Chisato Fujiyama

Reconstruction of the Urinary Bladder Mucosa in Three-Dimensional Collagen Gel Culture: Fibroblast-Extracellular Matrix Interactions on the Differentiation of Transitional Epithelial Cells

The purpose of this study was to reconstruct a urinary bladder mucosa in three-dimensional collagen gel culture conditions that included fibroblasts. Transitional epithelial cells and fibroblasts, isolated respectively from the epithelial and lamina propria layers of porcine urinary bladder, were cultured in monolayer. These fibroblasts were embedded and cultured within a collagen gel matrix to reconstruct a lamina propria. The isolated transitional epithelial cells were then seeded in vitro on this reconstructed lamina propria on which the transitional epithelial cells formed a stratified urothelium composed of basal, intermediate and superficial layers. Urothelial differentiation was observed only on the fibroblast-containing collagen matrix. Differentiation did not occur on a cell-free collagen matrix through use of fibroblast conditioned medium. Thus, urothelial differentiation depended upon a fibroblast-extracellular matrix interaction. The differentiated transitional epithelial cell layer thus produced closely resembled a urothelium in vivo. This culture model may provide a useful system in which to study various diseases of the urinary bladder.

Novel human PDCD4 (H731) gene expressed in proliferative cells is expressed in the small duct epithelial cells of the breast as revealed by an anti-H731 antibody.

The novel gene H731 (approved name: PDCD4 (programmed cell death 4)) has been isolated as an antigen gene of the monoclonal antibody Pr‐28 which recognized a nuclear antigen in proliferating cells. The gene is homologous to the mouse gene (MA‐3/Pdcd4/A7–1) which was associated with apoptosis and was shown to suppress tumor promoter‐induced neoplastic transformation. A polyclonal antibody against H731‐protein derived from an extract of Escherichia coli transformed with an H731 expression plasmid was prepared, and the H731‐protein expression in human normal and tumor cells using the antibody was studied. The staining patterns of asynchronous cultures of human normal fibroblasts (MRC‐5) were heterogeneous but the antigen was accumulated in the nuclei at the G0 phase. On the contrary, the antigen was overproduced and localized in the cytoplasm during the cell cycle in tumor cell lines. Immunohistological studies revealed that the H731‐protein was highly expressed in bladder carcinoma and breast carcinoma tissues compared with the normal tissues so far tested. These results indicated that expression of the H731‐protein was up‐regulated or induced in the proliferative cells. Immunohistological studies also revealed that the protein was abundantly expressed in the small duct epithelial cells of the normal mammary gland.

Hand-assisted retroperitoneoscopic nephroureterectomy for upper urinary tract urothelial tumors

PURPOSE We report our experience with hand-assisted retroperitoneoscopic nephroureterectomy for upper urinary-tract urothelial cancer. PATIENTS AND METHODS Our initial 10 cases of clinical stage T(1)N(0)M(0) renal pelvic and ureteral tumors treated with hand-assisted retroperitoneoscopic nephroureterectomy are included in the present report. Nephrectomy was conducted retroperitoneoscopically with hand assistance via a lower-abdominal midline incision. Resection of the lower ureter together with the bladder cuff was performed as open surgery and the specimen was removed en bloc via the same incision. RESULTS Hand-assisted retroperitoneoscopic nephroureterectomy was completed successfully in all 10 cases. The mean operating time was 456 +/- 90 minutes, and the mean estimated blood loss was 462 +/- 364 mL. The times to oral intake and walking were 1.5 +/- 0.5 days and 2.3 +/- 0.7 days, respectively. One case of renal vein injury, one case of pulmonary embolism, and three cases of wound infection were the complications. CONCLUSION This is the first report of hand-assisted endoscopic nephroureterectomy using the retroperitoneal approach. The surgical technique seems quite reasonable because the lower-abdominal incision can be utilized, not only as a route for hand assistance, but also as a window for open surgery when resecting the distal ureter as well as for extracting the surgical specimen.

Reconstruction of Prostatic Acinus-Like Structure from Ventral and Dorsolateral Prostatic Epithelial Cells of the Rat in Three-Dimensional Collagen Gel Matrix Culture

PURPOSE The study was carried out to reconstruct a prostatic acinus-like structure from prostatic epithelial cells in a new culture system which can provide a more physiological condition than conventional cell culture methods. MATERIALS AND METHODS Prostatic epithelial cells were isolated from ventral and dorsolateral prostates of the rat and were separately cultured in three-dimensional collagen gel matrix. The cultured cells were observed by photo-microscopy and transmission electron microscopy. Differentiation and proliferation of the cultured cells were examined by immunohistochemistry using PAP and PSA kits and a bromodeoxy-uridine (BrdU) kit. The cell area of acinus-like structures in the transverse sections was measured by computer with the Interaktive Bild-Analyse System. RESULTS Dissociated prostatic epithelial cells were organized into three-dimensional spherical or branching cellular aggregates in collagen gel matrix. Through cell proliferation and differentiation, the cellular aggregates formed prostatic acinus-like structures, which consisted of a small intercellular lumen enclosed by one layer of cuboidal epithelial cells. The epithelial cells were connected together by junctional complexes, had microvilli at the luminal surface, and the basal side was surrounded by a distinct basal lamina. In the lumina of acinus-like structures, secretory products were often found. CONCLUSIONS Prostatic acinus-like structures were reconstructed from dissociated prostatic epithelial cells in three-dimensional collagen gel matrix culture. The reconstructed acinus-like structures are similar to prostatic acini in vivo in structure and function. This three-dimensional collagen gel matrix culture model may provide a useful means for investigating prostatic diseases and influences that hormones and growth factors exert on prostatic epithelial cells.

Nonionic contrast media are less nephrotoxic than ionic contrast media to rat renal cortical slices.

BACKGROUND Nephrotoxicity induced by contrast media (CM) is well recognized. Nonionic CM with lower osmolality than that of conventional ionic CM have been developed in an effort to reduce toxicity. However, the nephrotoxic effects of nonionic CM have not been well evaluated. Although our previous experiments using rat renal cortical slices indicated that the direct cellular toxicity of nonionic CM is less than that of ionic CM, it was suggested that the less toxic effects of nonionic CM on the metabolic function of renal epithelial cells were in part attributable to the lower osmolality of nonionic CM. In the present experiment, the direct toxicity of nonionic CM on renal epithelial cells was compared with that of ionic CM under equiosmolar conditions, where the effects of osmotic pressure were excluded. METHODS Rat renal cortical slices were incubated with several kinds of CM at 37 degrees C for 120 min. Diatrizoate and iothalamate were employed as ionic CM. Iopamidol and iohexol were employed as nonionic CM. The activities of N-acetyl-beta-D-glucosaminidase (NAG), gamma-glutamyltransferase (GGTP), and lactate dehydrogenase (LDH) released from the renal slices into the incubation buffer were determined in order to evaluate renal epithelial damage caused by CM. Gluconeogenesis, p-aminohippuric (PAH) acid accumulation and ATP content in rat renal slices were determined with a view to examine the inhibitory effects of CM on the metabolic function of renal epithelial cells. The toxic effects of nonionic CM were compared with those of ionic CM under equiosmolar conditions, where mannitol was added to the experimental groups containing nonionic CM in order to exclude the effects of osmotic pressure. RESULTS A significant difference was generally not found with regard to enzyme release between ionic CM and nonionic CM plus mannitol. The inhibition of gluconeogenesis and PAH accumulation in rat renal slices by nonionic CM with mannitol was less than that by ionic CM. Although the ATP content was reduced by both ionic CM and nonionic CM plus mannitol, there was no significant difference between these two groups. CONCLUSIONS The present experiments demonstrated that nonionic CM were less nephrotoxic than ionic CM with regard to the function of renal epithelial cells, including gluconeogenesis and PAH accumulation, under equiosmolar conditions. These differences in nephrotoxicity between ionic and nonionic CM cannot be fully attributable to differences in osmotic pressure.

Case of mesothelioma of the tunica vaginalis testis with characteristic findings on ultrasonography and magnetic resonance imaging

Abstract A case of mesothelioma of the right tunica vaginalis testis in a 32‐year‐old man is reported. Trans‐scrotal ultrasonography revealed hydrocele and multiple nodular masses measuring 1.0–4.5 cm in size attached to the parietal vaginal layer. Magnetic resonance imaging demonstrated more clearly nodular masses with irregular surfaces lined on the hydrocele cavity. Histologic diagnosis of the tumor when orchiectomized was mesothelioma. The patient has been free of disease for approximately 3 years since the treatment.

Annual PSA tests are not necessary for men with a PSA level below 2 ng/mL: Findings of the Imari prostate cancer screening program

Background: Annual changes in prostate specific antigen (PSA) levels detected by the Imari prostate cancer screening program were evaluated to establish a more efficient and cost‐saving screening system, especially for men with low PSA levels.