Increased expression of DNA topoisomerase I gene and collateral sensitivity to camptothecin in human cisplatin-resistant bladder cancer cells.

We established three cis-diamminedichloroplatinum(II) (cisplatin)-resistant cell lines, T24/DDP5, T24/DDP7, and T24/DDP10, by the stepwise exposure of T24 human bladder cancer cells to increasing concentrations of cisplatin. The resistance to cisplatin of T24/DDP5, T24/DDP7, and T24/DDP10 cells was 2.2-, 5.2-, and 8.4-fold that of the parental T24 cells, respectively. The cisplatin-resistant cell lines also showed an increased resistance to vincristine, although their sensitivities to Adriamycin and etoposide resembled that of T24. In contrast, the cisplatin-resistant cells developed a collateral sensitivity to (4s)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyl oxy]dione hydrochloride trithydrate, a camptothecin derivative, and its active metabolite, 7-ethyl-10-hydroxy-camptothecin, that targets DNA topoisomerase I. Both a Northern blot analysis and an immunoblot analysis demonstrated increased cellular levels of DNA topoisomerase I mRNA in the resistant cell lines. However, the expression of DNA topoisomerase II in the three resistant cell lines did not differ significantly from that in the T24 cells. No significant differences in the glutathione S-transferase pi levels were observed, although the intracellular content of glutathione in the T24/DDP7 cells was slightly but significantly increased. In addition, the intracellular platinum concentration correlated negatively with the degree of cisplatin resistance and was found to be significantly decreased in T24/DDP10 at an external cisplatin concentration of 20 micrograms/ml. These results suggest that the increased levels of DNA topoisomerase I mRNA thus play an important role in cisplatin resistance and produce a collateral sensitivity to (4s)-4,11-diethyl-4-hydroxy-9[(4-piperidinopiperidino)carbonylo xy]dione hydrochloride trithydrate and 7-ethyl-10-hydroxycamptothecin in these cisplatin-resistant bladder cancer cell lines. In addition, the presence of decreased intracellular cisplatin accumulation may also contribute to the acquisition of resistance to cisplatin in these cell lines.

The mechanism of relaxation induced by atrial natriuretic peptide in the porcine renal artery.

1. The mechanisms underlying the relaxation of the porcine renal artery induced by atrial natriuretic peptide (ANP) were investigated, using front-surface fluorimetry with fura-2 and receptor-coupled permeabilization by alpha-toxin. 2. ANP decreased the cytosolic Ca2+ concentration ([Ca2+]i) and tension during the contraction induced by a high external K+ solution, in a concentration-dependent manner. This ANP-induced decrease in [Ca2+]i during the contraction induced by high K+ solution was composed of two phases, an initial rapid phase, followed by a maintenance phase. The initial rapid decrease in [Ca2+]i, but not the maintained decrease in [Ca2+]i, was inhibited when the tissue was treated with thapsigargin, a selective Ca2+ pump inhibitor of the sarcoplasmic reticulum. When the tissues were treated with thapsigargin and external Ca2+ was replaced by Ba2+, which cannot be transported by the Ca2+ pump, ANP did not induce a decrease in [Ba2+]i, even though the elevation of tension induced by Ba2+ was strongly inhibited. 3. In the absence of extracellular Ca2+, ANP inhibited the release of Ca2+ from the intracellular store induced by noradrenaline (NA). 4. The [Ca2+]i (abscissa scale)-tension (ordinate scale) relationship observed during the contraction induced by various concentrations of high external K+ solution was shifted downwards by the addition of 10(-8) M ANP, indicating that, at any given [Ca2+]i, the tension generated by high K+ solution was considerably inhibited by the addition of 10(-8) M ANP. The [Ca2+]i-tension curve of the contraction obtained by the cumulative application of external Ca2+ (0-3.75 mM) during depolarization with 118 mM K+ solution was shifted to the left by 3 x 10(-7) M NA. This NA-induced [Ca2+]i-tension relationship was shifted to the right by 10(-8) M ANP, indicating that the ANP-induced reduction of Ca(2+)-sensitivity operates during the contraction induced by NA. 5. In alpha-toxin-permeabilized preparations, ANP induced relaxation of tissues precontracted with a mixture of 3 x 10(-7) M Ca2+, 10(-5) M guanosine 5'-triphosphate (GTP) and 10(-6) M NA. Thus a component of ANP-induced relaxation took place by way of a reduction in the Ca2+ sensitivity of the myofilaments, independent of changes in [Ca2+]i. 6. These results indicate that ANP induces relaxation of the porcine renal artery by: (1) reducing [Ca2+]i mainly via the activation of the Ca2+ pumps located on the sarcoplasmic reticulum and sarcolemma, as well as via inhibition of agoinist-induced release of Ca2+ from the intracellular store; and (2) decreasing the Ca(2+)-sensitivity of the contractile elements.

Summary of clinical experience with cefpodoxime proxetil in adults in Japan.

Cefpodoxime proxetil is an oral cephem antibiotic of a new ester type, developed by Sankyo Co., Ltd in Japan. It has a broad antibacterial spectrum, which includes Staphylococcus, and a long half-life, allowing twice-daily administration. In Japan, clinical studies on this drug were performed in various fields, including internal medicine, surgery, urology, otorhinolaryngology, and obstetrics and gynaecology. Good or excellent clinical responses were observed in 2275 of 2902 patients analysed, giving a 78.4% efficacy rate overall. Side effects occurred in 98 patients (2.7%); these were mainly gastrointestinal and included diarrhoea, nausea, and vomiting. Abnormal laboratory test results observed included increased AST in 2.8% (55 of 1973), increased ALT in 3.2% (63 of 1965), and eosinophilia in 2.4% (36 of 1521).

Leiomyosarcoma in soft tissue: examination of p53 status and cell proliferating factors in different locations.

The biological behavior of soft tissue leiomyosarcoma varies with its location: tumors in deep soft tissue show a worse prognosis than those in superficial soft tissue. In this study, we analyzed cell proliferating factors (mitotic and Ki-67 indices) and the p53 status of both types of leiomyosarcoma (37 cases) to evaluate the possibility that these factors may serve as indicators in prognosis. Expression of both of the cell proliferating factors were higher in the superficial type, but these differences were not significant. The incidence of abnormal p53 detected by immunohistochemistry and mutation analysis was also higher in the superficial type, but, again, no significant difference was observed. Abnormal p53 and a high Ki-67 index in the deep type, and a large tumor size (>6 cm) in the superficial type may be useful as prognostic factors. Interestingly, all of the superficial cases who died of the disease showed abnormal p53. In conclusion, these two types of leiomyosarcoma are essentially the same, and their location remains a good prognostic factor. Furthermore, abnormality of the p53 gene was related to a poor prognosis regardless of the tumor's anatomic location.

Human papillomavirus DNA in condylomata acuminata from Japanese males.

We examined condylomata acuminata from Japanese males for the presence of human papillomavirus (HPV) genomes by Southern blot hybridization. HPV 6/11-related DNA was found in 91% (32/35) of the condylomata. HPV 6a DNA was found in 40% (14/35), HPV 6c DNA in 6% (2/35), and HPV 11a DNA in 37% (13/35). HPV 6-related DNA, which had an unusual PstI-cleavage pattern, was detected in one sample. Types and subtypes of HPV DNA in the samples studied (HPV 6a, 6c, and HPV 11a DNA) were not correlated with the patients' ages nor outcomes of the disease. HPV 16 DNA was not detected in any condyloma acuminatum.

New method for measuring bladder pressure.

We measured intramural pressure of the bladder using a pressure transducer embedded between bladder mucosa and muscle layers in female dogs. Intravesical pressure was recorded simultaneously by conventional transurethral catheter method. Results from the former showed slightly lower values than the latter, but both correlated well in all pressure ranges.

Transitional cell carcinoma of bladder in thirteen-year-old boy.

We report a rare case of primary epithelial bladder tumor in a thirteen-year-old boy. Histologically, the tumor was that of a solitary and papillary transitional cell carcinoma, grade 1. This tumor was detectable at cystoscopic examination, and cytologic procedures were without benefit for a diagnosis.

Extensive, obstructing sodium urate deposition on wall of isolated ileum for replacement of entire ureter.

We report a rare case of obstructing sodium urate stones occurring in the form of extensive deposition on the wall of isolated ileum for replacement of the entire ureter. The stones were disintegrated and removed with a rigid ureteroscope and an electrohydraulic lithotriptor. Hyperuriaciduria, excessive alkalization of urine and urinary stagnation due to reflux, mucous retention, and prostatic hyperplasia were considered as the etiologic factors in the stone formation.

Nonspecific granulomatous prostatitis.

Nonspecific granulomatous prostatitis is a relatively rare disorder of the prostate. We encountered 4 cases of this type of chronic inflammation, including 1 case of xanthogranulomatous prostatitis. In all cases the diagnosis was made by histologic examination of specimens obtained by transurethral resection, retropubic prostatectomy, or transrectal needle biopsy. Echography revealed a hypoechoic lesion in the case of xanthogranulomatous prostatitis, while the other cases showed no specific findings except for the associated adenomas. The major symptoms were frequency and dysuria caused by urinary tract infection or benign prostatic hyperplasia associated with the granulomatous prostatitis.

Intrasinusal pyelocalicotomy with lower pole nephrotomy for removal of renal stones.

A small renal sinus often limits exposure of the intrarenal pelvis and restrains the sinus approach for removal of renal stones. For 14 such cases, we used a lower pole nephrotomy combined with extended pyelolithotomy. Incision of the renal parenchyma through the intersegmental avascular plane greatly facilitated exposure of the intrarenal collecting system and enabled stone removal through the longitudinal incision of the collecting system. The indication for operative technique and results of the procedure are described.

Prognostic factors and value of adjunctive nephrectomy in patients with stage IV renal cell carcinoma.

We reviewed 57 cases of Stage IV renal cell carcinoma to clarify the factors influencing prognosis and to evaluate the value of nephrectomy. Cumulative survival from the initial diagnosis was analyzed with respect to the patients' age, sex, side of primary tumor, initial performance status (PS), site of metastasis, and nephrectomy. Overall survival for the patients was 51 percent at one year, 22 percent at three years, and 11 percent at five years. Age, sex, and side of primary tumor had no influence on survival. Improved survival was correlated with good PS, metastases limited to single organ, and removal of the primary tumor. With regard to histopathologic features in nephrectomized patients, low grade and stage were correlated with longer survival. These factors should be considered in the analysis of results of future clinical trials of metastatic renal cell carcinoma.

Antibiotic treatment of urinary tract infections.

Cefotaxime was compared with ceftezole for clinical efficacy and safety in the treatment of chronic complicated urinary tract infections. Cefotaxime was significantly superior to ceftezole in both overall clinical and bacteriological efficacy. There was no significant intergroup difference in adverse reactions or laboratory parameters. Cefotaxime, therefore, proved to be of considerable value in the treatment of chronic complicated urinary tract infections.

Early intravesical instillation of adriamycin with oral administration of 5-fluorouracil after transurethral resection for superficial bladder cancer: preliminary results.

In all, 199 patients were entered in this study by 21 collaborating hospitals. Patients with superficial transitional cell carcinoma of the bladder were randomized postoperatively into four groups. Group A received early (immediately and 2 days after transurethral resection) instillation of adriamycin (30 mg/30 mg); group B received early instillation of adriamycin with oral administration of 5-fluorouracil (200 mg/day); group C received delayed (7 days after transurethral resection) instillation of adriamycin (30 mg/30 ml); and group D received delayed instillation of adriamycin with oral administration of 5-fluorouracil (200 mg/day). All patients subsequently received instillations weekly for 2 more weeks, and then every 2 weeks for a further 14 weeks. After 4 months, they received one instillation per month for 8 months. 5-Fluorouracil was administered p.o. for 1 year. The postoperative follow-up period was 12 months. After 3 and 6 months there were significant differences in the non-recurrence rates between groups B and C. After 12 months the overall non-recurrence rates were 87.9% in group A, 83.5% in group B, 89.2% in group C, and 82.8% in group D, and there were no significant differences among the four groups. The number of patients entered and the follow-up period are not adequate for firm conclusions, and further studies are necessary. The main side effect was bladder irritation, which was observed in 38.8% of patients in the early instillation groups and in 26.3% of those in the delayed instillation groups. No severe systemic side effects were observed in this study.

Adjuvant chemotherapy with early intravesical instillation of adriamycin and long-term oral administration of 5-fluorouracil in superficial bladder cancer. The Kyushu University Urological Oncology Group.

A randomized controlled trial was performed to study the efficiency of adjuvant chemotherapy with early intravesical instillation of Adriamycin and long-term oral administration of 5-fluorouracil in 275 patients with superficial bladder cancer. All of the patients were randomized into four groups. Group A received early (immediately and 2 days after transurethral resection) instillation of Adriamycin alone; Group B received early instillation of Adriamycin with oral administration of 5-fluorouracil; Group C received delayed (7 days after transurethral resection) instillation of Adriamycin alone; and group D received delayed instillation of Adriamycin with oral administration of 5-fluorouracil. All patients subsequently received instillations weekly for 2 weeks and then every 2 weeks for a further 14 weeks. After 4 months, they received monthly instillations for 8 months. 5-Fluorouracil (groups B and D) was given daily p.o. for 1 year. Evaluation was possible in 187 patients. The postoperative follow-up period for determination of non-recurrence rates was 36 months, during which no significant difference was detected among the four groups. Moreover, no statistically significant difference was found between the early- and delayed-instillation groups. However, the non-recurrence rates obtained in the groups undergoing early instillation were higher than those determined in the delayed-instillation groups during the 36-month follow-up period, and this difference was especially significant at 4 and 5 months. In addition, the early-instillation groups showed significantly higher non-recurrence rates than did the delayed-instillation groups in terms of primary cases (P less than 0.01), tumor size of less than 1 cm (P less than 0.05), multiple tumors (P less than 0.01), pathological stage pTa (P less than 0.01), and histological grades G1 and G2 (P less than 0.05). Groups B and D, which were treated by intravesical instillation of Adriamycin with oral administration of 5-fluorouracil, showed no significant prophylaxis of recurrence during the 36-month follow-up as compared with groups A and C, which received intravesical instillations alone. The main side effect, which required discontinuation of the treatment, was bladder irritation. However, no significant difference in its incidence was found between the early- and delayed-instillation groups. No severe systemic side effect was encountered in this study. These results suggest that early as well as repeated intravesical instillation of Adriamycin is clinically tolerable and may be effective in preventing the recurrence of superficial bladder cancer.

Combination chemotherapy for infections due to methicillin-resistant Staphylococcus aureus with combination therapy by cefuzonam and fosfomycin or minocycline in the urologic field.

To determine the bacterial and clinical effects of methicillin-resistant Staphylococcus aureus (MRSA) on urinary tract infection (UTI), postoperative wound infection, and bacteremia, 22 strains of MRSA from infected patients were examined; minimal inhibitory concentration (MIC), fractional inhibitory concentration (FIC) index, coagulase typing, and change in MIC in the combination of cefuzonam (CZON) and fosfomycin (FOM) or minocycline (MINO), and the clinical effects of the combination therapy of CZON+FOM or CZON+MINO were investigated in 23 patients. MIC distribution was assessed for 11 drugs: methicillin (DMPPC), cefazolin (CEZ), cefotiam (CTM), cefuzonam (CZON), minocycline (MINO), vancomycin (VCM), arbekacin (ABK), imipenem (IPM), fosfomycin (FOM), ofloxacin (OFLX), and clarithromycin (CAM). For VCM and ABK, MICs ranged from 0.2 to 12.5 micrograms/ml. MINO showed a wide range of MIC, from 0.05 to 25 micrograms/ml. All strains were less sensitive to other antimicrobials. This MIC distribution was assessed in categories by coagulase typing. For CAM, type II strains revealed > or = 100 micrograms/ml of MIC50 and MIC90 compared with 0.2 and 3.13 micrograms/ml in type VII strains. For mixed combinations of CZON and MINO at ratios of 10 to 1 and 40 to 1, the rates of blood concentration for drugs 10 and 120 min after the intravenous injection, MIC distribution was observed between MINO and CZON. For mixed combinations of CZON and FOM at ratios of 1 to 1 and 1 to 4, MIC distribution was more sensitive than for FOM or CZON alone. With respect to the clinical effects of combination therapy by CZON+MINO and CZON+FOM, of seven cases of UTI with CZON+MINO, MRSA was eliminated completely in 4 patients (57.1%). In all patients who received CZON+FOM, MRSA was completely eliminated by this treatment regimen. In 4 of 7 (57.1%) patients with would infection and bacteremia, MRSA was eliminated by the combination of CZON+FOM or CZON+MINO. These results suggest that CZON+FOM is an effective combination in treating UTI and other MRSA infections in urology.

Phase II study of a new combined primary chemotherapy regimen, intravenous methotrexate and vincristine and intraarterial adriamycin and cisplatin, for locally advanced urinary bladder cancer: preliminary results.

A phase II study of a new combination therapy was performed using intraarterial (i.a.) cisplatin and Adriamycin in combination with i.v. methotrexate and vincristine for 27 patients with invasive urinary bladder carcinoma of stages T2-3NOMO, and the therapeutic effects were assessed. Methotrexate (20 mg/m2) was given i.v. on days 1,15, and 22, and vincristine (0.7 mg/m2) was injected i.v. on day 2 before i.a. infusion therapy and on days 15 and 22. The i.a. chemotherapy was performed after both superior gluteal arteries had been embolized using 3- or 5-mm stainless-steel coils. A mixture of cisplatin (50-70 mg/m2) and Adriamycin (20 mg/m2) was infused i.a. via both internal iliac arteries over a period of 20-30 min. Angiotensin II (mean dose, 21 micrograms) was simultaneously infused i.a. in 15 of 27 patients. In 24 of the 27 patients, at least 2 cycles of full-dose chemotherapy were completed. The dose was decreased in the remaining 3 patients because of their poor health status and advanced age. Among the 27 patients, 9 and 14 had complete (CR) and partial responses (PR), respectively; 3 manifested no change (NC), and 1 had progressive disease (PD). The objective response rate (CR+PR) was 85.2%. Among the 27 patients staged T2-3 NOMO, 6 (CR, 1; PR, 5) underwent total cystectomies and 18 (CR, 8; PR, 8; NC, 2) had transurethral resection of a bladder tumor (TUR-Bt) or partial resections following chemotherapy. The remaining 3 diminished-dose patients had no surgery. Of the 27 patients, 22 were alive after a median follow-up period of 21+ (range, 7-48+) months. No significant side effect was observed except for lower extremity paresthesias in 5 patients (18.5%). These results point to the effectiveness of this therapy and to the possibility of urinary bladder preservation in patients with invasive, advanced urinary bladder cancers.

Tosufloxacin in the treatment of nongonococcal urethritis, including Chlamydia trachomatis.

Sixty-three patients with nongonococcal urethritis were enrolled in an open, comparative study and assigned to 14 days of treatment with tosufloxacin 150 mg orally three times daily or doxycycline 100 mg orally twice daily. Chlamydia trachomatis was detected in 27 (42.9%) of the 63 patients: 13 (43.3%) of the 30 tosufloxacin-treated patients and 14 (42.4%) of the 33 doxycycline-treated patients. The clinical response between the two treatment groups was compared 7, 14, and 21 days after initiation of therapy. Clinical response to tosufloxacin was as good as that seen with doxycycline at the three follow-up visits. All C trachomatis-positive patients at baseline tested negative in both treatment groups at the three follow-up visits. Adverse reactions were observed in two patients treated with doxycycline. Although the number of study patients was small and the follow-up period short, these results indicate that tosufloxacin and doxycycline are equally effective in the treatment of nongonococcal urethritis.

Prophylactic intravesical chemotherapy with adriamycin plus verapamil for primary superficial bladder cancer: preliminary results. The Kyushu University Urological Oncology Group.

A prospective randomized trial was conducted to compare the prophylactic effect of intravesical installation of Adriamycin (ADM) plus verapamil (VR) with that of ADM alone for recurrence of superficial bladder cancer. A total of 226 patients were enrolled and randomized into 2 groups. Group A received intravesical instillation of ADM (30 mg/30 ml physiological saline) on 19 occasions during a 1-year period after transurethral resection, whereas group B received intravesical instillation of ADM (30 mg/24 ml physiological saline) plus VR (15 mg/6 ml saline) according to the same schedule used for group A. Evaluation was possible in 157 of the 226 registered patients (group A, 76; group B, 81). There was no significant difference in the patients' characteristics between the two groups, and there was no significant difference in the overall nonrecurrence rate determined over a 24-month follow-up period. However, group B showed a significantly higher nonrecurrence rate than did group A for tumors measuring less than 1 cm in diameter (P < 0.05) and for histological grade 2 tumors (P < 0.01) in spite of there being no significant difference in the other characteristics of each subgroup of patients. The incidence and severity of side effects were similar in both groups, and VR caused no significant systemic toxicity. Although further follow-up is necessary, these results suggest that intravesical instillation of ADM plus VR is clinically safe and may be more effective than instillation of ADM alone in preventing the postoperative recurrence of superficial bladder cancer (less than 1 cm in diameter, histological grade 2).

Prophylactic intravesical instillation chemotherapy against recurrence after a transurethral resection of superficial bladder cancer: a randomized controlled trial of doxorubicin plus verapamil versus doxorubicin alone. The Kyushu University Urological Oncology Group.

PURPOSE: We investigated whether verapamil (VR), a known chemosensitizing agent of P-glycoprotein-mediated multidrug resistance, could enhance the preventative effect of doxorubicin (Adriamycin, ADM) on both intravesical recurrence and disease progression after transurethral resection (TUR) of superficial bladder cancer. METHODS: The patients were randomized into two groups: one group received an intravesical instillation of ADM (30 mg) plus VR (15 mg) after TUR of superficial bladder cancer (19 times over 1 year), and the other group received ADM alone on the same treatment schedule. The nonrecurrence rate, the incidence of disease progression at the first recurrence and the side effects were compared over a median follow-up of 38.5 months. RESULTS: Of the 226 patients registered, 157 were evaluable. No significant differences were observed in the patients' characteristics between the two groups. Although the incidence of disease progression at the first recurrence was not significantly different between the two groups, the ADM plus VR instillation group did show a significantly higher nonrecurrence rate than the ADM-only instillation group, and such significance persisted even when any possible bias was allowed for in a multivariate analysis. In terms of side effects, the incidence and severity of bladder irritation symptoms were not significantly different between the two groups. CONCLUSIONS: Intravesical instillation chemotherapy with ADM plus VR was found to have a significantly greater beneficial effect than with ADM alone for preventing recurrence after TUR of superficial bladder cancer.

Treatment of advanced hormone-refractory prostate carcinoma with a combination of etoposide, pirarubicin and cisplatin.

A total of 20 patients with hormone-refractory prostate carcinoma entered a pilot study of combination chemotherapy based on the EAP (etoposide, Adriamycin and cisplatin) regimen, in which Adriamycin was replaced by pirarubicin, a less cardiotoxic derivative of Adriamycin. The response was assessed by criteria modified from those of the National Prostatic Cancer Project: prostate-specific antigen was employed instead of acid phosphatase. Of 18 evaluable patients, 6 achieved a partial response, 5 had stable disease, and in 7 the disease had progressed during therapy; thus, the overall response rate was 33.3% [95% confidence interval (CI) 11.5-55.1%]. Significant pain alleviation and performance status improvement were obtained in 5 of 12 patients (41.7%; CI 13.8-69.6%) and 3 of 13 patients (23.1%; CI 0.2-46.0%), respectively. Although myelosuppression was moderate to severe, no chemotherapy-related deaths or bacteriologically documented sepsis occurred; nor was there any clinical cardiotoxicity. All the responding patients received maintenance chemotherapy with etoposide thereafter. At present, the median duration of response is 33 weeks (range: 23-91 weeks) and the median survival period for all patients is 42 weeks (range: 27(+)-136 weeks), with 12 deaths. In spite of the small number of patients treated, these results suggest that this chemotherapy regimen is active in advanced hormone-refractory prostate carcinoma.