPDHS, ACTL7A, ACTL9, and REEP6, showed significantly reduced phosphorylation in KO testes compared to HET testes, despite unaltered total protein levels. We demonstrated that TSSK3 is essential for male fertility and crucial for phosphorylation of multiple infertility-related proteins. These studies and the pathways in which TSSK3 functions have implications for human male infertility and nonhormonal contraception.