Loss of heterozygosity on chromosomes 1p and 11p in sporadic pheochromocytoma.

We used 29 polymorphic DNA markers to analyze tumor DNA samples from six patients with sporadic pheochromocytoma for possible loss of chromosomal heterozygosity; four had benign disease and two had malignant disease. Loss of heterozygosity was observed on four chromosomes: 1p (three of four patients), 2p (one of one), 5q (two of six), and 11p (three of five). Chromosomes 1p and 11p frequently had allelic deletions in these tumors, and these deletions may play an important role in the development of pheochromocytoma.

Prognostic value of cadherin-associated molecules (alpha-, beta-, and gamma-catenins and p120cas) in bladder tumors.

Loss of E-cadherin-mediated adhesion is an important step in the progression of many carcinomas. In model systems, it has been shown that cadherin function requires not only proper E-cadherin expression but also its linkage to the cytoskeleton through catenins. Hence, defects in catenins may cause defective E-cadherin function, and catenins as well as E-cadherin might constitute prognostic indicators. Here, we extend our previous study on E-cadherin in bladder cancer (Cancer Res., 53: 3241-3245, 1993). We have evaluated the expression of E-cadherin-associated cytoplasmic molecules (alpha-, beta-, and gamma-catenins and p120cas) to clarify whether or not the pattern of their expression could provide additional prognostic information beyond that from E-cadherin alone. Forty-eight frozen bladder tumor specimens and 9 samples of normal urothelium were studied by immunohistochemistry. A discrepancy between the E-cadherin and catenin expression pattern was seen in 20.8% of cases. Abnormal expression of each molecule is significantly correlated with tumor grade (P < 0.01) and stage (P < 0.01). Reduced expression of all of the molecules correlates with poor survival (P < 0.01 for each variable). Proportional hazard regression analysis showed that beta-catenin, E-cadherin, and alpha-catenin have strong predictive value, whereas plakoglobin and p120cas have a somewhat lower predictive value. Within patients with invasive tumors, those with a normal staining for either E-cadherin, alpha-catenin, or beta-catenin show a trend toward better survival. However, the difference in survival is significant only for E-cadherin (P < 0.05). Thus, beta-catenin, E-cadherin, and alpha-catenin have similar prognostic values. Therefore, from a practical point of view, the expression of any of these proteins can be of prognostic value for patients with bladder cancer.

Lysosomal cysteine proteinases in rat epididymis.

To examine the precise localization of lysosomal cysteine proteinases, cathepsins B, H, and L in rat epididymal epithelial cells, immunohistochemistry and enzyme assay were applied to the epididymal tissue. Granular immunodeposits for cathepsins B and H were detected in epididymal epithelial cells, whereas faint or no immunoreactivity for cathepsin L was found. Moreover, immunoreactivity for cathepsin B appeared mainly in principal cells and was more intense in the head of the epididymis than in the tail, whereas that for cathepsin H appeared in both principal and clear cells and was more intense in the tail than the head. By enzyme assay, activities of cathepsins B and H showed a similar distribution to that of the immunoreactivity. The cathepsin L-specific activity was distributed evenly in each part of the epididymis and was also detected in epididymal fluids obtained from the body and tail parts. By immunoblotting, proforms of cathepsins B, H, and L were present in the seminal fluid. The results suggest that cathepsins B and H are involved in the intracellular degradation system of epididymal epithelial cells, and proforms of cathepsins B, H, and L may be secreted into the epididymal duct lumen.

A model of localized osteolysis induced by the MBT-2 tumor in mice and its responsiveness to etidronate disodium.

A new experimental method to test the effect of drugs on tumor-induced osteolysis using a bladder tumor in mice has been designed. The method consisted of inoculating MBT-2 tumor cells s.c. over the calvaria in mice, resulting in a local tumor causing fragmentation of the bone. This was accompanied by adjacent osteoplastic changes, which were evaluated by X-ray and histological examination. Etidronate disodium (EHDP), a diphosphonate derivative, at a dose of 3 mg/kg to 30 mg/kg s.c. protected the bone by decreasing the extent of osteolysis as judged by the same criteria. Therapy with EHDP prolonged the survival period. This inhibition was obtained with no apparent effect on the growth of the MBT-2 tumor.

Pharmacokinetics of tamsulosin hydrochloride in patients with renal impairment: effects of alpha 1-acid glycoprotein.

The pharmacokinetics of tamsulosin hydrochloride in patients with renal impairment were compared with those in healthy volunteers, and the factors that influenced plasma levels of tamsulosin were elucidated. A single oral dose of 0.2 mg of tamsulosin was given and blood and urine samples were obtained for 36 hours after administration. Unbound plasma concentration of tamsulosin was measured by a combination of equilibrium dialysis and liquid chromatography tandem mass spectrometry methods to examine the effect of protein binding on the pharmacokinetics of tamsulosin. Mean values for maximum concentration (Cmax) and area under the concentration-time curve (AUC) of total drug (Cmax,t and AUC1) in patients with renal impairment were 73% and 211% greater, respectively, than those in healthy volunteers. Mean Cmax and AUC of unbound drug (Cmax,u and AUCu), however, were almost the same in the two groups. A high correlation was found between alpha 1-acid glycoprotein (alpha 1-AGP) concentration and AUCt, but no correlation was found between alpha 1-AGP concentration and AUCu,0-36) or between creatinine clearance (ClCR) and AUCu,0-36). These results show that in patients with renal impairment, the pharmacokinetics of tamsulosin are affected by the change in protein binding that is associated with alteration of plasma alpha 1-AGP concentration, but are not largely affected by the decrease in the renal excretion. Although total tamsulosin levels increased as plasma protein binding increased, unbound tamsulosin levels (which are directly associated with the pharmacologic effects) remained unchanged in these patients.

Multielement analysis of kidney tissue with renal calculi.

Tissue distribution and concentration gradients of macro- and micro-elements in the papilla, medulla, and cortex of human kidney with renal calculi were measured with spectrometer. An uninvolved portion of the hypernephroma kidney was used as control. Cadmium (Cd), cobalt (Co), and molybdenum (Mo) were least in amount in renal papilla as compared with the cortex and medulla in renal stone kidney. Overall there was less Mo in stone kidney tissue, and also significantly less when compared with that of control renal papilla. Considering the reported fact that Mo was found in high concentrations in urinary tract stones and our data that Mo was less in stone kidney, it is speculated that Mo may play some unexplained but significant role in certain stage(s) of the stone formation.

Experience with Gleason histopathologic grading of prostatic cancer in Japan.

Histologic characteristics of prostate cancer in Japan were evaluated in a retrospective analysis of 267 cases. These specimens were graded by the Gleason histopathologic grading system, and the proportional distribution of histologic features and the death rate were compared with those of Gleason's results in the literature. The system demonstrates significant correlation with mortality rates for each grade group in our cases, and it was also found that the death rates as obtained by our figures were comparable to those of Gleason in each category. The results help to provide the basis for future comparative multinational trials of prostate cancer.

Studies on platelet function in patients with prostatic cancer. Preliminary report.

Platelet function was evaluated as an index of the thromboembolic tendency in patients with untreated, advanced prostatic cancer. Patients with benign prostatic hypertrophy (BPH) and similar age distribution served as a comparison group. Platelet aggregations were elevated in both groups, but not significantly different from each other. Platelet serotonin level in patients with prostatic cancer was lower than in patients with BPH (p less than 0.01), whereas plasma serotonin level in patients with prostatic cancer (within normal ranges in our series) was lower than in patients with BPH (p less than 0.001). Levels of 2 intraplatelet proteins, beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4) in the two groups of patients were similar. However, levels of beta-TG were elevated significantly in both groups of patients compared with those of healthy individuals. These studies revealed that the platelet serotonin levels in advanced prostatic cancer patients differed significantly from those in patients with BPH. The platelet serotonin level thus may provide an index of platelet activation in patients with prostatic cancer.

Pheochromocytoma with remission following phentolamine-induced shock.

A case of pheochromocytoma with spontaneous remission after recovery from shock induced by the phentolamine test is presented. Most of the surgically resected tumor demonstrated extensive necrosis.

Randomized clinical trial on chemoprophylaxis of recurrence in cases of superficial bladder cancer.

Several postoperative adjuvant therapeutic modalities have been adopted in attempts to reduce the recurrence rate of superficial bladder cancer. However, no definite conclusions on the effectiveness of intravesical chemoprophylaxis have been reached. A randomized clinical study on intravesical chemoprophylaxis was conducted by the Japanese Urological Cancer Research Group for Adriamycin to compare the recurrence rates among 575 patients with superficial transitional cell carcinoma of the urinary bladder. Group A received 30 mg/30 ml Adriamycin; group B received 20 mg/40 ml Adriamycin; group C received 20 mg/40 ml mitomycin C, and group D, no treatment (for control). Instillation was performed twice a week for 4 weeks after surgery. The postoperative observation period was 18 months. The overall recurrence rate in group D was 61.5%, which was statistically higher than in the other groups. The Adriamycin and Mitomycin C groups showed recurrence rates of 43%-48% and 57%, respectively. Intravesical Adriamycin and Mitomycin C appeared to be effective in the prophylaxis of recurrence during this observation period. The main side-effect was cystitis syndrome, which was observed in 10%-20% of the patients. There were no life-threatening adverse effects in this series of patients.

Effects of intravesical instillation of antitumor drugs on the induction of preneoplastic bladder lesions in rats.

The effects of adriamycin (ADR) and mitomycin C (MMC) as inhibitors of the development of bladder tumors in rats were studied. Six-week-old female F344 rats were divided into nine groups, five of which received 0.05% N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in their drinking water for the first 4 weeks, no treatment for 1 week, and then intravesical instillation once a week of ADR, MMC, or physiological saline or no instillation (no catheterization) for 12 weeks. The other four groups received no BBN for the first 5 weeks of the experiment and then received ADR, MMC, or physiological saline as above for 12 weeks. The bladders were examined by light microscopy 17 weeks after the beginning of the experiment. Results showed that development of preneoplastic lesions induced in the bladders of the rats by BBN was stimulated by subsequent instillation of ADR or MMC. This result suggests that ADR and MMC have promoting activities in bladder carcinogenesis of rats.

The effect of dose intensity on M-VAC therapy for advanced urothelial cancer.

M-VAC therapy (methotrexate, vinblastine, Adriamycin, and cisplatin) has improved the treatment results of urothelial cancer patients. However, it is sometimes complicated by drug toxicities, including bone marrow suppression. We analyzed the relative dose intensity in each patient undergoing M-VAC chemotherapy in relation to the chemotherapeutic effect and survival. In addition, the role of granulocyte colony-stimulating factor (G-CSF) in the dose intensity of M-VAC therapy was analyzed. Between June 1988 and March 1993, 29 patients with advanced urothelial cancer were treated with M-VAC therapy in our institution. Of 18 patients with evaluable lesions, 2 (11.1%) showed a complete response (CR) and 7 (38.9%) showed a partial response (PR), and the overall response rate was 50.0%. The median follow-up period for these 18 patients was 14.6 months and the median survival was 8.7 months, with 12 of the 18 patients being alive at the time of analysis. The relative dose intensity (RDI) for these 18 patients was 0.81 for methotrexate, 0.80 for vinblastine, 0.92 for Adriamycin, and 0.91 for cisplatin, for a mean RDI of 0.87. There was no correlation between the chemotherapeutic effect and the RDI. When we calculated the RDI for all 29 patients who underwent M-VAC therapy, G-CSF increased the RDI of Adriamycin significantly. The results of this retrospective study indicate that a dose intensity for M-VAC therapy in the range of 0.61-1.00 is unlikely to correlate with the chemotherapeutic effect, although G-CSF contributes to increasing the RDI of Adriamycin.

Intra-arterial chemotherapy for bladder cancer.

Intra-arterial infusion chemotherapy with adriamycin (ADM) was carried out in 32 patients with bladder cancer prior to total cystectomy. An oblique incision approximately 12 cm long was made in the gluteal region to expose either the superior or inferior gluteal artery, into which a Teflon catheter was inserted and fixed. The distal end of the catheter was taken out from under the skin in the precordial region. Via this catheter, a single dose of 10 mg ADM was injected twice a week. Superior-gluteal-artery infusion chemotherapy was performed in 7 patients; the 5-year survival rate was 14.3%, which was not as high as expected. Inferior-gluteal-artery infusion chemotherapy was performed in 25 patients. Cisplatin (CDDP) was used with ADM in 8 patients. Radiation and/or hyperthermia were used in 11 patients. The 5-year survival rate in these 25 patients was 58.4%, which was considered to be satisfactory. Of these 25 patients, 5 were stage-T4 cases; for these, the treatment was ineffective, and all 5 died within 2 years. Of the 6 patients at stage T2, 1 died, as did 1 patient with carcinoma in situ (CIS). Of the 13 patients with bladder cancer at stage T3, 3 died; lymph-node metastases were found in all 3 of these cases. Of the 25 patients who received inferior-gluteal-artery infusion chemotherapy, 9 died of cancer; all 9 died within 2 years due to distant metastases. There was no evidence of recurrence in any patient who survived for 2 years or more after total cystectomy. Therefore, inferior-gluteal-artery infusion chemotherapy may be effective as a preoperative adjuvant therapy with no serious side effects.

Prophylactic chemotherapy with intravesical instillation of adriamycin and oral administration of 5-fluorouracil after surgery for superficial bladder cancer. The Japanese Urological Cancer Research Group for Adriamycin.

The Japanese Urological Cancer Research Group for Adriamycin has conducted a series of clinical trials to investigate the efficacy and safety of prophylactic intravesical chemotherapy for superficial bladder cancer. In the third trial, reported herein, patients with recurrent bladder cancer or multiple primary cancer were selected and randomized to one of four groups using the envelope method after complete resection of the original tumors. Group A was given Adriamycin alone, group B received oral 5-fluorouracil (5-FU), group C was given Adriamycin and oral 5-FU, and group D served as the control group. Of the 544 patients registered, 331 were evaluable for the purpose of this study. The administration of 5-FU (group B) failed to prevent the recurrence of bladder tumors. Although group C (both Adriamycin and 5-FU) did not fare better than group A (Adriamycin only), Adriamycin was effective in preventing the recurrence of tumors, especially in high-risk patients with recurrent and multiple tumors. The risk of recurrence was reduced to 0.21 (95% confidence interval, 0.10-0.44) relative to the control group. There was no indication of a synergistic effect between 5-FU and Adriamycin. As side effects, cystitis syndrome was observed in 23%-30% of the patients in the Adriamycin groups and mild myelosuppression was observed in the 5-FU groups.

Comparative analysis of short-term and long-term prophylactic intravesical chemotherapy of superficial bladder cancer. Prospective, randomized, controlled studies of the Japanese Urological Cancer Research Group.

The Japanese Urological Cancer Research Group has conducted three randomized clinical studies on intravesical chemoprophylaxis of superficial bladder cancers. This paper presents a comparative analysis of the first and second of these. The protocol used in the first study was a so-called short-term schedule in which drugs (for group A, ADM 30 mg/30 ml; group B, ADM 20 mg/40 ml; group C, MMC 20 mg/40 ml; and group D, control) were administered twice a week for 4 weeks after transurethral resection (TUR), and in the second study a long-term schedule was used, in which drugs (for group E, ADM 30 mg/30 ml; group F, ADM 20 mg/40 ml; group G, MMC 20 mg/40 ml; and group H, control) were administered twice a week for 1 week, every 2 weeks for 7 weeks, monthly for 8 months, and finally once every 3 months for 1 year. In the first study 575 patients were evaluated and followed up for 5 years. The second study started 28 months later, and 607 patients were evaluated. A generally good prophylactic effect was obtained in the second study when the patients' backgrounds were adjusted in combination with the history and number of the tumors. The second study did not reveal any promoting or inhibitory effect on the progression of the recurrent tumors. There were no significant side effects in either study. The indications and the schedule for prophylactic intravesical chemotherapy should be more carefully studied.

Long-term results of intravesical chemoprophylaxis of superficial bladder cancer: experience of the Japanese Urological Cancer Research Group for Adriamycin.

Long-term results were analyzed in terms of tumor progression and survival in patients with superficial bladder cancer who were enrolled in the second intravesical chemoprophylactic study of the Japanese Urological Cancer Research Group for Adriamycin, which was started in July 1982. This study was a prospective, randomized, controlled trial conducted on primary tumors treated with a long-term instillation regimen that involved control versus intravesical instillations of Adriamycin or mitomycin C given once a week for the first 2 weeks, once every other week for 14 weeks, once a month for 8 months, and once every 3 months for 1 year, for a total of 21 instillations in 2 years. An analysis of the prophylactic effects of such treatment on bladder tumors after TUR has previously been performed, and the results have been published elsewhere. The present study represents a follow-up of the above trial. Of the 671 cases previously analyzed with regard to tumor prophylaxis, 158 cases (23.5%) were eligible to be followed for tumor progression and survival. A detailed comparison of the background factors between these 158 patients and the other 513 cases revealed no statistically significant difference. Thus, the 158 evaluable cases might reasonably be considered to represent all patients enrolled in the second study, and the results were thought to be reasonable enough to reflect the long-term efficacy of the long-term instillation regimen adopted in this study. The median follow-up for these 158 cases was 6.6 years. Tumor progression in terms of the disease stage and/or grade occurred in 43 of 127 patients who received prophylactic instillations and in 12 of 31 control cases. No significant difference in the incidence of tumor progression was found between the treatment and the control groups. In addition, no difference in survival was observed between the treatment group and the control group. Survival was also compared between patients who showed tumor progression and those who did not. All patients whose tumors did not progress survived, whereas the 7-year survival of those exhibiting tumor progression was less than 90%.

The 4th study of prophylactic intravesical chemotherapy with adriamycin in the treatment of superficial bladder cancer: the experience of the Japanese Urological Cancer Research Group for Adriamycin.

A multicentric randomised trial was conducted for the purpose of investigating the efficacy of intravesical chemoprophylaxis of superficial bladder cancers. A total of 443 patients (number of evaluable patients, 284) were registered from July 1987 to December 1989 and randomised into 3 groups. Group A received 21 intravesical instillations of Adriamycin (ADM) at 20 mg/40 ml physiological saline for 2 years after undergoing transurethral resection (TUR); group B was given the same dose as group A but received 6 intravesical instillations for 2 weeks before undergoing TUR; and group C served as a control and underwent TUR only. Better prophylactic effects were obtained in group A. The overall non-recurrence rates calculated for groups A and B differed significantly (P less than 0.05) on day 240, and those determined for groups A and C were also significantly different (P less than 0.01) on day 480. No benefit was obtained using intravesical instillation prior to TUR (group B). The major side effects encountered were pollakisuria and miction pain, which occurred in 32% of the patients in group A and in 52% of those in group B.

Evaluation of systemic chemotherapy with methotrexate, vinblastine, adriamycin, and cisplatin for advanced bladder cancer. The Japanese Urological Cancer Research Group for Adriamycin.

In a cooperative study of the Japanese Urological Cancer Research Group for Adriamycin, the usefulness of chemotherapy with methotrexate, vinblastine, Adriamycin, and cisplatin (M-VAC therapy) in treating advanced or recurrent bladder cancer was examined. Evaluation of the clinical responses obtained in 86 evaluable patients revealed 13 complete responses, 29 partial responses, 4 minor responses, 19 cases of no change, and 21 cases of progressive disease. The overall response rate was 48.8% (42/86). The rate of response to M-VAC therapy at each disease site was as low as 21.4% (3/14) in bone lesions but exceeded 40% in the primary lesion, the lymph nodes, the lung, the liver, and other lesions. The clinical response to M-VAC therapy was not significantly influenced by the performance status of the patients, the dose intensity, or previous therapy. The median duration of response for the 42 responders was 22.7 weeks (range, 8.1-134.1 weeks), and the median duration of survival for the 86 evaluable patients was 9.8 months. Side effects were frequently encountered; the patients experienced anorexia, nausea, vomiting, malaise, alopecia, and leukopenia, but all of these symptoms were tolerable.

Phase III trial of the Japanese Urological Cancer Research Group for Adriamycin: cyclophosphamide, adriamycin and cisplatinum versus cyclophosphamide, adriamycin and 5-fluorouracil in patients with advanced transitional cell carcinoma of the urinary bladder.

A non-randomized clinical study on systemic combination chemotherapy was conducted by the Japanese Urological Cancer Research Group for Adriamycin to compare the effectiveness of CAP (cyclophosphamide 200-500 mg/m2, adriamycin 30-50 mg/m2 and cisplatin 30-50 mg/m2) and CAF (cyclophosphamide 200-500 mg/m2, adriamycin 30-50 mg/m2 and 5-fluorouracil 250 mg/m2) in 123 patients (104 evaluable) with advanced and/or metastatic cancer of the urinary bladder. Among 96 patients who were non-randomly selected to receive CAP, 4 achieved complete remission, 12 achieved partial remission, 7 achieved minor response, 30 had stable disease, and 43 had disease progression. The response in the 8 patients who received CAF were: partial remission in 1 and progressive disease in 7. The overall response rate to CAP therapy was 17%, as against 13% for CAF therapy. The median duration of survival with CAP was 29 weeks and with CAF, 22 weeks. The differences between the two groups in duration of survival and response rate were not statistically significant. Complete and/or partial remissions were observed in the lymph nodes, lung and liver in 32%, 24%, and 57% of cases, respectively. There was no objective response in bone metastasis. The main side effects of CAP were anorexia (88%), nausea and/or vomiting (81%), alopecia (65%), leukopenia (72%), anemia (48%), and renal dysfunction (17%). No patients died as a result of toxicity of these combination chemotherapy modalities.

Long-term intra-arterial infusion chemotherapy with adriamycin for advanced bladder cancer.

Long-term intra-arterial infusion chemotherapy with Adriamycin (ADM) was performed in cases of bladder cancer prior to total cystectomy. This report describes the effects in 13 cases evaluated more than 3 weeks after infusion of 10 mg ADM once or twice weekly. An oblique skin incision approximately 10 cm long was made in the gluteal region to expose the gluteus maximus muscle. A teflon catheter was then inserted into the gluteal artery and fixed; the distal end was brought out from under the skin in the precordial region. A similar procedure was performed on the contralateral side. The catheter was inserted through the superior and inferior gluteal arteries in five and eight cases, respectively. In the former group, partial response was obtained in two cases, minimal response in two and no response in one, so that primary tumor remission was evident in 40% of the cases. In the latter group, all cases but one attained partial response, i.e., remission was seen in 87.5% of cases treated by inferior gluteal infusion. Skin erosion of the gluteal, perineal, and anal regions and sciatica-like pain were observed in some cases; however neither myocardial effect nor bone marrow suppression, which have been reported as side-effects of ADM, were observed in any of the cases. These results suggest that this therapeutic modality could be effective in the preoperative work-up of candidates for total cystectomy, and also that it could be useful in the treatment of patients in whom total cystectomy is contraindicated.