A nested variant of transitional cell carcinoma of the urinary bladder: a case report.

Nested variants of transitional cell carcinomas (TCC-NVs) are relatively rare neoplasms in the urinary bladder, but at least 25 cases have been described. This disease is characterized by the pathological finding of irregular nests and/or tubules of transitional carcinoma cells infiltrating the lamina propria without involvement of the mucosal layer. In our case, diagnosed by open biopsy, there were scattered tumor cells observed from the muscle layer to the subserosa, with a tendency toward increasing cellular anaplasia with the depth of invasion. Since the prognosis of TCC-NV is generally poor, comprehensive chemotherapy was performed. No changes were observed on computed tomography and the performance status (0) remained the same after 1 year, so the treatment was considered effective. We conclude that open biopsy should be carried out without hesitation when bladder cancer is suspected, even if there are negative findings of repeated urinary cytology examination and/or endoscopic cold cup biopsy. Immunohistochemical analysis may help in the diagnosis of TCC-NVs derived from epithelial cells. Diagnosis and treatment at an early stage should reduce the mortality of patients with TCC-NVs.

Involvement of calpain for apoptosis in dysfunction of the unaffected testis in rats with experimental testicular torsion.

PROBLEM: The dynamics of calpain and involvement of apoptosis in sperm formation disorder of the unaffected testis in rats with experimental testicular torsion were investigated. METHODS OF STUDY: Using 6-week-old Wistar rats, an experimental unilateral testicular torsion model was prepared. The bilateral testes were excised 1, 3, 5, 7, 14, 35, and 70 days after the left testis was twisted, and the unaffected testes were subjected to immunohistological staining, sodium dodecyl sulfate polyacrilamide gel electrophoresis and Western blotting using anti-calpain antibodies. Apoptosis was detected by the TdT-mediated dUTP biotin nick end-labeling (TUNEL) method. RESULTS: By immunohistological staining, positive immunostaining by anti-pro mu-calpain antibody was observed in the spermatocyte nucleus, but not with anti-pro m-calpain antibody. The staining was increased until 7 days after testicular torsion, then decreased with progression of sperm formation disorder. By Western blotting, the intensity of staining with anti-pro mu-calpain antibody was increased until 7 days after torsion of the testis. Apoptosis expression in the unaffected testis was significantly inhibited by addition of a calpain inhibitor. CONCLUSIONS: It was suggested that mu-calpain may be involved in apoptosis expression in sperm formation disorder of the unaffected testis in unilateral testicular torsion.

[A study on the mechanism of the spermatogenic damage after vasectomy in rats].

PURPOSE: Vasectomy may result in damage to spermatogenesis. There are several explanations for this damage, including an increase of pressure in the seminiferous tubules and an autoimmune reaction. Recently, vasectomy has been reported to induce germ cell death by apoptosis. However, the exact mechanism of this vasectomy-induced germ cell apoptosis is unclear. To elucidate this mechanism, we designed a vasectomized rat model and examined the testiscular alterations and apoptotic degeneration biochemically and microhistopathologically. Particularly, we analyzed the expression of nitric oxide synthase (NOS) and nuclear factor kappa B (NF kappa B), which plays a critical role in the induction of the iNOS gene, in the testis after vasectomy to gain insight into the association between germ cell apoptosis and these factors. MATERIALS AND METHODS: The testes of 40 Wistar rats (10-weeks old) were studied at 1, 2, 5 and 10 weeks after unilateral (left) vasectomy. Wistar rats weighting 290 to 310 g were divided into 2 groups and subjected to underwent either unilateral vasectomy or sham surgery under ether anesthesia. Bilateral testes were carefully observed biochemically and histopathologically. Apoptosis was detected by an in situ end-labeling technique (detection of cellular DNA fragmentation) and electron microscopy. Neuronal NOS (nNOS), endothelial NOS (eNOS) and inducible NOS (iNOS) protein was detected by Western blotting and immunohistochemical studies using each NOS monoclonal antibody. To confirm the co-localization of cellular DNA fragmentation in germ cells and each NOS, each set of consecutive testis sections (one stained for cellular DNA fragmentation and the others for each NOS) were examined. Expression of NF kappa B proteins was examined immunohistochemically using a NF kappa B p65 polyclonal antibody. RESULTS: At 5 and 10 weeks after vasectomy, the vasectomized left testis was significantly lighter than the unvasectomized right testis and sham-operated testis. At that time, the seminiferous tubules of vasectomized testes were highly damaged, presenting narrow tubular diameter, disorder of cellular arrangement, depletion of the germ cells, and local interstitial fibrosis. Vasectomized testes demonstrated a significantly increased number of apoptotic germ cells per cross-sectional area compared with sham-operated testes at 5 and 10 weeks after operation (p < 0.01). Electron microscopy revealed apoptotic germ cells each with a darkly stained nucleus. Western blotting and immunohistochemical studies demonstrated that iNOS proteins were more strongly expressed on vasectomized testes as time passed after vasectomy. Examination of consecutive sections from the vasectomized testis revealed that visibly apoptotic germ cells that exhibited positive staining for cellular DNA fragmentation were also intensely stained for eNOS and iNOS. NF kappa B p65 proteins were more strongly expressed in the nucleus of germ cells in the vasectomized testis than in the sham-operated testis. CONCLUSIONS: We found that vasectomy results in damage to spermatogenesis in adult rats, that may induce germ cell apoptosis, and that iNOS and NF kappa B may play a critical role in the germ cell apoptosis after vasectomy.

Developing germ cells in mouse testis express pheromone receptors.

Pheromone receptors are expressed in the accessory olfactory system, which is vital for non-specific chemical communication and for sexual behavior. Under the hypothesis that some of the pheromone molecules released from female reproductive organs might regulate sperm chemotaxis or chemokinesis, we examined whether the V1R type pheromone receptor mRNAs are expressed in developing germ cells. By a reverse transcription-PCR method, we obtained nine kinds of cDNA fragments belonging to the receptor family. In situ hybridization analysis in testicular sections using probes of testicular pheromone receptors (TVRs) revealed that TVR mRNAs were expressed by spermatids. TVRs were also expressed in the accessory olfactory organ. In the testis, hybridization signals were localized in subsets of the seminiferous tubules, suggesting that TVRs were expressed by selective subsets of the spermatids. In situ hybridization study suggests also that each sperm expresses multiple pheromone receptors. The testicular pheromone receptors might have an important role in the maturation and/or migration of sperm.

[Effects of the gene transfer into sperm mediated by liposomes on sperm motility and fertilization in vitro].

To clarify the problems of gene transfer in the fertility clinic, we investigated the safety and effectiveness of the gene transfer of mouse sperm using the lipofection method. We used the pCAGGS-lacZ plasmid connected with cytomegarovirus enhancer/chicken beta-actin promoter as an adventitious gene. After incubation of the DNA-liposome complex and mouse sperm of epididymis in the medium, we investigated the sperm motility after gene transfer, in vitro fertilization rate of the gene transferred sperm, and gene expression of the fertilized ovum and fetus. Sperm motility was lowered significantly with the increase in DNA density. The fertilization rate for in vitro fertilization was lowered significantly with the increase in DNA density. Gene expression was observed in some of the fertilized ova. In the fetus 12.5 days after embryo transfer, gene transfer was confirmed by the PCR method, but we could not find gene expression by X-gal staining. We proved that the sperm with a gene transferred by the lipofection method could fertilize the ovum and transfer adventitious gene to ova. We speculate that sperm motility and fertility rate were lowered because the introduction of DNA and the DNA-liposome complex destroyed parts of the cell. Development of a sperm fractionation method for DNA transfer and establishment of a method for effective and safe DNA transfer are awaited.

Results of closure of urethrocutaneous fistulas after hypospadias repair.

BACKGROUND: Urethrocutaneous fistulas are one of the major causes of morbidity after hypospadias repair. METHODS: During the last 2.5 years, 26 patients underwent repair of 41 urethrocutaneous fistulas. These fistulas were repaired by a 3-layered closure method, by using meticulous surgical techniques aided by optical magnification. In large fistulas, a dermal subcutaneous flap was created and brought over the surgically repaired urethral fistula. RESULTS: Twenty-four of the 26 patients with urethrocutaneous fistulas after hypospadias repair had fistula closure, with a 92% success rate. CONCLUSION: A high success rate was obtained with a multilayered closure using meticulous techniques to repair urethrocutaneous fistulas.

Asymptomatic intrarenal arteriovenous fistula accompanying severe renal vein dilatation detected 30 years after percutaneous renal biopsy.

We report on a patient with asymptomatic intrarenal arteriovenous fistula (AVF) accompanying severe renal vein dilatation detected 30 years after percutaneous renal biopsy. The patient was found to have microscopic hematuria. Three-dimensional computerized tomography confirmed the presence of an AVF, and arterial embolization was performed. Three-dimensional computerized tomography is very useful and effective to image the physiology of vascular lesions for the diagnosis of a renal AVF.

[Transurethral incision for ureterocele in children].

Transurethral incision (TUI) was performed as the initial treatment in 10 children with ureteroceles. Three patients had ureteroceles associated with a single ureter. TUI relieved hydronephrosis and preserved renal function in all 3 cases. Urinary tract infection developed in no patients. However, all the patients required an antireflux operation because of postoperative vesicoureteral reflux (VUR). Seven children had a total of 8 ureteroceles associated with a duplex system. TUI resulted in preservation of the upper pole function in 6 of the 8 ureteroceles. Urinary tract infections and VUR developed in 3 and 7 patients, respectively. Common sheath reimplantation was performed in 2 ureteroceles. TUI relieves obstruction before the onset of devastating infections although it carries the risk of postoperative VUR. We recommend TUI as the initial treatment for ureteroceles associated with both single and duplex systems.

Epidermoid cyst of the scrotum: a case report.

A 44-year-old male was admitted with the chief complaint of a huge mass in the right scrotum. Computed tomography (CT), magnetic resonance imaging (MRI) and ultrasonography demonstrated a homogeneous lesion in the right testis. Under the diagnosis of right testicular tumor, surgical resection was performed and the right testis itself was found to be essentially normal. The mass contained 500 ml of liquid. The pathologic diagnosis was an epidermoid cyst of the scrotum, a rare disease with only 11 cases reported in Japan.

Transinguinal laparoscopy for nonpalpable testis.

BACKGROUND: About 20% of all undescended testes are nonpalpable. Although surgical exploration was previously the method of choice for management, laparoscopy from the inferior edge of the umbilicus has been established as a useful method of examining a testis that is nonpalpable. However, conventional subumbilical laparoscopy is unnecessary when the testis or its remnant is located below the internal inguinal ring. We evaluated the efficacy of transinguinal laparoscopy after inguinal exploration performed for nonpalpable testis. METHODS: Twelve children with 14 nonpalpable testes (2 children had bilateral nonpalpable testes) were considered for a novel method of laparoscopy. We performed laparoscopy through the internal inguinal ring after opening the inguinal canal. RESULTS: Inguinal exploration revealed the testis or its remnant in 5 patients, and thus prevented unnecessary laparoscopy in these individuals. Transinguinal laparoscopy identified the anatomy in 7 patients. Four of the latter were found to have blind-ending vas deferens and spermatic vessels, while in 3 patients, the testes were located in the peritoneal cavity. CONCLUSION: Transinguinal laparoscopy may become the accepted method for managing nonpalpable testes, as inguinal exploration can eliminate the need for laparoscopy in some cases, and the visualization achieved with transinguinal laparoscopy is just as good as that with conventional laparoscopy.

Bladder squamous cell carcinoma with human papilloma virus type 6 [HPV 6].

A 91-year-old male was diagnosed with squamous cell carcinoma of the urinary bladder. Molecular biological examination using the polymerase chain reaction (PCR) method demonstrated the presence of HPV type 6. HPV type 6 is considered very rare in urinary bladder malignancies.