Genetic and epigenetic alterations on the short arm of chromosome 11 are involved in a majority of sporadic Wilms' tumours.

Wilms' tumour is one of the most common solid tumours of childhood. 11p13 (WT1 locus) and 11p15.5 (WT2 locus) are known to have genetic or epigenetic aberrations in these tumours. In Wilms' tumours, mutation of the Wilms tumour 1 (WT1) gene at the WT1 locus has been reported, and the WT2 locus, comprising the two independent imprinted domains IGF2/H19 and KIP2/LIT1, can undergo maternal deletion or alterations associated with imprinting. Although these alterations have been identified in many studies, it is still not clear how frequently combined genetic and epigenetic alterations of these loci are involved in Wilms' tumours or how these alterations occur. To answer both questions, we performed genetic and epigenetic analyses of these loci, together with an additional gene, CTNNB1, in 35 sporadic Wilms' tumours. Loss of heterozygosity of 11p15.5 and loss of imprinting of IGF2 were the most frequent genetic (29%) and epigenetic (40%) alterations in Wilms' tumours, respectively. In total, 83% of the tumours had at least one alteration at 11p15.5 and/or 11p13. One-third of the tumours had alterations at multiple loci. Our results suggest that chromosome 11p is not only genetically but also epigenetically critical for the majority of Wilms' tumours.

Prostate cancer cell growth is modulated by adipocyte-cancer cell interaction.

OBJECTIVE: To assess whether adipocytes (mesenchymal stromal cells thought to affect the proliferation and differentiation of epithelial cells) affect the behaviour of prostate cancer cells in vitro, as prostate cancer metastasizes to the bone, which is an adipocyte-rich environment. MATERIALS AND METHODS: The human bone-metastatic prostate carcinoma cell line PC3 was cultured with or without adipocytes in a three-dimensional collagen gel matrix. Histological and immunohistochemical assays were used to evaluate the proliferation and differentiation of PC3 cells. The cytokine expression of this culture assembly was also examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The proliferation and differentiation of cancer cells were clearly changed on co-culture with adipocytes compared with the control cultures. The mean (range) bromodeoxyuridine (BrdU) indices estimated (according to uptake) to evaluate the growth of the cultured cells were 36.3 (8.55)% in the co-culture and 26.95 (10.50) in the control (P < 0.02). PC3 cells in co-culture formed larger clusters than in the control, at 16.0 (11.0) vs 14.0 (10.0), respectively (P < 0.01). Cancer cells also showed pleomorphism, varying from cuboidal to spindle-shaped. The expressions of vascular endothelial and platelet-derived growth factor were greater in co-culture than in the control. CONCLUSION: These findings suggest that adipocytes modulate the growth, morphology and cytokine expression of prostate cancer cells. This specific mesenchymal stromal cell type is important in the biological behaviour of prostate cancer cells.

Bladder cancer cell implantation in reconstructed bladder in vitro: a model of tumour recurrence.

OBJECTIVE: To evaluate the involvement of umbrella cells in tumour adhesion and growth, by examining whether human urinary bladder carcinoma cells (HUBCC) can grow on reconstructed urinary bladder mucosa in vitro, as the implantation of tumour cells after resection is thought to be a cause of bladder tumour recurrence. MATERIALS AND METHODS: Normal transitional epithelial cells isolated from porcine bladder were cultured on reconstructed lamina propria using fibroblasts in type I collagen gel. The urothelium thus reconstructed was artificially injured either by a scalpel or by dilute acid, after which transitional epithelial cells began to grow in a stratified fashion within a few days of culture. A HUBCC line (HT-1197) was seeded onto this impaired mucosa to determine whether the cells could become implanted. Cultured cells on the reconstructed mucosa were evaluated by histological observation of vertical paraffin sections. RESULTS: The inoculated transitional epithelial cells grew in a stratified fashion and closely resembled urothelium in vivo. The superficial cells that were in contact with the medium solution differentiated into umbrella cells. HUBCC were unable to adhere to reconstructed mucosa which had not been injured, but these cells could adhere to and become implanted on the reconstructed mucosa after it had been injured either by a scalpel or by dilute acid. After acid injury, only the surface-covering cells were removed sporadically, while the lower epithelial cell layer remained intact. The bladder cancer cells adhered to and proliferated within these stripped regions. CONCLUSION: These results suggest that the urothelium, especially umbrella cells, seems to be important in preventing the adhesion and growth of urinary bladder tumour cells.

A novel imprinted gene, KCNQ1DN, within the WT2 critical region of human chromosome 11p15.5 and its reduced expression in Wilms' tumors.

WT2 is defined by a maternal-specific loss of heterozygosity on human chromosome 11p15.5 in Wilms' and other embryonal tumors. Therefore, the imprinted genes in this region are candidates for involvement in Wilms' tumorigenesis. We now report a novel imprinted gene, KCNQ1DN (KCNQ1 downstream neighbor). This gene is located between p57(KIP2) and KvLQT1 (KCNQ1) of 11p15.5 within the WT2 critical region. KCNQ1DN is imprinted and expressed from the maternal allele. We examined the expression of KCNQ1DN in Wilms' tumors. Seven of eighteen (39%) samples showed no expression. In contrast, other maternal imprinted genes in this region, including p57(KIP2), IMPT1, and IPL exhibited almost normal expression in these samples, although some samples expressed IGF2 biallelically. These results suggest that KCNQ1DN existing far from the H19/IGF2 region may play some role in Wilms' tumorigenesis along with IGF2.

Trans-mesosigmoid cutaneous ureterostomy.

BACKGROUND: A new method was developed in order to create a single stoma cutaneous ureterostomy in which both ureters traverse the abdominal cavity and yet are buttressed by the mesosigmoid and covered by its visceral peritoneum. METHODS: The long mesenterium which is attached to the most mobile part of the sigmoid colon was used for the bilateral ureteral pathway. Tunnels for the ureteral path were made just underneath the visceral peritoneum on the bilateral side of the mesosigmoid. Through the tunnels both ureteral ends were brought from the retroperitoneal space to the mesenterocolonic junction (MCJ) and the MCJ is then approximated and sutured to the inside of the ureteral tract through the abdominal wall. The ureters brought outside the skin, are conjoined and sutured to the V skin flap. Eight patients who carried a high risk for operation and/or had a bladder tumor judged to be incurable underwent this cutaneous ureterostomy. RESULTS: All cases except one with low urinary output could be managed without catheter indwelling during the follow-up period. Three patients suffered from paralytic ileus and one required laparotomy for mechanical ileus during the short postoperative period. Postoperative excretory urography evaluated 14 kidneys during the follow-up period from 2 to 61 months and showed normal upper urinary tract in 11 and a mildly hydronephrotic tract in three. CONCLUSIONS: Transmesosigmoid cutaneous ureterostomy provides a single catheterless stoma even when the available ureters are relatively short. It appears to be a good method for supravesical urinary diversion when indicated.

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

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.

Proliferation and differentiation of rat dorsal prostatic epithelial cells in collagen gel matrix culture, focusing upon effects of adipocytes.

BACKGROUND: Prostatic epithelial cells organize functional acinus structures under epithelial extracellular matrix and epithelial-stromal cell interactions. Recently, the adipose tissue, which surrounds and even exists within the prostate, has been suggested to affect the differentiation and proliferation of some cell types. Therefore, tissue fragments, which consist mainly of epithelial and fibromuscular stromal cells, were cultured in three-dimensional collagen gel matrix culture with adipocytes. METHODS: Tissue fragments of rat dorsal prostate, including both epithelial and fibromuscular stromal components, were cultured in collagen gel with or without adipocytes. Epithelial cell differentiation was evaluated with the reconstruction of acinus-like structures and with immunohistochemistry of rat dorsal prostate-specific proteins, dorsal protein-1 and probasin. The proliferation was examined by uridine uptake. RESULTS: Under coculture of the fragments and adipocytes, epithelial cells reconstructed more differentiated acinus-like structures surrounded by fibromuscular stromal cells than tissue fragment culture without adipocytes. Dorsal protein-1 and probasin expressions of epithelial cells in this coculture system were the same as in rat prostate in vivo. In the coculture, epithelial cells had a higher proliferation activity. CONCLUSION: These results indicate that adipocytes promote proliferation and differentiation of prostatic epithelial cells. Our new culture model with adipocytes suggests the importance of cell-cell interactions, including those of epithelial cells and adipocytes.

Significance of hematoma size for evaluating the grade of blunt renal trauma.

BACKGROUND: The hematoma size relative to the body size was measured on computed tomography films using a personal computer system in order to define whether that parameter is useful for decision-making in the management of blunt renal trauma. METHODS: From 1982 to 1997, 33 patients with intermediate or severe grade blunt renal trauma were retrospectively divided into three groups: group 1, managed conservatively without transcatheter embolization; group 2, managed by bedrest after selective transcatheter embolization; and group 3, managed operatively. In these three groups, the hematoma area (H) and the ratio of hematoma area to body area on CT (H/B) were measured and the chronological changes of the H/B in groups 1 and 2 were studied. RESULTS: The H and H/B of group 3 were significantly larger than those of group 1. The H/B was more clearly distinguished for each group compared with the H alone. Well-preserved kidney integrity, despite the presence of a large hematoma in group 2, allowed the conservative treatment following transcatheter embolization of the bleeding site. The H/B of all group 1 patients gradually decreased and on the 40th or 50th day after injury they reached a level equivalent to the ratio of contra-lateral normal kidney area to body area. CONCLUSION: The ratio of hematoma area to body area on CT was very useful in evaluating the grade of blunt renal trauma. In conservative treatment for blunt renal trauma changes of the hematoma size is a useful indicator for management.

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.

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.

[The management selection of renal angiomyolipoma].

BACKGROUND: Nephron-sparing surgery is ideal in the treatment of renal angiomyolipoma (AML). But, in fact, occasional cases are found in post-ruptured AML and/or in bilateral multiple AMLs, that is seen in tuberous sclerosis. And we cannot perform nephron sparing surgery so easily. We proposed the treatment selection in such complicated AML. METHODS: We experienced 10 cases (12 kidneys) for about ten years, and studied our treatment selection and prognosis on each cases retrospectively. RESULTS: Of the 5 kidneys with AML less than 4 cm in diameter, 4 have not encountered with rupture. Of the 7 kidneys with AML more than 4 cm in diameter, 4 kidneys had rupture. Of the 3 kidneys unruptured AML more than 4 cm in diameter, 2 kidneys were treated by enucleation and we performed preventive embolization for rupture in residual one kidney. The patients was suffered from tuberculosis sclerosis, and she had bilateral multiple AMLs. Of the 4 kidneys with ruptured AML, 2 kidneys were treated by enucleation, and the other 2 kidneys were entirely resected. We succeeded enucleation in 2 of 4 kidneys with ruptured complicated AML. In those cases, we did long term watching after rupture and in-situ perfusion technique at the operation. CONCLUSIONS: Active treatment of AML, that is more than 4 cm in diameter, might be recommended. Because most of those will be ruptured. The ideal treatment, nephron-sparing surgery is difficult in complicated situation, such as after rupture and bilateral multiple AMLs. In our opinion, the point of success of nephron-sparing surgery might be long term watching after rupture and in-situ perfusion technique at the operation.

Do human papillomaviruses have a role in the pathogenesis of bladder carcinoma?

PURPOSE: Since little is known of the associations between bladder carcinoma and human papillomaviruses (HPVs), data on the role of HPV in bladder carcinogenesis are controversial. We attempted to clarify whether HPVs are present in bladder carcinomas. MATERIALS AND METHODS: We examined 36 specimens of bladder carcinoma for HPV positivity by the polymerase chain reaction method. RESULTS: HPV-16 deoxyribonucleic acid was detected in 1 specimen (3%) of a transitional cell carcinoma from a 37-year-old woman who had concomitant squamous cell carcinoma of the uterine cervix with positive para-aortic lymph node metastasis. The cervical tumor, bladder tumor and para-aortic lymph node metastasis were all positive for the same type of HPV. CONCLUSIONS: On the basis of this low rate of HPV detection (3%), HPVs are not likely to have a prominent role in carcinogenesis of the bladder.

[Treatment of advanced renal cell carcinoma with a combination of interferon alpha and gamma].

A total of 29 patients with advanced renal cell carcinoma entered a pilot study of combination therapy with interferon alpha (IFN-alpha) and interferon gamma (IFN-gamma). IFN-alpha (HLBI: 3 x 10(6) IU, BALL 1:5 x 10(6) IU, IFN-alpha-2a: 9 x 10(6) IU or IFN-alpha-2b: 6 x 10(6) IU) was given intramuscularly every day and IFN-gamma (IFN-gamma-1a: 3 x 10(6) JRU) was given intravenously by drip infusion 3 times a week (every 2-3 days). The treatment was continued for 3 months as the induction therapy, and then the tumor response was evaluated. Of the 22 evaluable patients, 4 achieved a partial response (PR), 10 showed no change (NC), and in 8 the disease had progressed (PD) during the therapy. Thus, the overall response rate was 18.2% [95% confidence interval (CI) 2.1-34.3%]. A favorable response tended to be obtained in patients with good performance status or small pulmonary metastases, or in those who had no prior therapy with IFN-alpha, who received this treatment immediately subsequent to radical nephrectomy, or who received IFN-gamma as much as possible according to this regimen. Toxicity was evaluated in 28 patients: fever, general fatigue, anorexia, leukocytopenia and impaired liver function were frequently noted, and 3 patients were withdrawn from the study because of such adverse effects. In patients who had a PR or NC, the same dosage of IFN-alpha was continued to be given intramuscularly 2-3 times a week (every 2-4 days) as the maintenance therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

In situ perfusion by retrograde cannulation of a tumor artery for nephron-sparing surgery.

BACKGROUND: Although ice slush cooling or ex situ perfusion with bench surgery is most widely used for protecting ischemic renal damage which possibly accompanies complicated nephron-sparing surgery, each has its own disadvantages. The former does not allow excessively long ischemia and the latter requires complicated procedures as autotransplantation. In order to mitigate against these problems, we devised a novel method of in situ renal perfusion with intracellular hyperosmolar solution. METHODS: One renal segmental artery mainly supplying a tumor was isolated and cannulated with a small feeding tube. The tube was introduced through a small arteriotomy incision directed towards the proximal side, advanced until its tip remained in the main or first branch of the renal artery, and then it was anchored to that artery. After the main renal artery and vein were clamped, the kidney was perfused with cold Euro-Collins' solution through the tube, while the venous blood and perfusate were drained from the left gonadal vein or small venotomy incision of the right renal vein. RESULTS: In one case of renal cell carcinoma and three cases of angiomyolipoma, two of which ruptured, nephron-sparing surgery was carried out under in situ hyperosmolar perfusion. Ischemic time of these four cases was an average of 96 minutes, varying from 45 to 145 minutes. All the kidneys functioned well postoperatively. CONCLUSIONS: The method presented here is very simple, requires no unusual dexterity and safely allows for a long period of renal ischemia. This method is best indicated in cases where simple clamping of the renal pedicle with ice-slush cooling appears insufficient, yet ex situ surgery with autotransplantation seems excessive.

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.

[A case of recto-urethral fistula following transrectal hyperthermia treated by anterior anorectotomy approach].

We report a case of 73 years old male with recto-urethral fistula developing after transrectal hyperthermia for prostatic hypertrophy. This is the first case report of recto-urethral fistula probably caused by transrectal hyperthermia. Anterior anorectotomy approach for fistula closure provided a wide operative field and lead to a successful result.

[Megaureter in adults].

In comparison with megaureters in children, their reports in adult are not common. We had an opportunity to treat seven adults with megaureters during the last six years. They were all female and ages ranged from 46 to 67 years. Five patients with grade II and one with grade III (Pfister-Hendren's classification) were treated by reconstructive surgery, excision of the narrow segment, tapering of the dilated lower ureter and reimplantation through a submucosal tunnel. The outcome of all the grade II patients was excellent and the case with grade III showed mild improvement. The results suggested that surgical reconstruction could be equally effective for megaureters in adults compared to those in children.

[The prognostic value of mean nuclear area and mean nuclear volume in bladder cancer].

To evaluate the prognostic value of mean nuclear area (MNA) and mean nuclear volume (MNV) in bladder cancer, a retrospective study was performed comprising 67 bladder cancer patients who could be followed up for more than 3 years. Cosmozone 1SA, a Nikon image analyzing system was used for the morphometric study. The specimen of initially biopsied tumor tissue was set in an Olympus microscope at 400-fold magnification, and the image was superimposed on the monitor picture through a video camera attached to the microscope. MNA and MNV were measured by tracing the contour of the nucleus which were selected by the point-sampled intercept methods. The time required for measurement of the area and volume was about 15 minutes per case. MNA in cases with histological grade 1, 2 and 3 were 35 +/- 3 microns2 (mean +/- SD), 42 +/- 10 microns2 and 62 +/- 12 microns2 respectively. MNV with grade 1, 2 and 3 were 282 +/- 46 microns3, 371 +/- 148 microns3 and 644 +/- 182 microns3 respectively. The morphometric results were significantly related to histological grade. In cases with a value of MNA of 40 microns2 or more and/or a value of MNV of 370 microns3 or more, the proportion of cases who underwent cystectomy or died of cancer was significantly high and demonstrated poor survival. In contrast, those who showed MNA and MNV less than the above value had better prognosis. These results suggest that the measurements of MNV seems to be useful for objectively evaluating the malignant potential of bladder cancer.

Influence of extracellular matrix on the proliferation and differentiation of adrenocortical cells in culture.

Extracellular matrix is indispensable for cell differentiation in vivo. The aim of this study is to examine effects of extracellular matrix on adrenocortical cells embedded in collagen gel in terms of cellular proliferation and differentiation. Dissociated human or bovine adrenocortical cells were embedded in collagen gel; the gel was then immediately thin-coated on the bottom of culture dishes. The cortical cells were thus trapped in collagen and cultured in monolayer, a suitable means for observation of culture cells. To the authors' best knowledge, this "collagen gel-embedded" monolayer culture is described here for the first time. Cortical cells were spherical in shape and had the activities of proliferation and steroid production in collagen gel matrix of this culture method. Aldosterone production in particular tended to be maintained much longer than in conventional monolayer culture. This method seems to provide a physiological environment in which to study the differentiation of cortical cells.