[Collecting duct carcinoma (Bellini duct carcinoma) of the kidney with tumor extension into the inferior vena cava].

We report a case of collecting duct carcinoma (Bellini duct carcinoma) of the left kidney accompanied with a tumor thrombus in the inferior vena cava and the lymph node metastasis. A 69-year-old male presented with gross hematuria and left flank dullness. Computed tomography revealed an isodensity tumor in the left kidney with tumor extension into the inferior vena cava and the regional lymph node swelling. The T1-weighted magnetic resonance image displayed a slightly heterogeneous low-intensity-mass. Renal angiography revealed a hypervascular tumor. We performed left radical nephrectomy with tumor thrombectomy and regional lymphadenectomy. Histopathological examination revealed a collecting duct carcinoma (pT3bN1M0V2a). Seven months after surgery, multiple metastates in bone and liver developed. Then we performed systemic chemotherapy consisting of methotrexate and cisplatin. However, the patient died from the carcinoma 10 months postoperatively.

Transcript levels of aquaporin 1 and carbonic anhydrase IV as predictive indicators for prognosis of renal cell carcinoma patients after nephrectomy.

Since failure of differentiation has been suggested to be involved in the neoplastic process and progression of tumors, we evaluated whether the transcript levels of differentiation markers of proximal renal tubular cells, from which renal cell carcinoma (RCC) arises, could be used as prognostic markers. We used Northern blot analysis to study the expression of aquaporin 1 (aqp1) and carbonic anhydrase IV (ca4) genes in 66 paired samples of primary RCC and non-tumorous kidney tissues. Poor differentiation of tumor cells and non-clear cell-subtype RCC were significantly associated with low levels of aqp1 transcripts. When patients were divided into 2 groups according to level of aqpI transcript in RCC, a low level of aqp1 was significantly associated with unfavorable outcome. Among 18 patients with metastatic RCC and 40 patients with moderately differentiated RCC, those with RCC expressing low levels of aqpl mRNA demonstrated poorer survival than those with RCC expressing relatively high levels of aqp1. Similarly, decreased expression of ca4 mRNA in RCC was associated with poor survival. On multivariate analysis, transcript levels of aqpI and stage of the tumor were the independent factors predicting disease-specific survival. Transcript levels of aqp1 may serve as a new molecular prognostic marker in patients with RCC following nephrectomy.

Inhibitory effect of dexamethasone and progesterone in vitro on proliferation of human renal cell carcinomas and effects on expression of interleukin-6 or interleukin-6 receptor.

Interleukin-6 (IL-6) has been suggested as an autocrine growth factor of human renal cell carcinomas. Since steroids are known to inhibit IL-6 gene expression, we investigated their effects on the growth of renal cell carcinoma. Dexamethasone inhibited proliferation of 2 of 4 renal cell carcinoma cell lines in a dose-dependent manner. In one of these 2 cell lines, IL-6 gene expression was also inhibited, but not in the other. The inhibitory effect of dexamethasone on cell proliferation was not reversed by the exogenous IL-6. In 1 of the 2 remaining cell lines, the inhibition of IL-6 gene expression was observed, although there was no inhibition of cell proliferation. Thus, inhibition of growth by dexamethasone did not correlate with an inhibitory action of dexamethasone on IL-6 mRNA expression. Progesterone inhibited the growth of 1 cell line without concomitant inhibition of IL-6 gene expression. Expression of IL-6 receptor mRNA was not altered. A dose-dependent increase in mRNA expression of gp130, the transducer of IL-6 signal, was induced by dexamethasone and progesterone in 2 and 1 of the 4 cell lines, respectively. These data suggest that, in some renal cell carcinomas, steroids may inhibit cell proliferation by a mechanism independent of their effects on mRNA expression of IL-6 and IL-6 receptors. Dexamethasone may be useful, not only for palliation of paraneoplastic syndrome caused by overproduction of IL-6, but also for inhibition of growth of renal cell carcinomas.

Presence of alternative 5' untranslated sequences and identification of cells expressing ctk transcripts in the brain and testis.

From a mouse brain cDNA library, two species of the Csk-type tyrosine kinase (ctk) gene containing different 5' untranslated sequences were cloned. Using the common as well as specific nucleotide sequences of the two clones as probes, we examined the expression of ctk in various mouse tissues by Northern blot analysis. The results indicated that both species of ctk were expressed in the brain, testis and bone marrow. By in situ histochemistry of the brain, ctk transcript was detected in neurons throughout the entire brain, especially those of the cortex, the hippocampus and the cerebellum. This distribution pattern is similar to that of the Src family kinases including Yes, Src, Fyn and Lyn. In the testis, the major transcript (0.7 kb) was shorter than that expressed in the brain and the bone marrow (2.0 kb). A subsequent Northern blot analysis of fractionated germ cell populations and in situ histochemistry revealed that the short and long transcripts were expressed in germ cells and somatic cells, respectively, and that the expression level was quantitatively regulated during germ cell development. These results suggest that Ctk is involved in the regulation of neural function and differentiation of male germ cells through interactions with member(s) of the Src family kinases.

Induction of CL100 protein tyrosine phosphatase following transient forebrain ischemia in the rat brain.

Protein tyrosine phosphorylation is thought to play an important role in the regulation of neural function. To elucidate the role that protein tyrosine phosphatases (PTPs) may play in the postischemic brain, PTPs expressed in regions of the rat brain vulnerable to transient forebrain ischemia were examined. With the reverse-transcriptase polymerase chain reaction using degenerate primers, three PTPs, STEP, PTP delta, and SH-PTP2, were identified. They were expressed in the hippocampus 12 h after transient ischemia for 20 min. During the reperfusion period, the mRNA levels of these PTPs were not different from those in sham-operated rats. In contrast, a fourfold increase in the mRNA level of CL100 (3CH134), a PTP that is inducible by oxidative stress, was detected by Northern blotting in the hippocampus and cerebral cortex 1 h after the onset of reperfusion. In situ hybridization histochemistry showed a slight increase in the level of CL100 mRNA in neuronal cells in the hippocampus and cortex of postischemic rats compared to control rats. These findings suggest that PTPs play a role in the normal function of the hippocampus and cerebral cortex and demonstrate that ischemia induced CL100 expression.

Enhanced expression of multiple protein tyrosine phosphatases in the regenerating mouse liver: isolation of PTP-RL10, a novel cytoplasmic-type phosphatase with sequence homology to cytoskeletal protein 4.1.

To elucidate the role that protein tyrosine phosphatase (PTPs) may play in liver regeneration, PTPs expressed in the mouse liver after partial hepatectomy (PH) were investigated by a PCR-based cloning method. Sequencing of 115 cDNA clones identified 10 different sequences including MPTP (T cell PTP), PTP-1B, PTP-P19, mR-PTP mu, R-PTP alpha, PTP NE-3 (PTP-P1), R-PTP-kappa and the murine homologue of human LAR. The remaining two sequences, PTP-RL9 and PTP-RL10, encoded novel PTPs. PTP-RL10 cDNA contained an open reading frame of 1176 amino acids with no apparent membrane-spanning region. The amino-terminal region had sequence homology to those of human erythrocyte protein 4.1 and ezrin, cytoskeletal proteins. In the regenerating liver, the levels of five PTP gene mRNAs (MPTP, PTP-P19, R-PTP alpha, LAR homologue, and PTP-RL9) increased within 6 h, decreased to the normal level by 24 h, and increased again at 48 to 72 h after PH. The levels of PTP-1B and R-PTP-kappa mRNAs peaked within 6 h, decreased gradually, and returned to the normal level by 168 h after PH. In contrast, the levels of two PTP mRNAs (mR-PTP mu and PTP-RL10) peaked at 48 to 72 h, and returned to the normal level by 168 h after PH. No expression of PTP NE-3 was detected in the liver by Northern blotting. The differential expression of multiple PTPs during the pre-replicative and post-replicative stages of liver regeneration suggests that PTPs are involved in the regulation of growth and differentiation of liver cells.

[Bladder hernia associated with bladder diverticulum in a female].

A 77-year-old woman with a previous history of pelvic fracture had suffered from recurrent cystitis. In the excretory urography, post-void upright film revealed the bladder hernia associated with the bladder diverticulum. Transurethral incision and fulguration of the bladder diverticulum and left inguinal herniography was performed. There has been no recurrence since then.

Urinary excretion of mutagens and covalent DNA damage induced in the bladder and kidney after passive smoking in rats.

Using 32P-postlabeling assay, we studied the effect of sidestream smoke of cigarettes, so-called passive smoking, on the covalent DNA adduct formation in an animal model. Urine samples of 18 rats, 9 male and 9 female, before smoking resulted in an average of 2.4 adducts per 1 x 10(7) nucleotides per 24-h urine of a rat in the target plasmid DNA after incubation for 2 h in vitro. Urine samples of 4 out of 6 rats after exposure to sidestream smoke induced additional adducts in the target DNA. The incidence increased to 17.5 adducts per 1 x 10(7) nucleotides per 24-h urine of a rat. Without exposure to smoke, no increase in the adduct formation was observed. Adduct formations similar to those induced in vitro were detected in the bladder and kidney DNA, but not in the testicular DNA, of the four rats exposed to sidestream smoke. These observations suggest that passive smoking causes covalent DNA damage of the cells in the bladder and kidney by excreting chemicals in urine. Passive smoking as well as active smoking might contribute to the bladder and renal carcinogenic process.

Identification of protein tyrosine phosphatases expressed in murine male germ cells.

We cloned five cDNAs encoding putative protein tyrosine phosphatases (PTPs; protein-tyrosine-phosphatase phosphohydrolase, EC 3.1.3.48) from the murine testis by using degenerate primers and the polymerase chain reaction cloning technique. Two of them were identical to the mouse cytoplasmic PTP and PTP-1B. The remaining three were likely to represent the murine counterparts of human PTP delta, human PTP epsilon, and rat striatum-enriched PTP. The cells expressing these genes were determined either by in situ hybridization histochemistry or by Northern blot hybridization. Moreover, Northern blot hybridization revealed that the transcripts of PTP-1B, mouse cytoplasmic PTP and the murine homologue of human PTP delta were quantitatively and/or structurally regulated during germ cell development, suggesting their roles in spermatogenesis.

Application of polymerase chain reaction-single strand conformation polymorphism analysis to the diagnosis and screening of adenine phosphoribosyltransferase deficiency.

Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis is a rapid and sensitive method used to identify point mutations in a given sequence of genomic DNA. We applied this method to the diagnosis of adenine phosphoribosyltransferase (APRT) deficiency, which is an autosomal recessive hereditary disease leading to 2,8-dihydroxyadenine urolithiasis. Genomic APRT genes were amplified and labeled simultaneously with [alpha-32P]dCTP (cytidine triphosphate) by PCR. When run in a 6% polyacrylamide gel containing 10% glycerol, two types of mutant genes-APRT*QO and APRT*J-gave bands clearly distinct from those of the equivalent normal APRT genes. Using this method we diagnosed both homozygotes and heterozygotes for defective APRT genes. On screening 80 Japanese individuals for polymorphism or mutations by PCR-SSCP we did not find any alterations leading to a false positive diagnosis. These findings suggest that PCR-SSCP, in addition to being rapid and sensitive, is a useful diagnostic method which is highly specific in detecting mutant APRT genes in the Japanese population.

[Anti-interleukin-6 (IL-6) therapy of IL-6-producing renal cell carcinoma].

Interleukin-6 (IL-6) has been demonstrated to be an autocrine growth factor of renal cell carcinoma in vitro and released IL-6 has been thought to elicit the acute phase response in vivo. We investigated the possibility of anti-IL-6 therapy of IL-6-producing renal cell carcinoma. There was a dose-dependent decrease in the number of colonies formed in vitro of NC65 renal cell carcinoma cell line in the presence of dexamethasone which is known to inhibit the induction of IL-6 messenger RNA. IL-6 receptor antisense oligonucleotide and anti-IL-6 receptor antibody also showed growth inhibition of NC65 cells. IL-6 antisense oligonucleotide and anti-IL-6 antibody did not alter the proliferation of NC65 cells. These findings suggest that inhibitors of IL-6 production or IL-6 function may be useful for some renal call carcinoma patients.

[Comparative study of risk criteria for germ cell tumor].

The development of cisplatin-based chemotherapy has achieved a high cure rate in patients with advanced germ cell tumors (GCT) and it is more important to predict the prognosis of each patient before treatment and select the most suitable regimen of therapy. To date, 4 risk criteria for GCT are presented. From November, 1985 to April, 1991 our treatment protocol for GCT consisted of VAB-6 (vinblastine, actinomycin D, bleomycin, cisplatinum) or PVeBV (vinblastine, etoposide, bleomycin, high-dose cisplatin) as the induction chemotherapy and VIP (etoposide, ifosfamide, cisplatin) as the salvage chemotherapy. In total, 12 patients were entered on this protocol. They were divided into 2 groups based on the actual clinical course. Those who achieved complete remission within 3 cycles of chemotherapy were divided into "good response group" and others were into "poor response group". These results were compared with those classified by the 4 risk criteria. As a result of our study "The Indiana Staging System" seemed to be the most useful.

[Detection of mutant adenine phosphoribosyltransferase genes by polymerase chain reaction-single strand conformation polymorphism analysis].

Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis is a rapid and sensitive method to identify point mutations in a given sequence of genomic DNA. We tried to apply the PCR-SSCP to the diagnosis of adenine phosphoribosyltransferase (APRT) deficiency, which is an autosomal recessive hereditary disease leading to 2,8-dihydroxyadenine urolithiasis. Genomic APRT genes, with or without mutations, were amplified and labeled simultaneously with 32P-dCTP by PCR. When run in a 6% polyacrylamide gel containing 10% glycerol, two types of mutant genes, APRT*Q0 and APRT*J, gave bands clearly distinct from those of the respective normal APRT genes. Since heterozygotes as well as homozygotes for these mutant APRT genes can be detected in 2 days, PCR-SSCP should be a valuable method in the diagnosis of APRT deficiency and in screening a large population for APRT mutant genes.

Enhanced expression of interleukin-6 in primary human renal cell carcinomas.

We have demonstrated interleukin-6 (IL-6) production by human renal carcinoma cells. The IL-6 gene expression was detected by Northern blot analysis in 22 of 43 primary renal cell carcinoma tissues and in five of seven renal cell carcinoma cell lines. Immunohistochemical analysis confirmed the expression of IL-6 by the tumor cells. Patients with a high-level expression of IL-6 had significantly greater incidences of lymph node metastasis and a larger increase in serum C-reactive protein than those without it. We have also probed for the presence of IL-6 receptor by Northern blot analysis; we detected this receptor in 11 of the 43 primary renal cell carcinoma tissues but in none of the seven renal cell carcinoma cell lines. However, by use of the complementary DNA-polymerase chain reaction, the IL-6 receptor transcript was detected in all specimens, including the seven cell lines. No expression of the interleukin-3 (IL-3) gene was identified in any of the 43 primary renal cell tumors. These data provide evidence that IL-6 and its receptor may play a role in promoting the transformation and/or proliferation of renal cell carcinomas as well as in teh development of symptoms.

Adhesion of mouse mast cells to fibroblasts: adverse effects of steel (Sl) mutation.

Mouse bone marrow-derived cultured mast cells proliferate on +/+ mouse embryo-derived 3T3 fibroblasts, but not on Sl/Sld mouse embryo-derived 3T3 fibroblasts, in the absence of IL-3 and IL-4 (Fujita et al: Proc. Natl. Acad. Sci. U.S.A. 86:2888-2891, 1989). To further characterize the mast cell-fibroblast interactions and the effects of Sl mutation, we tried to analyze the adhesion of cultured mast cells to 3T3 fibroblasts in vitro. Mast cells plated onto NIH/3T3 fibroblasts showed marked adhesion within 30 min, which reached a plateau after 3 h. The numbers of adhered mast cells were linear over the range of 10(3) to 5 x 10(5) cells inoculated into each (2 cm2) of 24 multiwells. Adhesion required active energy production and the presence of divalent cations. It was not inhibited by an RGD-containing peptide, an anti-LFA-1 antibody, or asialofetuin. Mast cells adhered efficiently to the eight 3T3 cell lines derived from +/+ mouse embryos, but not to the eight 3T3 cell lines derived from Sl/Sld mouse embryos. Adhesion to +/+ mouse spleen-derived fibroblasts lacking mast cell-supporting activity was comparable to that to Sl/Sld/3T3 cells. The failure of mast cells to adhere to fibroblasts with the Sl mutations was not due to a production of a diffusible inhibitor by the latter. These results indicate that production of wild type Sl gene product by fibroblasts is mandatory for adhesion/migration, as well as for proliferation of mast cells on them, and that the coculture system should be useful for the biochemical and molecular analysis of these interactions.

[Adjuvant chemotherapy after cystectomy of bladder tumor].

Adjuvant chemotherapy mainly consisting of cisplatin, adriamycin and mitomycin C was administered to 17 patients after cystectomy for bladder tumor (chemotherapy group). Seventeen concurrently treated patients did not receive adjuvant chemotherapy (control group). From the comparison of survival curves of these two groups, the following results were obtained. 1) Survival curves of the patients in all stages did not differ significantly between chemotherapy and control groups. 2) Among patients in stage pT3, pT4 and/or N+, survival of the chemotherapy group seemed to have some advantage over that of the control group, but the survival curves did not differ significantly between these two groups. 3) Among patients in N+, survival of the chemotherapy group was far better than that of the usual cases. Review of the literature on adjuvant chemotherapy after cystectomy for bladder tumor revealed the necessity of randomized study to determine whether adjuvant chemotherapy is effective.

[Clinical analysis of 40 total cystectomies for bladder tumor].

Between June, 1983 and December, 1988, 40 patients with primary bladder tumor underwent total cystectomy. Of the 40 patients, 33 (82.5%) were treated by total cystectomy at first presentation. Only 7 patients (17.5%) had prior history of bladder tumors. The mean time from the onset of symptoms to consultation was 11.5 months. In 68.5% of the evaluable 35 patients, gross hematuria was the first symptom. In 74.4% of the 39 evaluable patients, preoperative urine cytology was positive. If class III (suspicious) was included, the positive urine cytology rate was 82.1%. The operative mortality rate was 15%. Early complications occurred in 52.5% of the 40 patients. Late complications occurred in 35.3% of the 34 patients. The 1-, 2- and 3-year actual survival rates of the 40 patients were 69.6%, 65.5% and 65.5% respectively. The 2-year survival rate according to pathologic stage was 14.3% for patients in pT4, 43.8% in pT3b, 75% in pT3a, 100% in pT2, 78.8% in pT1 and 100% in pT0 + pTis. Of the 28 patients who underwent pelvic lymphadenectomy, 8 (28.6%) had positive nodes, including 1 of 1 (100%) in pT4, 5 of 7 (71.4%) in pT3b, 1 of 6 (16.7%) in pT3a, 1 of 4 (25%) in pT2 and 0 of 10 (0%) in pT1 + pT0 + pTis. The prognosis of the 7 patients who had prior history of bladder tumor was poor. The selection of initial therapy and the clinical follow-up should be done carefully.

[A case of renal cell carcinoma which was left untouched for five years].

We report a case of bilateral synchronous renal cancers for which only right radical nephrectomy was performed at another hospital. Five years later we performed partial nephrectomy (enucleation) for the left renal cancer. By computed tomographic scans we studied the radiological change of this tumor which occurred during a five-year period. We found a slow doubling time of this tumor. Three years after enucleation, tumor recurrence was noticed and we performed left partial nephrectomy with a normal parenchymal margin. We reviewed the literature about enucleation of renal cancer.

[A study of 22 cases of renal injury].

Between June, 1984 and May, 1988, 22 cases of renal injury were treated. There were 13 males and 9 females. The most frequent causes were traffic accidents (13 cases) and the next were falls (7 cases). Associated injuries were seen in 9 cases, including bone fractures, head injuries, liver lacerations, hemothorax and splenic laceration. Out of the 22 patients, there were 13 cases of renal contusion, 6 cases of renal laceration, 1 case of renal rupture and 2 cases of pedicle injury. Evaluation was made by IVP, CT, arteriography and so on. Arteriography was the most useful method whether immediate surgery should be done or not. Three cases were treated surgically (2 nephrectomies for rupture and pedicle injury, 1 drainage for laceration). The other cases were treated conservatively and no complications have been seen.

[A case of urothelial tumor involving renal pelvis, ureter and bladder in a patient receiving hemodialysis].

A 59-year old male who had undergone hemodialysis for 5 years, visited our hospital with a complaint of asymptomatic gross hematuria. Urinary cytology was positive and random biopsy revealed invasive transitional cell carcinoma of the bladder. The patient underwent total cystectomy and ureterocutaneostomy. Also right nephrectomy was performed because pathological examination during the operation revealed that the right ureteral margin had carcinoma in situ. Carcinoma in situ involved the right renal pelvis, the right ureter and the bladder in the resected specimen. Frequency of urothelial tumor in patients who undergo hemodialysis is still unknown, and the number of reported cases is too small to acknowledge the natural history of the urothelial tumor in patients on hemodialysis.