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INTRODUCTION AND OBJECTIVE:In cryptorchid boys, germ cell development failure occurs during infancy. Thus, the establishment of serum markers for predicting infertility is warranted. Serum inhibin ...

INTRODUCTION AND OBJECTIVE:We previously detected differentially expressed genes (EEF1A1 and TPT1) in the testis of cryptorchidism (Mizuno K, et al., J Urol. 2009). Although these proteins were spe...

We elucidated the mechanism of spermatogonial stem cell disturbance of cryptorchidism and investigated the expression of miRNAs and their target genes in undescended testes.Using microarray analysis we compared total miRNA expression in unilateral undescended testes with that in contralateral descended and normal testes in a rat model of cryptorchidism. The model was derived by administering flutamide to pregnant Sprague Dawley® rats. We identified mRNA targets of miRNAs by bioinformatic analysis, followed by in situ hybridization and immunohistochemistry to localize candidate miRNAs and mRNAs, respectively. We also investigated whether miRNAs could inhibit target protein expression in vitro.Microarray analysis and subsequent quantitative reverse transcriptase-polymerase chain reaction showed that only miR-135a was expressed at a lower level in undescended testes. We identified its target as FoxO1, which is essential for stem cell maintenance. miR-135a and FoxO1 localized to spermatogonial stem cells. FoxO1 localized to the spermatogonial stem cell nucleus less frequently in undescended testes, indicating that the activity of FoxO1, which acts as a transcription factor, is altered in undescended testes. Finally, miR-135a transfection into spermatogonia in vitro resulted in down-regulation of FoxO1 expression.In cryptorchid testes there is a decreased number of spermatogonial stem cells in which FoxO1 is activated, indicating that failure of spermatogonial stem cell maintenance results in spermatogenesis alteration. We also noted interaction between miR-135a and FoxO1, and propose that miR-135a contributes to spermatogonial stem cell maintenance through modulation of FoxO1 activity.

INTRODUCTION AND OBJECTIVES: Cisplatin is a widely used chemotherapeutic agent for the treatment of various genitourinary tumors. Although it is a very effective, its use is limited by its renal and testicular toxicity. Mesenchymal stem cells (MSCs) are pluripotent undifferentiated cells that have the ability to differentiate and divide into another cell types and can self renew. Our aim was to investigate the possible protective role of MSCs against Cisplatin induced testicular apoptosis and damaged oxidant/antioxidant balance in rats. Furthermore, we explored the involvement of apoptotic markers in the pathogenesis of testicular toxicity and their role in monitoring treatment. METHODS: A total of 30 male Sprague Dawley rats were used for the experiment. They were divided into 3 groups: Group 1 served as control group received intraperitoneal (IP) saline, Group 2 (Cisplatin group) received a single IP dose of Cisplatin 7mg/kg and Group 3 (Stem Cells group) received the same IP single dose of Cisplatin and was administered BM-MSCs (2x10 cells) by penile vein injection 24h after the Cisplatin injection. Testicular weight and plasma testosterone level were determined along with testicular malondialdehyde (MDA) and reduced glutathione (GSH) levels as oxidative stress markers. Testicular gene expressions of Caspase-3 and mitogen activated protein kinase (MAPK) were estimated by real time-PCR. Histopathological examination for testicular tissues was also done. RESULTS: Cisplatin induced a significant reduction in testicular weights, plasma testosterone and testicular GSH levels in addition to significant elevation of testicular MDA levels and gene expressions of Caspase-3 and MAPK when compared with control group (p

INTRODUCTION AND OBJECTIVES: The UCiTrainer is a low cost, portable laparoscopic trainer that utilizes an electronic tablet for video and optics; it can be printed with a consumer-level three-dimensional (3D) printer. We report our initial experience on the feasibility of using 3D printing technology as a novel method of manufacturing and distribution for low cost, surgical education tools on a global on demand basis. METHODS: We created computer-aided designs (CAD) using Solidworks Software (Dassault Systemes Solidworks Corp., Velizy-Villacoublay, France) for nine separate components which could then be assembled into a laparoscopic training device including an instrument port mold. These files were provided to two remote institutions to 3D print and assemble the trainer. The CAD files were uploaded to Makerware software (MakerBot Industries, New York City, NY) to be sliced into layers and prepared for printing. All pieces were printed using a Flashforge Creator 3D printer (Flashforge, Jinhua, China) in five separate prints. The following printer settings were used: raft layer, 15% infill, 2 shells, 0.10 mm layer height, 230 ? C extruder, 110 ? C build plate, 90 mm/s extrusion and 150 mm/s travel. In this process, successive layers of a heated, acrylonitrile butadiene styrene filament were extruded and laid on top of one another in both the horizontal and vertical axis. The printed mold was used to create two instrument ports using a liquid silicone rubber. Both remote institutions were provided oral and written instructions for assembly of the components. RESULTS: Initial attempts of 3D printing of the larger pieces (Figure 1b) were not successful due to deformation of the plastic. Additionally, initial calibration and leveling of the 3D printer were challenging to novices. Accordingly, the build plate was increased to 120 ? C and a full enclosure was made for the 3D printer to retain heat during the printing process. With these adjustments, successful printing of the components was accomplished. Both institutions were subsequently able to completely assemble and use the training device using only the oral and written instructions provided. CONCLUSIONS: 3D printing technology can be used successfully to disseminate a training device for use in surgical education. The device is inexpensive and can be made on an as-needed basis.

Laparoscopic extravesical ureteral reimplantation for vesicoureteral reflux has gained acceptance as a feasible treatment associated with minimal morbidity. However, ureteral advancement with this technique has not been attempted. We examined the usefulness of ureteral advancement via laparoscopy.A total of 30 patients with 51 refluxing ureters underwent laparoscopic extravesical ureteral reimplantation for treatment of vesicoureteral reflux between August 2009 and September 2011. Mean ± SD patient age was 60.8 ± 48.6 months. During the procedure 15 patients underwent ureteral advancement (advancement group), while 15 did not (nonadvancement group). We compared operative times and postoperative rates of urinary tract infections and persistent reflux between the groups.There was no significant difference in operative times in unilateral (mean ± SD 110 ± 25 vs 125 ± 42 minutes) and bilateral cases (mean ± SD 214 ± 52 vs 203 ± 40 minutes) between the nonadvancement vs advancement groups. All patients underwent voiding cystourethrography 3 to 4 months postoperatively. Reflux resolution rate for ureters was significantly higher in the advancement group (100%) than in the nonadvancement group (85%, p0.05). No patient in the advancement group had postoperative urinary tract infection. Fixation of the ureter with the bladder muscularis at the proximal limit of the detrusor defect and/or a percutaneous hitch stitch placed in the ventral side of the proximal limit of the detrusor defect facilitated ureteral advancement.Ureteral advancement is a simple and feasible procedure in laparoscopic ureteral extravesical reimplantation and may improve the resolution rate of vesicoureteral reflux.

Four disorders, including poor semen quality, testicular cancer, cryptorchidism and hypospadias, are thought to represent testicular dysgenesis syndrome and have been hypothesized to share a common etiology. We predicted testicular function in prepubertal boys with a history of cryptorchidism and/or hypospadias by measuring serum hormone levels.A total of 82 prepubertal boys who underwent orchiopexy and/or hypospadias repair in childhood were enrolled in the study. Patients were surgically treated for cryptorchidism (23 in group 1), hypospadias (49 in group 2), cryptorchidism and hypospadias (10 in group 3), and hydrocele testis (7 in control group 4). Serum hormones, including luteinizing hormone, follicle-stimulating hormone and total testosterone, were measured separately by age less than 12.5, 12.5 to 13.5 and greater than 13.5 years, and by Tanner pubertal stage.Follicle-stimulating hormone in group 3 was significantly higher than in groups 1, 2 and 4 at ages 12.5 to 13.5 and greater than 13.5 years, and for Tanner stages 2 and 3 (p0.05). However, luteinizing hormone and testosterone did not differ among the groups regardless of age or Tanner stage. Group 3 patients had significantly higher follicle-stimulating hormone regardless of the severity of cryptorchidism or hypospadias.Data suggest that testicular function in patients with cryptorchidism plus hypospadias is more severely impaired than that in patients with cryptorchidism or hypospadias, lending clinical support to the testicular dysgenesis syndrome hypothesis of a common origin.

To evaluate the validity of 1-stage Fowler-Stephens orchiopexy, we performed this procedure for intra-abdominal testes in an experimental cryptorchid rat model and assessed postoperative spermatogenesis.Cryptorchidism in rats was induced by injecting flutamide into the abdomen of pregnant Sprague Dawley rats for 7 days (days 14 to 20 of gestation). Four-week-old cryptorchid rats were divided into the 4 groups of sham operation (group 1, 7 rats), orchiopexy only (group 2, 7), spermatic vessel ligation only (group 3, 7), and spermatic vessel ligation and orchiopexy (1-stage Fowler-Stephens orchiopexy, group 4, 7). The testes were removed 12 weeks after birth in all groups, and testicular weight, testicular histopathological findings and presence of sperm in the epididymis were examined.All operated testes had reticulated vessels on the tunica albuginea. There were no statistically significant differences in the testicular weight between groups 1 (mean +/- SD 0.47 +/- 0.04 gm) and 4 (0.30 +/- 0.19), suggesting that Fowler-Stephens orchiopexy did not induce significant testicular atrophy. However, seminiferous tubule diameters in group 4 were significantly smaller than in group 1 (p0.001), and no sperm was observed in the epididymis of group 4 rats, suggesting that Fowler-Stephens orchiopexy reduced seminiferous tubule diameters and did not improve spermatogenesis.Although Fowler-Stephens orchiopexy is a good procedure to maintain testicular size and correct cosmetic deformity of an absent scrotum, it may not significantly contribute to the improvement of spermatogenesis.

We examined 3-dimensionally the process of external genitalia formation in the experimental hypospadiac rat model.We administered 7.5 mg flutamide daily, a blocker of androgen receptor, into the abdomen of naturally pregnant female Sprague-Dawley rats from gestational days 14 to 20 to produce a hypospadiac rat model. The control group consisted of male offspring not exposed to flutamide. The fetal phallus was obtained at gestational days 17.5, 19.5 and 21.5. We observed them by scanning electron microscopy.In the 17.5-day-old embryo the projection on the ventral side of the phallus was observed from the base of the phallus to the coronary sulcus in the control rat. This finding demonstrated that the urethra develops from the base of the phallus to the coronary sulcus. On the other hand, this projection was not observed in the hypospadiac rat and the urethra was not seen on the ventral side of the phallus. In the 19.5-day-old embryo the ventral preputial closure appeared in the proximal phallus of the control rat but not of the hypospadiac rat. In the control rat the scrotum was discerned in the perineum and divided by a median fold (the raphe). The raphe reached the base of the phallus, where it was concurrent with the preputial fold that covers the urethra. On the other hand, the hypospadiac rat did not have a raphe and the hollow at the base of the phallus appeared to give rise to the future orifice of the urethra. In the 21.5-day-old embryo the control rat prepuce completely surrounded the distal phallus, whereas that of the hypospadiac rat was dorsally hooded with a ventral cleft and the external appearance resembled the morphology of human penoscrotal hypospadias.This hypospadiac rat model is considered valuable for further studying penile growth and differentiation, and the molecular mechanisms in external genitalia formation in hypospadias.

INTRODUCTION AND OBJECTIVES: Male infertility is a serious complication of cryptorchidism. In this study, we focused on early stage spermatogenesis and spermatogonial stem cell (SSC) activity in undifferentiated spermatogonia in cryptorchid testes to elucidate the mechanism of infertility caused by cryptorchidism. METHODS: Histological findings and expression patterns of the stem cell marker, undifferentiated embryonic cell transcription factor 1 (UTF1), were examined in a unilateral cryptorchid rat model induced by flutamide and in cryptorchid patinents. We removed both unilateral descended testis (UDT) and contralateral descended testis (DT), respectively, from cryptorchid and normal rats (control) 18 days postcoitum (dpc) to 144 days postpartum (dpp). Human testicular tissues were also excised from 5 boys (8–20 months old) during orchiopexy between January 2003 and December 2008. Hematoxylin and eosin staining, real-time RT-PCR, western blot analysis, and immunohistochemical analysis with UTF1 were performed to assess early stage spermatogenesis and SSC activity. RESULTS: Histopathological analysis showed that gonocyte differentiation into early A spermatogonia occurred at 9 dpp; however, this transformation was disturbed in UDT. Quantitative study revealed that UTF1 expression decreased with testicular development, but did not disappear in adulthood. Immunohistochemical analysis showed that all gonocytes and some early type A spermatogonia expressed UTF1 in rat DT. Furthermore, the UTF1-negative early A spermatogonia/positive early A spermatogonia ratio was significantly higher in the UDT group (0.69 0.04) than in the control (0.46 0.10; p 0.037) and DT (0.44 0.05; p 0.022) groups, indicating decreased early A spermatogonia with SSC activity in cryptorchid testes. In humans, UTF1-negative Ad spermatogonia/positive Ad spermatogonia ratios in UDT were also significantly higher than in the DT (0.55 0.36, 0.16 0.07, respectively; p 0.006). CONCLUSIONS: In cryptorchid testes, the differentiation from gonocytes into early A spermatogonia, as well as the stem cell activity of early A spermatogonia, were altered during the early stage of spermatogenesis, suggesting that the loss of SSC activity in cryptorchid rats resulted in altered spermatogenesis, thus interfering with fertility.

INTRODUCTION AND OBJECTIVES: Androgens positively affect penile growth in children. Androgen stimulation is often used prior to hypospadias surgery in an attempt to increase penis size when the penis is too small to repair. Although recent publications have shown negative effects of androgens on normal skin healing, its influence on the foreskin of hypospadias is not clear. We histologically evaluated the foreskin to elucidate the effects of androgen stimulation before hypospadias surgery. METHODS: Of 134 boys who underwent the first step of hypospadias repair at our institute from January 2005 to August 2011, we performed preoperative androgen treatment in 10 boys with small penises. These cases received as many as 3 intramuscular injections of testosterone enanthate (25 mg) every 4 weeks. We obtained excess foreskin that had been excised during surgery and compared histological findings in hormonal treatment and non-treatment cases. We also performed immunohistochemistry using anti-VEGF and collagen type I and type III antibodies. RESULTS: The mean age at surgery was 31.3 and 20.8 months in treatment and non-treatment groups, respectively. Testosterone enanthate treatment significantly increased penile length from 20.6 5.8 mm to 30.1 4.3 mm (p 0.0001) and the glans width from 10.3 2.1 mm to 12.9 2.4 mm (p 0.008). We histologically evaluated the foreskin of 117 out of 134 cases. In the treatment group (n 9), we observed edema, fibrosis, and inflammation in 6 (66.7%), 6 (66.7%), and 7 (77.8%) cases, respectively. In the non-treatment group (n 108), we observed edema, fibrosis, and inflammation in 19 (17.9%), 17 (16.0%), and 46 (43.4%) cases, respectively. These histological findings were significantly increased in the foreskin after testosterone treatment (p 0.005, p 0.005, p 0.078, respectively). VEGF expression was higher in the foreskins of the treatment group than in the non-treatment group. CONCLUSIONS: Testosterone administration prior to hypospadias surgery in small penile cases is useful because it significantly enlarges penile size. In the present study, significant histological changes such as edema and fibrosis were found in the treatment group foreskins compared to the non-treatment group, suggesting that the effects of testosterone on the foreskin stroma via the androgen receptor facilitates these changes. High VEGF protein expression in the treatment group suggests that topical testosterone led to angiogenesis via VEGF expression and subsequently induced distinctive histological reactions of the foreskin.

INTRODUCTION AND OBJECTIVES: Spermatogenesis, a fundamental process in the male reproductive system, requires a series of tightly controlled epigenetic and genetic events in germ cells ranging from spermatogonia to spermatozoa. We recently reported that important differentiation occurs in the testes of rats at approximately 9 days. We observed through microarray gene expression analysis that Jumonji, AT rich interactive domain 1A (Jarid1a) was highly expressed in the undescended testes compared with normal testes at 9 days. The purpose of this study was to examine the effects of Jarid1a, a key epigenetic regulator expressed in the testis, on spermatogenesis. METHODS: We injected flutamide (7.5 mg/day per body) into the abdomen of pregnant rats for 7 days from the 14th to 20th day of gestation to obtain offspring with undescended testes. Nine-day-old male rats were divided into 3 groups: (1) f-UDT, offspring with undescended testes, (2) f-DT, offspring with descended testes, and (3) nD9, normal testes of normal rats. Jarid1a gene expression in these 3 groups was assessed by real-time polymerase chain reaction (RTPCR) and western blot. The localization of expression was assessed by immunohistochemistry. Moreover, these groups were assessed by western blot using the anti-histone H3 lysine 4 (H3K4) antibody and methylated antibody to evaluate the state of H3K4 modification. RESULTS: RT-PCR study showed that Jarid1a was significantly expressed in the f-UDT compared to f-DT or nD9. Similar findings were observed by western blot study analysis. Immunohistochemical analysis showed that Jarid1a was expressed in the nuclei from Ad spermatogonia to spermatids. Furthermore, western blot analysis of H3K4 methylation showed that levels of dimethylated H3K4 (H3K4me2) and tri-methylated H3K4 (H3K4me3) were higher in f-UDT than in f-DT or nD9. CONCLUSIONS: Jarid1a belongs to the Jumonji family, whose members share the Jumonji C (JmjC) domain and suppress transcription by means of demethylating triand di-methylated H3K4. Because Jarid1a was expressed in rat testes in widespread spermatogenesis and was especially high in early spermatogenesis, the state of methylation of H3K4 highly correlates with spermatogenesis. Jarid1a demethylase-regulated histone modification is likely to inhibit spermatogenesis via a low level of methylated H3K4 in undescended testes.

isothiocyanate (AI), a TRPA 1 agonist, on the single unit mechanosensitive afferent activities (SAAs) primarily originating from the bladder in the rat. METHODS: Female Sprague-Dawley rats were used. Under urethane anesthesia, SAA primarily originating from the bladder were identified by electrical stimulation of the left pelvic nerve and by bladder distension. Nerves with conduction velocity (CV) 2.5 m/sec were determined as myelinated A -fibers and those with 2.5 m/sec as unmyelinated C-fibers. Before drug administration, the baselines of SAA were recorded during cystometry with saline instillation at a rate of 0.08 ml/min until intravesical pressure reached 30cmH2O. Then, AIsolution (10 M) was instilled intravesically three times. As the vehicle of AI, 0.01% ethanol was used. The SAAs are expressed as a percentage of baseline activity, integrated during the whole filling phase, which is based on pressure and volume. RESULTS: Twenty-three SAAs were isolated in 13 rats. Ten units corresponded to criteria for A -fibers (CV: 9.09 9.15 m/sec.), and 13 for C-fibers (CV: 1.03 0.55 m/sec.). Neither AI-instillation nor vehicle-instillation changed bladder compliance significantly (data not shown). SAA of both A and C-fibers significantly increased after AI-instillation, but not after vehicle-instillation (Figure). The difference between the groups was statistically significant. CONCLUSIONS: The results of the present study demonstrate that AI can stimulate the afferent activities of both mechanosensitive A and C-fibers in the rat bladder. To our knowledge, this is the first direct demonstration of the stimulatory effect of the TRPA 1 agonist on bladder mechanosensitive afferent activities.

proximal penile) in 4.6%, and hypospadias NOS (not otherwise specified) in 16%. The prevalence of hypospadias increased during the surveillance period: 66.9 (1998–02) vs 81.0 (2003–7) (p 0.0003). While the prevalence rates for every degree of hypospadias also increased, the prevalence of hypospadias NOS decreased from 14.8 to 9.0 during these periods. Increased prevalence rates were associated with Caucasian ethnicity, higher order births, low birth weight, prematurity, and primaparous births. The central region of the state had the highest prevalence rate with 92 as compared to the northwest 72.3, northeast 69.3, southeast 59, and southwest region 60.2. CONCLUSIONS: Hypospadias prevalence in Arkansas is on the rise and is significantly higher than other states in the surveillance group. In light of these factors and the existing buccal tissue repository, there is an opportunity for genetic probing to identify causative factors for hypospadias. Improvement in the categorization of degrees of hypospadias also may have an impact on the reported prevalence rates.

ture (2). The remaining 22 cases have no ventral curvature and early excellent cosmetic results, with a slitlike meatus placed in the glans. CONCLUSIONS: Our results suggest that the use of deep ventral transverse corporeal incisions for correction of severe curvature allows SPGU and concurrent CVL. The early cosmetic and functional results with this approach are satisfactory with an acceptable complication rate.

labeled hAFSC can be tracked using bioluminescence. Moreover, we show that hAFSC provide a protective effect, ameliorating ATN in the acute injury phase as reflected by decreased creatinine and BUN blood levels and by a decrease in the number of damaged tubules and apoptosis therein, as well as by promoting proliferation of tubular epithelial cells. We show significant immunomodulatory effects of hAFSC, over the course of ATN. CONCLUSIONS: We therefore speculate that AFSC could represent a novel source of unmodified stem cells that may function to modulate the kidney immune milieu in renal failure caused by ATN and improve recovery of the kidney during acute damage.