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Summary Varicoceles are the most common and treatable cause of male infertility. The pathophysiology of varicoceles primarily includes elevated temperature, adrenal hormone reflux, gonadotoxic metabolite reflux, altered testicular blood flow, antisperm antibody formation and oxidative stress. The diagnosis of a varicocele is mainly clinical. However, a Doppler ultrasound is used to obtain clinical data and to more accurately measure testicular size. Acoustic radiation force impulse (ARFI) is an additional technique to simultaneously show different areas with different densities in a colour‐coded image and a B‐mode or greyscale image. This can be used for structural analysis of testicular tissue and has become an additional method for detecting pathologic tissue alterations. We enrolled 30 patients who had clinically diagnosed with left varicoceles and male infertility (Group 1). All patients were evaluated by history taking, physical examination, a spermiogram and an endocrine profile. Thirty control patients (Group 2) were randomly chosen from patients who had applied to an andrology clinic for infertility; their physical examinations and laboratory results showed normal findings. Mean elastography results were significantly different between the groups, and significantly lower in patients who had varicoceles. The relationship between hormonal profiles and elastography parameters was calculated as statistically significant negative correlations between FSH and elasticity. Additionally, a negative correlation was determined between varicocele grade and elasticity of testes. In conclusion, our prospective study showed that ARFI imaging may be more useful than palpation for determining early damage of testicular structure by varicoceles.

Lymphatic filariasis is a disease of considerable socioeconomic burden in the tropics. Presently used antifilarial drugs are able to strongly reduce transmission and will thus ultimately lower the burden of morbidity associated with the infection, however, a chemotherapeutic principle that directly induces a halt or improvement in the progression of the morbidity in already infected individuals would constitute a major lead. In search of such a more-effective drug to complement the existing ones, in an area endemic for bancroftian filariasis in Ghana, 33 microfilaremic and 18 lymphedema patients took part in a double-blind, placebo-controlled trial of a 6-wk regimen of 200 mg/day doxycycline. Four months after doxycycline treatment, all patients received 150-200 microg/kg ivermectin and 400 mg albendazole. Patients were monitored for Wolbachia and microfilaria loads, antigenemia, filarial dance sign (FDS), dilation of supratesticular lymphatic vessels, and plasma levels of lymphangiogenic factors (vascular endothelial growth factor-C [VEGF-C] and soluble vascular endothelial growth factor receptor-3 [(s)VEGFR-3]). Lymphedema patients were additionally monitored for stage (grade) of lymphedema and the circumferences of affected legs. Wolbachia load, microfilaremia, antigenemia, and frequency of FDS were significantly reduced in microfilaremic patients up to 24 mo in the doxycycline group compared to the placebo group. The mean dilation of supratesticular lymphatic vessels in doxycycline-treated patients was reduced significantly at 24 mo, whereas there was no improvement in the placebo group. Preceding clinical improvement, at 12 mo, the mean plasma levels of VEGF-C and sVEGFR-3 decreased significantly in the doxycycline-treated patients to a level close to that of endemic normal values, whereas there was no significant reduction in the placebo patients. The extent of disease in lymphedema patients significantly improved following doxycycline, with the mean stage of lymphedema in the doxycycline-treated patients being significantly lower compared to placebo patients 12 mo after treatment. The reduction in the stages manifested as better skin texture, a reduction of deep folds, and fewer deep skin folds. In conclusion, a 6-wk regimen of antifilarial treatment with doxycycline against W. bancrofti showed a strong macrofilaricidal activity and reduction in plasma levels of VEGF-C/sVEGFR-3, the latter being associated with amelioration of supratesticular dilated lymphatic vessels and with an improvement of pathology in lymphatic filariasis patients.

Aim Aim of this study is to evaluate the effects of micronised purified flavonoid fraction (Daflon ® ) on pain, semen analysis and color Doppler parameters in patients with painful varicocele. Materials and methods Forty varicocele patients whom have normal sperm concentration (>20 million/ml) were involved in the study. The patients were divided into two groups such as Daflon ( n  = 20) and placebo ( n  = 20) group. Pain score, semen analyses and Doppler sonography were performed in all patients before and after the treatment. Results In the first group, mean pain scores at 1, 3, 6 and 12 months were 1.80 ± 1.32, 1.15 ± 0.93, 1.05 ± 0.95 and 0.95 ± 0.89, respectively, all were significantly lower ( P  < 0.001 for each) than baseline (5.25 ± 1.07). While semen volume, total sperm count, sperm concentration and morphology were not changed significantly, the motility of sperm increased significantly ( P  = 0.015) due to decrease in grade 1 sperms at the 6th month in the first group. Reflux time of left spermatic vein during the Valsalva maneuver decreased significantly ( P  < 0.001). Conclusions Results of this study suggest the safety and efficacy of Daflon in the treatment of varicocele-associated pain. However, these results of the present study must be confirmed by randomized placebo-controlled studies by using different drug doses and durations before making any recommendation for the use of Daflon.

Using slit and nonslit mesh in laparoscopic totally extraperitoneal preperitoneal (TEPP) inguinal hernia repair are well-known approaches. The aim of this prospective, randomized, clinical study was to assess testicular perfusion after these procedures. In the study period, 40 male patients with unilateral inguinal hernia were assigned into 2 equal groups as follows: slit (S) and nonslit (NS). TEPP hernia repair was performed in all patients. In the 2 groups, testicular arterial blood flow and testis volumes were measured by Doppler ultrasonography preoperatively, on the 5th postoperative day, and 6 months postoperatively, respectively. No statistically significant difference was found between the preoperative, 5th day postoperatively, and 6-month arterial resistance index (ARI) results when comparing the S and NS groups in ultrasonographic testicular blood flow studies. There was no statistically significant difference of testicular volume between the preoperative period, the 5th postoperative day, and 6 months postoperatively in the 2 groups. According to the results, no statistically significant difference was found in terms of testicular perfusion and volume between those 2 methods of TEPP repair for inguinal hernia.

Purpose The two most common procedures for open tension-free groin hernia repair with prosthetic mesh are the Lichtenstein operation and the mesh plug (Rutkow–Robbins) technique. Our study evaluated these two techniques on testicular blood flow and volume, and sperm function in young adults. Methods We randomized operation types with a systematic sampling method, and handled consecutive patients of age 20–30 years having unilateral inguinal hernia repair at our institution from March to August 2008. The study subjects were divided into the Lichtenstein group (LG) and the mesh plug group (MPG). All subjects received color Doppler ultrasonography to determine testicular volume and resistive index (RI) the day before surgery and 3 months postoperatively by a physician blinded for the type of planned or performed operation. Spermiograms done preoperatively and at 3 months postoperatively measured sperm concentration and the rate of progressive motility. Results Sixty-four patients met the study criteria, with 32 patients each in the LG and MPG. RI levels were elevated postoperatively in both the LG ( P  = 0.027) and MPG ( P  = 0.012); there was no significant alteration in terms of testicular volume and spermiogram in the LG and MPG. Conclusion The Lichtenstein and mesh plug techniques in unilateral inguinal hernia increase the RI level significantly in the early postoperative period, but do not have a significant effect on sperm concentration and the rate of progressive motility.

The evolutionarily conserved peripheral benzodiazepine receptor (PBR), or 18-kDa translocator protein (TSPO), is thought to be essential for cholesterol transport and steroidogenesis, and thus life. TSPO has been proposed as a biomarker of neuroinflammation and a new drug target in neurological diseases ranging from Alzheimer’s disease to anxiety. Here we show that global C57BL/6- Tspo tm1GuWu(GuwiyangWurra) -knockout mice are viable with normal growth, lifespan, cholesterol transport, blood pregnenolone concentration, protoporphyrin IX metabolism, fertility and behaviour. However, while the activation of microglia after neuronal injury appears to be unimpaired, microglia from GuwiyangWurra TSPO knockouts produce significantly less ATP, suggesting reduced metabolic activity. Using the isoquinoline PK11195, the ligand originally used for the pharmacological and structural characterization of the PBR/TSPO, and the imidazopyridines CLINDE and PBR111, we demonstrate the utility of GuwiyangWurra TSPO knockouts to provide robust data on drug specificity and selectivity, both in vitro and in vivo , as well as the mechanism of action of putative TSPO-targeting drugs. The 18-kDa translocator protein (TSPO) has been implicated in steroid biogenesis and neuroinflammation. Here, the authors create viable and fertile global TSPO knockout mice, challenging the assumption that TSPO is essential for mouse development but suggesting that it may have a role under certain disease conditions.

Classical histology struggles to preserve three-dimensional spatial context, prompting the emergence of optical tissue clearing techniques that enable imaging of intact specimens at cellular or subcellular resolution. These techniques have revolutionised fields like cell biology, developmental biology, and neuroscience. However, their application in reproductive biology remains unexplored - particularly in studying the complexities of testicular development. We developed a novel, efficient and affordable toolbox for studying intact testicular tissues, PT-CLEAR3D, that stands for primate testis - whole mount staining, tissue clearing and three-dimensional imaging. Intact testicular tissues from humans (transgender model), common marmosets and macaques underwent antibody labelling, clearing with organic solvents, and three-dimensional imaging using light sheet fluorescence microscopy. Marker specificity was confirmed by immunofluorescence staining of 3 and 25 µm testicular sections, followed by imaging with confocal microscopy. The testicular structure was evaluated using several markers: spermatogonia (melanoma-associated antigen 4), least differentiated spermatogonia (Piwi-like protein 4), Sertoli cells (vimentin and SRY-Box transcription Factor 9), peritubular myoid cells and vasculature (alpha-smooth muscle actin), and NucSpot as a nuclear dye. PT-CLEAR3D efficiently achieved optical transparency while a commercial kit that was run in parallel was inefficient. This study presents a pioneering three-dimensional visualization of intact testicular samples of up to 50 mm3 in size and imaging depth of up to 4.5 mm across three primate species. Remarkably, PT-CLEAR3D revealed critical details at both tissue and cellular levels such as the spatial distribution of germ and somatic cells, cellular bridges, and vasculature. Furthermore, PT-CLEAR3D enabled three-dimensional reconstructions that effectively reduces confirmation bias enhancing our observation of spermatogonial clones organized as single cells, pairs, and quartets. Importantly, it adeptly identified testicular pathology and the persistence of germ cell clones in select tubules within the transgender testis following hormonal suppression of spermatogenesis. This technological development offers a versatile toolbox with benefits such as applicability across multiple species, fluorophore multiplexing, compatibility with different fixatives and deep tissue volumetric imaging with cellular resolution. Overall, PT-CLEAR3D establishes a foundation for spatial evaluation of testicular development, presenting substantial potential for advancing our understanding of the intricate kinetics of spermatogenesis in health and disease.

PurposeTo assess the role of 3.0 T Diffusion Tensor Imaging (DTI) and Fiber Tractography (FT) of the testes in the work-up of nonobstructive azoospermia (NOA).MethodsThis prospective study included consecutive NOA men and controls. A 3.0 T scrotal MRI was performed, including DTI. The testicular apparent diffusion coefficient (ADC) and fractional anisotropy (FA) were calculated. FT reconstructions were created. The Kruskal–Wallis test, followed by pairwise comparisons, assessed differences in testicular ADC and FA between NOA histologic phenotypes (group 1: hypospermatogenesis; group 2: maturation arrest; and group 3: Sertoli cell-only syndrome) and normal testes. The Mann–Whitney-U test compared ADC and FA between NOA testes with positive and negative sperm retrieval. Visual assessment of the testicular fiber tracts was performed. Fiber tracts fewer in number, of reduced thickness, disrupted and/or disorganized were considered “abnormal”. Chi-square tests and binary logistic regression analysis assessed variations in testicular fiber tracts morphology.ResultsTwenty-nine NOA men (mean age: 39 ± 5.93 years) and 20 controls (mean age: 26 ± 5.83 years) were included for analysis. Higher ADC (p < 0.001) and FA (p < 0.001) was observed in NOA testes compared to controls. Differences in FA were found between groups 1 and 3 (0.07 vs 0.10, p = 0.26) and groups 2 and 3 (0.07 vs 0.10, p = 0.03), but not between groups 1 and 2 (p = 0.66). An increase in FA was observed in NOA testes with Sertoli cell-only syndrome compared to hypospermatogenesis and maturation arrest. FA was higher in NOA testes with negative results for the presence of sperm compared to those with positive results (0.09 vs 0.07, p = 0.006). FT showed “abnormal” fiber tracts in NOA testes (p < 0.001).Conclusion3.0 T DTI and FT provide an insight into deranged spermatogenesis in NOA testes.

Mass spectrometry imaging (MSI) is a promising analytical method to visualize the distribution of lipids in biological tissues. To clarify the relationship between cellular distribution and lipid types in a tissue, it is crucial to achieve both an improvement in ion detection sensitivity and a reduction in the ionization area. We report methods for improving the efficiency of ion transfer to a mass spectrometer and miniaturizing the extraction area of a sample for tapping-mode scanning probe electrospray ionization (t-SPESI), atmospheric pressure sampling, and ionization methods. To verify the efficacy of the new t-SPESI measurement system, MSI was performed on mouse testes with a pixel size of 5 µm. Lipid images of the testes from wild-type (WT) and lysophospholipid acyltransferase 3 (LPLAT3) knockout mice revealed the characteristic distribution of docosahexaenoic acid–containing phospholipids (DHA-PLs). A comparison of the ion images obtained by MSI and optical images of the same tissues stained with hematoxylin and eosin suggested that the distribution of DHA-PLs was significantly altered by spermatogenesis in the WT mouse testes. Graphical abstract

Background To investigate the correlation between testicular fat content (TFC) and sex hormone levels in patients with cryptorchidism and its value in assessing postsurgical testicular function. Methods Pelvic MRI with the mDIXON Quant sequence was performed on 23 cryptorchidism patients and 15 normal controls. The TFC before and after surgery was measured and compared. The correlations between cryptorchid TFC and testosterone (TSTO), follicle-stimulating hormone (FSH), and estradiol (E2) levels were analyzed, as was the specificity of TFC and each hormone for assessing testicular function after surgery. Results The preoperative cryptorchid TFC (3.06% ± 0.74) was higher than that of the normal controls (1.36% ± 0.49). TSTO was negatively correlated with the cryptorchid TFC (r = −0.698), while FSH and E2 were positively associated with the cryptorchid TFC (r = 0.658, 0.676). Cryptorchid TFC after surgery (2.01% ± 0.55) was lower than the preoperative TFC, but hormone levels were not significantly different. The TFC after surgery (0.864) had a larger AUC value than did TSTO (0.639), FSH (0.597), and E2 (0.586). Conclusion Noninvasive quantification of cryptorchid TFC using the mDIXON Quant sequence is more specific than hormone levels for assessing postsurgical changes in testicular function. Impact The cryptorchid testicular fat content is significantly higher than the normal testicular fat content. Cryptorchid testicular fat content is negatively correlated with presurgical serum TSTO levels and positively correlated with presurgical FSH and E2 levels. Pre- and postoperative changes in cryptorchid testicular fat content change are more sensitive than changes in TSTO, FSH, or E2 levels. Noninvasive cryptorchid testicular fat content quantified by the mDIXON Quant sequence is more specific than serum TSTO, FSH, and E2 levels for assessing changes in testicular function after cryptorchidism surgery.