We have measured androgen receptor in whole cell, cytosol and nuclear fractions of cultured skin fibroblasts originated from the genital area. Samples were obtained from the following patients: 10 boys with phimosis, 5 boys with cryptorchism, 4 boys and a man with hypospadia, 2 infertile men and 3 normal men. Androgen receptor was measured using methyltrienolone (R1881) as the ligand according to the assay technique of Eil et al. using dispersed cell method. The results were as follows: 1. There were no significant statistical differences in the dissociation constant among the diseases. 2. Hypospadia cases showed lower maximum binding capacities (Bmax) of whole cell than phimotic boys but the difference was not statistically significant. 3. The Bmax of normal men were statistically higher than those of phimotic boys and about 80% of these receptors were seen in the nuclear fraction. We also experienced low Bmax levels in an adult hypospadia case and two infertile men. These resutls suggest that this method is a simple and rapid method to study both nuclear and cytosol androgen receptor at the same time. It is also suggested that there are some cases with reduced androgen receptor levels in hypospadia and infertile cases.
The effect of dietary protein load on the incidence of nephrolithiasis was studied in rats and men. Three groups of adult male Wister rats were fed with a standard protein diet, a high protein diet, or a low protein diet for 4 weeks. In the high protein group, calcium excretion was significantly increased and citrate excretion was remarkably decreased. This group also exhibited low grade metabolic acidosis due to catabolism of excess amino acids, and increases in urinary cyclic AMP excretion and bone resorption. These findings indicate that protein-induced hypercalciuria is due to low grade metabolic acidosis, which directly affects renal handling of calcium. Long-term calcium loss in the urine may lead to negative calcium balance and hyperfunction of the parathyroid gland may induce bone resorption. The influence of 40g animal protein load on urinary risk factors of calcium stone formation was investigated in 23 healthy males and 26 patients with nephrolithiasis. All subjects were given control diets each day containing 60g protein for a week and during the next week each received an additional 40g animal protein. In the controls, added dietary protein resulted in decreased urinary citrate and increased urinary uric acid, with no change in urinary calcium or cyclic AMP excretion. In contrast, the patients showed increased urinary calcium and cyclic AMP as well as decreased urinary citrate. Further examinaiton of the patients revealed that the significant increases of calcium and cyclic AMP excreton occurred only in hypercalciuric patients, who seemed to be classified into renal hypercalciuria. These results suggest that even 40g animal protein affects citrate and uric acid metabolism in normal subjects and patients, and affects calcium metabolism in hypercalciuric patients. These findings indicate the importance of diet guidance to patients with nephrolithiasis, with special regard to the correction of excessive animal protein intake.
We evaluated 107 patients who were diagnosed as benign prostatic hypertrophy preoperatively and underwent transurethral resection of the prostate. We obtained the maximum cross section of the adenoma by transabdominal scanning using a real-time large-sized convex scanner and a 3.5MNz. transducer. Regarding the adenoma as an ellipsoid with revolution around its transverse axis, 2 diameters (breadth and height) were measured with transverse scan (a and b, respectively). We then calculated the values of the estimated size of the adenoma according to the formula for a spheroid, V=(π/6)ab2. Thus the calculated weight were correlated with the resected weight of the specimen after a complete transurethral resection of the prostate. As a result, the sonographic estimation provided a high correlation coefficient: r=0.956. The result suggestd that our method of transabdominal ultrasonography in the preoperative evaluation of the prostatic size was rapid, simple and noninvasive, was well adapted for screening sufferers from prostatic deseases, and gave preoperative information for transurethral resection of the prostate and other surgical operation.
We report a novel way of obtaining a monoclonal antibody to renal cell carcinoma (RCC). BALB/c mice were immunized with RCC cells (ACHN, ATCC CRL1611) and hyperimmunized spleen cells were fused with Sp/2 murine myeloma cell line by PEG 2000. Many hybridoma supernatants were screened by enzyme linked immunosorbent assay (ELISA). After the cloning by limiting dilution, we established a hybridoma reactive to RCCs and named it A25. It belonged to the IgG1 subclass of immunoglobulins. According to our results using ELISA, 4 of 5 RCC cell lines were reactive to A25, while the remaining 23 non RCC cell lines did not react. The supernatant from A25 was used as a primary antibody preparation for avidin-biotin complex immuno peroxidase staining of multiple cases of RCC, normal tissue, and other tumors. This antibody reacted with 3 of 3 grade 1 RCC, 10 of 11 grade 2 RCC and 0 of 3 grade 3 RCC. Proximal tubules of the kidney shared this antigen. However, cross reactivity of this antibody was observed to pyloric glands of the stomach and adenocarcinoma of the colon. The epitope of A25 seemed to originate from normal kidney tubules. Low grade tumors preserved this epitope well, but this character of the original tissue seemed to disappear as tumor grade increased.
Positive cases of serum human chorionic gonadotropin (hCG) have been increased in patients with pure seminoma due to the progress of methods of measuring serum hCG. But the positive ratio and the value of serum hCG in pure seminoma cases are lower than those of non-seminoma cases. The relationship between the pathological type of testicular tumor and the hCG levels in both serum and tumor tissue was investigated, and clinical usefulness of the serum hCG levels as a tumor marker of pure seminoma was discussed. The materials were 19 pure seminomas, 14 non-seminomas and 11 normal testes as controls. HCG and hCG-β in tumor tissue extracts and in the serum of blood from the spermatic and peripheral veins were measured by radioimmunoassay (RIA). The tissue content of hCG and hCG-β were 276.7±136.1mIU/g·tissue and 16.5±3.20ng/g·tissue, respectively, in pure seminoma; 224, 376±91, 619mIU/g·tissue and 4, 608±1, 817ng/g·tissue, respectively, in non-seminoma including choriocarcinoma component; 1, 807±1, 428mIU/g·tissue, and 37.0±11.2ng/g·tissue, respectively, in non-seminoma including seminoma component but not including choriocarcinoma component; 20.4±3.1mIU/g·tissue and 1.27±0.38ng/g·tissue, respectively, in normal testes, (each M±SE). The tissue content of hCG-β in pure seminoma is significantly higher than that in normal testis (p<0.01) and significantly lower than that in non-seminoma including choriocarcinoma component. There was no significant difference in hCG and hCG-β in tumor tissue extracts between pure seminoma and non-seminoma including seminoma component, but not including choriocarcinoma component, and between normal control and non-seminoma (yolk sac tumor, mature treatoma, epidermoid cyst) excluding both seminoma and choriocardinoma component. In all patients with pure seminoma, serum hCG or hCG-β of the spermatic vein was higher than those of the peripheral vein. These results strongly suggest that all pure seminomas produce hCG-β like substances. With more precision in measuring hCG, serum hCG may be a clinically useful tumor marker of all pure seminoma. Furthermore, measurement of hCG in the tumor tissue may assist pathological diagnosis of testicular tumors.
Subrenal capsule assay as a chemosensitivity test was performed on 8 renal cell carcinomas and 7 transitional cell carcinomas using athymic mice. Twelve of the 15 trials were considered suitable for evaluable assay. In 2 of the 3 non-evaluable trials the histological examination showed fibroblasts without cancer cells in the implants of the control group while the implants grew macroscopically. No host cell infiltration was observed microscopically in any implant of the control groups of evaluable assays. Mice were treated with several anticancer agents and sensitivities were evaluated by the tumor growth inhibition rate (TGIR) on the day 10 or 12 and by histological changes. No correspondence was observed between TGIR and histological changes. The charasteristic histological finding of the treated groups of transitional cell carcinomas was cystic degeneration and it seemed to be a factor of the discordance between TGIR and the histological changes.
The purpose of this study is to evaluate the effect of sodium pentosan polysulfate (SPP) and sodium copper chlorophyllin (SCC) on the formation, growth and aggregation of calcium oxalate crystals in vivo, and to measure the number and the volume of crystals formed in the rat kidney, quantitatively, with a Coulter counter TA-II. The deposition of calcium oxalate crystals in the rat kidney was induced by intraperitoneal injection of 2. 5g per Kg of body weight of hydroxy-L-proline and administration of 0.4% ethlene glycol as the drinking fluid ad libitum for 7 days. Daily excretions of urinary oxalate, calcium (ratio to urinary creatinine) and urinary volume were measured. Both kidneys were removed after protocol. The kidneys were homoginized with 0.2M Tris-buffer (pH8.0) and subsequently digested in soluene-100. After calcium oxalate crystals were collected, they were suspended in saline saturated with calcium oxalate. The crystal size distribution was measured with a Coulter counter TA-II. In addition, the renal calcium content was measured by atomic absorption spectrometry, and the kidneys were examined by optical microscopy and scanning electron microscopy. The crystals formed in the rats' kidneys were analyzed by infrared spectroscopy. The results were as follows: 1. There was no deposition of crystals in the kidney of the rats which were not treated. There was intratubular depositon of crystals in the kidneys of the rats injected with hydroxy-L-proline and administered 0.4% ethylene glycol. They consisted of calcium oxalate monohydrate. 2. Renal calcium content was significantly higher in the groups with induced crystals than the control group. 3. The crystal volume and the mean size of crystals were statistically increased in the groups with induced crystals. In the groups with induced crystals, there was statistical difference between the crystal-forming group and sodium pentosan polysulfate or sodium copper chlorophyllin treated groups in the crystal volume and the mean size of the crystals. The results suggest that it is valuable to measure the crystals induced in rat kidneys, quantitatively, with Coulter counter, and that SPP and scc have a strong inhibitory effect on calcium oxalate crystal growth in vivo.
Serum concentration of laminin was measured radioimmunologically in 19 patients with renal cell carcinoma and 71 normal controls. The serum laminin levels of renal cell carcinoma patients were significantly higher than those of normal controls. After nephrectomy the elevated serum levels of laminin showed marked decreases in most of the patients without metastasis. By the immunofluorescent technique with LAM-1, a monoclonal antibody which is specific to laminin A and B chanins, strong fluorescence was found in the extracellular matrix in renal cancer tissue. Taken all these facts together, renal cell carcinoma may be a laminin synthetic tumor. The average serum laminin level of the patients with a disseminated disease were significantly higher than that of the patients with a localized disease. The elevated serum level of laminin may play an important role in the process of tumor metastasis. Furthermore, our data suggest that elevated laminin levels may predict the subsequent course of a tumor condition. The monitoring of serum laminin level may be valuable for following-up of the course of renal cell carcinoma.
The prognosis of paraquat poisoning has been assessed from serum paraquat levels using e. g. produfoot survival curve. The prognosis of paraquat poisoning has not been investigated from urinary paraquat levels. The purpose of this study is to clarify whether the prognosis of paraquat poisoning can be determined by urinary paraquat levels or not. Urinary severity index of paraquat poisoning (U-SIPP)was employed. U-SIPP is calculated as hours between ingestion of paraquat and start of intensive treatment multiplied by the paraquat level in the urine at the same time. Urinary excretion of paraquat is correlated with renal function. The prognosis of paraquat poisoning using U-SIPP was further discussed, based on creatinine clearance (Ccr) and the relation between serum and urine paraquat levels. The following results were obtained. 1) When creatinine clearance was greater than 20ml/min, patients who died of ciculatory failure were with U-SIPP level over 1250, patients who died of respiratory failure were with U-SIPP level between 1250 and 250, and survivors were with U-SIPP below 250. 2) Survival duration was closely correlative with U-SIPP in cases with Ccr greater than 20ml/min(r=-0.8123, p<0.025). 3) When Ccr was greater than 20ml/min, urinary paraquat levels were correlated very well with the serum levels at admission, even during direct hemoperfusion (DHP) treatment. 4) When diuretic agents such as furosemide was effective, the assessment of the prognosis of paraquat poisoning using urinary paraquat levels seemed to be possible. Urinary paraquat level was approximately measured by an applied qualitative analysis method with using a dilution method.
Renal scarring in 271 kidneys of 172 children with primary vesicoureteral reflux (VUR) was evaluated by 99mTc-DMSA renoscintigraphy. 58% of refluxing kidneys were with renal scar by the initial DMSA renoscintigraphy. Only 52% of these kidneys showed good correlation between the findings on IVP and DMSA renoscintigram. Of the 144 refluxing kidneys with normal IVP, 41% had renal scarring on DMSA renoscintigram. DMSA renoscintigram revealed widespread renal scarring in 28% of kidneys with only calyceal clubbing and in 60% of those with segmental cortical thinning on IVP. It is realized that IVP was an in-sensitive method to evaluate renal scarring of refluxing kindeys and such kidneys with segmental renal scar on IVP accompanies more widespread scar on DMSA renoscintigram. These cases were allocated to 2 age groups, younger than 3 years and older than 4 years. In the former group less than 10% of kidneys with low grade VUR and about 40% with high grade UVR had widespread renal scarring. On the contrary, in the latter group severe renal scar was recognized in more than 20% of kidneys with low grade VUR and in about 60% with high grade UVR.
A luteinizing hormone-releasing hormone (LH-RH) analogue was administered for 3 to 32 months to 15 prostatic cancer patients in stage B-D (B:3, C:8, D:4). Intratesticular, intratubular and prostatic androgen levels were measured by radioimmunoassay before and after LH-RH analogue therapy. The measurement of serum prostatic acid phosphatase (PAP) and prostatic specific antigen (PA) levels was also conducted. Thereafter, we assessed the effect of the LH-RH analogue on androgen levels and the relation of prostatic tissue 5α-dihydrotestosterone (DHT) level to the clinical response. The results were as follows: 1) Johnsen's mean germinal epithelium count was significantly decreased from 7.7±2.1 (mean±S.D.) to 4.3±2.3, and the wall thickness of seminiferous tubules was increased from 5.93±1.31 to 11.9±3.64μm. 2) Plasma testosterone (T), intratesticular and intratubular androgen levels were significantly decreased (plasma T: from 4.40±1.84 to 0.61±0.32ng/ml, intratesticular T: from 335.3±170.3 to 4.6±3.8ng/g.t.w., DHT: from 25.3±11.7 to 3.7±2.7ng/g.t.w., intratubular T: from 50.8±36.6 to 0.10±0.99ng/g.t.w. and DHT: 7.54±3.20 to 0.63±0.90ng/g.t.w.). 3) Crude nuclear DHT levels in prostatic tissue fell from 15.3±9.3 (N=8) to 0.37±0.54pg/mg protein (N=3) and the level was non-detectable in 5 of 8 cases. 4) Complete remission was achieved in 1 patient, partial response in 5, objective stable in 8, and objective progression in 1 patient, according to Shimazaki's criteria. Moreove, the clinical response in patients with high pretreatment prostatic tissue DHT levels (over 10pg/mg protein or 2ng/g.t.w.) is good to LH-RH analogue therapy. Therefore, the LH-RH analogue seems to have a strong inhibitory effect on both intratesticular and intratubular androgen levels and appears to be an appropriate agent for treating prostatic cancer patients with high prostatic tissue DHT levels.
We report a 63 year-old female with solid, nonpapillary, and smooth-surfaced bladder tumescences of approximately 30 and 7mm in diameter. Biopsy of the tumor and other apparently normal epithelium revealed dense lymphocytic infiltrations with formation of many lymph follicles. Because of the relatively large nodular form of the lesion we regarded this to be pseudolymphoma of the bladder rather than follicular cystitis. Pseudolymphoma in the bladder has not yet been reported in English literatures to our knowledge. The tumorous changes have disappeared by 6 months after the initial 3 week chemotherapy.
The patient was a 70-year-oold male with complaint of macrohematuria at the first visit to our clinic on June 10, 1986. At that time, cystoscopy revealed a thumb sized papillary tumor and a rice sized non papillary tumor, and the biopsy specimen was pathologically diagnosed as undifferentiated carcinoma. But, he refused admission. On January 30, 1987, he came back to our clinic with complaints of dyspnea, general fatigue and weight loss. Moderate lt. gynecomastia was found and the level of serum hCG-β was detected as high as 101ng/ml. Excretory urogram and enhanced CT revealed a large mass in the bladder. In the seventeeth day after admission, he died of lung edema and heart failure. The findings of autopsy showed a large light greenish to light brownish tumor of 10×10×3cm in the bladder. Distant metastases were observed in internal, common iliac and paraaortic lymph ndes, but without other distant metastasis. In histological and immunohistochemical studies, the final diagnosis is choriocarcinoma of the bladder, containing syncytiotrophoblastic giant cells with hCG-β granules as an undifferentiated carcinoma. To our knowledge this case is the eighth described in Japan. Herein we report a new case of primary choriocarcinoma of the bladder and make a brief review of the literatures.